Clinical and Microbiological Study on Local Application of an Ozonated Olive Oil Gel in the Periodontal Pockets: a Randomized Double-blind Trial.
Phase: NA
Conditions: Periodontal Pocket
Sponsor: University of Bari

 
Clinical and microbiological study on local application of an Ozonated 
Olive Oil gel in the periodontal pockets: a randomized double-blind 
trial.
16/05/2025

 
FLOW CHART: The study was conducted in full compliance with Good Clinical Practice and in
accordance with the Declaration of Helsinki.
First visit and tartar ablation session
for all the patients
one or two session if necessary a week apart
15 days after the last tartar ablation session --> enrolment in the study (t ) - All patients will have a periodontal
0
survey + microbiological sampling and finally
Control Group : placebo application
Interventional Group : product application
After seven days(t1) - Compliance t1
Control Group : placebo application
Interventional Group : product application
After seven days(t2) - Compliance t2
Control Group : placebo application
Interventional Group : product application
After seven days(t ) - Compliance t + periodontal survey + microbiological sample
3
3
All the patients enrolled
Evaluation of blinded clinical outcomes

 
In this study have been enrolled 16 (sixteen) patients 8 male and 8 female who were randomized in
two groups: Control group has randomized enrolled 5 males and 3 females and Interventional
Group has randomized enrolled 5 females and 3 males. The average age of the patients enrolled in
the study is 51 years old.
Table 1 shows demographic data of patients enrolled in the study, divided into
control group and intervention group
Group Control
Age
Group Intervention
Pz A
Gender
Pz B
Age
Gender
1
3
4
8
1
1
1
1
A
29 years
61 years
30 years
68 years
63 years
44 years
28 years
28 years
43 years
F
2B
59 years
81 years
61 years
55 years
51 years
59 years
53 years
49 years
F
A
M
M
F
5B
M
F
A
6B
A
7B
M
F
2A
4A
5A
6A
M
M
M
F
9B
10B
11B
13B
F
F
M
Control
group
average
age
Intervention 
58 years 
Average
age 51 years
group
patients
average age
enrolled
(control
+
intervention)
P value was calculated between the age of the two 0,0654
groups
All statistical tests were conducted at a significance level of .05 using Microsoft Excel Software.
The significance value (p-value) was also calculated using data analysis with the t-test.

 
The International Standards Organisation Designation System (ISO) numbering drawn up by the
WHO is used to indicate the tooth involved, while the following letters are used to indicate the site
involved: the letter V to indicate the site located on the buccal side, the letter P for the site located
on the palatal side, the letter L for the site located on the lingual side, the letter M for the mesial site
and the letter D for the distal site. These letters can combine to indicate a specific site (e.g. MV17 -
mesiovestibular site of 17). Table shows the PPD values, divided into the two groups Control and
Intervention, found during the first and second survey in the three deepest sites surveyed. In
addition, a further table showing the mm of PPD reduction from the beginning of the study to the
end of the study, possibly expressing an improvement in the periodontal index, can be derived.
Patients treated in group Control and those treated in group Intervention will be differentiated in
such a way as to show significant differences between the two groups.
Table 2 shows PPD detections at the 3 selected sites measured with the periodontal survey at study
enrolment (t ) and at the end of the study (t ) divided into groups: Control and Interventional
0
3
Group Control
Group Intervention
Pz A
Site 1 PPD
Site 2 PPD
Site 3 PPD
Pz B 
Site 1 PPD 
Site 2 PPD 
Site 3 PPD
Before - after
Before - after
Before - after
Before -
after
Before -
after
Before - after
1A
3A
4A
8A
MV17
DL46
DV36
2B
5B
MP14
MP18
DP28
5
6
1
mm – 5mm
5mm – 4mm
5mm – 4mm
8 mm – 6 mm 
9 mm – 7 mm
7mm – 7mm
MV27
DP17
DV47
V14
DV45
MV24
mm – 5mm
7 mm – 7 mm
9 mm – 7 mm
5 mm – 4 mm 
4 mm - 3 mm
6mm – 4mm
DV47
DL36
DV16
6B
MV36
DV16
DV17
0mm – 6mm
5 mm – 3 mm
7 mm – 3 mm
5 mm – 3 mm 
5 mm – 3 mm
5mm – 4mm
DP27
DV36
DL37
7B
MP16
V27
V44
4
5
7
4
5
mm – 3mm
4 mm – 3 mm
4 mm – 3 mm
6 mm – 3mm 
4 mm – 3 mm
4mm – 2mm
12A
14A
15A
16A
DV27
DL37
MP16
9B
DV34
P14
DP17
mm – 6mm
6 mm - 6 mm
5 mm – 5 mm
7 mm – 2 mm 
8 mm – 2 mm
8mm – 3mm
MP13
DP26
MV26
10B
11B
13B
MV16
DV26
MV31
mm – 7mm
10 mm – 8 mm
12 mm – 10 mm
8 mm – 2 mm 
5 mm – 3 mm
6mm – 5mm
ML47
DL36
MV27
DP13
DV24
DV11
mm – 3mm
5 mm – 4 mm
5 mm – 5 mm
5 mm – 3mm
4 mm – 3mm
4mm – 2mm
ML45
ML46
MV16
DP16
DL44
DV33
mm – 4mm
5 mm – 4 mm
6 mm – 5 mm
7 mm – 2 mm 
5 mm – 2mm
5mm – 2mm

