A Randomized, Double-Blind, Placebo-Controlled Study to Determine the Efficacy SALI-10 Oral Probiotics in Experimental Gingivitis
Phase: PHASE2
Conditions: Gingivitis
Sponsor: Ostia Sciences

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Clinical Trial Study Protocol 
 
A Randomized, Double-Blind, Placebo-Controlled Study to 
Determine the Efficacy SALI-10 Oral Probiotics  in 
Experimental Gingivitis 
 
 
Protocol Date: October 11, 2025 
 
Ostia Sciences 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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1. INTRODUCTION 
Gingivitis is an oral disease condition affecting 50% to 90% of adults globally (Albandar and 
Rams, 2002). Not only is it common in the adult population, but it is a precursor to periodontal 
disease, which is a more severe condition of gum disease that permanently damages the soft tissues 
and the bone supporting the teeth (Löe et al., 2009). It is an issue that the investigators do not want 
to overlook since it has also been proven that periodontitis exacerbates different systemic diseases, 
such as diabetes and atherosclerotic vascular disease (Lockhart et al., 2012; Mealy et al. 2006). 
Previous studies and clinical research have shown that combining antimicrobial treatment with 
non-surgical periodontal therapy, such as scaling and root planning, is an efficient way to tackle 
periodontal disease (Kapoor et al. 2012). However, modern practitioners are looking for an 
alternative therapy, as antibiotics are innocuous drugs by nature and can lead to several side effects, 
damage to our commensal bacterial flora and favour the development of antibiotic-resistant 
bacteria (De Nies et al. 2023; Soares et al. 2012). Knowing the complexities associated with the 
treatment and management of periodontal disease, the primary strategy for success lies in its 
prevention. Therefore, it is also imperative to identify effective methods for treating and preventing 
gingivitis. 
 
 
The gingivitis pathology can be reversed by reduction or removal of microbial plaque that 
accumulates on hard and soft tissues and is considered standard of care in the industry (Berezow 
Alex and Darveau Richard, 2010, van der Weijden and Slot, 2011). Regular oral hygiene in 
combination with therapeutics that delivers an anti-microbial benefit is thought to mitigate the 
onset of gingivitis (van der Weijden and Slot, 2011, Gunsolley, 2006). However, testing 
therapeutics for prophylaxis benefit to mitigate development of gingivitis has not been fully 
examined.  
 
The classical model of experimental gingivitis (EG) was developed in 1965 by Loe and Silness 
who convincingly demonstrated the causative relationship between the accumulation of dental 
plaque and the development of clinically evident gingivitis in healthy young adults abstaining from 
all oral hygiene practices for a 21-day period. Furthermore, on resuming customary oral hygiene 
practices, all study participants demonstrated a return to gingival health within two weeks (Loe et 
al., 1965). To understand how the participants of this study returned so quickly to gingival health 
we can reflect on our current understanding that our clinically healthy state is an active and 
dynamic process.  
 
Neutrophils, a type of white blood cell (leukocyte), represent a key component of the innate 
defence system that protects periodontal tissue from both gingivitis and periodontitis. Not only are 
they the first line of cellular defence, but they are among the most abundant leukocytes within the 
periodontal tissues (Hirschfeld, 2020). For example, gingivitis is associated with a significant 
increase in the number of neutrophils that migrate to periodontal tissue. In contrast, individuals 
with too few neutrophils brought about by either congenital deficiencies in neutrophil numbers, or 
transit (LAD 1 and 2) or have an induced neutropenia by chemical induction with antimitotic 
agents such as cyclophosamide invariably develop periodontitis. Likewise, studies in KO mice that 
are defective in neutrophil transit also develop periodontitis. Consistent with the key contribution 
of neutrophils to both gingivitis and periodontitis, neutrophil transit to gingival tissue is highly 
regulated. The periodontium contains a highly orchestrated expression of select innate host defense 

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mediators that facilitate the transit of neutrophils from the highly vascularized gingival tissue to 
the gingival crevice, where they form a “wall” between the host tissue and the dental plaque 
biofilm. 
 
However, the prolonged presence of neutrophils in gingival tissue is not tolerated in the healthy 
state. For example, the failure to down regulate orchestrated neutrophil transit  0results in an 
increase in neutrophil numbers in gingival tissue and a significant increase in per 
iodontal bone loss. Therefore, neutrophil homing to the gingival crevice is highly regulated such 
that under conditions of periodontal health the appropriate amount of neutrophils are present to 
maintain control of dental plaque bacterial growth and yet not elicit tissue damage. Evaluation of 
oral and blood neutrophils during experimental gingivitis showed that people with uniquely high 
inflammatory response had an exaggerated polymorphonuclear neutrophil response both in the 
oral cavity as well as in the blood (Wellappuli 2017).  
 
Gingivitis is a reversible inflammatory condition caused by the accumulation of dental plaque and 
the associated disruption of the host–microbial homeostasis. During gingivitis, the microbial 
community transitions from being dominated by gram-positive health-associated bacteria, such 
as Streptococcus species, to gram-negative periopathogens, including species of the genera 
Porphyromonas, Tannerella, Treponema and Prevotella. This dysbiotic shift triggers 
inflammatory responses, leading to tissue damage and, in some cases, progression to 
periodontitis. Those pathogenic bacteria also contribute to the production of volatile sulfur 
compounds; as a result, people with gingivitis have an increased probability of having halitosis by 
3.6 times (Lee et al. 2023).  
 
A recent study (Kerns, 2023) on human experimental gingivitis identified three distinct host 
response phenotypes—high, low, and slow responders—based on clinical, inflammatory, and 
microbial parameters: 
1. High Responders: Rapid plaque accumulation accompanied by a significant increase in gram-
negative periopathogens and elevated inflammatory markers, such as interleukin-1β (IL-1β). 
2. Low Responders: Similar plaque accumulation to high responders but lower inflammation, 
suggesting a more muted host response to bacterial dysbiosis. 
3. Slow Responders: Delayed plaque accumulation and microbial succession, with prolonged 
dominance of health-associated Streptococcus species. This group exhibited delayed or reduced 
inflammation, demonstrating a more resilient microbial community and host response. 
 
The microbial analysis revealed that the persistence of beneficial Streptococcus species, such as 
S. sanguinis and S. oralis, in slow and low responders correlates with a protective effect against 
the emergence of periopathogens and the associated inflammatory cascade. Conversely, the loss 
of these beneficial bacteria in high responders was linked to more severe inflammation, 
highlighting the critical role of the oral microbiome in modulating gingivitis severity. 
 
Lantibiotic salivaricins are polycyclic peptides containing lanthionine and/or β-
methyllanthionine residues that are produced by certain strains of Streptococcus salivarius, 
which almost exclusively reside in the human oral cavity. These molecules importance stems 
from their antimicrobial activity towards relevant oral pathogens which has been applied through 
the development of salivaricin-producing probiotic strains. However, salivaricins may also prove 

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to be of great value in the development of new and novel antibacterial therapies in this era of 
emerging antibiotic resistance (Barbour, Wescombe, & Smith, 2020). In a study by Barbour & 
Philip 2014, they found that the bacteriocin, levan-sucrase production and basic safety features 
of S. salivarius strains isolated from healthy Malaysian study participants demonstrating their 
potential for use as probiotics. A new bacteriocin production medium was developed with 
potential scale up application for pharmaceuticals and probiotics from S. salivarius generating 
different lantibiotics. This is relevant for the clinical management of oral cavity and upper 
respiratory tract in the human population. Appendix 1 includes a summary table that lists the 
relevant clinical trials that utilized S. salivarius. 
 
Proposed Solution: S. salivarius SALI-10 to be used as a probiotic to help prevent gingivitis 
 
We propose using a novel strain, Streptococcus salivarius SALI-10, as a targeted microbial 
intervention to modulate the oral microbiome and prevent gingivitis. S. salivarius SALI-10 is 
hypothezised to:  
 
• Maintain a stable population of beneficial streptococci during plaque accumulation. 
• Inhibit the growth of periopathogens such as Porphyromonas, Tannerella and Prevotella 
through competition and production of Salivaricin 10. 
• Delay or suppress the dysbiotic shift to gram-negative dominance, thereby reducing the 
inflammatory response. 
By preserving microbial homeostasis, SALI-10 may emulate the microbial resilience observed in 
slow responders, offering a novel strategy for gingivitis prevention. 
 
 
2. STUDY OJECTIVES 
1) The aim of this study is to evaluate the efficacy of S. salivarius SALI-10 in modulating 
the oral microbiome and preventing gingivitis, using a placebo-controlled experimental 
gingivitis model. Specifically, the investigators aim to: 
Microbiome Modulation: 
- 
Assess the ability of SALI-10 to maintain higher levels of beneficial streptococci (S. 
sanguinis, S. oralis, S. parasanguinis and SALI-10) during plaque accumulation. 
- 
Determine whether SALI-10 prevents or delays the increase of gram-negative 
periopathogens (Tannerella, Prevotella, and Porphyromonas) and dysbiosis. 
2) To evalute the effects of probiotic Sali-10 intake regarding clinical parameters of 
periodontal health during an experimental gingivitis.    
3) To assess differences in the onset of gingivitis, the extent of the condition, and its resolution 
among experimental and control groups. 
4)  To evaluate the capability to establish or increase oral colonization by Streptococcus 
salivarius SALI-10. The expected outcomes will be attributed to the effect of antimicrobial 
and immunomodulation metabolites (mainly, the natural product Salivaricin 10) produced 
by this strain present in the probiotic. 
5) To analyze the level of sulfured compounds emitted from the oral cavity with a halimeter 
(Halimeter PLUS) from each study participant to assess the level of halitosis objectively, 
then evaluate the difference between the control and test groups as they go through the 
experimental gingivitis clinical trial. 

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3. STUDY OVERVIEW 
This research study is designed to evaluate a new strain of Streptococcus salivarius containing 
probiotic properties for its effect on gingivitis including the oral neutrophil phenotype and oral 
microbiome modulation during an induced gingivitis state. A 7-week (8 appointments, 7 weeks) 
clinical study will be conducted to evaluate a probiotic for therapeutic effects that aim to delay the 
onset of experimental gingivitis and hasten the resolution of the disease.  
 
4. RECRUITMENT 
4.1 Test Site and Investigator 
Site: University of Toronto, Canada 
Study Originator: Dr. Michael Glogauer (Osteia Sciences) 
Principal Investigator: Dr. Michael Goldberg 
Co-Investigator: Dr. Howard Tenenbaum 
 
4.2 Study Population 
Advertisements (Appendix 2) will be placed at University of Toronto downtown campus to invite 
volunteers who are interested to participate. All study participants will undergo informed consent 
process, prior to enrollment in the study. Categorization of participants and sites will be based on 
the clinical assessments. Other than the study-related oral health history recorded, there is no 
intention to obtain information from the participants‟ other dental or medical records. Sixty (60) 
qualifying volunteers that agree to participate and sign all corresponding forms will be enrolled.  
 
The first participant will be assigned a number, and all the following enrolled participants will be 
assigned consecutive numbers by accession. The following inclusion and exclusion criteria will be 
utilized in this study. 
 
4.3 Inclusion Criteria: 
1) Male or female volunteers aged 18-70 years  
2) In good general health, ASA I 
3) No clinical signs of gingival inflammation at >90% of sites observed  
4) Absence of Periodontal Pockets, Probing Depth (PD) < 3.0 mm on all teeth/site 
5) Absence of Clinical Attachment Loss (CAL) = 0 mm  
6) No periodontal disease history 
7) Have at least 20 gradeable teeth 
8) Non-smokers  
9) Fluent in English  
10) For study participants of childbearing potential, both men and women, at least one of the 
following birth control measures must be used: abstinence, hormonal birth control (oral, 
injectable, transdermal, intra-vaginal), intrauterine devices, confirmed successful 
vasectomy, or condoms. 
 
 
4.4 Exclusion Criteria: 
1) Presence of orthodontic bands.  

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2) Presence of partial or full  dentures.  
3) Tumour(s) of the soft or the hard tissues of the oral cavity.  
4) Cavitated carious lesions requiring immediate restorative treatment.  
5) History of allergy to a consumer or personal care products or dentifrice ingredients as 
determined by the dental profession monitoring the study.  
6) Participation in any other clinical study or test panel within one month before entering the 
study.  
7) Medical condition which requires pre-medication before dental visits/procedures  
8) Current use of anti-inflammatory, antibiotics, or antimicrobial drugs or within the last 30 
days of enrolment.  
9) History of periodontal disease.  
10) History of systemic inflammatory, immune conditions and immunocompromised 
conditions 
11) Pregnant or nursing women or those planning to get pregnant  
12) Use of tobacco products.  
13) Long-term antibiotic or anti-inflammatory therapy.  
14) Medication or Natural Health Products (NHPs) that could affect the gingiva like calcium 
channel blockers, anti-epileptic therapy etc.  
15) Medical condition or any current usage of medication that the investigator considers may 
compromise the study participant's safety as well as the quality of the study results 
16) Allergy to any of the following ingredients: Streptococcus Salivarius, Sorbitol, Isomalt, 
Calcium phosphate dibasic, Potato starch, Mint, Glyceryl dibehenate, Stevia, Maltodextrin 
17) Use of other probiotics 
18) Taking anticoagulant medications and those suffering from blood and bleeding disorders 
19) Recently experienced or will be experiencing dental, oral or any type of surgery. 
20) Use of anti-plaque/ anti-gingivitis products 
21) Presence of active infections 
22) Participants who are experiencing nausea, fever, vomiting, bloody diarrhoe or severe 
abdominal pain.  
23) Use of any antibiotics. 
 
4.5 Continuing Criteria 
Each study participant must meet continuance criteria at each visit in order to continue in the 
study. Study participants will be released from further participation for the following reasons:  
• use of non-study dentifrice during treatment periods (acceptable dentifrice will be provided 
during the study) 
• use of oral care products (other than those permitted) during the treatment periods including 
mouth rinse products and chewing gum. 
• non-compliance with study procedures 
• actively participating in another clinical study unless permission has been granted by the 
Principal Investigator 
• development of a medical condition or taking a medication which would affect the study 
outcome (such as an antibiotic) as detailed in the exclusion criteria. 
• Subject fails to substantially comply with the protocol requirements. 
• Subject fails to report for a scheduled examination. 

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• Subject is treated with medications during the course of the study, which may interfere 
with the parameters of the study. 
• Subject develops an adverse reaction.  The study Investigator will immediately notify the 
study monitor and information will be recorded on Adverse Reaction Form. 
• Subject receives emergency dental or medical treatment, which may interfere with the 
parameters of the study. 
• Sponsor elects to terminate the study. 
• Subject elects to terminate participation in the study. No pregnant women will intentionally 
be enrolled in this study. All female subjects being considered for this study will be asked 
about their pregnancy status.  the clinical/research investigator if they become pregnant 
during the course of the 9-week study. In the event a woman enrolled in this clinical 
research study becomes pregnant at any time during this study, participation in this study 
will be terminated upon the clinical staff’s notification of the event. 
•  
 
4.6 Concomitant Therapy 
If a study participant takes concomitant medications as a matter of necessity for the treatment of 
a medical condition, then such medication may be permitted for the duration of the study at the 
discretion of the investigator.  However, it is the responsibility of the investigator to disqualify 
from entering the study any study participant who, upon screening, is using medication or 
consumer product that might obscure the interpretation of study results. All medications 
currently used by the study participant at enrollment, or any time through the end of the study, 
will be recorded on the visit report form (Appendix VI). Study participants may receive 
medication to treat adverse events as deemed necessary by the investigator or the study 
participant’s physician. 
 
4.6 Missed Doses 
If a study participant misses their dose, they will be asked to take it if no more than 3 hours has 
passed since they brushed their teeth. If more than 3 hours have passed, they will be asked to 
skip the dose completely. Do not double up on the next dose. Do not exceed 2 lozenges per day.  
 
