Production of Polymeric Membranes Functionalized with Vitreous Humor and Secretome from Dental Pulp Stem Cells for Use in Epithelial Regeneration

Abstract

This study investigated the development of porous membranes based on a matrix of bacterial cellulose (BC) and sodium alginate (ALG) incorporated with proteins extracted from the tuna vitreous humor and complete conditioned medium (CCM) obtained from the in vitro culture of dental pulp stem cells. These bioactive components were integrated into BC-ALG membranes, which were then evaluated for morphology, physicochemical properties, in vitro cytotoxicity, and regenerative potential in vitro. The membranes exhibited high porosity (72% to 89%), and remarkable swelling capacity reaching up to 3376 ± 0.78% in 72 min, indicating excellent moisture retention essential for wound healing in vivo. Cytotoxicity assay showed that none of the formulations were cytotoxic to the tested mammalian cells, confirming their cytocompatibility. Membranes containing CCM showed enhanced cell viability at lower concentrations (500 μg/mL and 250 μg/mL). In vitro scratch assays demonstrated that membranes with vitreous humor (VH) and CCM reduced scratch closure time from 18 to 12 h, suggesting their potential for accelerating skin regeneration. This work underscores the promise of using sustainable, waste-derived materials for creating bioactive membranes with applications in tissue engineering and regenerative medicine.