Abstract
We aimed to evaluate the effect of activated platelet-rich plasma (PRP) on proliferation and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Six mature male rabbits were included in this study. PRP was obtained by two-step centrifugation from whole blood, and it was activated using CaCl2 solution. BMSCs were isolated and proliferated from bone marrow of rabbits and characterized by flow cytometry. Passage 3 BMSCs were cultured in high-glucose Dulbecco's modified Eagle's medium (HG-DMEM) with the four different compositions for consecutive 7 days, including 10% fetal bovine serum, 5% PRP, 10% PRP, and 15% PRP. Cell counting assays were performed to evaluate the cell proliferation of BMSCs. BMSCs (5 × 105 cells/well in 6-well plates) were induced in four conditions for 21 days to chondrogenic differentiation evaluation, including commercial chondrogenic medium (control), 5% PRP (HG-DMEM+5% PRP), 10% PRP (HG-DMEM+10% PRP), and 15% PRP (HG-DMEM+15% PRP). The gene expression levels of ACAN, COL2A1, and SOX9 in pellets were detected. Morphological and pathological assessments were performed by the blind observer. After purifying, the percentages of cells with CD105(+)/CD34(−) and CD44(+)/CD45(−) were 96.5% and 92.9%, respectively. The proliferation of BMSCs was enhanced in all groups, and 10% PRP revealed more significant outcome than the others from day 5. The levels of ACAN, COL2A1, and SOX9 were lower in the three PRP groups than control group, but the levels of ACAN and SOX9 were higher in 10% PRP group than 5% and 15% PRP groups. Histological examinations showed that 10% PRP-treated pellets had more regular appearance, larger size, and abundant extracellular matrix than 5% or 10% PRP groups, but still inferior to commercial chondrogenic medium. In conclusion, our results show that PRP may enhance the proliferation of rabbit BMSCs. However, PRP have limited effect on chondrogenic differentiation in comparison with commercial chondrogenic medium in pellets culture.
Highlights
Repairing massive articular cartilage defect remains a challenging issue
Bone marrow-derived mesenchymal stem cells (BMSCs) have the potential for chondrogenic differentiation, self-renewal, and proliferation with less loss of phenotypes, which have been considered as an ideal cell source for tissue-engineered cartilage formation [4]
We aimed to evaluate the effects of different concentrations of platelet-rich plasma (PRP) on the proliferation and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) cultured in 3D pellets in vitro
Summary
Repairing massive articular cartilage defect remains a challenging issue. Current available surgical techniques include osteochondral transplantation and autologous chondrocyte implantation [1, 2]. Despite of relative satisfying results, these techniques have been limited by morbidity of the donor sites and loss of the chondrogenic phenotype during in vitro expansion [3]. Whether tissue-engineered cartilage can effectively repair massive articular cartilage defects has been a novel focus in this field. Bone marrow-derived mesenchymal stem cells (BMSCs) have the potential for chondrogenic differentiation, self-renewal, and proliferation with less loss of phenotypes, which have been considered as an ideal cell source for tissue-engineered cartilage formation [4]. The extremely low percentage of BMSCs in bone marrow means that in vitro expansion is the first step to obtain enough cells
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