Expression of collagen type III in healing tympanic membrane

Abstract

Objectives: Type III collagen plays significant role in skin wound healing, forming provisional matrix guiding the inflammatory cells and fibroblasts into the wound site. Our preliminary study performed on rat's tympanic membrane (TM) using Rat Wound Healing RT2 Profiler PCR Array revealed up-regulated expression of collagen type III α1 chain mRNA also during healing of TM. This study was undertaken to confirm and evaluate collagen type III protein expression and distribution during TM healing process.

Methods: Sixty rats were used, of which 10 served as controls and the others had their TM perforated. The experimental animals were divided into five subgroups on the basis of time points (03, 06, 09, 14, 20 day after injury). Videootoscopy and histology were employed to assess morphology of the healing process. The expression of collagen type III was evaluated using Western blot analysis and its tissue localization was determined by the immunohistochemical method.

Results: The expression of collagen type III remained on the same level as in control TM up to day 06. On day 09 abrupt (p = 0.01) increase of the collagen type III expression was observed and it maintained on the same level to the end of observation period. In perforated TM collagen type III was detected in the healing area along the perforation border and around dilated blood vessels. On day 14 and 20 collagen type III was found in the connective tissue filling up the TM previous defect.

Conclusions: Taking into consideration our recent and previous data, as well as results obtained by other authors, is seems possible that the increase of collagen type III expression in the late stage of TM healing contributes to proper scar formation.

Keywords: Collagen type III; Healing process; Perforation; Rats; Tympanic membrane.