纳米晶胶原基骨修复骨缺损

骨髓间充质干细胞复合音猬因子/纳米

晶胶原基骨修复骨缺损

(作者： __________ 单位：___________ 邮编： ___________ )

作者：史坚强孙广友，马鹏，徐成振钱栋徐晓峰

【摘要】目的：探讨骨髓间充质干细胞(marrow mesenchymalstem cells )复合音猬因子(sonic hedgehog,Shh )修饰的纳米晶胶原基骨(nano Hydroxyapatite/collage n, nHAC) 修复大鼠股骨缺损的效

果。方法：SD大鼠MSCs经成骨诱导后，种植于支架材料形成细胞支架复合物。SD大鼠30只制成股骨干5 mm节段性缺损，分为空白对照组：骨缺损处不植植入物，MSCs/nHA组：植入MSCs/nHA复合物，

MSCs/Sh拟nHAC组：植入MSCs/Sh拟nHAC复合物。通过大体观察、X线放射学、组织学对比各组骨缺损修复情况。结果：SD大鼠MSCs

在体外与nHACShhU.nHAC复合培养第7天，电镜扫描证实Shh以nHAC 更有利于MSCs黏附、增殖。术后第6、12周放射学检查评价新骨生成各组间差异有统计学意义(P0.05)。术后组织学检查各组新骨生成速度、生成量差异有统计学意义(P0.05)。结论：MSCs/Sh^' nHAC 复合物能够修复SD大鼠股骨缺损，修复效果明显优于MSCs/nHACE 合物。

【关键词】骨髓间充质干细胞；纳米晶胶原基骨；骨缺损；音猬因

子

[Abstract ] Objective: To explore the repair effect of the

segme ntal bone defects in rats with nano Hydroxyapatite/collage n modified by bone marrow mese nchymal stem cells co ^cultured with sonic hedgehog(Shh ) .Methods : MSCs obtained from SD rats bone marrow were induced and proliferated.After their osteoblastic phe no type had bee n dem on strated, MSCs were seeded on to the scaffolds.A bone

defect(5 mmin length) was created in each rat and treated with three kinds of implantations」n blank control group , defect was filled with none; in MSCs/nHACgroup with MSCs/nHAC

compo und;in MSCs/Shh)；nHAC group with MSCs/Shh)；nHAC compo un d.The repair effect of bone defect of every group was evaluated by gen eral observatio n, radiographic exam in atio n and histologic analysis. Results : MSCs of SD rats were

co 拟cultured in vitro with nHAC, Shh 拟nHAC separately for 7

days.The sca nning through electro n microscope con firmed that Shh以nHAC was more con ducive to adhesi on and proliferati on of

MSCs.6 and 12 weeks after operatio n, radiology exam in ati on disclosed a significant differenee of new bone formation

betwee n the groups(P0.05); histological exam in ati on disclosed a

sig ni fica nt differe nee of new bone formati on in rate and

qua ntity betwee n the groups(P0.05). Con clusi on : The MSCs/Shh；〔nHAC compo und can repair the femoral bone defect of SD rats, and its effect of reparatio n surpassed the MSCs/ nHAC compo und obviously.

［Key words ］marrow mesenchymal stem cells; nano Hydroxyapatite/ collage n; bone defects; sonic hedgehog 严重

创伤、炎症、肿瘤、先天性畸形等导致的节段性骨缺损是骨科医师面临的临床难题之一。利用组织工程技术构建组织工程骨治疗骨缺损是具有临床应用价值的治疗策略。骨髓间充质干细胞(marrow mesenchymalstem cells,MSCs)是骨组织工程最重要的种子细胞，纳米晶胶原基骨(nano Hydroxyapatite /collagen, nHAC) 已经作为人

工合成生物支架材料替代骨缺损应用于临床［1］,研究表明其复合细

胞后具有体内成骨能力］2］。

音猬因子(sonic hedgehog, Shh)具有促进造血前体细胞增殖的作用［3］。在牙胚发育过程中,Shh与BMP FGF和Wnt协同作用，调控牙齿的发育和萌出［4］。本实验以SD大鼠MSCs为种子细胞，以nHAC 及Shh以nHAC为支架材料体外共同培养，并采用构建的组织工程骨对SD 大鼠股骨缺损进行修复，采用大体观察、X线、病理切片等指标评价修复效果，为骨缺损治疗中新型植入材料的选择提供实验依据。

1材料和方法