5. TEST PRODUCTS/INTERVENTIONS 
Each participant randomized to the intervention group will receive mint-flavoured SALI-10 
lozenges. They will be instructed to take a lozenge twice daily after brushing by letting it 
dissolve in the mouth. The participants will be using the SALI-10 lozenges throughout the study. 
 
5.1 SALI-10 Lozenges Manufacturer Information 
The placebo and probiotics lozenges will be manufactured by   
Lallemand Health Solutions,  
17 975 rue des Gouverneurs 
Mirabel, Québec, Canada  
J7J 2K7 
Tel : 450-433-9139 
 
Content of the placebo lozenge:  

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Content of the probiotic lozenge:  
 
 
5.2 Dispensing, Storage, and Accountability 
All product(s) are distributed by the study coordinator at the clinical test site.  The study 
participant will follow provided instructions on when and how to brush their teeth.  These 
instructions are explained in the procedure section.  
 
One of the study personnel, Dr. Martin Lambert, will be responsible for dispensing and 
collecting the containers of the SALI-10 lozenges at each visit.   
 
6. STUDY PROCEDURES 

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6.1 Randomized Assignment and Product Blinding 
Qualified study participants will be randomly assigned to one of two study groups, either Group 
A or Group B, using randomization software. To maintain blinding, the manufacturer will produce 
both probiotics and placebo lozenges, dispense them in identical, opaque bottles, and label them 
as either Group A or Group B. The assignment of probiotics to one of these groups will be 
determined by the manufacturer and disclosed only at the study's conclusion. Each bottle will be 
labeled with a code (either letter A or B, followed by a number) and emergency contact 
information. The products will be distributed in sealed boxes that look identical to prevent any 
differences in appearance or packaging between study groups. Each product inside the box will be 
overwrapped and include instructions for at-home use and safety information, including 
emergency contact details. 
 
To monitor study participant compliance, the investigators will use a pharmaceutical scale to 
ensure the number of lozenges remaining in the bottle matches the expected count at each study 
visit. A scale will be used to avoid the operator from seeing the lozenges, in order to preserve 
blindness in this study (placebo and probiotic lozenges are not perfectly identical).  
 
Study participants will be assigned a unique identification number in chronological order (e.g., 
from 1 to 60) as they enroll in the study. These identification numbers have been pre-assigned to 
a study group based on a computer-generated randomization list provided by an independent 
statistician. Cohabitant study participants will be assigned to the same study group and receive the 
same products. Study participants will be instructed to avoid using any other oral hygiene products. 
There will be no dietary restrictions during the study. 
 
6.2 Product Dispensing and Use at Home 
Study participants will be instructed to brush their teeth twice daily (morning and evening) for two 
minutes each time with a toothpaste provided by the study and toothbrush and floss. The 
participants will be informed as to when home oral hygiene should be performed and the times 
they will be instructed to refrain from using oral hygiene products during certain times during the 
study. Study participants There will be no restrictions regarding diet habits during the course of 
the study. 
The SALI-10 lozenges and placebo lozenges will be pre-packed when given to each 
participant.  
 
6.3 Telephone Pre-Screening 
Potential study participants who contact the clinical trial coordinator for more information about 
the study via email will then be contacted via telephone. At this stage the Recruitment 
Questionnaire (Appendix 3) will be completed to determine whether they are eligible for the 
screening phase of the study. If the study participant is eligible for the study the screening visit 
will be scheduled.  
 
6.4 In-Person Screening 
Before any participant screening takes place, they will be given the Informed Consent Form to 
read and ask any questions they may have and sign. If signed, the remainder of the visit  entails 
completion of the Personal Information Form (Appendix 4), and the Health Questionnaire form 

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(Appendix 5) and completion of an oral exam of both the soft and hard tissues (Screening Oral 
Assessment Form – Appendix 6). Female participants will be asked to take a urine pregnancy 
test. This is done to determine if the study participant is eligible for the study. The study 
personnel will then complete the screening form (hard copy).. The screening will be carried out 
until sixty (60) study participants are enrolled.  
 
6.5 Pre-Induction Phase (Health Baseline – Study participant maintain dental hygiene at 
home for 14 days) 
 
6.5.1 Visit 1 (t-2) Day -14 
Study participants will undergo an oral hard and soft tissue exam as well as measurements of 
probing depth (PD), attachment level (AL), visible plaque index (VPI), gingival index (GI) and 
bleeding on probing (BOP) and recorded on the Oral Assessment Form (Appendix 7). The 
biospecimens collection will include gingival crevicular fluid (GCF), sub-gingival plaque, saliva 
and a saline oral rinse.  All study participants will receive a prophylaxis (cleaning) which 
involves scaling and polishing to remove all supra- and subgingival plaque and calculus. Study 
participants will receive their assigned study product for use at home with directions. The 
participants will take two lozenges daily, one after their morning brushing and one after evening 
brushing. They will allow the lozenges to dissolve without biting or swallowing and avoid eating 
and drinking for one hour. The participants will be instructed on proper oral hygiene and will be 
instructed to continue their home oral hygiene in addition to use of their assigned product for 14 
days (2 weeks). The participants will be instructed to return the lozenges’ containers at each visit 
for protocol compliance check. Researchers will query participants on their use of NHPs, as well 
as any adverse effects they experienced. 
 
6.5.2 Visit 2 (t-1)  (Day -7) 
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. Researchers will query participants on their use of NHPs, as well as 
any adverse effects they experienced. 
 
6.6 Induction Phase (No Oral Hygiene – for 21 days) 
 
6.6.1 Visit 3 (t0) (Day 0)  
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. Additionally, study participants will be asked to refrain from any 
oral hygiene for 21 days and to continue using the lozenges they were assigned, one during the 
morning, and one during the evening.   Researchers will query participants on their use of NHPs, 
as well as any adverse effects they experienced. 

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6.6.2 Visit 4 (t1) (Day 7) 
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. The participants will again be reminded to refrain from all forms of 
oral hygiene. Researchers will query participants on their use of NHPs, as well as any adverse 
effects they experienced. 
 
6.6.3 Visit 5 (t2) (Day 14) 
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. The participants will again be reminded to refrain from all forms of 
oral hygiene. Researchers will query participants on their use of NHPs, as well as any adverse 
effects they experienced. 
 
6.7 Resolution (Study participants resume to regular dental hygiene home care for the rest 
of the study) 
 
6.7.1 Visit 6 (t3) (Day 21) 
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. All study participants will receive a prophylaxis (cleaning) which 
involves scaling and polishing to remove all supra- and subgingival plaque and calculus. The 
study participants will be instructed to continue using the lozenges they were assigned and 
resume home oral hygiene for the next 14 days (2 weeks). Researchers will query participants on 
their use of NHPs, as well as any adverse effects they experienced. 
 
6.7.2 Visit 7 (t4) (Day 28) 
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. The study participants will be reminded to continue at home oral 
hygiene. Researchers will query participants on their use of NHPs, as well as any adverse effects 
they experienced. 
 

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6.7.3 Visit 8 End of Study (t5) (Day 35) 
The study participants will return the containers containing the lozenges to a member of the 
study team. Study participants will then undergo an oral hard and soft tissue exam as well as 
measurements of probing depth (PD), attachment level (AL), visible plaque index (VPI), 
gingival index (GI) and bleeding on probing (BOP) and recorded on the Oral Assessment Form 
(Appendix 7). The biospecimen collection will include gingival crevicular fluid (GCF), plaque, 
saliva and a saline oral rinse. The study participants will be informed that this is the end of the 
study. They will be given their compensation and told that they may resume their regular oral 
hygiene. Once the study has been completed, the manufacturer will reveal to which group was 
the probiotic assigned to (either A or B). Afterwards, the investigators will inform participants to 
which group they were assigned and will ask for their feedback in regards to their experience. 
Researchers will query participants on their use of NHPs, as well as any adverse effects they 
experienced. 
 
Please refer to figure 1. Illustrating the breakdown and timeline of the study as well as the 
relevant study procedures at each time point and the objectives.  
 
Figure 1. Study Flow Infographic 
 

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7. STUDY PROCEDURES AND CLINICAL REGISTRATIONS 
 
7.1 Abbreviated periodontal health assessment.  
Clinical data will be documented based on probing depth (PD), attachment level (AL), visible 
plaque index (VPI), gingival index (GI) and bleeding on probing (BOP). All clinical measurements 
will be conducted using a manual periodontal probe.  
 
7.2 Gingival Bleeding Index  
Bleeding on Probing (BOP): Absence or presence of bleeding after gentle probing  will be recorded 
within 20 seconds of probing. All six sites of the all the teeth will be probed to determine BOP. 
 

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7.3 Visible Plaque Index 
The surfaces of the study teeth will be scored and given either a “0” no plaque or “1” plaque 
assessment: 0 = No plaque; 1 = Visible Plaque  
 
7.4 Loe and Silness Gingival Index (GI) 
The surfaces of the study teeth will be evaluated using light air and a periodontal probe. A score 
is given based on the following scale: 0 = Normal gingiva; 1 = Mild inflammation, a slight change 
in colour, slight edema, no bleeding on probing; 2 = Moderate inflammation, moderate glazing, 
redness, bleeding on probing; 3 = Severe inflammation, marked redness and hypertrophy, 
ulceration, a tendency to spontaneous bleeding.  
 
7.5 Gingival Crevicular Fluid (GCF) Samples 
Before GCF collection, the plaque index will be recorded. The sites to be sampled will be isolated 
with cotton rolls and gently air-dried. GCF samples will be collected with sterile paper strips 
(Periopaper strips, Oraflow Inc.) that will be inserted into the gingival crevice until mild resistance 
is felt and left in place for 30 seconds. GCF will be collected from the mesiobuccal and 
mesiopalatal surfaces of teeth (#’s 3, 4, 5, 12, 13, 14). Paper strips visibly contaminated with saliva 
and blood will be excluded from the study. All samples will be collected in microcentrifuge tubes. 
They will be transported on ice and stored within 24 hours at -80ºC until processing. At our lab at 
Dental Sciences building, at University of Toronto, Faculty of Dentistry, these samples will 
undergo 40-plex chemokine analysis, oral neutrophil count and targeted proteomics 
 
7.6 Sub-gingival Plaque Samples 
Sub-gingival plaque will be collected using sterile paper points inserted in the gingival sulcus for 
30 seconds from the mesiobuccal and mesiopalatal surfaces of teeth numbers 14, 15, 16, 24, 25, 
and 26). Plaque samples will be placed in microcentrifuge tubes then transported on ice to -80ºC 
freezer in our lab at Dental Sciences building, at University of Toronto, Faculty of Dentistry. There 
they will be processed and analyzed by shotgun metagenomics and targeted proteomics.  
7.7 Saliva and Oral Rinse Samples 
Saliva samples (5 ml) will be collected at all visits and placed on ice to -80ºC freezer. Samples 
will be sent our lab at Dental Sciences building, at University of Toronto, Faculty of Dentistry. 
There the saliva samples will be processed and undergo 16S rRNA sequencing and targeted 
proteomics analyses. The oral rinse samples will undergo oral neutrophil count.  
 
7.8 Halitosis Measurement 
The Halimeter PLUS is a specialized instrument designed and manufactured by Interscan 
Corporation to measure the concentration of volatile sulfur compounds (VSC), particularly 
hydrogen sulfide, methyl mercaptan, and dimethyl sulfide, in a person's breath. These 
compounds are often associated with oral malodor, commonly known as bad breath. The 
Halimeter contains electrochemical sensors that detects the concentration of VSCs in breath. 
When a person exhales into the device, the sensor measures the levels of these sulfur 
compounds, providing a quantifiable reading. Elevated levels of VSCs indicate the presence of 
oral bacteria and potential oral health issues. 
 

Protocol_V8_October_11, 2025 
 
15 
Since all research projects will be done on de-identified samples, it will not be possible to return 
individual results to participants. Unless participants ask to withdraw from the study, samples 
may be retained indefinitely. Because all research will be done on de-identified, aggregated data, 
it will not be possible to withdraw the data from a specific participant when samples are 
withdrawn from the study.  
8.0 STATISICAL ANALYSES  
Correlations between tissue and blood neutrophil subsets and standard periodontal disease 
parameters from oral exam will be assessed. Data will be complied in SPSS version 20. Changes 
in the clinical parameters of experimental gingivitis such as bleeding on probing, gingival index, 
plaque index and probing depths during day 0, 7, 14, 21, 28 and 35 will be compared using one-
way analysis of variance (ANOVA) followed by Tukey multiple comparison. 
 
Oral neutrophil counts as well as neutrophil subset formation during day 0, 7, 14, 21, 28 and 35 in 
oral rinse samples as well as blood samples will be compared using one-way analysis of variance 
(ANOVA) followed by Tukey multiple comparison.  
 
Multiple sites within each study participant will be evaluated using a Gingival Bleeding Index 
(BOP) at baseline and 3-week post product use exams. The gingivitis bleeding scores will be 
averaged to provide a mean score per study participant for each exam period. Product means and 
standard deviations will be calculated for each time period. Analysis of Variance techniques 
(possibly including Analysis of Covariance using the baseline scores as the covariant) will be used 
to compare the difference between the two test treatments using the 95% confidence level (p ≤ 
0.05). The correlation of baseline to post-treatment results will be assessed graphically. If there is 
evidence of correlation, the baseline result will be included in the ANOVA model to adjust for 
baseline differences. 
 
9.0 REPORT OF ADVERSE EVENTS 
Study participants will be told of any possible adverse reactions or side effects from exposure to 
this method or product.  Oral irritation is possible.  If side effects occur, it is expected to be mild 
and temporary.  Any event will stop when the study participant stops being exposed to the 
instrument or returns to their normal oral hygiene.  At all examinations, each participant will be 
asked if they have experienced any discomfort or oral irritation. The investigators will record any 
and all adverse reactions and report this documentation to the Product Safety Assurance 
Department.  The nature of the reaction and any correlation with product usage will be assessed.  
If the evidence indicates that the adverse reaction may be due to product usage, the study 
participant will be instructed to discontinue product use and appropriate treatment will be provided.  
In the event of an adverse experience, emergency or other problems or questions regarding your 
participation in this study participants can contact the following investigators: 
 
Dr. Michael Golderg at (416) 979-4928 ext. 4408 or the Research Oversight and Compliance 
Office - Human Research Ethics Program at ethics.review@utoronto.ca or Tel: 416 946-3273 
In the event of a medical emergency, please contact your physician 
 
9.1 Definitions 
Adverse Events (AEs) and Serious Adverse Events (SAEs) are defined by the ICH Guideline for 
Good Clinical Practice (ICH GCP) as follows: 

Protocol_V8_October_11, 2025 
 
16 
 
Adverse Event:  Any untoward medical occurrence in a study participant or clinical 
investigations study participant administered a pharmaceutical product and which does not 
necessarily have to have a causal relationship with this treatment.  An adverse event (AE) 
can therefore be any unfavorable and unintended sign (including abnormal laboratory 
finding), symptom, or disease temporally associated with the use of a medicinal product, 
whether or not considered related to the medicinal product. 
 
AEs include any clinically significant deterioration of a study participant’s medical status, after 
being enrolled and signing an Informed Consent Form.  The AE may involve any organs or systems 
and can be represented by the new onset or the deterioration of a disease, a syndrome, a symptom, 
a physical sign, as well as by findings and results of instrumental examinations and laboratory 
tests.  Any medically relevant and untoward change from study enrollment, including frequency 
or pattern changes for a fluctuating condition (e.g., migraine), occurring after signing an informed 
consent is an adverse event.  All such occurrences must be recorded and reported accordingly, 
whether they appear causally related to the study medication, or not. 
 
