Evaluation of JDBM porous scaffold coated with DCPD in promoting angiogenesis and repairing bone defects

doi:10.3969/j.issn.1674-8115.2021.06.005

Abstract

Abstract: Objective

·To evaluate the biological effects of JDBM (Mg-Nd-Zn-Zr) scaffold coated with DCPD (CaHPO₄·2H₂O) on angiogenesis and repairing bone defects in vivo and in vitro.

Methods

·The JDBM-DCPD and JDBM-MgF₂ scaffolds were constructed by using template replication method and chemical deposition method, and the characteristics of the scaffolds were observed by micro-CT and scan electron microscope. The bone marrow mesenchymal stem cells (BMSCs) were seeded on the scaffolds and the biocompatibility of scaffolds was evaluated by CCK-8 experiment and cell adhesion experiment. Transwell cell migration experiment and tube formation experiment were used to detect the effects of scaffold extracts on the migration and tube-forming ability of endothelial cell line Ea.hy926 cells, and immunofluorescence was used to further observe the secretion of vascular endothelial growth factor (VEGF). Alkaline phosphatase staining and alizarin red staining were used to detect the effect of the extracts on the osteogenic ability of BMSCs. Furthermore, the model of critical bone defect of femoral condyle was constructed in SD rats. The JDBM-DCPD and JDBM-MgF₂ scaffolds were implanted into the defects, respectively, and the effects of scaffolds on the osteogenesis and vascularization were assessed by Microfil vascular perfusion, micro-CT scanning, and tissue section staining after 8 weeks of operation.

Results

·The main spherical pore size of JDBM-DCPD scaffold was 400?450 μm and the calcium and phosphorus particles were evenly distributed on the pore wall with the size of 15?25 μm. BMSCs adhered and grew well on the surface of JDBM-DCPD scaffold. Compared with the control group and the JDBM-MgF₂ scaffold extract, the JDBM-DCPD scaffold extract could significantly promote the migration, tube formation and VEGF expression of Ea.hy926 cells, and significantly enhance the early and late osteogenic differentiation of BMSCs in vitro. After 8 weeks of implantation, the JDBM-DCPD scaffold dramatically facilitated the regeneration of new bone and new vessels in the defect area compared with the JDBM-MgF₂ scaffold.

Conclusion

·The JDBM-DCPD scaffold exhibits excellent vascularization effects both in vivo and in vitro, especially early vascularization effect after implantation and bone regeneration promotion in vivo.

Key words: Mg-Nd-Zn-Zr alloy (JDBM), DCPD coat, angiogenesis, osteogenic differentiation, bone regeneration, bone tissue engineering

CLC Number:

R318.08

Qing WANG, Wei WANG, Da-jun JIANG, Wei-tao JIA. Evaluation of JDBM porous scaffold coated with DCPD in promoting angiogenesis and repairing bone defects[J]. JOURNAL OF SHANGHAI JIAOTONG UNIVERSITY (MEDICAL SCIENCE), 2021, 41(6): 732-740.

Figures/Tables 7

Fig 1 Comparison of JDBM-DCPD and JDBM-MgF2 porous scaffolds in characteristics

Fig 2 Comparison of JDBM-DCPD and JDBM-MgF2 in biocompatibility

Fig 3 Comparison of the effects of JDBM-DCPD and JDBM-MgF2 scaffold extracts on Ea.hy926 cell migration and tube formation

Fig 4 Comparison of the effects of JDBM-DCPD and JDBM-MgF2 scaffold extracts on the expression of VEGF in Ea.hy926 cells

Fig 5 Comparison of effects of JDBM-DCPD and JDBM-MgF2 scaffold extracts on osteogenic differentiation of BMSCs

Fig 6 Osteogenesis and angiogenesis of JDBM-DCPD and JDBM-MgF2 scaffolds in vivo detected by radiography 8 weeks after implantation

Fig 7 Histological observation of the bone defect areas in JDBM-DCPD group and JDBM-MgF2 group rats 8 weeks after implantation

TrendMD

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