Angiogenesis and in Vitro Bone Differentiation of Hmscs on Pcl Nanofiber Scaffolds Containing Homogenized Platelet-Rich Fibrin-Chitosan Nanoparticles: An in Ovo Cam Model Publisher



Alizadeh M ; Mahmoodi M
Source: Progress in Biomaterials Published:2025

Abstract

Bone regeneration remains a major clinical challenge, as current scaffolds rarely replicate the optimal microenvironment required for effective tissue repair, particularly in stimulating vascularization. This study aimed to develop and evaluate a polycaprolactone (PCL) nanofiber scaffold incorporating chitosan (CH) nanoparticles loaded with homogenized platelet-rich fibrin (PCL/CH-hPRF) for bone tissue engineering. Scaffolds were fabricated via electrospinning and characterized by dynamic contact angle measurements, mechanical testing, degradation test, and in vitro bioactivity. Also, biological performance was evaluated by protein adsorption, MG-63 cell biocompatibility, and osteogenic differentiation of human mesenchymal stem cells (HMSCs) through alkaline phosphatase activity, calcium deposition, and Alizarin red staining. Furthermore, angiogenic potential was investigated using the in ovo chorioallantoic membrane (CAM) assay. Incorporating CH-hPRF nanoparticles into PCL nanofibers reduced the average fiber diameter from 230 nm in PCL scaffolds to 189 nm in PCL/CH-hPRF scaffolds. The PCL/CH-hPRF scaffold showed an increased degradation rate relative to other scaffolds, which can be attributed to enhanced fluid infiltration driven by its increased porosity and lower contact angle. The PCL/CH-hPRF scaffolds exhibited a significantly higher elastic modulus (45.35 MPa) compared to PCL scaffolds (35.53 MPa). Moreover, the hPRF release from the PCL/CH-hPRF scaffold reachesd 31.25% after 14 days, indicating a gradual and prolonged release of hPRF from the nanofibrous scaffold. After 28 days of immersion in simulated body fluid (SBF), the PCL/CH-hPRF scaffold exhibited higher bioactivity and enhanced biomineralization compared to the PCL scaffold. The PCL/CH-hPRF scaffold demonstrated excellent cell compatibility and promoted rapid osteogenic differentiation of HMSCs, with substantial calcium deposition observed over 14 days. Furthermore, the in ovo CAM assay revealed that the PCL/CH-hPRF scaffold induced approximately threefold higher blood vessel formation compared to the CH-hPRF scaffold. These results suggest that the PCL/CH-hPRF composite nanofiber scaffold possesses superior osteogenic, angiogenic, and biological properties, making it a promising candidate for bone tissue engineering applications. © 2025 The Author(s).