Evaluation of the influence of soft liners on the integrin alpha 5 beta 1 of keratinocytes

Abstract

Soft liner materials have been used for reconditioning of denture-bearing mucosa that has inflamed and distorted by ill-fitting dentures, in maxillofacial prostheses and obturators, in patients with chronic tissue soreness, traumatized mucosa, avitaminosis or other debilitating diseases, and to provide aftercare of an immediate denture and during implant therapy. It is also known that, in the healing process, the integrins and their ligands play an important role in the regulation of cell proliferation, differentiation and migration. Thus, the objective of this study is to evaluate the influence of two silicone-based soft liner materials (Ufigel P and Sofreliner S), 2 acrylic-based (Durabase Soft and Trusoft) and two tissue conditioners (Softone and Coe Comfort) on integrin alpha5Beta1 of HaCat cells, after direct contact for 24 or 48 hours. Specimens of each material (10 X 1 mm) will be made and placed, individually, in the wells of a culture plate and the cells (1 × 105 cells/mL) will be seeded directly. Wells containing sterile glass coverslips with the same diameter of the specimens will be used as controls. Thereafter, the plates will be incubated at 37ºC in a 5% CO2, 95% air atmosphere for 24 or 48 hours. To analyze the integrin alpha5Beta1, the Kit Human Total Integrin alpha5Beta1 (Duo Set IC - R & D Systems, Inc) will be used according to the manufacturer's instructions. The optical density will be determined immediately using a microplate reader set to 450 nm and 570 nm. Experiments will be conducted in triplicate and the average of these readings for each standard and specimen will be calculated, and then the average zero standard optical density will be subtracted. Initially, a standard curve will be created by plotting the mean absorbance for each standard on the y-axis against the concentration on the x-axis and drawing a best fit curve through the points on the graph. The integrin concentration in the samples will be quantified by interpolation from the standard curve. (AU)