 
Table 3 shows  PPD at the three selected sites measured with the periodontal
survey at the time of study enrolment (t ) and at the end of the study (t ) divided
0
3
into Control Group and Intervention Group.
Group Control
Group Intervention
Pz



Pz



PPD
PPD
PPD
PPD
PPD
PPD
Site 1
Site 2
Site 3
Site 1
Site 2
Site 3
A
B
1
3
4
A
MV17
DL46
DV36
-1 mm
2B
5B
6B
MP14
-2 mm
MP18
-2 mm
DP28
0 mm
-1 mm
0 mm
A
MV27
DP17
mm
DV47
-2 mm
V14
DV45
-1 mm
MV24
0
-1 mm
-1 mm
-2 mm
DV17
-1 mm
A
DV47
DL36
2 mm
DV16
-4 mm
MV36
DV16
-2 mm
-
-4 mm
DP27
1 mm
DV27
1 mm
MP13
mm
ML47
1 mm
ML45
1 mm
Average 
-0.875
mm
-2 mm
MP16
-3 mm
DV34
-5 mm
MV16
-6 mm
DP13
-2 mm
DP16
-5 mm
-3.25
8
1
1
1
1
A
DV36
-1 mm
DL37
0 mm
DP26
DL37
-1 mm
MP16
0 mm
MV26
-2 mm
MV27
0 mm
MV16
-1 mm
-1.375
mm
7B
9B
V27
V44
-
-1 mm
P14
-2 mm
DP17
-5 mm
MV31
-1 mm
DV11
-2 mm
DV33
-3 mm
- 2
2A
4A
5A
6A
+
-6 mm
DV26
-2 mm
DV24
-1 mm
DL44
-3 mm
-2.25
mm
10B
11B
13B
0
-2 mm
DL36
-1 mm
ML46
-1 mm
-1 mm
-
-
Average
PPD
PPD
mm
mm
Table 3 shows the results of a difference made between the values in the previous Table, clearly
showing whether there was a reduction or an increase or no change in PPD
( PPD). For each site identified in each group, the values of  PPD were averaged, resulting in:
-
-
an average reduction of 0.875 mm for site 1 of the control group;
an average reduction of 1 mm for site 2 of the control group;

