A S Sotnichenko
University of the Ministry of Healthcare of the Russian Federation, Russia
Title: Development of method for obtaining the dermal extracellular matrix
Biography
Biography: A S Sotnichenko
Abstract
Statement of the Problem: Despite the achievements of modern surgery in the treatment of cutaneous injuries, the search for new methods for faster and more effective wound healing remains topical. Tissue engineering is undoubtedly of interest for the development of such technologies.
Aim: of the study was to determine the optimal protocol for obtaining a decellularized dermal matrix for the subsequent development of tissue-engineered skin.
Materials and Methods: The experimental animal was 1 pig of the Landrace breed. After skin pretreatment with a dermatome, the samples were taken of a thickness of 0.3 cm. Two decellularization protocols were examined: protocol No.1 based on the using of Triton X-100 and deoxycholate, protocol No.2 only based on deoxycholate. Total processing cycles for 2 protocols were 5. The cellular matrices after treatment were examined with: histological analysis, quantitative determination of DNA content in wet tissue. Further, static matrix recellularization of porcine dermis fibroblasts was carried out. After that, the matrices were assessed for cytotoxicity using an XTT test and a test for the differential staining of living and dead cells.
Results: The comparative analysis of two protocols for decellularization of porcine dermis showed that both protocols effectively remove cells and nuclear material, while maintaining the architectonic of the intercellular substance intact, since fibrous structures are not destroyed. But when analyzed the functional properties of matrices on the basis of cell viability analysis according to the XTT test and cell adhesion to the matrix, the matrix processed according to protocol No.1 demonstrates the advantages.
Conclusion: In this study, a decellularization protocol based on Triton X-100 and deoxycholate was noted. The results are the first stage for the further development of tissue-engineered skin.