Methacrylated collagen hydrogel formulation extracted from tilapia skin functionalized with proanthocyanidin for endodontic regeneration

Abstract

Regenerating pulp tissue is challenging due to anatomical irregularities and individual variations, leading to limited efficacy of conventional treatments. This highlights the need for new, minimally invasive therapeutic approaches and the development of adaptable materials that not only precisely fit the defect site but also promote the migration and differentiation of resident pulp and apical papilla cells, a process referred to as cell homing therapy. This project aims to develop injectable scaffolds with a macroporous and interconnected structure, enriched with proanthocyanidin, a polyphenol known for its bioactive, pro-angiogenic, and antimicrobial properties capable of releasing bioactive dosages to stimulate pulp regeneration. Initially, the bioactive and pro-angiogenic concentrations of the polyphenol (proanthocyanidin) on dental papilla cells (SCAPs), endothelial cells (HUVECs), and antimicrobial effect (Enterococcus faecalis) will be determined (Phase 1). Then, ColMA concentrations will be tested to identify the ideal formulation that favors pulp regeneration, focusing on collagen deposition and angiogenic structure formation by apical papilla and endothelial cells, including the physicochemical, mechanical, and biological characterization of the hydrogels (Phase 2). Subsequently, the formulation and functionalization of ColMA with proanthocyanidins will be conducted, selecting the incorporation method to release bioactive concentrations, with evaluation of the bioactive and antimicrobial potential in vitro (Phase 3). Finally, the injectable hydrogels will be implanted subcutaneously in nude mice (nu/nu) after filling the root space of root fragments, to evaluate the potential to induce the formation of vascularized tissue and collagen-rich matrix deposition (Phase 4). In the end, qualitative data will be descriptively analyzed, and quantitative data will be subjected to specific statistical analysis.