Fourty-four implants (cp titanium) were placed in the alveolar bone of 11 systemically healthy human subjects missing 2 or more teeth.Following anesthesia(Lidocaine 2% with epinephrine (1:100,000)), two separate crestal incisions approximately 6mm in length were made and full thickness mucoperiosteal flaps were raised. Each subject received two TIO and two OS implants. Two osteotomies were prepared per site to allow placement of one implant of each surface (1 TiO and 1 Os). This paired surgical design permitted the retrieval of the first 2 implants (1 of each surface) at the earlier time point without disturbing the healing of the other 2 implants. Drilling was completed using a 2.0 mm twist osteotomy drill under profuse irrigation with sterile saline. Single use osteotomy drills were used on each subject. All implants were placed by a self-tapping procedure and primary stability at the time of implant placement was insured. A submerged implant installation technique was used and the mucosal tissues were closed primarily with 4-0 chromic gut sutures
Growth protocol
11 Human subjects missing 2 or more teeth were included.Following anesthesia(Lidocaine 2% with epinephrine (1:100,000)), two separate crestal incisions approximately 6mm in length were made and full thickness mucoperiosteal flaps were raised. Each subject received two TIO and two OS implants. Two osteotomies were prepared per site to allow placement of one implant of each surface (1 TiO and 1 Os). This paired surgical design permitted the retrieval of the first 2 implants (1 of each surface) at the earlier time point without disturbing the healing of the other 2 implants. Drilling was completed using a 2.0 mm twist osteotomy drill under profuse irrigation with sterile saline. Single use osteotomy drills were used on each subject. All implants were placed by a self-tapping procedure and primary stability at the time of implant placement was insured. A submerged implant installation technique was used and the mucosal tissues were closed primarily with 4-0 chromic gut sutures.
Extracted molecule
total RNA
Extraction protocol
At 3 and 7 days following surgery, one surgical site was chosen at random, re-entered and the paired (One TIO and one OS) implants were removed by reverse threading. The implants were then immediately rinsed in cold PBS and then placed into 1000 ul of Tri-reagent (Invitrogen, Carlsbad, CA). Cell lysates were snap frozen, and stored at -80°C until further use. Total RNA was isolated from the lysates using the standard Tri-reagent protocol and collected by ethanol precipitation. This was followed by purification using RNeasy MinElute Clean up kit (Qiagen, Valencia, CA, USA). RNA was assessed for quality and quantity using a bioanalyzer (Agilent, Santa Clara, CA, USA) and nanodrop ND-1000 spectrophotometer (Nanodrop, Wilmington, DE) respectively.
Label
Biotin
Label protocol
Samples were processed and hybridized to the Affymetrix Human gene 1.1 ST Array at the UNC core facility following the manufacturer’s recommended protocols and reagents (Affymetrix, San Clara,CA). Labeling used Nugen Encore Biotin Module
Hybridization protocol
The Encore Biotin Module V2-NuGen/Affymetrix Human gene 1.1 ST Array
Scan protocol
Affymetrix GeneChip Scanner 3000 7G Plus with Autoloader
Data processing
The data was analyzed using GeneSpring software v.12.0
