异位成骨材料修复新西兰兔桡骨干骨缺损
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摘要
目的探索异位成骨材料修复新西兰兔桡骨干骨缺损的可行性。方法13只新西兰白兔(4周龄,体质量1.5 kg)作为实验动物。取新西兰白兔骨髓间充质干细胞进行成骨诱导,2周后予碱性磷酸酶染色确认其成骨活性。取诱导后的间充质干细胞悬液与β-磷酸三钙生物陶瓷共培养,3天后植入4只新西兰白兔股骨骨膜旁。于植入后第8周收取异位成骨材料,行骨组织切片检查。取8只新西兰兔,制作桡骨干骨缺损动物模型,并将其均分为实验组(4只)和对照组(4只),分别植入异位成骨材料和β-磷酸三钙生物陶瓷修复骨缺损,于术后4、8、12周行手术侧X线摄片,术后12周取修复处骨,行组织切片检查。结果异位成骨植入第8周见植入物成骨完整,获取的成骨材料组织切片显示与股骨干骨细胞结构相似。骨缺损修复动物实验术后X线摄片显示,实验组修复效果显著优于对照组;术后12周修复处骨组织切片显示,实验组新生骨与周边骨组织边界消失,融合度好,骨小梁密度高,新生血管较多。结论异位成骨材料可用于兔桡骨干骨缺损修复。
Objective To investigate the feasibility of ectopic osteogenic compound repairing radial bone defects in New Zealand white rabbits.Methods Thirteen healthy New Zealand white rabbits(4 weeks old,1.5 kg)were selected as experimental animals.Bone marrow-derived mesenchymal stromal cells(BMSC)were isolated.Two weeks after osteoinduction of BMSC,alkaline phosphatase(ALP)staining were undertaken to evaluate the bone formation.After osteoinduction,BMSC suspension was added to the biological ceramic containingβ-tricalcium phosphate(β-TCP)for co-culture for 3 days before they were implanted besides the femoral periosteum.The ectopic osteogenic compound was harvested at 8 weeks,and examined under microscope.Eight New Zealand white rabbits were used to established radial bone defect models and randomly divided into the experimental group(ectopic osteogenic compound,n=4)and the control group(β-TCP,n=4),respectively.The X-ray examination was carried out at 4,8 and 12 weeks.The regenerated tissue was harvested for examination at 12 weeks.Results The ectopic osteogenic compound was harvested successfully at 8 weeks after implantation,and its cellular structure was similar to the normal one of the adjacent femur shaft.In terms of the repair outcome of bone defects,the X-ray demonstrated that the experimental group was significantly superior to the control group;at 12 weeks,the regenerated bone in the experimental group was integrated better with the adjacent normal bone,with higher bone density and more blood vessels,compared with the control group.Conclusion The ectopic osteogenic compound can be used to repair radial bone defects in New Zealand white rabbits.
Objective To investigate the feasibility of ectopic osteogenic compound repairing radial bone defects in New Zealand white rabbits.Methods Thirteen healthy New Zealand white rabbits(4 weeks old,1.5 kg)were selected as experimental animals.Bone marrow-derived mesenchymal stromal cells(BMSC)were isolated.Two weeks after osteoinduction of BMSC,alkaline phosphatase(ALP)staining were undertaken to evaluate the bone formation.After osteoinduction,BMSC suspension was added to the biological ceramic containingβ-tricalcium phosphate(β-TCP)for co-culture for 3 days before they were implanted besides the femoral periosteum.The ectopic osteogenic compound was harvested at 8 weeks,and examined under microscope.Eight New Zealand white rabbits were used to established radial bone defect models and randomly divided into the experimental group(ectopic osteogenic compound,n=4)and the control group(β-TCP,n=4),respectively.The X-ray examination was carried out at 4,8 and 12 weeks.The regenerated tissue was harvested for examination at 12 weeks.Results The ectopic osteogenic compound was harvested successfully at 8 weeks after implantation,and its cellular structure was similar to the normal one of the adjacent femur shaft.In terms of the repair outcome of bone defects,the X-ray demonstrated that the experimental group was significantly superior to the control group;at 12 weeks,the regenerated bone in the experimental group was integrated better with the adjacent normal bone,with higher bone density and more blood vessels,compared with the control group.Conclusion The ectopic osteogenic compound can be used to repair radial bone defects in New Zealand white rabbits.
引文
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[2]裴国献.组织工程学:21世纪面临的机遇与挑战[J].中华创伤骨科杂志,2006,8(1):2-4.
[3]Florczyk SJ,Leung M,Li Z,et al.Evaluation of three-dimensional porous chitosan-alginate scaffolds in rat calvarial defects for bone regeneration applications[J].J Biomed Mater Res A,2013,101(10):2974-2983.
[4]Service RF.Tissue engineering.Technique uses body as‘bioreactor’to grow new bone[J].Science,2005,309(5735):683.
[5]Dong Y,Chen X,Wang M,et al.Construction of artificial laminae of the vertebral arch using bone marrow mesenchymal stem cells transplanted in collagen sponge[J].Spine(Phila Pa 1976),2012,37(8):648-653.
[6]Yang CR,Wang YJ,Chen XF.Preparation and evaluation of biomimetric nano-hydroxyapatite-based composite scaffolds for bone-tissue engineering[J].Chin Sci Bull,2012,57(21):2787-2792.
[7]Lin RZ,Moreno-Luna R,Li D,et al.Human endothelial colony-forming cells serve as trophic mediators for mesenchymal stem cell engraftment via paracrine signaling[J].Proc Natl Acad Sci USA,2014,111(28):10137-10142.
[8]Xie F,Teng L,Wang Q,et al.Ectopic osteogenesis of allogeneic bone mesenchymal stem cells loading onβ-tricalcium phosphate in canines[J].Plast Reconstr Surg,2014,133(2):142e-153e.
[9]Victorelli G,Aloise AC,Passador-Santos F,et al.Ectopic implantation of hydroxyapatite xenograft scaffold loaded with bone marrow aspirate concentrate or osteodifferentiated bone marrow mesenchymal stem cells:apilot study in mice[J].Int J Oral Maxillofac Implants,2016,31(2):e18-e23.
[10]Caplan AI.New era of cell-based orthopedic therapies[J].Tissue Eng Part B Rev,2009,15(2):195-200.
[11]陈欣,张春林.组织工程骨修复骨缺损的研究进展及临床应用[J].中国组织工程研究与临床康复,2010,14(24):4486-4490.
[12]Zhang ZY,Teoh SH,Hui JH,et al.The potential of human fetal mesenchymal stem cells for off-the-shelf bone tissue engineering application[J].Biomaterials,2012,33(9):2656-2672.
[13]Lapi S,Nocchi F,Lamanna R,et al.Different media and supplements modulate the clonogenic and expansion properties of rabbit bone marrow mesenchymal stem cells[J].BMC Res Notes,2008,1:53.
[14]Kavalkovich KW,Boynton RE,Murphy JM,et al.Chondrogenic differentiation of human mesenchymal stem cells within an alginate layer culture system[J].In Vitro Cell Dev Biol Anim,2002,38(8):457-466.