MicroRNA and MicroRNA Inhibitors Socket Study, Pilot Clinical Trial

Study Overview
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Brief Summary
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To evaluate the efficacy of locally delivering plasmid DNAs encoding microRNAs, and/or microRNA inhibitors, in the promotion of osteogenesis and modulation of the inflammatory response on the basis of different clinical, radiographic, histologic and biomolecular outcomes in post-extraction socket defects in humans.

Detailed Description
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The purpose of this study is to evaluate the efficacy of locally delivering plasmid DNAs encoding microRNAS and/or microRNA inhibitors, a naturally occurring microRNA molecule, in the promotion of bone formation and attenuation of local inflammation in a tooth socket model in humans. Patients requiring tooth extractions and future tooth replacement therapy with a dental implant will be eligible for the study. Recruited subjects will be randomly assigned to either a control (tooth extraction and 10µg empty vector in bovine collagen sponge) or one of 3 experimental groups: Experimental group 1: Tooth extraction and bovine collagen sponge containing 10µg of pSil-miR200c Experimental Group 2: Tooth extraction and bovine Collagen sponge containing 10µg of PMIS miR200a Experimental Group 3: Tooth extraction and bovine Collagen sponge containing 5µg of pSil-miR200c and 5µg of PMIS miR200a Subjects will be clinically re-evaluated at 1, 2, 3 and 4 weeks. In each one of these visits, a fluid sample will be obtained from the healing site in a minimally invasive manner to assess local biomolecular profiles. Blood samples will be drawn at 1, 2, 3, 4 and 14 week visits to assess for miR-200c and PMIS-miR 200a expression and liver function. Photos and/or videos will also be obtained. A CBCT scan and a saliva sample (approx. 2 ml) will also be obtained at baseline and at 14 weeks to assess bone volume and intraoral biomolecular profiles, respectively. Implant placement surgery will be performed at 16 weeks from the time of tooth extraction. A bone core biopsy will be harvested at this time for histologic analysis. A periapical radiograph (small dental x-ray image) will be obtained at baseline (before tooth extraction) and at 14 weeks (prior to implant placement) to assess bone height changes. Follow-up visits will occur at 6 months and 12 months post implant at which time measurements and photos will be taken.

Official Title
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Efficacy of Local Delivery of MicroRNAs and MicroRNA Inhibitors in Promoting Osteogenesis and Modulating Local Inflammation: A Pilot Clinical Trial Using the Tooth Socket Model

Conditions
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Tooth Extraction Status Nos

Intervention / Treatment
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* Procedure: tooth extraction
* Radiation: CBCT scan
* Drug: Anesthesia
* Other: clinical measurements
* Drug: Biodegradable sponge (type I bovine collagen)
* Drug: 10µg of pSil-miR200c
* Procedure: cross mattress suture
* Drug: 10µg of PMIS miR200a plasmids
* Drug: 5µg of pSil-miR200c and 5µg of PMIS miR200a
* Procedure: Blood
* Other: Photos/videos
* Procedure: Wound fluid
* Procedure: saliva
* Radiation: periapical xray
* Other: PVS impression


Participation Criteria
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Eligibility Criteria
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INCLUSION CRITERIA Age: 25 to 65 years. Gender: No restriction. Subjects must require a single-rooted tooth extraction (Tooth deemed as periodontally, endodontically and/or restoratively hopeless). Subjects must be able and willing to follow instructions related to the study procedures. Subjects must have read, understood and signed an informed consent form. EXCLUSION CRITERIA Reported allergy or hypersensitivity to any of the products to be used in the study. Severe hematologic disorders, such as leukemia. Active severe infectious diseases that may compromise normal healing. Liver or kidney dysfunction/failure. Currently under cancer treatment or history of cancer of any kind. Subjects who have a long-term history of oral bisphosphonate use (i.e. 4 years or more). Subjects with a history of IV bisphosphonates. Subjects with uncontrolled diabetes. Subjects with severe metabolic bone diseases, such as Paget's disease of bone, will be excluded. Pregnant women or nursing mothers. Smokers: Within 6 months of study onset. Concomitant medications: Subjects on concomitant drug therapy for systemic conditions, such as antibiotics or patient taking non-steroidal anti-inflammatory (NSAID) agents that may affect the outcomes of the study will not be included in the study. Subjects taking biologics or disease modifying agents will also be excluded. Occasional, short-term use (7-14 days) of analgesics or common cold medications is permitted. Any other non-specified reason that from the point of views of the investigators will make a candidate not a suitable subject for the study (e.g. limited mouth opening).