Serious Adverse Event: Any adverse event occurring at any dose that results in any of the 
following outcomes: 
• Death 
• Life threatening adverse event 
• Instudy participant hospitalization, or prolongation of existing hospitalization 
• Persistent or significant disability/incapacity 
• Congenital anomaly/birth defect 
• Temporary loss of daily function 
Important medical events that may not result in death, be life-threatening, or require 
hospitalization may be considered a serious adverse experience when, based upon 
appropriate medical judgment, they may jeopardize the study participant or study participant 
and may require medical or surgical intervention to prevent one of the outcomes listed in this 
definition. 
 
Examples of such important medical events include allergic bronchospasm requiring intensive 
treatment in an emergency room or at home, blood dyscrasias or convulsions that do not result in 
instudy participant hospitalization, or the development of drug dependency or drug abuse. 
 
IMPORTANT NOTES: The concepts of Adverse Event / Experience1 (AE) and Serious Adverse 
Event / Experience (SAE) represent regulatory instruments used to evaluate and monitor the 
safety of clinical trial study participants.  Therefore, these terms only apply in light of their 
regulatory definition.  The term “serious”, in a regulatory sense, does not necessarily mean 
“severe”.  All adverse events (serious and non-serious) reported during a study will be taken into 
account when analyzing the study data and establishing the safety profile of the investigational 
drug, and will be included in the final study report.  The SAE concept is primarily used to identify, 
 
 

Protocol_V8_October_11, 2025 
 
17 
during the conduct of the trial, those adverse events that may require an expedited reporting 
procedure to regulatory authorities. 
 
Death: The outcome of death requires that the AE that resulted in death be reported as an SAE.  
Death, in and of itself, is not an AE; it is only an outcome.  The cause of death is the AE; therefore, 
the investigator should make every effort to obtain and document the cause of death for all study 
participants who die during the study.  If, despite all efforts, the cause of death remains unknown, 
the AE it should be documented as “unspecified fatal event”. 
 
Life-threatening Adverse Event:  Any adverse event that places the study participant or study 
participant, in the view of the investigator, at immediate risk of death from the reaction as it 
occurred (i.e., it does not include reaction that had it occurred in a more severe form, might have 
caused death). 
 
Hospitalization:  It should be noted that hospitalization, in and of itself, does not represent an 
SAE.  It is the adverse event leading to the study participant’s hospitalization that becomes 
“serious” when it requires instudy participant care.  Consequently, SAE should not be reported in 
case of pre-planned hospitalizations for pre-existing conditions that did not worsen during the 
study. 
 
Disability: A substantial disruption of a person’s ability to conduct normal life functions. 
 
9.2 Documenting and Reporting Adverse Events 
 
9.2.1 General Procedures for All Adverse Events 
All clinical complaints, symptoms, or signs that meet the adverse event definition will be recorded 
on the Adverse Reaction Form (Appendix VIII) using a recognized medical term or diagnosis that 
accurately reflects the event.  Source documentation should be maintained that allows for clear 
identification of each adverse event and the following parameters required for the form: 
• AE description 
• Date of onset 
• Date of resolution 
• Outcome 
• Severity 
• Seriousness 
• Relationship to study drug (causality) 
• Actions taken 
Adverse events will be assessed by the investigator or designee for severity, relationship to the 
study product, possible etiologies, and whether the event meets the criteria as a serious adverse 
event and therefore requires immediate notification of the sponsor. 
 
For data collection purposes, the outcome of all adverse events recorded on the Adverse Reaction 
Form will be designated as of the completion of the final evaluation or examination.  However, 
the investigator is responsible for following all adverse events until resolution or until no longer 
of clinical concern, and providing these data to the sponsor. 

Protocol_V8_October_11, 2025 
 
18 
 
9.2.2 Reporting Procedures for Serious Adverse Events 
Any adverse event that is serious or potentially serious requires additional detailed reports and 
follow-up.  A serious adverse event must be reported via telephone to the sponsor’s representative 
immediately (within 24 hours) so as to facilitate discussion and implementation of necessary 
follow-up measures, and to enable the sponsor to submit necessary reports to regulatory authorities 
and other investigators.  Following the initial telephone notification, the investigator must 
complete and submit a Serious Adverse Event Report Form to the sponsor within one calendar 
day.  Serious Adverse Event  
Report Forms will be provided to the investigator upon initiation of the study.  Once the sponsor 
reviews the Serious Adverse Event Report Form, additional information may be requested from 
the investigator to allow appropriate medical evaluation and determine the regulatory reporting 
requirements. 
The investigator is responsible for following all adverse events, especially those deemed “serious”, 
until resolution or until the event is no longer of clinical concern, and for providing these data to 
the sponsor in an agreed-upon format.  The investigator is also responsible for reporting all serious 
adverse events to the Institutional Review Board (IRB) overseeing the conduct of the study at the 
respective study center, according to the rules and procedures established by the IRB. 
 
Safety will be evaluated by oral examinations, clinical laboratory evaluations, descriptive 
analysis of AEs (including incidence, severity, seriousness, and relatedness), and immune 
response, over the entire period of the study.  
 
9.2.3 Reporting Adverse Events to Health Canada 
All Adverse Events will be reported to Health Canada as per the Natural Health Product 
Regulations. This report must be submitted to Health Canada by the researchers no later that 
7 days after they have been notified. The online form is available at: 
 
https://www.canada.ca/en/health-canada/services/drugs-health-products/medeffect-canada/adverse-reaction-
reporting/natural-health-products.html 
 
 
10. COMPENSATION 
Participants of the study will receive monetary compensation of 1000 CAN$ after completion of 
all trial visits. In addition, they will receive 2 professional dental cleaning treatments prior and 
subsequent to stopping all oral hygiene procedures. They will also receive proper oral hygiene 
instructions and 3 kits for at-home daily hygiene during the trial.  
 
11. QUALITY ASSURANCE/CONDUCT OF THE STUDY 
This clinical research study will be conducted in compliance with this protocol and U.S. Federal 
Regulations governing informed consent (21 CFR 50), Institutional Review Board (21 CFR 56), 
applicable regulations governing Investigator conduct (21 CFR 312) and/or any local regulatory 
agency (where applicable). 
 
It is the responsibility of the Investigator to ensure that all study participant data are collected 
and reported according to the study protocol. Study participant records will indicate study 

Protocol_V8_October_11, 2025 
 
19 
participant and examination information such as visit dates, examiners, etc. that is unique to this 
study and the study participants. Proper documentation of all adverse events and final resolutions 
will be maintained. Case report forms will be used for recording all clinical data. All CRFs used 
in this study will be provided by the study sponsor.  The Investigator will be responsible for 
maintaining original consent forms, case report forms (CRFs), and other source documentation. 
 
These are examples of case report forms that can be used to collect information/data while 
conducting this study: 
• Study participant Assignment Log (Randomization chart) 
• Demographic Log 
• Inclusion/Exclusion Criteria Form 
• Continuance Criteria   
• Adverse Event Form  
• Oral Hard & Soft Tissue Exam Form 
Forms may be added or deleted, if deemed necessary, by agreement between the PI (principal 
investigator) and study sponsor prior to the study start date. 
 
12. DATA MANAGEMENT RESPONSIBILITIES 
Data collection is the responsibility of the clinical research site staff under the direction of the 
Principle Investigator. The Investigator will keep a copy of every document (clinical and 
laboratory) related to the research study. 
 
Data generated at the clinical site will be transferred to the sponsor by the Principal Investigator 
via mail or electronic email for statistical analyses. The principal Investigator and study manager 
are responsible for the review and interpretation of the analyzed data.  
 
12.2 Study participant Termination/Procedures 
A genuine effort will be made to determine the reason(s) why a study participant fails to return for 
the necessary visit(s) or is dropped from the study. Study participants could be dropped from the 
study if any of the following occur: 
1. Study participant fails to substantially comply with the protocol requirements. 
2. Study participant fails to report for a scheduled examination. 
3. Study participant is treated with medications during the course of the study, which may 
interfere with the parameters of the study. 
4. Study participant develops an adverse reaction.  The study Investigator will immediately notify 
the study monitor and information will be recorded on Adverse Reaction Form. 
5. Study participant receives emergency dental or medical treatment, which may interfere with 
the parameters of the study. 
6. Sponsor elects to terminate the study. 
7. Study participant elects to terminate participation in the study. 
12.3 Removal of Study participants From Study 
Study participants will be dropped from the study if they receive emergency dental 
treatment, which in the opinion of the monitor could influence the parameters of the study. 
Any study participant treated with an antibiotic during the study will be dropped from the 
study. Either the investigator or the sponsor may terminate the study at any time for well 

Protocol_V8_October_11, 2025 
 
20 
documented reasons, provided written notice is submitted at a reasonable time in advance 
of the intended termination. 
 
12.4 Pregnancy 
No pregnant women will intentionally be enrolled in this study. All female study participants being 
considered for this study will be screened with a urine pregnancy test before the start of the trial.  
All enrolled female study participants will be required to report to the clinical/research investigator 
if they become pregnant during the course of the 9-week study. In the event a woman enrolled in 
this clinical research study becomes pregnant at any time during this study, participation in this 
study will be terminated upon the clinical staff’s notification of the event. The study participant’s 
medical records used in this study will be updated to reflect the pregnancy and there will be follow- 
up contact until the end of the pregnancy to record the outcome in the clinical file. 
 
12.5 Ethical and Regulatory Requirements/Human Study participant Protection 
Participants may opt out of the study at any time and request the destruction of all stored samples. 
Sample destruction and disposal will be completed and properly documented by Coriell. All 
samples and data will be relayed to researchers in the absence of identifying information; therefore, 
no personal information or results will be relayed back to participants. The testing done in this 
study is for research purposes only and not intended to make any medical diagnosis. 
 
12.6 Protocol Approval, Study Monitoring and Compliance 
Prior to initiation of the study, the Investigator will obtain approval from the UofT Health Sciences 
Research Ethics Board for the study protocol, the informed consent document, study instructions, 
and any forms of advertising in compliance with regulations. The reviewers will also review any 
change(s) in the protocol before the change is initiated.  
 
12.7 Adherence to Protocol/Amendment(s) 
The Investigator will be required to adhere to the final protocol. Any changes to the protocol, 
except those necessary to eliminate apparent hazards, will require prior approval by the local 
reviewers through the submission of a protocol amendment. These changes to the protocol must 
be implemented only through formal written protocol amendments and only upon joint approval 
by the sponsor and investigator.  If a protocol amendment requires changes to the informed consent 
form, the revised consent form must also be approved by any local board. 
 
Departures from eligibility requirements may be allowed on a case-by-case basis by the medical 
monitor or other authorized sponsor representative.  Such departures must be medically and 
scientifically justified, must be pre-authorized, and must be documented in the case report form 
and tracked as official eligibility waivers. 
 
12.8 Institutional Review Board 
Prior to initiation of the study, the Investigator will obtain Institutional Review Board approval for 
the study protocol, the informed consent document, study instructions, and any forms of 
advertising in compliance with regulations. The IRB will review the investigation at least once a 
year and will review any significant change in the protocol before the change is initiated. The 
Investigator will maintain all original correspondence. IRB approval for this study will be obtained 

Protocol_V8_October_11, 2025 
 
21 
from the U of T Health Sciences Research Ethics Board Tel: +1 416 946-3273 
ethics.review@utoronto.ca 
 
12.9 Advertising 
No newspaper, radio, or television advertising will be used for recruitment purposes. A flyer was 
created to recruit participants and will be approved by an Institutional Review Board/Ethics Board 
prior to posting or dissemination (Appendix 1). 
 
12.10 Informed Consent Process 
Written informed consent will be obtained from all study participants prior to their enrollment into 
the study. The purpose and description of the study in lay language, possible adverse reactions, 
risks and benefits of participation and the study participant’s right to withdraw without prejudice 
at any time must be explained to each study participant in the presence of a witness.  The study 
participant must read, understand and sign the informed consent form provided (Appendix 10)/The 
informed consent form and any other written information for study participants should meet local 
requirements of language and interpretation (i.e., non-English speaking study participants must be 
presented with informed consent forms in a language that they can understand). The consent form 
will comply with all applicable regulations governing protection of the participating study 
participants in the study, and include basic elements specified in the U. S. Code of Federal 
Regulations, 21 CFR 50.25(a) and 50.27 and ICH-GCPs, Chapter 4, subpart 4.8. The Informed 
Consent form will be reviewed and approved by the clinical site’s IRB.  
 
Each study participant will be given unlimited time to read the consent form and ask questions. 
Study participants who agree to participate will be asked to sign and date an IRB-approved 
informed consent form. A copy of the signed and dated consent form will be given to each study 
participant prior to their participation in the study. The original signed and dated informed consent 
document will be retained by the Investigator. All informed consent forms will be documented in 
a log by date and study participant ID; the log will be kept as source documentation. All study 
procedures must be explained in non-technical terms. Study personnel will assure that participants 
are clearly informed regarding their roles and obligations to protect vulnerable study participants 
and ensure they are not under coercion or undue influence.  Study participants have the right to 
withdraw consent at any time.  
 
12.11 Confidentiality 
All records of study participant participation in this study are confidential and these records are 
available only to the investigator, specifically trained site personnel, supervising dentist/examiner, 
potentially the sponsoring company and Ethics/Institutional Review Board (IRB). In addition, the 
identity of participating study participants must be protected. 
 
Only investigators (and specifically trained clinical site staff) will collect and have access to a 
study participant’s private information (e.g., name, medical records, etc.). Investigators will assign 
a study number to study participants which will be used to conceal their identity on all case report 
forms and other documents prior to their sharing with the broader study team, including the 
sponsor. Documents that identify the study participant by name (e.g., the signed informed consent 
form or health questionnaire) will not be transferred or submitted to the sponsor and will be 
maintained in strict confidence by the investigator, except to the extent necessary to allow auditing 

Protocol_V8_October_11, 2025 
 
22 
by the sponsor personnel to verify study participant, product safety and study compliance. The 
results of the study may be published in a scientific journal or a government public clinical 
database. If any publication occurs, only the study participant’s study number/ID, gender and/or 
age may be used.  
 
12.12 New Findings 
Study participants will be informed of any significant new findings related to study products or 
procedures when they become known during the course of this clinical research study. Such 
information may affect the study participant’s decision to continue participation in the study. 
ADMINISTRATIVE ASPECTS 
 
12.13 Curriculum Vitae 
The investigator will complete the FDA form 1572 and provide the sponsor with copies of his/her 
curriculum vitae and those of all sub-investigators listed on the form, at sponsor’s request. 
 
12.14 Data Collection in the Case Report Form 
All study data will be recorded in the case report form supplied by the sponsor. All entries will be 
written clearly in black ink. Only the principal investigator, sub-investigators, or study 
coordinators may make entries in the case report forms. If erroneous data are entered on the case 
report forms, corrections to the data must be made by crossing out the incorrect entry with a single 
line (such that the initial entry remains legible) and entering the correction. All corrections on a 
case report form will be initialed and dated by the investigator, sub-investigator, or study 
coordinator making the correction. 
 
12.15 Documentation of Consent and Storage of Study Documents 
All informed consent forms will be documented in a log by date and study participant ID; the log 
will be kept as source documentation. Informed consent forms will be stored in a secure locked 
room designated for research charts storage in the clinical site. The Investigator will provide each 
study participant with a clear and understandable consent letter regarding the processing in 
connection with the study of personal data (i.e., any information relating to an identified or 
identifiable individual) by the investigator, sponsor and other persons involved in the study 
(Appendix II). Each study participant will be given a copy of the consent letter/form which will be 
referenced as part of the informed consent process. 
 
Any deviation to the consent letter as regards processing by the sponsor (including its vendors, 
monitors and other representatives) must be agreed with the sponsor. In addition, the Investigator 
is responsible for ensuring that all study study participants understand and complete the consent 
letter/form, and that it includes any local requirements regarding data security and privacy laws 
and regulations applicable to the study and the study participants. Should the consent letter/form 
fail to meet any applicable local requirements, the Investigator is responsible for amending the 
consent letter/form to bring it into compliance with local applicable regulations and the 
Investigator (including Principal and sub-Investigators) agrees to indemnify and be liable to the 
Sponsor for any damages resulting from such non-compliance. Study participants' study charts 
will be stored in a secure locked room designated for research charts storage in the clinical site. 
The chart room is a limited- access area and only delegated study personnel will have access to 
the study charts and study participants' data. 

