Advanced 3D-Printed Multiphasic Scaffold with Optimal PRP Dosage for Chondrogenesis of BM-MSCs in Osteochondral Tissue Engineering

Abstract: In osteochondral tissue engineering (OCTE), simultaneously regenerating subchondral bone and cartilage tissue presents a significant challenge. Multiphasic scaffolds were created and manufactured using 3D printing to address this issue. Excellent interfacial mechanical properties and biocompatibility enhance the growth and chondrogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs). The subchondral bone bottom layer is mimicked by incorporating varying concentrations of graphene oxide (GO) (0%, 1%, and 2% w/v) into a bioink composed of alginate (Alg) and gelatin (Gel). Based on evaluations of mechanical and biocompatibility properties, 1% GO is selected for further studies. Subsequently, the GO concentration is kept constant while varying the platelet-rich plasma (PRP) dosage in the multiphasic scaffolds. Different PRP dosages (0%, 1%, 2%, and 3% w/v) are integrated into the Alg-Gel bioink to simulate cartilage tissues. Results indicate that 3D-printed scaffolds containing 1% or 2% PRP exhibit favorable biomechanical properties, with no significant differences observed. However, BM-MSCs exposed to 2% PRP demonstrate enhanced adhesion, growth, and viability. Additionally, real-time PCR and Alcian blue staining confirm increased chondrogenic expression and glycosaminoglycans (GAGs) synthesis. This work highlights the promising potential of 3D-printed multiphasic frameworks in the development of OCTE.