Ages Eligible for Study
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Minimum Age: 25 Years
Maximum Age: 65 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Health Services Research
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: control group<br>CBCT scan limited to the dental arch that includes the study site will be obtained. All subjects will receive local infiltrative anesthesia, following which minimally invasive tooth extraction will be performed. After tooth extraction clinical measurements of the site will also be obtained and recorded for both groups (i.e. keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness). A biodegradable sponge (type I bovine collagen) to stabilize the blood clot will be placed.The site will be stabilized with a simple external, cross mattress suture. Blood , wound fluid, and saliva samples will be taken periodicallly. In addition, photos/videos, periapical xrays and PVS impressions will be obtained. | Procedure: tooth extraction<br>* The study tooth will be removed<br>Radiation: CBCT scan<br>* a CBCT scan limited to the dental arch that includes the study side will be obtained<br>Drug: Anesthesia<br>* all subjects will receive local infiltrative anesthesia, prior to extraction of the tooth<br>* Other names: local infiltrative anesthesia;Other: clinical measurements<br>* After tooth extraction, clinical measurements of the site will be obtained and recorded (keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness<br>Drug: Biodegradable sponge (type I bovine collagen)<br>* control group subjects will receive a biodegradable sponge (type I bovine collagen) to stabilize the clot<br>* Other names: Bovine collagen;Procedure: cross mattress suture<br>* The site will be stabilized with a simplet external, cross mattress suture<br>Procedure: Blood<br>* Subjects will have venipuncture performed to obtain a small blood sample (approx. 2 ml) to assess for miR-200c and PMIS-miR-200a expression and liver function (AST, ALT, bilirubin levels)<br>* Other names: venipuncture;Other: Photos/videos<br>* subjects will have photos and/or videos of the extraction site and/or implant taken at each visit.<br>Procedure: Wound fluid<br>* a small sample of wound fluid will be obtained from the extraction site in a minimally invasive manner using a paper point at 1, 2, 3 & 4 weeks post extraction<br>Procedure: saliva<br>* approximately 2 mls of saliva will be obtained in a minimally invasive manner at the time of extraction and at 14 weeks post extraction.<br>Radiation: periapical xray<br>* periapical xrays will be obtained at screening, 16 weeks and at 12 month followup time point<br>Other: PVS impression<br>* PVS impressions will be taken to plan the implant placement surgery. This will be done at the screening visit and also at 14 weeks post extraction.<br>|
| Active Comparator: Experimental group 1<br>CBCT scan limited to the dental arch that includes the study site will be obtained. All subjects will receive local infiltrative anesthesia, following which minimally invasive tooth extraction will be performed. After tooth extraction clinical measurements of the site will also be obtained and recorded for both groups (i.e. keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness). 10µg of pSil-miR200c plasmids in a biodegradable sponge (type I bovine collagen) will be locally delivered. The site will be stabilized with a simple external, cross mattress suture. Blood, wound fluid, and saliva samples will be taken periodicallly. In addition, photos/videos, periapical xrays and PVS impressions will be obtained. | Procedure: tooth extraction<br>* The study tooth will be removed<br>Radiation: CBCT scan<br>* a CBCT scan limited to the dental arch that includes the study side will be obtained<br>Drug: Anesthesia<br>* all subjects will receive local infiltrative anesthesia, prior to extraction of the tooth<br>* Other names: local infiltrative anesthesia;Other: clinical measurements<br>* After tooth extraction, clinical measurements of the site will be obtained and recorded (keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness<br>Drug: 10µg of pSil-miR200c<br>* subjects in the experimental group will receive a 10µg of pSil-miR200c plasmids in a biodegradable sponge (type I bovine collagen) to stabilize the clot<br>* Other names: pSil-miR-200C plasmids;Procedure: cross mattress suture<br>* The site will be stabilized with a simplet external, cross mattress suture<br>Procedure: Blood<br>* Subjects will have venipuncture performed to obtain a small blood sample (approx. 