Protocol_V8_October_11, 2025 
 
23 
 
12.16 Study Management 
Under the direct supervision of the Principal Investigator, certain duties may be delegated during 
the course of the study. These responsibilities will be documented on the Transfer of 
Responsibilities form maintained in the Investigator’s clinical file for the study. 
 
12.17 Study Monitoring 
Appropriate phone calls or visits will be made by the monitor to the investigative site in order to 
review the protocol and to ascertain that the facility is adequate for satisfactory conduct of the 
study, as well as to discuss the obligations of both the sponsor and the investigator. 
 
The investigator will permit a representative of the sponsor or the sponsor’s designate and the 
local regulatory authorities, if requested, to inspect all case report forms and corresponding 
portions of the study study participants’ original office, hospital and/or clinic medical records at 
regular intervals throughout the study. These inspections are for the purpose of assessing the 
progress of the study, verifying adherence to the protocol, determining the completeness and 
exactness of the data being entered on the case report forms and assessing the status of study drug 
storage and accountability.  During on-site visits, case report forms will be examined by the study 
monitor(s) and the data verified by comparison with corresponding source data (such as clinic, 
hospital and/or office records).  Due to the geographic distance of the clinical site from the 
sponsor, phone conferences or teleconferences may be used to complete the monitoring 
requirement. 
 
12.18 Final Report 
Following the completion of the study, the Investigator shall prepare a final study report. The final 
report will include a general description of the conduct of the study including protocol deviations, 
study participant withdrawals, discussion of any adverse events, safety and efficacy data, 
laboratory data, and statistical analysis of the data if available. This report will be approved and 
signed by the Principal Investigator. 
 
12.19 Record Retention and Access to Source Data/Documents 
Source documents must be kept for at least fifteen (15) years after terminating the study. The 
Investigator will maintain all study documentation for all study participants entered into the study 
in a secure area ensuring the confidentiality of the information collected. Securing records includes 
placing written forms in locked file cabinets and/or sealed and labeled storage boxes in a locked 
room. Access will be denied to all persons with the exception of the Principal Investigator and his 
designees.  The Sponsor will be notified before any destruction of study documents occurs. 
 
12.20 Publication 
All manuscripts or presentations based upon this study, including press statements and internal 
public notices and memoranda must be submitted to the sponsor for review and approval prior to 
release for publication or presentation.  This review period will be up to 60 days in duration.  
Review of an abstract may be expedited in some circumstances. 
 
13. References 
 

Protocol_V8_October_11, 2025 
 
24 
Albandar, J. M. & Rams, T. E. (2002) Global epidemiology of periodontal diseases: an 
overview. Periodontol 2000 29, 7-10. 
 
Barbour, A., & Philip, K. Variable characteristics of bacteriocin-producing Streptococcus 
salivarius strains isolated from Malaysian Study participants. PLoS One 2014, 9(6), e100541. 
 
Barbour, A., Wescombe, P., & Smith, L. Evolution of Lantibiotic Salivarcins: New Weapons to 
Fight Infectious Diseases. Trends in Microbiology 2020 28(7), 578-593. 
 
Berezow Alex, B. & Darveau Richard, P. (2010) Microbial shift and periodontitis. Periodontol 
2000 55, 36-47. doi:doi:10.1111/j.1600-0757.2010.00350. 
 
Gallagher, H., Ramsay, S. C., Barnes, J., Maggs, J., Cassidy, N. & Ketheesan, N. (2006) 
Neutrophil labeling with [(99m)Tc]-technetium stannous colloid is complement receptor 3-
mediated and increases the neutrophil priming response to lipopolysaccharide. Nucl Med Biol 
33, 433-439. doi:10.1016/j.nucmedbio.2005.12.014. 
 
Gunsolley, J. C. (2006) A meta-analysis of six-month studies of antiplaque and antigingivitis 
agents. J Am Dent Assoc 137, 1649-1657. 
 
Kerns KA, Bamashmous S, Hendrickson EL, Kotsakis GA, Leroux BG, Daubert DD, et 
Localized microbially induced inflammation influences distant healthy tissues in the human oral 
cavity. Proc Natl Acad Sci. 2023 Oct 10;120(41):e2306020120. 
 
Loe, H., Theilade, E. & Jensen, S. B. (1965) Experimental Gingivitis in Man. J Periodontol 36, 
177-187. doi:10.1902/jop.1965.36.3.177. 
 
van der Weijden, F. & Slot, D. E. (2011) Oral hygiene in the prevention of periodontal diseases: 
the evidence. Periodontol 2000 55, 104-123. doi:10.1111/j.1600-0757.2009.00337.x. 
 
Wellappuli N, Fine N, Lawrence H, Goldberg M, Tenenbaum H & Glogauer M (2017) Oral and 
Blood Neutrophil Activation States during Gingivitis. JDR Clinical & Translational Research 
Vol 3, Issue 1, pp. 65 – 75 
 
 
 
 
 
 
 
 
 
 
 
APPENDICES 
 

Protocol_V8_October_11, 2025 
 
25 
Appendix 1. Table Clinical Trials Utilizing S. Salivarius 
 
Study Design 
Population 
Dose and 
Duration 
Safety-
Related 
Outcomes 
Reference 
Prospective, 
randomized, 
single-blind, 
placebo-
controlled 
pilot study 
Children (M & F; 6 to 11 y) who had at 
least 3 episodes of microbiologically 
documented infections with clinical 
symptoms suggesting Group A beta-
hemolytic Streptococcus (GABHS) 
pharyngitis. 
 
Initial n = 84 
Final n = 82 
Test: S. salivarius 
24SMB and S. 
oralis. The mix 
suspension 
consisted of a 
minimum of 125 × 
109 (CFU/mL) in 
10 mL of saline. 
Control: matched 
placebo 
 
2 puffs once a day, 
with an oral spray 
that provided 2 × 
109 CFU per puff. 
 
For 3 months 
All study 
participants 
were compliant 
to the assigned 
intervention to 
them (> 80% 
compliance). 
Only four 
study 
participants 
reported an 
adverse 
event—three in 
the probiotic 
group and one 
in the control 
group. A mild 
cough was 
reported by 
two study 
participants in 
the probiotic 
group versus 
one study 
participant in 
the control 
group. Only 
one study 
participant in 
the probiotic 
group reported 
nausea.  
 
(Andaloro 
et al., 
2019) 
Double-blind, 
randomized, 
two-arm 
parallel-group  
Healthy adults (M & F; 20 to 24 y) 
 
Initial n = 31 
Final n = 30 
Test: S. salivarius 
K12 (>1 x 109 CFU 
in 1 tablet) 
Control: matched 
placebo lozenge 
 
1 lozenge per day 
 
For 4 weeks 
One participant 
from the 
probiotic group 
was lost in 
follow-up due 
to sickness not 
related to the 
intervention 
(COVID-19). 
No adverse 
effects were 
registered. 
(Babina et 
al., 2022) 
Randomized, 
open-label, 
controlled, 
parallel 
Children (M & F; 4 to 10 y) with black 
teeth stains 
 
initial n = 58 (29/group) 
final n = 54 
Test: no less than 
1×10⁹ CFU/day 
Control: no 
intervention was 
administered to the 
Four 
participants 
(one in the test 
group and 
three in the 
(Bardellini 
et al., 
2020) 

Protocol_V8_October_11, 2025 
 
26 
control group 
 
For 3 months 
control group) 
were excluded 
from the study 
because they 
started 
antibiotic 
therapy. 
However, the 
study authors 
did not report 
any adverse 
events 
observed by 
the 
participants. 
Randomized, 
triple-blind, 
placebo-
controlled, 
parallel 
Participants (M & F; 10 to 30 y) wearing 
orthodontic braces 
 
initial n = final n = 64 
Test: 7.2×10⁹ 
CFU/day 
Control: matched 
placebo lozenge 
 
For 1 month 
According to 
the study 
authors, no 
adverse events 
were observed 
or recorded 
during the trial. 
(Benic et 
al., 2019) 
Randomized, 
double-blind, 
placebo 
controlled 
Healthy adults performing regular 
exercise training (M & F; 20 to 25 y) 
 
Initial n = 24 
Final n = 20 
Test: 5x109 
CFU/day 
Control: matched 
placebo tablet 
 
For 30 days 
The study 
authors did not 
report any 
adverse events 
observed by 
the 
participants. 
Four study 
participants 
abandoned the 
study for 
personal 
reasons. The 
authors 
showed an 
adherence rate 
of 94%. 
(Bertucciol
i et al., 
2023) 
Randomized, 
parallel. 
Participants 
were blinded. 
Blinding of 
investigators 
was not 
reported. 
Healthy adults (18 y and older; average 
age 19 y; gender not reported) 
 
initial n = 75 
final n = not reported 
Test 1: 1×10⁶ 
CFU/day 
Test 2: 1x10⁷ 
CFU/day 
Test 3: 1x10⁸ 
CFU/day 
Test 4: 1x10⁹ 
CFU/day 
 
For 28 days 
The study 
authors did not 
provide 
information on 
any adverse 
events that 
may have been 
experienced by 
the study 
participants. 
(Burton, 
Wescombe 
et al., 
2013) 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Children (M & F; 5 to 10 y) with a 
history of dental caries 
 
initial n = 100 
final n = 83 
Test: 7.2×10⁹ 
CFU/day 
Control: matched 
placebo lozenge 
 
For 3 months 
11 participants 
withdrew for 
various 
reasons, 
including not 
liking the taste 
of the lozenges 
(Burton et 
al., 2013) 

Protocol_V8_October_11, 2025 
 
27 
(n=6), protocol 
deviations 
(n=1), and 
being lost to 
follow-up 
(n=4). In 
addition, data 
from six 
participants 
were excluded 
from the 
analysis 
because they 
did not comply 
with the study 
requirements, 
consuming less 
than 75% of 
the prescribed 
lozenges per 
month. 
Open-label, 
single-arm 
Healthy adults (M & F) 
n = 14 
4×10⁹ CFU/day, 3 
days 
No adverse 
effects 
(Burton et 
al., 2006) 
Open-label, 
observational 
Healthy adults (M & F)  
n = 23 
Test: >1×10⁹ 
CFU/lozenge 
Control: placebo 
lozenge 
 
3-day regimen of 
CHX rinsing, 
followed by intake 
of lozenges (test, 
control) at 2h 
intervals over 8h 
for 3 days (>4×10⁹ 
CFU/day). 
Subsequently, 
study participants 
in the test group 
(n=13) took the 
lozenge twice daily 
(morning & night) 
for 2 weeks. Two 
of these study 
participants 
continued to take 2 
lozenges/day for 28 
days (>2×10⁹ 
CFU/day). 
 
For 3 days, 2 
weeks (test group 
only), 28 days (2 
study participants 
in test group only) 
The study 
authors did not 
report any 
adverse events 
observed by 
the 
participants. 
(Burton, 
Chilcott et 
al., 2006) 

Protocol_V8_October_11, 2025 
 
28 
Randomized, 
parallel 
 
Blinding not 
reported 
Healthy adults (M & F; mean age 19 y) 
 
initial n = 100 
final n was not reported 
Test 1: 1.5×10⁹ 
CFU/day 
Test 2: 1.1×10⁸ 
CFU/day 
Test 3: 2×10⁷ 
CFU/day 
Test 4: 1×10⁶ 
CFU/day 
Test 5: 7.5×10⁴ 
CFU/day 
 
for 14 days 
The 
participants did 
not report any 
adverse 
reactions. 
(Burton et 
al., 2010) 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Healthy adults (M & F; age 20 to 60 y) 
 
initial n = 56 
final n = 53 
Test: 1.1×10¹⁰ 
CFU/day 
Control: matched 
placebo 
 
For 28 days 
There was no 
significant 
difference 
found between 
the test and 
control groups 
in terms of oral 
health and 
gastrointestinal 
symptoms 
assessed using 
a 10-point 
VAS. No 
serious adverse 
events 
occurred in 
either group 
and the 
proportion of 
participants 
reporting any 
adverse events 
was similar in 
both groups. 
Hematology 
and clinical 
chemistry 
parameters 
showed no 
significant 
difference 
between the 
groups. 
However, the 
specific gravity 
of the urine 
was slightly 
higher in the 
placebo group 
compared to 
the test group, 
but still within 
normal limits. 
(Burton et 
al., 2011) 

Protocol_V8_October_11, 2025 
 
29 
Double-blind, 
randomized 
placebo-
control, 
prospective 
trial 
Healthy adults (M & F; age 21 to 45 y) 
who 
practice good oral hygine 
 
Initial n = 25 
Final n = 24 
Test: powder 
sachet (1.5 g) 
contained 
approximately 7.77 
x 109 CFU of L. 
acidophilus DDS-
1®, 
8.25 x 109 CFU of 
B. lactis UABla-
12, and 2 x109 CFU 
of S. salivarius 
BLIS K12TM 
Control: matched 
placebo 
 
one daily dose (one 
1.5 g sachet) of 
probiotic powder 
mixed with any 
liquid. 
 
For 2 weeks 
The authors 
found that the 
treatment was 
well tolerated 
by the study 
population 
except for one 
participant 
who 
experienced 
hives during 
supplementatio
n which 
disappeared 
when 
supplementatio
n ceased. All 
other 
participants 
were 
compliant. The 
overall attrition 
rate for this 
study was 4%. 
(Cernioglo 
et al., 
2021) 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Infants (M & F, 7 to 13 months) with 
high risk of acute otitis media 
 
initial n = 224 
final n = 202 by 2-month visit, 166 by 
12-month visit 
Test: follow-up 
formula containing 
proB (S. 
thermophilus NCC 
2496, S. salivarius 
DSM 13084 [K12], 
L. rhamnosus LPR 
CGMCC 1.3724) 
and preB 
[Raftilose/Raftiline
]) 
Control: follow-up 
formula only 
 
Formula contained 
2.5×10⁷ CFU/g S. 
salivarius (1×10⁹ 
to 2×10⁹ CFU/day 
according to GRN 
807). 
 
For 12 months 
The study 
authors 
reported that 
both the test 
and control 
formulas were 
well-tolerated. 
The main 
reason for 
discontinuation 
was non-
compliance 
with the study 
protocol, 
specifically, 
missing three 
consecutive 
days per month 
with less than 
300 mL of 
milk consumed 
per day. The 
majority of the 
reported 
adverse events 
(93.1%) were 
not considered 
related to the 
study. There 
were five 
adverse events 
considered 
(Cohen et 
al., 2013) 

Protocol_V8_October_11, 2025 
 
30 
likely related, 
with four in the 
test group and 
one in the 
control group. 
These included 
lack of appetite 
for milk, 
regurgitation, 
dry skin, 
chronic 
diarrhea, and 
abdominal 
pain. One 
adverse event, 
constipation, 
was considered 
related, but no 
further details 
were provided. 
Open-label 
 
Study was not 
randomized 
and not 
blinded. 
Healthy study participants (M & F; 19 to 
64 y) 
 
Initial n = Final n = 23 
Test: 
Streptococcus 
salivarius 24SMBc 
and Streptococcus 
oralis 
89a nasal spray 
(10⁹ CFU/dose) 
 
two bilateral spray 
injections 
into each anterior 
nostril/day  
 
For 7 days. 
 
