Three-Dimensional Printing of Customized Scaffolds with Polycaprolactone-Silk Fibroin Composites and Integration of Gingival Tissue-Derived Stem Cells for Personalized Bone Therapy.

Abstract

Regenerative biomaterials play a crucial role in the success of maxillofacial reconstructive procedures. Yet today, limited options are available when choosing polymeric biomaterials to treat critical size bony defects. Further, there is a requirement for 3D printable regenerative biomaterials to fabricate customized structures confined to the defect site. We present here a 3D printable composite formulation consisting of polycaprolactone (PCL) and silk fibroin microfibers and have established a robust protocol for fabricating customized 3D structures of complex geometry with the composite. The 3D printed composite scaffolds demonstrated higher compressive modulus than 3D printed scaffolds of PCL alone. Furthermore, the compressive modulus of PCL-Antheraea mylitta (AM) silk scaffolds is higher than that of the PCL-Bombyx mori (BM) silk scaffolds at their respective ratios. Compressive modulus of PCL-25AM silk scaffolds (73.4 MPa) is higher than that of PCL-25BM silk scaffolds (65.1 MPa). Compressive modulus of PCL-40AM silk scaffolds (106.1 MPa) is higher than that of PCL-40BM silk scaffolds (77.7 MPa). Moreover, we have isolated, characterized, and integrated human gingival mesenchymal stem cells (hGMSCs), an effective autologous cell source, onto the 3D printed scaffolds to evaluate their bone regeneration potential. The results demonstrated that PCL-silk microfiber composite scaffolds of Antheraea mylitta origin showed much higher bioactivity than the Bombyx mori ones because of arginine-glycine-aspartic acid (RGD) sequences present in the Antheraea mylitta silk fibroin protein favoring cell attachment and proliferation. By day 14, the metabolic activity of hGMSCs was highest in PCL-40AM (4.5 times higher than that at day 1). In addition, to show the translational potential of this work, we have fabricated a patient defect-specific model (mandible) using the CT scan obtained by the micro-CT imaging to understand the printability of the composite for fabricating complex structures to restore maxillofacial bony defects with precision when applied in a clinical scenario.