2 ml) to assess for miR-200c and PMIS-miR-200a expression and liver function (AST, ALT, bilirubin levels)<br>* Other names: venipuncture;Other: Photos/videos<br>* subjects will have photos and/or videos of the extraction site and/or implant taken at each visit.<br>Procedure: Wound fluid<br>* a small sample of wound fluid will be obtained from the extraction site in a minimally invasive manner using a paper point at 1, 2, 3 & 4 weeks post extraction<br>Procedure: saliva<br>* approximately 2 mls of saliva will be obtained in a minimally invasive manner at the time of extraction and at 14 weeks post extraction.<br>Radiation: periapical xray<br>* periapical xrays will be obtained at screening, 16 weeks and at 12 month followup time point<br>Other: PVS impression<br>* PVS impressions will be taken to plan the implant placement surgery. This will be done at the screening visit and also at 14 weeks post extraction.<br>|
| Active Comparator: Experimental group 2<br>CBCT scan limited to the dental arch that includes the study site will be obtained. All subjects will receive local infiltrative anesthesia, following which minimally invasive tooth extraction will be performed. After tooth extraction clinical measurements of the site will also be obtained and recorded for both groups (i.e. keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness). 10µg of PMIS miR200a plasmids in a biodegradable sponge (type I bovine collagen) will be locally delivered. The site will be stabilized with a simple external, cross mattress suture. Blood, wound fluid, and saliva samples will be taken periodicallly. In addition, photos/videos, periapical xrays and PVS impressions will be obtained. | Procedure: tooth extraction<br>* The study tooth will be removed<br>Radiation: CBCT scan<br>* a CBCT scan limited to the dental arch that includes the study side will be obtained<br>Drug: Anesthesia<br>* all subjects will receive local infiltrative anesthesia, prior to extraction of the tooth<br>* Other names: local infiltrative anesthesia;Other: clinical measurements<br>* After tooth extraction, clinical measurements of the site will be obtained and recorded (keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness<br>Procedure: cross mattress suture<br>* The site will be stabilized with a simplet external, cross mattress suture<br>Drug: 10µg of PMIS miR200a plasmids<br>* subjects in the experimental group will receive a 10µg of PMIS miR200a plasmids in a biodegradable sponge (type I bovine collagen) to stabilize the clot<br>* Other names: PMIS miR200a plasmids;Procedure: Blood<br>* Subjects will have venipuncture performed to obtain a small blood sample (approx. 2 ml) to assess for miR-200c and PMIS-miR-200a expression and liver function (AST, ALT, bilirubin levels)<br>* Other names: venipuncture;Other: Photos/videos<br>* subjects will have photos and/or videos of the extraction site and/or implant taken at each visit.<br>Procedure: Wound fluid<br>* a small sample of wound fluid will be obtained from the extraction site in a minimally invasive manner using a paper point at 1, 2, 3 & 4 weeks post extraction<br>Procedure: saliva<br>* approximately 2 mls of saliva will be obtained in a minimally invasive manner at the time of extraction and at 14 weeks post extraction.<br>Radiation: periapical xray<br>* periapical xrays will be obtained at screening, 16 weeks and at 12 month followup time point<br>Other: PVS impression<br>* PVS impressions will be taken to plan the implant placement surgery. This will be done at the screening visit and also at 14 weeks post extraction.<br>|