No severe side 
effects were 
recorded in any 
enrolled study 
participants 
after the 
treatment. 
Only 
10% of the 
study 
participant 
suffered from 
allergic cold, 
and 80% did 
not show any 
problems 
breathing. 
However, 90% 
of people 
reported nasal 
dripping 
immediately 
after the 
treatment 
administration. 
(De 
Grandi et 
al., 2019) 
Open-label 
 
Study was 
randomized 
but not 
blinded. 
Children (M & F; 6 to 17 y) at high risk 
for dental caries 
 
initial n = final n = 76 
Test: no less than 
1×10⁹ CFU/day 
Control: no 
intervention was 
administered to the 
control group 
 
For 90 days 
No participants 
withdrew. S. 
salivarius M18 
was found to 
be safe with no 
treatment-
related side 
effects, and no 
participants 
dropped out. 
The majority 
(Di Pierro, 
Zanvit et 
al., 2015) 

Protocol_V8_October_11, 2025 
 
31 
of participants 
(35 out of 38) 
rated the 
tolerability of 
the treatment 
as "good" or 
"very good," 
and the 
remaining 
three 
participants 
rated it as 
"acceptable." 
Open-label 
 
Study was not 
randomized 
and not 
blinded. 
Children (M & F; 3 to 12 y) with and 
without recurrent streptococcal 
pharyngitis and/or tonsillitis 
 
initial n = 82 
final n = 78 
Test: 5×10⁹ 
CFU/day 
Control 1: no 
intervention was 
administered to 
controls with 
recurrent illness 
Control 2: no 
intervention was 
administered to 
controls without 
recurrent illness 
 
For 90 days 
The test tablet 
was reported to 
be well-
tolerated, and 
there were no 
observed side 
effects. 
However, four 
participants in 
the test group 
were excluded 
from the 
analysis due to 
non-adherence 
to the study 
protocol, 
specifically, 
missing over 
20 days of 
treatment. 
(Di Pierro 
et al., 
2012) 
Open-label 
 
Study was not 
randomized 
and not 
blinded. 
Adults (M & F; 18 to 65 y) with 
recurrent oral streptococcal pharyngitis 
 
initial n = final n = 40 
Test: 5×10⁹ 
CFU/day 
Control: no 
intervention was 
administered to the 
control group 
 
For 90 days 
All 20 study 
participants 
who received 
the test tablets 
successfully 
completed the 
study, and no 
participants 
dropped out. 
The study 
authors 
reported that 
the test tablet 
was well 
tolerated, and 
no side effects 
related to the 
treatment were 
reported. 
(Di Pierro 
et al., 
2013) 
Open-label 
 
Study was 
randomized 
Children (M & F; 3 to 13 y) with 
recurrent oral streptococcal disorders 
 
Test: no less than 
1×10⁹ CFU/day 
Control: no 
intervention was 
The study 
authors 
reported that 
the test tablet 
(Di Pierro 
et al., 
2014) 

Protocol_V8_October_11, 2025 
 
32 
but not 
blinded. 
initial n = 61 
final n = 60 
administered to the 
control group 
 
For 90 days 
was well 
tolerated and 
did not cause 
any notable 
side effects. 
However, one 
study 
participant 
dropped out of 
the study 
immediately 
after enrolment 
due to the poor 
taste of the test 
product. 
Open-label, 
single-arm 
Children (M & F; 3 to 9 y) with recurrent 
secretory otitis media 
 
initial n = final n = 22 
Test: no less than 
1×10⁹ CFU/day 
 
For 90 days 
The safety 
profile of S. 
salivarius K12 
was reported as 
very good, 
with no 
treatment-
related side 
effects and no 
study 
participant 
drop out. 
Tolerability 
was rated as 
good or very 
good in 20 of 
the 22 study 
participants, 
with the 
remaining 2 
study 
participants 
rating it as 
acceptable. 
(Di Pierro, 
Di 
Pasquale 
et al., 
2015) 
Open-label 
 
Study was 
randomized 
but not 
blinded. 
Children (M & F; 3 to 10 y) with 
recurrent streptococcal 
pharyngotonsillitis 
 
initial n = final n = 124 
Test: no less than 
1×10⁹ CFU/day 
Control: no 
intervention was 
administered to the 
control group 
 
For 90 days 
According to 
the study 
authors, the 
use of S. 
salivarius K12 
was well-
tolerated and 
highly 
compliant 
among the 
participants, 
and there were 
no reported 
side effects 
observed 
during the trial. 
(Di Pierro, 
Colombo 
et al., 
2016b) 

Protocol_V8_October_11, 2025 
 
33 
Open-label 
 
Study was 
randomized 
but not 
blinded. 
Healthy children (M & F; 33 to 45 
months) 
 
initial n = final n = 222 
Test: no less than 
1×10⁹ CFU/day 
Control: no 
intervention was 
administered to the 
control group 
 
For 180 days 
All children 
who were 
enrolled in the 
study 
completed it. 
The study 
authors noted 
that there were 
no observable 
side effects in 
the treatment 
group, both 
during the 
treatment 
period and the 
follow-up 
period. 
(Di Pierro, 
Colombo 
et al., 
2016a) 
Retrospective 
observational 
Children (M & F; 3 to 14 y) with 
recurrent non-streptococcal infection 
 
n = 133 
Test: no less than 
1×10⁹ CFU/day 
 
For 90 days 
The study 
authors 
reported that 
compliance 
and tolerability 
were excellent. 
They also 
noted only one 
side effect, 
which occurred 
in a 6-year-old 
boy who 
experienced a 
mild episode of 
bronchospasm 
after a few 
days of 
treatment with 
S. salivarius 
K12. However, 
the study 
participant was 
able to 
continue with 
the study 
without any 
further 
incidents. 
(Di Pierro 
et al., 
2018) 
 Randomized, 
placebo 
controlled, 
parallel 
 
Blinding not 
reported 
Children at high risk of acute rheumatic 
fever (M & F; 5 to 14 y) 
 
initial n = 1314 
final n = 1137 
Test: 2.5×10⁹ 
CFU/day 
Control: matched 
placebo lozenge 
(*b) 
 
209 days 
The study 
authors did not 
provide any 
information 
about adverse 
events 
experienced by 
the 
participants. 
However, they 
reported that 
(Doyle et 
al., 2018) 

Protocol_V8_October_11, 2025 
 
34 
most children 
found the 
lozenges to be 
well accepted, 
with only two 
children 
refusing to take 
them regularly. 
Retrospective 
observational 
Children (M & F; 3 to 7 y) with recurrent 
group A beta-hemolytic streptococci 
pharyngo-tonsillar infections 
 
n = 130 
Test: 1×10⁹ 
CFU/day 
Control: no 
intervention was 
administered to the 
control group 
 
For 90 days 
All children 
were able to 
complete the 
study without 
stopping the 
test tablet 
before the 
intervention 
period ended. 
(Gregori et 
al., 2016) 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Adults (M & F; 18 y and older) with 
severe acute pharyngotonsillitis 
 
initial n = 60 
final n = 53 
Test: 4×10⁹ 
CFU/day 
Control: matched 
placebo tablet 
 
10 days 
Seven 
participants 
were excluded 
from the study 
due to 
noncompliance 
with the 
treatment, but 
the authors did 
not mention if 
any adverse 
events were 
observed by 
the 
participants. 
(Gilbey et 
al., 2015) 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Adults (M & F; 23 to 44 y) with tongue-
coating associated halitosis 
 
initial n = 33 
final n = 28 
Test: 2×10⁹ 
CFU/day 
Control: matched 
placebo tablet 
 
For 30 days 
 
All participants 
in the study did 
not experience 
any adverse 
events. 
However, five 
participants 
were excluded 
from the study, 
with two 
participants in 
the control 
group and 
three in the test 
group, one of 
which was 
using 
antibiotics, and 
four were lost 
to follow-up. 
(He et al., 
2020) 
Not applicable 
(single study 
participant) 
Single healthy adult (M), 40 y old 
4×10¹⁰ CFU/day 
 
For 3 days 
No adverse 
events were 
reported 
(Horz et 
al., 2007) 

Protocol_V8_October_11, 2025 
 
35 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Adults (M & F; >18 y) with oral 
candidiasis 
 
initial n = 56 
final n = 49 (safety-analyses) 
Test: ≥2×10⁹ 
CFU/day 
Control: matched 
placebo lozenge 
 
For 4 weeks 
The study did 
not report any 
severe adverse 
events. 
However, AEs 
were reported 
by 6 study 
participants in 
the test group 
and 8 study 
participants in 
the control 
group. One 
study 
participant in 
the K12 group 
reported 
borborygmus 
and pharyngeal 
discomfort, 
which was 
considered a 
possible drug-
related adverse 
event by the 
study authors. 
(Hu et al., 
2019) 
Randomized, 
double-blind, 
placebo 
controlled, 
parallel 
Adults (M & F: 18 y and older) with 
spondyloarthritis 
 
initial n = final n = 63 
Test: powder 
containing 1×10⁸ 
CFU/g of S. 
salivarius K12, 
4×10⁸ CFU/g of B. 
lactis LAFTI B94, 
and 4×10⁸ CFU/g 
of L. acidophilus 
LAFTI L10 
Control: matched 
placebo powder 
 
Participants were 
told to take 1 
spoonful of powder 
(ca. 0.8 g) by 
mouth twice daily, 
corresponding to 
ca. 1.6×10⁸ 
CFU/day of S. 
salivarius K12 
 
For 12 weeks 
All participants 
in the study 
completed it 
successfully. 
Minor and self-
limiting 
adverse events 
were reported 
by 43.8% 
(14/32) of the 
test group and 
38.7% (12/31) 
of the placebo 
group, with no 
significant 
difference 
between them. 
The most 
common AE in 
both groups 
was a change 
in bowel habit, 
reported by 7 
participants in 
the test group 
and 6 in the 
placebo group. 
No serious 
AEs occurred 
(Jenks et 
al., 2010) 

Protocol_V8_October_11, 2025 
 
36 
during the 
study. 
At the end of 
the study, there 
was no 
significant 
difference 
between the 
test and control 
groups in terms 
of fecal 
calprotectin or 
change in 
bowel 
symptom 
questionnaire 
scores. 
Double-
blinded, 
placebo 
controlled 
human clinical 
trial 
Healthy adults (M & F; 18 to 75 y) 
 
Initial n = 60 
Final n = 53 
Test: 1 x 1010 
CFU/g BLIS K12 
Control: matched 
placebo 
 
single 1 g dose 
mixed with 30 mL 
water and 
swallowed directly. 
 
For 1 day 
The dose 
chosen for use 
in this study 
was relatively 
high, but safe, 
dose to 
administer. No 
adverse 
reactions were 
reported by the 
authors. 
(Laws et 
al., 2021) 
Open-label, 
single-arm 
Study participants with rheumatoid 
arthritis taking stable doses of 
sulfasalazine (M & F; mean age = 56 y) 
 
initial n = final n = 12 
A powder blend 
(BioRestore™️) 
containing S. 
salivarius K12 at 
1×10⁸ CFU, L. 
acidophilus L10 at 
4×10⁸ CFU, B. 
lactis B94 at 4×10⁹ 
CFU. 
 
The powder was 
taken twice a day 
for total S. 
salivarius K12 of 
2×10⁸ CFU/day. 
 
For 7 days 
During the 
intervention 
period, four 
study 
participants 
reported 
adverse events 
(AEs), with 
three reporting 
gastrointestinal 
disturbances 
and one 
experiencing a 
flare-up of 
rheumatoid 
arthritis. The 
reported AEs 
were of mild to 
moderate 
severity. 
(Lee et al., 
2010) 
Randomized, 
non-blinded, 
controlled, 
parallel 
Adults with oral lichen planus (M & F; 
22 to 79 y) 
 
initial n = final n = 40 
Test: no less than 
2×10⁹ CFU/day 
Comparator: 
topical 0.1% 
triamcinolone 
acetonide dental 
paste 
No adverse 
reactions were 
observed. 
(Li et al., 
2020) 

Protocol_V8_October_11, 2025 
 
37 
 
4 weeks 
Randomized, 
double-blind, 
placebo-
controlled 
study 
Healthy adults (M & F; 18 to 65 y) 
Initial n = 64 
Final n = 60 
Test: S. salivarius 
DB-B5 at 109 CFU 
Control: matched 
placebo  
 
1 sachet in 
approximately 4 
ounces of bottled 
water daily  
 
For 4 weeks. 
The safety was 
confirmed. 4 
individuals did 
not complete 
the study: One 
participant in 
the test group 
discontinued 
from the study 
because they 
changed their 
mind about 
participation. 
The remaining 
3 participants 
were from the 
placebo group, 
and they were 
discontinued 
from the study 
for the 
following 
reasons: 
physician 
decision due to 
elevated 
eosinophils in 
the baseline 
blood sample 
(n = 1), 
occurrence of 
adverse event 
(n = 1), and 
lost to follow-
up (n = 1). 
There was a 
total of 15 
adverse events 
reported in 6 
participants in 
the study. In 
the test group, 
2 participants 
reported a total 
of 5 adverse 
events 
throughout the 
study that were 
considered 
“possibly 
related” to the 
interventions. 
All the events 
were mild in 
(Li et al., 
2021) 

Protocol_V8_October_11, 2025 
 
38 
nature and 
resolved on 
their own. 
Moreover, a 
similar 
frequency of 
adverse events 
was reported in 
the placebo 
group, in 
which 4 
participants 
reported a total 
of 10 adverse 
events, 
including one 
individual 
discontinued 
from the study 
due to the 
occurrence of 
mild urticaria.  
A prospective, 
single-open, 
multi-centre 
study 
Caucasian children with recurrent 
respiratory infections (RRIs) (M & F; 1 
to 12 y) 
 
Inicial n = 100 
Final n = 91 
Test: 
Streptococcus 
salivarius 24SMBc 
and Streptococcus 
oralis 
89a nasal spray 
(10⁹ CFU/dose) 
 
2 puffs for nostril 
twice/day for 7 
days/months.  
 
For 3 months. 
 
Although a 
good 
tolerability 
profile was 
reported, 9 
children 
experienced 
burning nose, 
leading to 
interruption of 
therapy. 
However, none 
of the 
children were 
withdrawn 
from the study 
because of 
adverse events. 
The authors 
suggest that 
this treatment 
is safe and 
seems to be 
effective on 
short-term in 
the treatment 
of RRIs 
(Manti et 
al., 2020) 
Open-label 
 
Study was 
randomized. 
Blinding not 
reported. 
Adults (M & F; 67 to 83 y) who are 
denture wearers. 
 
initial n = final n = 50 
Test: BactoBlis™️ 
containing S. 
salivarius K12 (10⁹ 
CFU/day) 
Control: no 
intervention was 
administered to the 
The study 
authors did not 
provide 
information on 
whether any 
adverse events 
were reported 
(Passariell
o et al., 
2020) 

Protocol_V8_October_11, 2025 
 
39 
control group 
 
For 30 days 
by the study 
participants. 
Open-label, 
single-arm 
Infants (age and sex not reported) prone 
to otitis media scheduled to undergo 
ventilation tube placement 
 
n = 19 
Test: powdered 
formulation with S. 
salivarius K12 
(reported as 1×10¹⁰ 
to 3.4×10¹⁰ 
CFU/day in GRN 
581) 
 
1 teaspoon was 
placed on the 
child's tongue 
twice daily. 
 
For 10 days 
The study 
authors did not 
provide 
information on 
whether any 
adverse events 
were reported 
by the study 
participants. 
(Power et 
al., 2008) 
Open-label 
 
Study was 
randomized. 
Microbiologic
al analyses 
were blinded. 
Children (M & F; 1 to 6 y) attending 
daycare centers 
 
initial n = 121 
final n reported as number of biological 
samples collected at 1-month and 2- 
month time period 
Test (children ≤3y 
old): 
powdered 
formulation with S. 
salivarius K12 
(5×10⁹ 
CFU/sachet) 
Test (older 
children): 
Chewable tablet 
containing S. 
salivarius K12 
(1×10⁹ CFU/tablet) 
Control: no 
intervention was 
administered to the 
control group 
 
Daily dose 
provided was 
5×10⁹ CFU/day 
(powder) and 
1×10⁹ CFU/day 
(tablet). 
 
