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摘要
The treatment of long bone defects and nonunions remains a major challenge for orthopaedic surgeons. This study evaluates the ability of porous nano-hydroxyapatite/polyamide66 (nHA/PA66) scaffolds combined with MSCs, gene-modified by Nell-1, to repair a fracture with severe periosteal damage that is known to result in nonunion. The nHA/PA66 scaffolds were prepared and rat MSCs were transduced with Nell-1 gene via lentivirus. The prolonged expression of Nell-1 gene was confirmed by western blotting and RT-qPCR. The osteogenic differentiation after transduction was verified by ALP activity, alizarin red staining and RT-qPCR. At 7 days after induction, the lentiviral-transfected MSCs and untransfected MSCs were seeded on scaffolds. Cell adhesion and proliferation were assayed using MTT assay and scanning electron microscopy (SEM). Then the hybrid scaffolds and scaffolds alone were implanted in rat femur fracture sites with severe periosteal damage. At 4 and 8 weeks after implantation, new bone formation was assayed by radiograph, Micro-CT and histological analysis respectively. Overall, the Nell-1 gene was successfully expressed for up to eight weeks and LvNell-1 transduction promoted the osteogenic differentiation of MSCs. The nHA/PA66 scaffolds possesses good biocompatibility and improve the efficiency of new bone formation combined with LvNell-1-transduced MSCs. These results demonstrate that LvNell-1 gene modified rat MSCs seeded on nHA/PA66 scaffolds can serve as an ideal tissue-engineered bone to repair fractures with severe periosteal damage, which will predictably lead to nonunion.
- 出版日期2016-5
- 单位哈尔滨医科大学