| Active Comparator: Experimental group 3<br>CBCT scan limited to the dental arch that includes the study site will be obtained. All subjects will receive local infiltrative anesthesia, following which minimally invasive tooth extraction will be performed. After tooth extraction clinical measurements of the site will also be obtained and recorded for both groups (i.e. keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness). 5µg of pSil-miR200c and 5µg of PMIS miR200a plasmids in a biodegradable sponge (type I bovine collagen) will be locally delivered. The site will be stabilized with a simple external, cross mattress suture. Blood, wound fluid, and saliva samples will be taken periodicallly. In addition, photos/videos, periapical xrays and PVS impressions will be obtained. | Procedure: tooth extraction<br>* The study tooth will be removed<br>Radiation: CBCT scan<br>* a CBCT scan limited to the dental arch that includes the study side will be obtained<br>Drug: Anesthesia<br>* all subjects will receive local infiltrative anesthesia, prior to extraction of the tooth<br>* Other names: local infiltrative anesthesia;Other: clinical measurements<br>* After tooth extraction, clinical measurements of the site will be obtained and recorded (keratinized mucosa width, horizontal ridge width, facial and lingual bone thickness<br>Procedure: cross mattress suture<br>* The site will be stabilized with a simplet external, cross mattress suture<br>Drug: 5µg of pSil-miR200c and 5µg of PMIS miR200a<br>* subjects in the experimental group will receive a 5µg of pSil-miR200c and 5µg of PMIS miR200a plasmids in a biodegradable sponge (type I bovine collagen) to stabilize the clot<br>* Other names: 5µg of pSil-miR200c and 5µg of PMIS miR200a plasmids;Procedure: Blood<br>* Subjects will have venipuncture performed to obtain a small blood sample (approx. 2 ml) to assess for miR-200c and PMIS-miR-200a expression and liver function (AST, ALT, bilirubin levels)<br>* Other names: venipuncture;Other: Photos/videos<br>* subjects will have photos and/or videos of the extraction site and/or implant taken at each visit.<br>Procedure: Wound fluid<br>* a small sample of wound fluid will be obtained from the extraction site in a minimally invasive manner using a paper point at 1, 2, 3 & 4 weeks post extraction<br>Procedure: saliva<br>* approximately 2 mls of saliva will be obtained in a minimally invasive manner at the time of extraction and at 14 weeks post extraction.<br>Radiation: periapical xray<br>* periapical xrays will be obtained at screening, 16 weeks and at 12 month followup time point<br>Other: PVS impression<br>* PVS impressions will be taken to plan the implant placement surgery. This will be done at the screening visit and also at 14 weeks post extraction.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Percent of mineralized tissue upon histomorphometric analysis of bone core biopsies | compared using exact Wilcoxon rank sum tests | at 16 weeks postoperatively | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Bucco-lingual width changes of the alveolar ridge (in mm) | Fisher's exact tests will be used to compare the treatment groups | up to 16 weeks postoperatively | 
| Mid-buccal height changes of the alveolar ridge (in mm) | compared using exact Wilcoxon rank sum tests | up to 16 weeks postoperatively | 
| Mid-lingual height changes of the alveolar ridge (in mm) | compared using exact Wilcoxon rank sum tests | up to 16 weeks postoperatively | 
| Volumetric reduction of the alveolar ridge (in cc) via CBCT scan analyses | compared using exact Wilcoxon rank sum tests | at 16 weeks postoperatively | 
| Expression of different biomarkers (VEGF, PDGF, TGF-b, IL-1b, TNF-a) in wound fluid expressed in pg/ml | compared using exact Wilcoxon rank sum tests | up to 4 weeks postoperatively |