For 30 days 
The 
administration 
of S. salivarius 
K12 did not 
cause any 
changes in the 
diversity of the 
nasopharyngea
l or saliva 
microbiome. 
However, there 
was a 
temporary 
increase in the 
proportion of 
S. salivarius in 
the saliva of 
children who 
received the 
product 
containing S. 
salivarius K12. 
(Sarlin et 
al., 2021) 
Randomized, 
double-blind, 
placebo 
controlled 
Adults (M & F; mean age 53.5 y in 
placebo, 53.3 y in test) who had received 
radiotherapy in the previous 6 months 
 
initial n = 17 
final n = 13 
Test: 3.5×10⁹ 
CFU/day 
Control: matched 
placebo lozenge 
 
For 4 weeks 
Four 
participants 
dropped out of 
the study: three 
from the 
placebo group 
and one from 
the test group. 
Two 
participants 
were lost to 
follow-up, one 
received 
(Vesty et 
al., 2020) 

Protocol_V8_October_11, 2025 
 
40 
antibiotics, and 
one did not 
take the 
lozenges as 
instructed. The 
study authors 
did not 
mention 
whether any 
adverse events 
were reported 
by the 
participants. 
randomised 
double-blind 
 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
41 
Appendix 2. Recruitment Flyer/Advertisement 
 
NOVO PROBIOTICS SALI-10 FOR THE 
PREVENTION OF GINGIVITIS: A 
CLINICAL TRIAL 
 
Why are the investigators doing this? 
Gum disease is a common condition in the mouth. This study is looking to see if 
using probiotics can help prevent gum disease in adults. 
 
Who can participate? 
Healthy adults (male/female) who don’t have gum disease 
(gingivitis/periodontitis). 
 
What participants must do? 
Take the lozenges (tablets) provided daily. Participants will either have the 
probiotic or placebo lozenges. Study volunteers will come once a week (for a total 
of 7 weeks) to the Faculty of Dentistry for a clinical exam and biospecimen 
collection (saliva, dental plaque). Two weeks into this clinical trial, participants 
will refrain from oral hygiene (brushing + flossing) for three weeks. They will also 
get a free professional dental cleaning (scaling + prophylaxis) twice: at the 
beginning of the study and after the three weeks of oral hygiene abstinence.  
 
Where will this study take place? 
Faculty of Dentistry, University of Toronto 
 
Will I be compensated? 
Yes! Financial compensation of 1000$ will be provided for those who complete 
the clinical trial!   
 
How do I sign up? 
Please email us at martin.lambert@mail.utoronto.ca expressing your interest in 
taking part in this study* 
 
 
*No personal information  
should be shared via email 

Protocol_V8_October_11, 2025 
 
42 
Appendix 3. Recruitment Questionnaire  
 
Recruitment Questionnaire 
 
1. Name __________________________________  
 
2. Age (in years) _______________  
 
3. Profession _______________________________  
 
4. Oral Hygiene Habits:  
Do participant brush your teeth? Yes No 
If so, how many times a day? _________  
Do participant use dental floss? Yes No 
If so how often? ___________________  
 
5. Dental Visit History: 
Have participant visited a dentist before? Yes No 
When was your last visit to the dentist (if applicable)? ___________________ 
At your last dental visit what was done (if applicable)? ___________________ 
 
6. Smoking History: 
Do participant smoke cigarettes? Yes No 
Do participant use any other tobacco product(s)?  Yes No 
Do participant smoke anything other than cigarettes? Yes No 
 
7. General Health:  
Do participant suffer from any major illness? Yes No 
If so what are they? _______________________________________________ 
Do participant take any medications? Yes No 
If so what are they? _______________________________________________ 
Do participant have any allergies? 
If so what are they? _______________________________________________ 
 
8. Are all your teeth missing? Yes No 
 
9. Do participant wear a complete denture? Yes No 
 
10. Do participant wear a partial denture? Yes No 
 
11. Are participant pregnant (if applicable)? Yes No 
 
12. Are participant currently nursing (if applicable)? Yes No 
 
13. Are participant currently using a method of birth control? Yes No 
 

Protocol_V8_October_11, 2025 
 
43 
14. English Language Proficiency: 
Do participant require English translation? Yes No 
Are participant able to read in English? Yes No 
 
15. Participants in this trial will be asked to be available for 8 visits over a 7-week span. Do 
participant think participant will be able to follow through with such a request? Yes No  
 
16. Participants in this trial will be asked not to practice any kind of oral hygiene for a 3-weeks. 
This will include no tooth brushing, no mouth rinse, no dental floss or any other oral hygiene 
procedures. Do participant think participant will be able to follow through with such a 
request? Yes No  
 
17. To those who are found eligible for the initial screening visit please be aware that in the case 
participant are not eligible to participate in the trial, participant will not be compensated for 
the screening visit. However, a full intra-oral examination* will be performed during that 
visit. Do participant agree to this? Yes No 
* Intra-oral examination will include:  
• assessment of soft tissues (including lips, gums, gi, hard palate, soft palate, back 
of the throat, tongue, and floor of the mouth),  
• evaluation of gum health via measure gum pockets 
•  and dental assessment to evaluate the condition of the teeth, filling, crowns, 
bridges and/or implants 
 
 
Eligible for the initial screening visit? Check One 
 
YES  
 
 
NO 
 
Reason(s) (violation of inclusion/exclusion 
criteria): 
 
 
 
 
 
 
 
 
 
 
 
 
 
If YES, scheduled date of initial screen (Month/Day/Year): _____________________________ 
 
 

Protocol_V8_October_11, 2025 
 
44 
Name of Assessor: _________________________________________ 
 
 
Signature of Assessor: ______________________________________ 
 
 
Date: ____________________________________________________ 
 
 
 

Protocol_V8_October_11, 2025 
 
45 
Appendix 4. Personal Information Form 
 
 
 
 
 
* This form will be filled out once participants have consented to participate in this study. 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
46 
Appendix 5. Health Questionnaire 
Confidential Medical History 
Health Questionnaire Form 
Study Participant ID No…….....................…  
 
 
        
 
Before beginning any study, the investigators need participant to complete and return this form. 
Please tick the appropriate response 
A yes answer does not necessarily mean participant will not be able to do the study. 
1. Are participant attending or receiving treatment from a doctor? ... ............ YES___NO___ 
2. Are participant taking or using any medicines, pills, tablets, ointments, injections or 
    any other drug, either from your doctor or on your own accord?...........................
 
YES___NO___ 
3. Are participant allergic to or have participant ever had any unfavorable reaction to any 
    medicine, food or any other substance?...............................................................
 
YES___NO___ 
4. Have participant had any serious illnesses as a child or 
adult? .......................................YES___NO___ 
5. Have participant ever been a hospital in-patient or ill at home for a long period? ...
 
YES___NO___ 
6. Do participant have or have participant ever had any heart or blood pressure problems? ....... 
YES___NO___ 
7. Have participant ever had rheumatic fever or chorea (St. Vitus` dance)? .......................
 
YES___NO___ 
8. Do participant have a heart murmur? ..............................................................................
 
YES___NO___ 
9. Do participant have a heart pacemaker? ........................................................................
 
YES___NO___ 
10. Have participant ever had any heart surgery? ..............................................................
 
YES___NO___ 
11. Do participant have any chest or breathing problems? .................................................
 
YES___NO___ 

Protocol_V8_October_11, 2025 
 
47 
12. Do participant suffer from eczema, asthma or any form of allergy? ..............................
 
YES___NO___ 
13. Do participant suffer from fainting attacks, fits or seizures? ..........................................
 
YES___NO___ 
14. Have participant ever suffered from hepatitis, jaundice, liver or kidney 
disease? …......YES___NO___ 
15. Are participant diabetic? ................................................................................................
......................................................................................................................... YES___NO___ 
16. Have participant had any problems arising from a blood sample, a blood donation or  
 
transfusion……………………………………………………………………………..YES___
NO___ 
17. Do participant carry a warning card from your doctor or specialist? .............................
 
YES___NO___ 
18. Is there anything else concerning your health, such as a joint replacement, 
      participant think the investigators should know 
about?........................................................................ ...................................... YES___NO___ 
19. Following extraction, surgery or injury have participant or any other member of your 
      family bled for such a time as to cause participant to be worried?.................................
 
YES___NO___ 
20. Do participant suffer from a dry mouth when eating food i.e. do participant have to drink  
 
liquids to swallow easily?...................................................................................
 
YES___NO___ 
 
21.  Do participant currently suffer from an active infections, or are experiencing nausea, fever, 
vomiting, bloody diarrhoea or severe abdominal pain? ..................................  
YES___NO___ 
 
 
 
Please inform us immediately if there is any change in this information 
To the best of my knowledge this information is correct. I understand this information may be 
inspected by authorized personnel and will be treated in strict confidence 
 

Protocol_V8_October_11, 2025 
 
48 
Please sign Here ……………………..                                    Date …..………………… 
Checked by..................................................……                     Date ……………………… 
Confidential Medical History Continued  
Please write information on questions overleaf in the box below: 
 
Medical History Review Record 
 
 
 
 
 
 
 
 
 
 
Question 
Number 
Medication 
Description 
Study 
participant 
Initials 
Date 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Visit 
Date 
Any change in 
MH- details 
Staff to initial 
and date 
Study 
participant to 
initial and date 
Study 
Dentist to  
sign and 
date * 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
49 
 
 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
50 
Appendix 6. Screening Oral Assessment Form 
 
Screening Oral Assessment Form 
 
Study participant ID No: ___________________________ 
 
 
Male                              Female     
 
 
 
Visit Number: ___________________ 
 
Tick the appropriate box 
 
 
AREA  
 
 
 
NORMAL 
 
   ABNORMAL 
 
1. 
Perioral area/lips 
 
 
 
 
 
2. 
Buccal mucosa 
 
 
 
 
 
 
 
 
 
3. 
Labial  mucosa 
 
 
 
 
4. 
Sublingual mucosa 
 
 
 
 
5. 
Gingiva free/attached  
 
 
 
6. 
Tongue 
 
 
 
 
 
7. 
Palate hard/soft 
 
 
 
 
8. 
Uvula  
 
 
 
 
 
9. 
Oropharynx 
 
 
 
 
 
10. 
All other soft/hard tissues 
 
 
 
DESCRIBE ANY IRREGULARITIES:  
 
 
 
 
 
 
 
______________________________________________________________________________
______________________________________________________________________________
________________________________________________________________________ 
   
Name of Assessor: _________________________________________ 
 
*Only if there has been a change in medical history 

Protocol_V8_October_11, 2025 
 
51 
 
Signature of Assessor: ______________________________________ 
 
 
Date: ____________________________________________________ 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
52 
Appendix 7. Oral Assessment Form 
Oral Assessment Form 
 
Study Participant ID No: ___________________________ 
 
Visit Number: ___________________ 
 
 
Male                              Female     
 
*Changes in the medical history or changes in the oral examination will be recorded.  
 
Tick the appropriate box 
 
 
AREA  
 
 
 
NORMAL 
 
   ABNORMAL 
 
1. 
Perioral area/lips 
 
 
 
 
 
2. 
Buccal mucosa 
 
 
 
 
 
 
 
 
 
3. 
Labial mucosa  
 
 
 
4. 
Sublingual mucosa 
 
 
 
 
5. 
Gingiva free/attached  
 
 
 
6. 
Tongue 
 
 
 
 
 
7. 
Palate hard/soft 
 
 
 
 
8. 
Uvula  
 
 
 
 
 
9. 
Oropharynx 
 
 
 
 
 
10. 
All other soft/hard tissues 
 
 
 
DESCRIBE ANY IRREGULARITIES:  
 
 
 
 
 
 
 
______________________________________________________________________________
______________________________________________________________________________
________________________________________________________________________ 
 
 
 

Protocol_V8_October_11, 2025 
 
53 
 
 

Protocol_V8_October_11, 2025 
 
54 
Appendix 8. Adverse Events Form 
 
Ostia Sciences Non-Serious Adverse Event/ Adverse Reaction Form 
 
Instructions: Do not leave any field blank. Please indicate if information is unknown, not provided or not available 
(refused). Please note that this form is expandable. 
 
Date of Awareness (dd/mmm/yyyy):       
Protocol #:       
Protocol Title:       
Indication of use protocol (if applicable):       
Investigator:       
Study Originator/Director:       
Type of study:  
Clinical 
Consumer 
Panel 
Product Category: 
Fabric care 
Household Surface care 
Oral Care 
Personal care 
Other:       
Phase of study the earliest event (s) occurred during: 
After consent  
 Pre-randomization 
 
Randomization: 
no 
product 
exposure 
 
Randomization: 
product 
exposure 
Other:       
 
Study participant/Study participant information: 
ID 
Initials 
Sex 
Age 
Weight 
Ethnic group 
       
       
       
       
       
       
Product information: 
Product name (PIM#):       
Start date (dd/mmm/yyyy):       
Stop Date or Duration (dd/mmm/yyyy):      
Dosage:       
Frequency:       
Randomization group:       
Reaction/Event information: 
Onset date 
(dd/mmm/yyyy) 
Stop date or duration 
(dd/mmm/yyyy): 
Severity  
(mild, moderate, severe) 
Relationship to product 
(Possibly related, Related, 
Unrelated, Unknown) 
      
      
      
      
 
Describe reaction(s)/event(s) in detail: 
      
 
 

Protocol_V8_October_11, 2025 
 
55 
 
 
Outcome: 
Resolved, Date (dd/mmm/yyyy): 
      
Resolving 
Unknown 
Not resolved 
Resolved with sequelae 
Other:       
Action taken with the product: 
    Continued 
    Reduced 
    Discontinued 
    Unknown 
    Temporarily discontinued 
    Other:       
Did the event(s) abate after product was stopped or dose reduced (yes/no)?       
Did the event(s) reappear after product was reintroduced (yes/no)?       
    Protocol Continued 
    Protocol Discontinued 
 
Treatment rendered for the event(s):       
 
 
 
Relevant Medical History Data: 
 Yes (List below)  
 None    
 Not provided    
 Unknown 
(Medical history with onset dates) 
      
 
 
Relevant Concomitant Medications: 
 Yes (List below)  
 None    
 Not provided    
 Unknown 
(Medication name, dose, frequency, start/stop dates [dd/mmm/yyyy] or duration of therapy) 
      
 
 
Relevant Lab data: 
 Yes (List below)  
 None    
 Not provided    
 Unknown 
(Lab test, results, dates [dd/mmm/yyyy]) 
      
 
 

Protocol_V8_October_11, 2025 
 
56 
Serious Adverse Events Report Form for Studies on Humans 
 
Instructions: Please complete all fields of this form. Please indicate if ‘Not provided,’ ‘Not Available’ or ‘Unknown.’ 
(Date eg: dd/mmm/yyyy 01/Dec/2009) 
 
Case-ID-No:       
 
A) Study site details 
Study No.:       
Study type : clinical / consumer test / panel test 
If clinical, please define:       
Study product:       
Dosage form and strength:       
Indication if appropriate:       
Centre Name:       
Investigator:       
Address:       
 
Country of occurrence:      
 
 
B)  Reporter Information 
Sender / Reporter 
Name:       
Address:       
 
 
Tel.:       
Fax:       
e-mail:       
 
Health professional: YES 
  NO 
 
Profession (Speciality):       
 
Date of Awareness (dd/mmm/yyyy):       
 
Date of report:(dd/mmm/yyyy)        
 
C)  General information 
 
Which product is concerned?  
 
 Test Product           
 Comparator (Control product)           
 Placebo              
 Unknown 
Specify when the event occurred during the study? 
 After consent
 Pre-randomization      
 Randomized with no product initiated                     
 Randomized with product initiated 
Study participant concerned: 
Study participant’s ID-number: 
Sex: 
 Male    
 Female        Year of birth (dd/mmm/yyyy):       
In case of presumption or intoxication:  Weight (kg):      
If Female, Pregnancy: 
 NO    
 YES, For how many months?       
 