 
Table 4 shows the summary values of the mean reduction of PPD at the three
sites identified for each group e il p value was calculated on the mean
reduction of PPD at the three sites identified for each group.
- an average
reduction of 1.375
mm for site 3 of the
control group;
Group Control
Group Intervention
- an average
Mean
Site 1
PPD
Mean  PPD
Mean  PPD
Mean  PPD
Mean  PPD
Mean  PPD 
reduction of 3.25 mm
for site 1 of the
Site 2
Site 3
Site 1
Site 2
Site 3
intervention group;
-0.875 mm 
-1 mm
-1.375 mm
-3.25 mm
-2.25 mm
-2 mm
0,0015
- an average
Mean  PPD 
-1.08 mm 
Mean  PPD of all
of all the sites 
the 
sites 
of
-2.5 mm
p value
reduction of 2.25 mm
for site 2 of the
intervention group;
of
control
intervention
group
group
- an average
reduction of 2 mm for site 3 of the intervention group.
Table 4 shows the summary values, listed above, of the mean reduction of PPD at the three sites
identified for each group, then the mean reduction of PPD at all sites was calculated, which
corresponds to -1.08 mm for the control group and -2.5 mm for the intervention group. For
statistical purposes, the significance value (p value) on the mean reduction of PPD at the three sites
identified for each group was calculated and it is 0.0015 therefore, it is less than 0.05 and this
establishes that the mean reduction of PPD at the three sites identified of the intervention group is
statistically significant compared to the mean reduction of PPD at the three sites of the control
group.

 
Table 5 shows the data on the total microbial load found in the first and second
microbiological sampling. The cell next to it shows the changes in percentage
increase or decrease in the total microbial load detected by the two tests
Group Control
Group Intervention
Pz
Total
microbial
load in the 
in the second
Total
%
Pz
Total
microbial
Total microbial
load 
in 
the
%
microbial load
A
B
load in the 
second test
first test
first test
test
1
3
4
8
1
1
1
1
A
315089
365101
15,87% 
2B
-5,62% 
5B
9191825
3960222
112536
883461
52360
-90,39%
-98,68%
-89,25%
-95,91%
-21,24%
-86,39%
-99,85%
-94,89%
-84,58%
A
343899
324567
A
442355290 7470638
-83,11% 
6B
-94,96% 
7B
12093
A
645010
38298
4349829
501307
177747
394825
904892
12578
2A
4A
5A
6A
29928098
62634388
14584057
30809235
17246417 
-42,37% 
9B
79553873 
27,01% 
10B
6649643
8403042
9010221
14323174 
-1,79%
11B
75890416 
146,32% 13B
460648
Mean
Mean % -4,72%
Control
%
Intervention
gruppo
group

 
Table shows the data on the total microbial load found in the first and second microbiological
sampling. The cell next to it shows the changes in percentage increase or decrease in the total
microbial load detected by the two tests obtained by the following calculation:
[(total charge test 2 - total charge test 1) X 100]/ total charge test 1
It is clear from the percentage value obtained whether there is a decrease or increase in the total
microbial load detected. The average value of percentage increase or decrease in group A results in
an average reduction of 4.72%, while in group B an average reduction of 84.58% in the total
microbial load was obtained.
As shown in the flow chart, compliance was collected at each follow-up meeting one week apart for
3 weeks. No side effects from the administration of the gel were recorded.
The evaluation of the study's limitations is important in order to identify any bias. The sample of 16
patients examined may be small in order to highlight any statistically significant differences sought
by the investigator, which is why the study aims to be a pilot study and on the basis of the results
that have been highlighted it may be a starting point for other investigators, also by virtue of the
second limitation identified, namely monocentricity. In fact, the trial was carried out in a single
centre.
Results show a significant reduction of PPD (Control Group -1,08mm versus Intervention Group -
2,5 mm; p value = 0,0015). Due to low number of subject statistical analysis of total microbial load
was not performed (Control Group -4,8% versus Intervention Group -84,6%).
In conclusion, the gel based on ozonated EVO olive oil showed effectiveness as adjuvant treatment
of periodontal pockets in non-diabetic young adults patients, anyway our study was underpowered
because of slowness of inclusion.