Effect of Porous Chitosan Microspheres Loaded with Platelet-Rich Plasma and Bone Marrow-Derived Mesenchymal Stem Cells on Regeneration of Tibia Defect

Abstract

Objective. Repair of bone defects represents a grave clinical challenge because of the tremendous difficulties in the recovery of bone function and regeneration of bone loss. Therefore, we investigated the effects of platelet-rich plasma-loaded (PRP) porous chitosan microspheres (PCMs) on the differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) and the proliferation and differentiation potential of BMSCs loaded by PCMs in vitro. We also established the model of bone defect repair in rat tibia to further explore the effects of PCMs loaded with PRP and BMSCs on bone regeneration. Methods. MTT assay was used to detect the proliferative ability of BMSCs after hypoxia/reoxygenation (H/R) treatment and the proliferative ability of BMSCs loaded by PCMs; polymerase chain reaction (PCR) was used to detect the expression of alkaline phosphatase (ALP), type I collagen (Col I), and type II collagen (Col II) in BMSCs after hypoxia and in BMSCs induced by PRP-loaded PCMs; PCR was used to detect the expression of Runt-associated transcription factor 2 (Runx2) and osteocalcin (OC) in the newly generated bone tissue; micro-CT scanning was applied to measure the bone mineral density and bone volume of the newly generated bone tissue in rats. Results. BMSCs still have the normal potential of proliferation and differentiation after H/R treatment. PCMs can provide a larger surface for the attachment of BMSCs, facilitating cell proliferation. Loaded by PCMs, PRP can be slowly released, effectively stimulating the differentiation of BMSCs. PCM/PRP/BMSC composites increased the expression levels of Runx2 and OC in the newly generated bone in rat tibia defect and the bone mineral density. Moreover, the composites improved the rate of regenerated bone volume. Conclusion. The application of PCM/PRP/BMSC composites is promising in the repair of tibia defects.