Protocol_V8_October_11, 2025 
 
57 
D) Information on SAE 
Reason for serious adverse event report 
 Death                                                  
 Hospitalisation/Prolonged Hospitalization      
 Life Threatening                                  
 Congenital Anomaly 
 Disability/Incapacity                             
 Other, specify:       
 Suspected transmission via a medicinal product of an infectious agent     
If hospitalisation, give dates (dd/mmm/yyyy)   
 Hospitalization ongoing 
From:       
 
To:       
 
 Diagnosis / Symptoms 
Please, indicate diagnosis or main symptom (s) and list serious most significant adverse event first: 
1.       
2.       
3.       
4.       
Date of primary symptom (dd/mmm/yyyy) :        
Duration of taking the product:       
Outcome of serious adverse event : 
   
 Not resolved 
 Resolving 
  
 Resolved with sequelae 
   
 Resolved, date:       
              (dd/mmm/yyyy) 
 Unknown/ lost to follow up 
  
 *Death, date:       
         (dd/mmm/yyyy) 
Cause of death if known:       
Autopsy:  
 YES         
 NO         
 Unknown       
 Outcome:       
Action taken: 
 Protocol continuation                                   
 Clinical study product dose reduced, new dose:       
 Clinical study product discontinued             
 Medical intervention 
 Exclusion of the study participant                                
 Other:        
 
Event abated after drug stopped or dose reduced? 
 YES           
 NO                         
 N.A. 
Event reappeared after drug reintroduction? 
 YES           
 NO                         
 N.A. 
 
 
 

Protocol_V8_October_11, 2025 
 
58 
 
Causality 
  Related 
 Possibly related 
 Not related*              
 Unknown 
*If not related, please provide an alternative causality:       
Code broken (unblinded) 
 YES           
 NO                         
 
 N.A. 
 
E) Case narrative 
Please provide full details of the serious adverse event, dechallenge/rechallenge information and vital 
signs. 
Attach any relevant reports from the source document or hospitalisation file. In case of death, report cause 
and attach a copy of the autopsy report, if performed.  
           
 
 
 
 
 
 
 
Information enclosed:  
    NO         
 YES,   specify:       
 
F)  Relevant medical history 
 
Description 
Start Date 
(dd/mmm/yyyy) 
Stop Date 
(dd/mmm/yyyy) 
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
 
G)  Concomitant medication 
       Please report the medication taken in the last 4 weeks prior to the SAE(s) 
Drug 
Dose [unit] Route 
Frequency 
Indication 
Start Date 
(dd/mmm/yyyy) 
Stop Date 
(dd/mmm/yyyy) 
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
 
 

Protocol_V8_October_11, 2025 
 
59 
 
H)  Clinical study products 
       Please indicate if Test product, Comparator (control product) or Placebo 
Product 
Lot. No. 
Dose 
[unit] 
Route 
of 
administration 
Frequency 
Start Date 
(dd/mmm/yyyy) 
Stop Date 
(dd/mmm/yyyy) 
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
 
 
I)  Relevant laboratory data & other test procedure for SAE 
Test 
Date 
(dd/mmm/yyyy) 
Result (normal, abnormal, clinically significant) 
      
      
      
      
      
      
      
      
      
      
      
      
      
      
      
 
 
 
 
This report has to be sent within 1 Calendar day (No later than 1 business day): 
 
 
 
- 
By the Investigator to the Study Manager: 
• 
Address 
• 
Phone 
• 
E-mail 
 
 
- 
By the Study Manager to contact@osia-sciences.com 
• 
Email:  
                                                                                                
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
60 
Appendix 9. Study participant Withdrawal Form 
 
Study Participant Withdrawal Form 
This form should be completed for all study participants who do not complete full participation 
of the study. 
 
 
Study participant ID Number 
 
Date of Withdrawal 
 
Person completing withdrawal form 
 
Date informed of withdrawal 
 
Name of person reporting the withdrawal 
 
 
Withdrawal was made at the request of (tick one :) 
 
Study participant _______ Investigator _______ Study Dentist _______ Assessor _______ 
 
REASON FOR WITHDRAWAL (Tick all that Apply) 
 
 
PROTOCOL VIOLATION (Give details) 
 
NON-COMPLIANCE (Give details) 
 
ADVERSE EVENT (Please attach copy of completed AE form) 
 
MEDICAL REASONS AND/OR EXCLUSION CRITERIA (Give details) 
 
STUDY PARTICIPANT'S PERSONAL REASONS (Give details) 
 
OTHER (Give details) 
 
Person reporting withdrawal ______________________________  Date_________________ 
Investigators signature          ______________________________  Date_________________ 
Product returned  
Yes             No   
 NA    
(circle as appropriate) 
COMMENTS: ____________________________________________________ 
 
 
 
 

Protocol_V8_October_11, 2025 
 
61 
Appendix 10. Study Termination Form 
Study Termination Form 
 
STUDY PARTICIPANT ID No_____________________________________ 
 
MEDICATION HISTORY 
 
 
Does the study participant currently take any medications (including OTC products)?  Yes 
 
No (Circle One) 
 
If YES, please list all medications below: 
 
Medication 
 
Total Daily 
Dose 
 
Date Started 
 
Date Stopped 
(circle C if continuing) 
 
Indication 
 
 
 
C 
 
 
 
 
C 
 
 
 
 
C 
 
 
 
 
C 
 
 
ORAL EXAMINATION 
   
Tissue Examined  
Normal Abnormal If abnormal, comment below: 
 
1.  Perioral Area/Lips  
 ______ ________    _________________________ 
 
2.  Labial mucosa/Buccal  
 ______ ________ 
___________________________  
     mucosa 
 
3.  Mucolabial fold/  
______  ________ 
___________________________ 
     Mucobuccal fold 
 
4.  Gingiva/Free and attached ______ ________  ______________________________ 
      
5.  Palate/Hard and soft  
______  ________ 
____________________________  
 
6.  Oropharynx/Uvula  
______ 
________   ____________________________ 
 
7.  Tongue  
______  ________ 
____________________________  
 
8.  Floor of mouth  
______ 
_______ 
____________________________ 
 
 
9.  Other  
______  ________ 
____________________________  
 
10.  Hard Tissues 
______  ________ 
____________________________ 

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62 
 
 
 
______________________ 
______________________________ 
Date 
 
 
 
 
 
 
Clinical Examiner's Signature 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Protocol_V8_October_11, 2025 
 
63 
Appendix 11. Consent Form 
 
 
 
 
 
CONSENT TO PARTICIPATE IN A RESEARCH STUDY (CLINICAL TRIAL) 
Title 
Clinical study to assess the efficacy of SALI-10 Probiotics 
Experimental Gingivitis 
 
Principal Investigator 
Dr. Michael Goldberg  
 
University of Toronto, Faculty of Dentistry  
 
Tel: (416) 979-4928 ext. 4408 
 
Sponsor 
Ostia Sciences  
 
Tel: (647) 643-7547 
 
 
INTRODUCTION 
Participant are being invited to take part in a type of research study called a clinical trial. The 
information below will tell participant about the study, what will be asked of participant and if 
there are any risks and benefits to you. It is up to participant to decide whether participant would 
like to participate in the study. Please read the consent form very carefully and make sure that all 
of your questions are answered before participant consent to take part. Before participant make 
your decision, feel free to talk about this study with anyone participant wish. Participation in this 
study is voluntary. 
BACKGROUND AND PURPOSE 
Clinical trials are necessary to gain or expand knowledge about the efficacy and tolerability of 
medicinal products. The clinical trial, which the investigators are presenting to participant here, 
has been reviewed and approved by the Health Sciences Research Ethics Board, University of 
Toronto and Health Canada, to be conducted in Canada. The study was initiated and is funded by 
Ostia Sciences the sponsor of this study. 

Protocol_V8_October_11, 2025 
 
64 
We are conducting this clinical trial with a new oral probiotic in the form of dissolvable oral 
lozenges (like a tablet) that is being evaluated for improving oral health and promoting healthy 
gum tissue. Gingivitis (gum disease), or the inflammation of the gums, is commonly due to 
infection of the gum tissues that is caused by bacteria that attach to the teeth. When these 
bacteria are not properly removed (by brushing and flossing), the bacterial accumulation may 
cause the inflammation of the gums. Clinical signs of gum disease include (but are not limited 
to) redness or swelling of the gums, and bleeding from the gums, usually after irritation such as 
brushing or flossing. Gum disease is a completely reversible condition with no long-term 
complications if adequate daily hygiene is performed. However, it is currently estimated that 
80% of adolescents and adults have gum disease.  
The purpose of the study is to evaluate an oral probiotic named SALI-10 for its effect on 
preventing gum disease. This new probiotic has been harvested, isolated, and then sanitized from 
the oral cavity of a healthy individual 
The results from this study will provide additional information for the use of SALI-10 oral 
lozenge probiotic in study participants to help prevent gum disease. 
The focus of this study is to analyze and prevent induced gum disease caused by bacteria in 
dental plaque of healthy individuals participant have been given this information sheet to read 
because participant are in good general and oral health; with no braces or dentures; and a non-
smoker. Under these circumstances, participant may be a good candidate to participate in the 
study, which will be confirmed after a clinical exam. 
STUDY DESIGN 
This study is described as a Randomized, Double Blind, Single-center and controlled study to 
investigate the effect SALI-10 oral lozenge probiotic on the resolution of gum disease compared 
to a placebo (a lozenge with no SALI-10 probiotic or any form of active ingredient). Gingivitis 
(gum disease) is a reversible condition causing inflammation of the gums. In this study, gum 
disease will be induced by having study participants refrain from using any form of oral hygiene, 
including any treatments from a dental professional, i.e., dentist, hygienist. A description of these 
terms is given below:  
Randomized Trial: Sometimes because the investigators do not know which way of treating 
study participants is best, the investigators need to make comparisons. Participants in this study 
will be put into 2 groups and then compared. The groups are selected by a computer, which has 
no information about the individual. Therefore, participant have an equal chance to be in either 
group - like the toss of a coin. Participants will receive either the probiotic lozenges or the 
placebo lozenges, which they will have to take daily. The placebo lozenge will look identical to 
the probiotic one, but it will not contain the probiotic (active ingredient) that the investigators are 
testing. 
Single center: The study is planned at only one center, the Faculty of Dentistry, University of 
Toronto, and it will involve a total of around 60 people in it. 
Double-blind trial: In a blind trial participant will not know which treatment group participant 
are in. If the trial is a double-blind trial, neither participant nor the researcher and sponsor will 
know in which treatment group participant are (although, if the researcher needs to find out i.e. 
in an emergency, he/she can do so). 

Protocol_V8_October_11, 2025 
 
65 
STUDY VISITS AND PROCEDURES 
If participant decide to participate, you’ll be asked to sign the consent form. At the start of the 
study, you’ll be asked to fill out a health questionnaire, and your mouth, teeth and gums will be 
examined to determine if participant are eligible to participate in this study.  
For the clinical trial, the health questionnaire will inquire about any known illnesses, current 
medications, and any allergies that participant may have. 
The intra-oral examination using a mirror, a dental probe and a dental explorer will include:  
• assessment of soft tissues (including lips, mucosa, gingiva, hard palate, soft palate, oro-
pharynx, tongue, and floor of the mouth),  
o This will involve a visual examination 
• evaluation of gum health (via measuring the gum pockets),  
• dental assessment (to evaluate the condition of the teeth and any restorations i.e., fillings, 
crowns, bridges, Implants).  
• and plaque levels where the dental explorer will pick-up (if visible) plaque from different 
tooth surfaces and a plaque index score will then be given. 
The professional dental cleanings will be done by Dr. Martin Lambert, a member of the study 
team, who is a licensed dentist and periodontics resident at the Faculty of Dentistry, University 
of Toronto. The professional dental cleanings will be done to ensure all study participants start at 
the same gum health prior to when the gum disease is induced and also done at the end of the 
study to restore study participants’ gum health to normal and healthy.  
To measure bad breath, similar to a breathalyzer, participant will breathe through a tube 
connected to a machine (Halimeter PLUS) that will quantify the degree of bad breath. By 
measuring the level of sulfur compounds produced (responsible for the unpleasant odour) from 
the mouth, the investigators can evaluate how the probiotic SALI-10 battles against the bacteria 
responsible for foul breath. 
All examinations and treatments are performed with sterile instruments (sterilized by an 
autoclave at the Faculty of Dentistry of the University of Toronto).  
If participant are accepted into the study, participant will be required to follow a specific set of 
instructions. participant will agree to use only the oral lozenges given to participant for the 
duration of the study, as well as only the toothpaste provided. participant will not perform or 
receive any oral hygiene (other than that described below) or dental care throughout the study, 
except emergency care, until this study is completed. If participant must receive emergency 
dental treatment, participant will notify the principal investigator Dr. Micheal Goldberg at (416) 
979-4928 ext. 4408.  There will be no restrictions regarding your dietary habits. 
Study participants who qualify for the study will be randomized in one of the two possible 
groups in a 1:1 ratio. As a result, your chance of receiving the SALI-10 probiotic lozenge is 
about 50%. The study researcher and participant will not know to which group participant are 
assigned. 
All examinations and procedures associated with the study will take place in a dental chair, 
similar to when participant visit your dentist, within a dental operatory at the Faculty of 
Dentistry, University of Toronto.  

Protocol_V8_October_11, 2025 
 
66 
Your participation in the clinical study will last up to 7 weeks as described below: 
Visit 1 In-Person Screening 
Here participant will read over and complete the consent form. Female participants will also be 
asked to take a urine pregnancy test to confirm they are not pregnant. Then participant will 
complete a health questionnaire form and an oral exam will be done to see if participant are eligible 
to take part in the study. If participant are deemed eligible to take part in the study participant will 
be invited to the next visit of the study. 
Visit 2 (Day -14) 
Participant cannot eat/drink for 2 hours before the visit. participant will undergo an oral soft and 
hard tissue assessment and the results will be recorded on the appropriate form. A clinical 
assessment will be carried out to assess the level of gum disease and the level of plaque in your 
teeth. Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be collected. 
participant will receive a professional dental cleaning to remove all soft (dental plaque) and hard 
(tartar) deposits from your teeth. participant will be assigned the lozenges and instructed on how 
to use them as well as provided with at-home oral hygiene instructions. Researchers will query 
participants on their use of NHPs, as well as any adverse effects they experienced. 
Visits 3  (Day -7)  
Participant cannot eat/drink for 2 hours before the visit. participant will undergo an oral soft and 
hard tissue assessment and the results will be recorded on the appropriate form. A clinical 
assessment will be carried out to assess the level of gum disease and the level of plaque in your 
teeth. Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be collected. 
participant will resume at-home oral hygiene. Researchers will query participants on their use of 
NHPs, as well as any adverse effects they experienced. 
Visits 4, 5, 6 (Day 0, 7, 14)  
Participant cannot eat/drink for 2 hours before the appointment. participant will undergo an oral 
soft and hard tissue assessment and the results will be recorded on the appropriate form. A 
clinical assessment will be carried out to assess the level of gum disease and the level of plaque 
in your teeth. Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be 
collected. Bad breath level with be measured using a non-invasive intra-oral device called the 
Halimeter PLUS. At this point participant will be instructed to continue the use of the lozenges 
assigned to participant but will be told to refrain from all forms of home oral hygiene for the next 
21 days. Researchers will query participants on their use of NHPs, as well as any adverse effects 
they experienced. 
Visit 7 (Day 21) 
Participant cannot eat/drink for 2 hours before the appointment. participant will undergo an oral 
soft and hard tissue assessment and the results will be recorded on the appropriate form A 
clinical assessment will be carried out to assess the level of gum disease and the level of plaque 
in your teeth Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be 
collected. Bad breath level with be measured using a non-invasive intra-oral device called the 
Halimeter PLUS.  participant will receive a professional dental cleaning to remove all soft 
(dental plaque) and hard (tartar) deposits from your teeth. participant will continue taking the 
lozenges assigned to you. After this visit participant can return to doing at-home oral hygiene. 

