gender: female subject: 140 time: Day 1 tissue: palate
Treatment protocol
Participants between 18-35 years of age were screened using a survey-form, which contained information regarding demographics and general health. Individuals were ineligible for the study if they: currently used tobacco products, had an oral disease needing emergency treatment, had a known skin disease other than acne, or had a medical problem that made them a high surgical risk. Furthermore, females were ineligible for the study if they were currently pregnant, trying to become pregnant, or breast-feeding. All wounding procedures were performed by a periodontist. Wounds were placed between 10-11 a.m. to minimize variability. Individuals were asked to refrain from alcohol and non-prescription drugs for a period of 24 hours prior to the experimental sessions. One skin and one mucosal wound were placed on each subject. For oral wounds, the side of the mouth was chosen by coin toss. The wounding site was anesthetized using 2% lidocaine. A mucosal wound measuring 1x5 millimeters (mm) was placed between the first and second molars, approximately 3 mm from the marginal gingiva using a Bard Parker handle, with two blades affixed 1 mm apart to assure uniformity of the wounds. A single blade scalpel was then used to make a relatively uniform 1.5 mm deep wound, removing the biopsy tissues. The wounds were not dressed and subjects were instructed not to change their normal oral hygiene procedures, with the exception of avoiding the use of alcohol-based mouthwash. A 2x5 mm longitudinal excisional biopsy was made using a 2 mm two blade scalpel to collect oral wound biopsies. The dermal wound site was on the ventral side of the non-preferred forearm, approximately 4 centimeters below the elbow. The biopsy site was anesthetized with 2% lidocaine. Dermal wounds were created in a similar fashion to mucosal wounds, a 1x5 mm longitudinal wound approximately 1.5 mm deep. Wounds were dressed for one day with a standard adhesive bandage. Dermal and oral wounds were biopsied at 6 hours, Day 1, Day 3, and Day 7 post-wounding. Unwounded tissue (time 0) was also processed. A separate wound was used for each biopsy.
Extracted molecule
total RNA
Extraction protocol
Trizol followed by Qiagen RNeasy column-based kit purification was performed according to the manufacturer's instructions.
Label
biotin
Label protocol
Affymetrix standard eukariotic 3' labeling protocol
Hybridization protocol
10ug of fragmented cRNA was hybridized to HG-U133 Plus 2.0 microarrays
Scan protocol
Affymetrix Standard protocol
Description
1x5mm wound, 2x5mm collection
Data processing
R/bioconductor, affy package, RMA normalization, Custom CDF hgu133plus2hsensgcdf_22.0.0 from BrainArray
