ABSTRACT
Objective: To separate peripheral blood mesenchymal stem cells (PBMSC) and peripheral blood endothelial progenitor cells (PBEPC) from peripheral blood, and investigate the biological characteristics of composite cell sheets of PBMSC and PBEPC. Methods: The peripheral blood of healthy adult New Zealand white rabbits was extracted and PBMSC and PBEPC were separated by density gradient centrifugation. Morphological observation and identification of PBMSC and PBEPC were performed. The 3rd generation of PBMSC and PBEPC were used to construct a composite cell sheet at a ratio of 1∶1, and the 3rd generation of PBMSC was used to construct a single cell sheet as control. The distributions of cells in two kinds of cell sheets were observed by HE staining. In addition, the expression of alkaline phosphatase (ALP), osteocalcin (OCN), and vascular endothelial growth factor (VEGF) in the supernatants of cell sheets were observed by ELISA at 1, 5, and 10 days after osteogenic induction. Results: The morphology of PBMSC was spindle-shaped or polygonal, and PBMSC had good abilities of osteogenic and adipogenic differentiation. The morphology of PBEPC was paved stone-like, and the tube-forming test of PBEPC was positive. Two kinds of cell sheets were white translucent. The results of HE staining showed that the composite cell sheet had more cell layers and higher cell density than the single cell sheet. The expressions of ALP, OCN, and VEGF in the supernatant of the two groups of cell sheets increased with the time of induction. The expression of OCN in the group of composite cell sheet was significantly higher than that in the group of single cell sheet on the 5th and 10th day, ALP on the 10th day was significantly higher than that in the group of single cell sheet, VEGF expression on the 1st, 5th, and 10th day was significantly higher than that in the group of single cell sheet, all showing significant differences ( P0.05). Conclusion: PBMSC have stable differentiation ability, and they have good application prospects because of their minimally invasive access. Composite cell membranes constructed by co-culture of two kinds of cells and induction of membrane formation provides a new idea and exploration for tissue defect repair.
ABSTRACT
Objective This study aimed to explore the effect of peripheral blood mesenchymal stem cells combined with porous absorbable gelatin sponge/self assembling peptide composite scaffolds on SD rat femoral con-dyle bone defect reconstruction and provide a new strategy for the repair of bone defects. Methods 30 female SD rats,8W age,were randomly divided into 3 groups,10 every group.The group A was blank control group,group B was porous absorbable gelatin sponge/self assembling peptide composite scaffold group,and group C was periph-eral blood mesenchymal stem cells combined with porous absorbable gelatin sponge/self assembling peptide compos-ite scaffold group. The effect of osteogenesis was observed by paraffin section,hematoxylin eosin staining,X-ray examination,and Micro-CT scanning in 3 dimensional reconstruction of femoral condyle defect. Results Imaging examination showed that the experimental group had better osteogenesis effect. Histological examination showed that a lot of new bone tissue was found in group C,while only a small amount of new bone was found in the group of A and B. Conclusions The experiment shows that peripheral blood mesenchymal stem cells as the seed cells for tissue engineering,combined with porous absorbable gelatin sponge-self assembling peptide has better ability to repair bone defects,and has good application prospect,which is worthy of further research.