Protocol_V8_October_11, 2025 
 
67 
Researchers will query participants on their use of NHPs, as well as any adverse effects they 
experienced. 
Visit 8 (Day 28) 
Participant cannot eat/drink for 2 hours before the appointment. participant will undergo an oral 
soft and hard tissue assessment and the results will be recorded on the appropriate form A 
clinical assessment will be carried out to assess the level of gum disease and the level of plaque 
in your teeth Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be 
collected. Bad breath level with be measured using a non-invasive intra-oral device called the 
Halimeter PLUS. participant will continue taking the lozenges assigned to participant as well as 
continue at-home oral hygiene. Researchers will query participants on their use of NHPs, as well 
as any adverse effects they experienced. 
Visit 9 (Day 35) 
Participant cannot eat/drink for 2 hours before the appointment. participant will undergo an oral 
soft and hard tissue assessment and the results will be recorded on the appropriate form. A 
clinical assessment will be carried out to assess the level of gum disease and the level of plaque. 
Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be collected. Bad 
breath level with be measured using a non-invasive intra-oral device called the Halimeter PLUS.  
After completion of the study, participant will receive compensation, and a study personnel will 
reveal to participant what lozenges participant had been assigned and ask for your feedback with 
regards to your experience in the study. Researchers will query participants on their use of 
NHPs, as well as any adverse effects they experienced. 
Any questions or concerns participant may have will be addressed in this visit and participant 
will then be formally exited from the study 
While participating as a study participant in this clinical research, participant cannot participate 
as a study participant in any other clinical studies. In addition, participant cannot use drugs, other 
than over the counter analgesics, during the course of the study. participant agree to inform the 
investigator about any new medications participant are planning to take, including but not limited 
to antibiotics, antiseptics, decongestants and antihistamines. 
This product falls into the regulatory category of natural health products (NHP). There may be 
unknown risks with taking the investigational NHP. Study participants agree to inform the study 
researchers of any new medications and natural health products that they begin taking during the 
study. Study participants do not give up any legal rights by participating in this trial and they do 
not release the Investigators from liability for negligence. Records identifying the study 
participants will be kept confidential for 15 years. 
 
Unscheduled visits 
If the researcher believes that participant should have additional visits for your safety, for 
example in the event of a new symptom or side effect, participant may be asked to come for an 
additional visit. If necessary, additional tests related to such a safety concern may be performed. 
If the safety concern goes beyond the scope of dentistry, study participants will be referred to the 
appropriate healthcare provider. 
Missed Doses 

Protocol_V8_October_11, 2025 
 
68 
If a study participant misses their dose, they will be asked to take it if no more than 3 hours has 
passed since they brushed their teeth. If more than 3 hours have passed, they will be asked to 
skip the dose completely. Do not double up on the next dose. Do not exceed 2 lozenges per day.  
 
 
Description of sample collection 
Samples of dental plaque, saliva, oral rinse, and fluid from your gums will be collected on visits 
2-9 and are described as follows: 
 
• Dental Plaque: sterile paper points (these made of thin paper) will be placed in the space 
between the gum and the tooth for 30s and then removed. Six teeth will be sampled. 
• Saliva: you’ll be asked to chew on a gum and then spit into a tube (up to 15ml). 
• Oral Rinse: you’ll be asked to rinse with 10ml of a salted solution for 30 seconds and then 
spit into a tube (30ml). 
• Fluid from Gums: sterile paper strips (germ-free tiny piece of paper) will be placed in the 
space between the gum and the tooth for 30s and then removed. Six teeth will be sampled. 
 
 
 
Calendar of Visits 
 
Visit 
Day 
Health 
Questionnaire 
(Pregnancy test) 
Clinical 
Exam 
Oral soft/ hard 
tissue 
assessment 
Clinical 
Data 
Collection 
Sample 
collection 
Professional 
dental 
cleaning 
Time 
(mins) 
Visit 1 
Screening 
X 
X 
 
 
 
 
60 
Visit 2 
-14 
 
X 
X 
X 
X 
X 
90 
Visit 3 
-7 
 
X 
X 
X 
X 
 
60 
Visits 
4,5.6 
 
0, 7, 14 
 
X 
X 
X 
X 
 
60 
Visit 7 
21 
 
X 
X 
X 
X 
X 
90 
Visits 
8, 9 
 
28, 35 
 
X 
X 
X 
X 
 
60 
RISKS RELATED TO PREGNANCY  
Pregnant women must not take part in this study, neither should women who plan to become 
pregnant during the study. Pregnancy status will be checked through a urine pregnancy test 
before the start of the trial. Study participants of child bearing potential, both men and women, 
must practice medically acceptable methods of birth control, which are: abstinence, hormonal 
birth control (oral, injectable, transdermal, intra-vaginal), intrauterine devices, confirmed 
successful vasectomy, or condoms. Any woman who finds that she has become pregnant while 
taking part in the study should immediately stop taking the investigators product immediately  
and inform the study researcher.  In the event a woman enrolled in this clinical research study 

Protocol_V8_October_11, 2025 
 
69 
becomes pregnant during the course of the study, participation in this study will be terminated 
upon the clinical staff’s notification of the event. Your medical records used in this study will be 
updated to reflect the pregnancy and participant will have follow-up contact until the end of the 
pregnancy to record the outcome in your file. Pregnant and nursing women have an elevated 
level of hormones (oestrogen and progesterone), which favour gum disease and are therefore 
excluded from this study.    
VOLUNTARY PARTICIPATION 
Participant are under no obligation whatsoever to participate in this study and that your 
participation in this study is strictly voluntary. participant may withdraw or discontinue 
participation at any time and may decline to answer any question or participate in any parts of 
the procedure. There will be no consequences if participant choose to withdraw from the study 
other than the following compensation effects:  
• No financial compensation will be awarded if participant have not completed the trial 
period  
• In case participant choose to withdraw before visit 7, participant will not receive the 
second dental cleaning treatment  
Participant may also withdraw consent for the use of your data, but participant understand that 
participant must do this in writing. participant understand that the investigator has the right to 
withdraw participant from the study at any time because it would not be in your best interest to 
stay in it. The study researcher can stop treatment even if participant are willing to stay in the 
study. Any data that has already been sent to the sponsor of the study cannot be withdrawn 
because there may not be any identifiers with the data. 
If participant decide to pull out of the study for any reason, participant may be asked to return for 
at least one additional visit for safety reasons. 
COMPENSATION 
Participation in research is voluntary. Compensation of $1000 will be given to study participants;  
$500 will be given after the 21 days of experimental gingivitis (gum disease) and the remaining 
$500 will be given at the end of the study. This amount will be given to also cover costs that 
study participants’ may have while taking part in the study i.e., travel costs. It is the 
responsibility of the participant to comply with the Income Tax rules with respect to this 
monetary compensation.  
Participant will receive 2 professional dental cleaning treatments for free prior and subsequent to 
stopping all oral hygiene procedures (approximate value of $650). participant will also receive 
proper oral hygiene instructions. 
BENEFITS 
Participation in this study may not benefit participant personally. The results of the study may 
allow improved methods to detect and treat gum disease and disease-causing plaque in the early 
stages and the investigators would like to thank participant for considering participating in this 
study. 
ALTERNATIVES TO PARTICIPATING IN THIS STUDY 

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70 
There are no alternatives to participating in this study as participation in this study is on a 
voluntary basis and participant may choose to not participate in this study. 
DISCOMFORTS AND RISKS 
Summary of Risks: 
• Bleeding gums 
• Discomfort during dental examination and probing of the gums 
• Gum pain or sensitivity due to the inflammation 
• Bad breath 
• Temporary tooth staining 
• Dental plaque build-up on the gums 
• Allergic reaction to the SALI-10 probiotic oral lozenges 
These events are expected to be mild and temporary and will stop when participant receive the 
professional dental cleaning the end of the study.   
In the event that study participants experience any fever, vomiting, bloody diarrhea or severe 
abdominal pain, or symptoms of digestive upset such as diarrhea lasting more than 3 days, 
participants should consult the study doctors and the decision to discontinue participation in the 
study will be taken on a case by case basis.  
Additionally, at each examinations participant will be asked if participant have experienced any 
discomfort, pain, irritation in the mouth  and/or any other concern(s) that participant may have. 
The investigators will record any adverse reactions and report this documentation to the sponsor. 
The nature of the reaction and any correlation with product usage will be assessed. If the 
evidence indicates that the adverse reaction may be due to product usage, participant will be 
instructed to discontinue product use and appropriate treatment will be provided. In the event of 
an adverse experience, emergency or other problems or questions regarding your participation in 
this study participant can contact the principal investigator Dr. Micheal Goldberg at:  (416) 979-
4928 ext. 4408 or the Research Oversight and Compliance Office - Human Research Ethics 
Program at ethics.review@utoronto.ca or 416-946-3273. In the event of a medical emergency, 
please contact your physician. 
As mentioned above, gum disease is a completely reversible condition, as was demonstrated in 
many studies. Despite the inflammation of the gums, study participants experience no pain and 
no irreversible pathology. Experimental gum disease is a common and safe clinical tool which is 
used in many fields of oral research.  
Bad breath may occur during the 21 day-period of no oral hygiene. Yet, mouthwash and gum 
chewing are not allowed until the end of the study. 
Participant will be under close monitoring, and if any unexpected disease arises, it will be treated 
as soon as it is diagnosed. participant can also call the 24-hour number at 581-991-0184 to 
address any concerns as they arise. 
No discomfort or side effects are expected from the collection of dental plaque, saliva, oral rinse 
oral fluid and measurement of bad breath using the Halimeter PLUS.  
NEW FINDINGS 

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71 
Sometimes during the course of a research study, new information becomes available about the 
treatment/drug that is being studied. participant will be informed of any significant new findings 
related to study products or procedures as soon as they are known. If this happens, the study 
researcher will tell participant about it and discuss with participant whether participant want to 
continue in the study. If participant decide to continue in the study, participant will be asked to 
sign an updated consent form. 
Also, on receiving new information the study researcher might consider it to be in your best 
interests to withdraw participant from the study, and explanations will be given accordingly. 
CONFIDENTIALITY 
If participant agree to join this study, the study researcher and his study team will look at your 
personal health information and collect only the information they need for the study. Personal 
health information is any information that could be used to identify participant and may include 
your name, address, date of birth, and new or existing medical records, that includes types, dates 
and results of medical tests or procedures. 
The results of this study may be published and/or submitted to Health Canada and/or local 
regulatory agencies in other countries, including US FDA. Your identity will be kept 
confidential, only your study participant number/ID, gender and/or age may be used participant 
in connection with any such publication of the study results.  No documents that identify 
participant by name (e.g., the signed informed consent form and health questionnaire) will be 
transferred or submitted to the study sponsor (Ostia Sciences) but will be maintained in strict 
confidence by the investigator in a locked filing cabinet in a locked room at the Faculty of 
Dentistry. However, the investigator must allow auditing/monitoring by the sponsor personnel to 
verify study participant enrollment, product safety and study compliance. If any sponsor 
monitoring occurs, study documents can be looked at, but no private information will be copied 
or removed from the clinical site. 
The following people may come to the Faculty to look at the study records and at your personal 
health information to check that the information collected for the study is correct and to make sure 
the study followed proper laws and guidelines: 
• The study sponsor or its representatives/partner companies; 
• Representatives of the University of Toronto Research Ethics Board; 
Some study information will be sent outside of the Faculty to the Sponsor. Prior to disclosing 
such personal data to the larger study team, including the sponsor of the study, investigators and 
their team will code your personal data. Therefore, any information about participant that is sent 
out of the Faculty will have a code and will not show your name or address, or any information 
that directly identifies you. 
All information collected during this study, including your personal health information, will be 
kept confidential and will not be shared with anyone outside the study unless required by law. 
participant will not be named in any reports, publications, or presentations that may come from 
this study. 
There will be no future use of data that is collected from participant for the purposes of this 
study, 

Protocol_V8_October_11, 2025 
 
72 
If participant decide to leave the study, the information about participant that was collected 
before participant left the study will still be used. No new information will be collected without 
your permission. 
What will happen to the results of the research study? 
The results of this study may be published online, in a medical journal and shown at medical 
meetings. participant will not be identified (by name or any other personal means) in any of these 
publications. 
If participant would like to know the study results, the study researcher will inform participant 
when the results are available. 
CONFLICT OF INTEREST  
Ostia Sciences is sponsoring this clinical trial. 
The Institution and study researcher will be paid to run this study. 
All of these people have an interest in completing this study. Their interests should not influence 
your decision to participate in this study. participant should not feel pressured to join this study. 
QUESTIONS ABOUT THE STUDY 
If participant have any questions, concerns or would like to speak to the study team for any 
reason, please contact Dr. Martin Lambert through email: martin.lambert@mail.utoronto.ca. 
Please note that the security of e-mail messages is not guaranteed. Messages may be forged, 
forwarded, kept indefinitely, or seen by others using the Internet. Do not share personal 
information via email. 
 
If participant have any questions about your rights as a research participant or have concerns 
about this study, please contact the Research Oversight and Compliance Office - Human 
Research Ethics Program at ethics.review@utoronto.ca or call 416 946 3273. The Research 
Ethics Board is a group of people who oversee the ethical conduct of research studies. These 
people are not part of the study team. Everything that participant discuss will be kept 
confidential. 
 
INFORMED CONSENT 
Participant will be asked to sign the consent form at the end of this study participant information 
sheet. If participant take part in this study, participant should follow the study procedures and 
attend all study visits. 
 
I, _____________________________________________________have read and understand the  
                               
 
(PRINT NAME) 
information provided above and have been given an opportunity to ask questions and they were 
satisfactorily answered.  I have been provided with a copy of this informed consent. By my 
signature below, I consent to participate in the study and agree to keep confidential any and all 
information disclosed to me, either directly or indirectly, about this study, including but not 

Protocol_V8_October_11, 2025 
 
73 
limited to the purpose of the study, any procedures and any toothpaste.  I agree that I will follow 
the instructions. 
 
(Participant) 
By signing this form, I confirm that: 
• This research study has been fully explained to me and all of my questions have been answered 
to my satisfaction 
• I understand the requirements of participating in this research study 
• I have been informed of the risks and benefits, if any, of participating in this research study 
• I have been informed of any alternatives to participating in this research study 
• I have been informed of the rights of research participants 
• I have read each page of this form 
• I authorize access to my personal health information and research study data as explained in this 
form 
• I have agreed to participate in this study. 
 
 
_________________________ 
__________________________ 
________________  
Participant’s Name (Print)                             Signature 
 
 
 
Date 
 
 
(Person obtaining consent) 
By signing this form, I confirm that: 
• This study and its purpose has been explained to the participant named above 
• All questions asked by the participant have been answered 
• I will give a copy of this signed and dated document to the participant 
 
 
 
_________________________ 
__________________________ 
_______________ 
Person Obtaining Consent  
 
Signature 
 
 
 
Date 
(Print) 
 
 
 

Protocol_V8_October_11, 2025 
 
74 
Appendix 12. Contact Information for Study Withdrawal 
 
Study Withdrawal Contact Form 
 
Title 
Clinical study to assess the efficacy of SALI-10 Probiotics 
Experimental Gingivitis 
 
Principal Investigator 
Dr. Michael Goldberg  
 
University of Toronto, Faculty of Dentistry  
 
Tel: (416) 979-4928 ext. 4408 
 
Sponsor 
Ostia Sciences  
 
Tel: (647) 643-7547 
 
If participant choose to withdraw from the study, please contact one of the study research 
personnel, Dr. Martin Lambert, via email: 
 
martin.lambert@mail.utoronto.ca 
 
Please indicated in your email if participant want to be contacted by phone or email. 
 
Please do not share personal information via email.  
 
After receiving your email, a member of the study will contact participant based on the 
preference participant gave in the email a formal withdrawal interview with you.