In vitro analysis of cell proliferation and inflammatory mediators' production on keratinocytes stimulated by different adhesive systems used to fabricate implant-retained maxillofacial prosthesis

Abstract

The implant-retained maxillofacial prosthesis can be supported by skin and mucosa. The sub-products produced by the materials used to fabricate these prostheses may act as an irritant factor and cause allergic reaction when in contact with the skin or mucosa. The aim of the present study is to evaluate the cell proliferation and inflammatory mediators' production on keratinocytes stimulated by different adhesive systems used to bond the acrylic resin and the facial silicone during implant-retained maxillofacial prosthesis fabrication. A total of 21 round shape samples (10 x 1 mm) made of resin and silicone bonded or not with primer and adhesive will be fabricated. Samples will be divided into 7 groups: Resin (R), Silicone (S), Resin + Silastic Medical Adhesive Type A + Silicone (RAS), Resin + DC 1205 Primer + Silicone (RDCpS), Resin + Sofreliner Primer + Silicone (RSpS), Resin + DC 1205 Primer + Silastic Medical Adhesive Type A + Silicone (RDCpAS), and Resin + Sofreliner Primer + Silastic Medical Adhesive Type A + Silicone (RSpAS). Elution samples of the materials will be prepared by placing three samples of each experimental group in 9 ml of DMEM sterile tissue culture medium (Dulbecco's Modified Eagle's) and incubated at 37 ºC during 24 hours. The medium will be tested for toxicity by an assay of cell survival/proliferation (MTT test) in cultures of human keratinocytes (HaCaT). The levels of IL-1, IL-6 and TNF-± and the chemokine MIP-1± will be evaluated by ELISA (Enzyme-Linked Immunoabsorbent Assay). The mRNA expression for MMP-9 and TGF-² will be analyzed by the RT-PCR (Real time polymerase chain reaction). Data will be submitted to the analysis of variance. The results will be also evaluated according to the ISO-standard 10993-5, that describes the in vitro methods to evaluated cytotoxicity. (AU)