An Evaluation of antimicrobial property of orthodontic adhesive modified with silver nanoparticles : An In Vivo Study

•This will be a split mouth in-vivo study. 25 patients undergoing orthodontic treatment will be selected for the study. Each participant will be treated with  0.022 x 0.028” slot metal MBT bracket system.

•Participants will be allocated into 2 sides using lottery method ; so the brackets on one side of the patient will be bonded with orthodontic adhesive with silver nanoparticles and the other side brackets will be bonded with orthodontic adhesive without silver NP. Control side will be the adhesive without silver NP. Experimental side will be the silver

containing adhesive side.

 â€¢The experiment will be a  single blinded study  conducted by keeping the technician blinded about the groups to prevent the bias.

Instructions given to patient :

•Brush twice a day

•All patients will be instructed not to use any chemical plaque control method.

Sample collection :  Dental floss will be cleaned with a recombigen swab and plaque sample will be collected after 21 days of bonding from the cervical area of tooth and some amount of plaque will be scraped with a scaler and stored in a saline containing test tube and will be sent for microbiology test in Genexis Biotech Pvt. Ltd. Lab.

•To check the Antimicrobial activity, plaque sample collected on 21st day will be inoculated in Luria Broth for isolation of  S.mutans and Lactobacillus acidophilus.   Grown culture will be used for serial dilution, plated on chrome agar plates for specific identification of  bacterial growth. The different colour colonies will be further inoculated in Luria Broth and incubate overnight at 37°C for growth.  Bacterial isolate will be used for genomic DNA extraction, G DNA will be isolated using Genomic DNA extraction kit, followed by 16srdna using universal primers for 16srDNA,  PCR samples will be confirmed on (0.8%) agarose gel electrophoresis. PCR sample will be purified using PCR clean up kit. PCR cleanup product will be sent for sequencing.  Protocol used for pcr amplification will be followed accordingly. Thermocycle conditions are 94°C for 3 minutes,  94°C for 45 seconds,  50°C for 60 seconds,  72°C for 90 seconds,  Repeat steps 2-4 35 times,  72°C for 10 minutes, 4°C HOLD

ʉۢFor Results - Colony Forming Units (CFU) will be counted and antimicrobial property of orthodontic adhesive modified with silver nanoparticles will be compared with the adhesive without silver nanoparticles.