Bioactive Nanofibrous Scaffolds Incorporating Decellularized Cell-Derived Extracellular Matrix for Periodontal Tissue Engineering

Periodontium is a complex structure that supports the teeth, composed of gingiva, cementum, periodontal ligament, and alveolar bone. Destruction of the alveolar bone can occur due to periodontitis, trauma, or following tumor resection. Current reconstructive treatments are based on the use of bone grafts with limited efficacy and predictability, lacking bioactive signals to induce tissue repair and coordinated periodontal regeneration. Thus, alternative strategies are needed to improve clinical outcomes. Cell-derived extracellular matrix (ECM) has been combined with biomaterials to enhance their biofunctionalities for various tissue engineering (TE) applications. In this work, bioactive cell-derived ECM-loaded electrospun polycaprolactone/chitosan (PCL/CTS) nanofibrous scaffolds were developed by combining polymer solutions with lyophilized decellularized ECM (dECM) derived from human periodontal ligament stem/stromal cells (PDLSCs). This work aimed to fabricate and characterize cell-derived ECM electrospun PCL/CTS scaffolds in terms of morphological, physicochemical, thermal, and mechanical properties and assess their ability to enhance the osteogenic differentiation of PDLSCs, envisaging potential applications in periodontal TE, particularly focused on the regeneration of alveolar bone defects. PDLSCs-derived dECM was characterized regarding morphology, protein expression, DNA removal efficiency, glycosaminoglycans and collagen contents. Osteogenic differentiation of PDLSCs was performed on PCL, PCL/CTS, and PCL/CTS/ECM electrospun scaffolds for 21 days. The obtained results demonstrate that PCL/CTS/ECM scaffolds promoted cell proliferation compared to PCL and PCL/CTS scaffolds while maintaining similar physical and mechanical properties of PCL/CTS scaffolds. PCL/CTS/ECM scaffolds enhanced the osteogenic differentiation of PDLSCs, confirmed by increased alkaline phosphatase activity, calcium deposition, and bone-specific marker gene expression. Moreover, PCL/CTS scaffolds showed higher levels of cell mineralization than did PCL scaffolds. Overall, this work describes the first use of lyophilized cell-derived ECM-loaded electrospun scaffolds for periodontal TE applications, highlighting the potential of this strategy for innovative therapeutic strategies, in particular, for alveolar bone regeneration.