PROLIFERATION AND VIABILITY OF KERATINOCYTES AND FIBROBLASTS CULTURED ON QUITOSANA SCAFFOLDS AND SUBMITTED TO LASERTHERAPY
Tissue engineering; skin; biomaterials; laser therapy; cell proliferation
Wound repair is one of the most complex biological processes that occurs during life, where multiple biological pathways are activated and synchronized as response to an injury. In the skin, some injuries may remain for a long time or even be permanent and cause physical, psychological and social consequences. Recent advances in biotechnology may provide new tools for the therapy of these lesions, highlighting in this regard tissue engineering, a science that combines the use of biomaterials, cells and stimulatory factors in order to repair injured organs. Low-level laser has been used in experiments to stimulate the proliferation of different cell types, representing a potential tool for use in tissue engineering techniques. In order for the cells to proliferate and differentiate in vitro it is necessary to develop a favorable microenvironment, which can be provided by a biomaterial that mimics the natural extracellular matrix, and for this purpose the chitosan is distinguished by its mechanical and biological properties. The aim of this study is to evaluate the influence of laser therapy on the proliferation and viability of keratinocytes and fibroblasts cultured on chitosan scaffolds. HaCaT (keratinocytes) and NIH-3T3 (fibroblasts) will be cultured separately on chitosan films and the scaffolds will be divided into two groups: C - control without irradiation; and L - cells irradiated with diode laser (InGaAlP, wavelength of 660 nm, potency of 30 Mw, in a single dose of 1 J/cm²). Cell viability and proliferation will be assessed at 24, 48 and 72 hours after irradiation through the Alamar Blue and Live/Dead assays. Cell morphology will be evaluated by SEM at 72 hours. The quantitative data will be submitted to statistical tests, with significance level of 5%.