Decellularized matrix of porcine skeletal muscle (dECM-pSM) and its in vitro biocompatibility as a graft approach for oral mucosa regeneration

Oral mucosa regeneration represents a significant clinical challenge due to the limited availability of autologous grafts and associated morbidity. This study focused on developing and characterizing decellularized matrices (dECM-pSM) for their potential use in oral mucosa restoration. A perfusion decellularization protocol was implemented on porcine skeletal muscle segments, utilizing a combination of physical (perfusion), chemical (SDS), and enzymatic (DNase-1) agents over 18 days. The effectiveness of the process was evaluated macroscopically, through histological stains (H&E, DAPI, Masson's Trichrome), scanning electron microscopy (SEM), DNA quantification, and FTIR spectroscopy. The thermal properties (TGA, DSC), swelling, and biocompatibility of the dECM-pSM with human gingiva fibroblast cells (HGF) were analyzed, including adhesion and proliferation assays. The results showed successful decellularization, with significant removal of nuclear material (0.7 ng/mg residual DNA) with preservation of the three-dimensional architecture of ECMs and physicochemical properties, as confirmed by histological integrity of the fiber and porous structures, preserved characteristic bands of amide groups, and stable thermal properties after the decellularization process. The dECM-pSM maintained their swelling capacity (300% after 5 minutes) and demonstrated excellent biocompatibility, promoting the adhesion at 92% after 2 days, and proliferation of 80% after 4 days of HGF compared to the control, without evidence of cytotoxicity. These results suggest that the developed protocol yields decellularized matrices with suitable properties for tissue engineering applications.