BACKGROUND:Previous study has proved that implantation of the tissue-engineering bone with vascular bundles or sensory nerve tracts can promote bone formation, which may be related to the expression of sensory neuropeptide receptors. OBJECTIVE:To investigate the effect of implantation of tissue-engineered bone containing vascular bundles and sensory nerve tracts on the middle-and long-term expression of calcitonin gene-related peptide type Ⅰ receptor (CGRP1R) in the repair of rabbit large femoral defects. METHODS:Thirty-six New Zealand white rabbits were enrol ed and modeled into 1.5 cm femoral defects, and then randomized into three groups. Bone marrow mesenchymal stem cells (BMSCs) were subjected to osteoblastic induction for 7 days, and then seeded onto the β-calcium phosphate scaffold to construct the tissue-engineered scaffold.In sensory nerve group, the tissue-engineered scaffold was implanted into the defect region, and autologous sensory nerve bundles were implanted into the lateral groove of the tissue-engineered bone;in vascular bundle group, the tissue-engineered scaffold was implanted, and autologous femoral blood bundles were implanted into the lateral groove of the tissue-engineered bone;in blank control group, only the tissue-engineered scaffold was implanted. X-ray examination, immunohistochemistry and real-time PCR were performed at 24 and 48 weeks postoperatively. RESULTS AND CONCLUSION:The X-ray scores in the sensory nerve and vascular bundle groups were significantly higher than that in the control group (P<0.05), but no significant difference was found between sensory nerve and vascular bundle groups. Real-time PCR found that the expression level of CGRP1R mRNA in the vascular bundle group was significantly higher than that in the other two groups (P<0.05), but showed no significant difference between sensory nerve and blank control groups.Immunohistochemistry findings showed that CGRP1R positive expression rate in the sensory nerve and vascular bundle groups was higher than that in the blank control group. These results reveal that implantation of the tissue-engineered bone containing vascular bundles can promote the CGRP1R expression.%背景:前期实验证实,植入含血管束或感觉神经的组织工程骨促成骨程度与其内的感觉神经肽受体表达相关.目的:观察植入含感觉神经束和血管束的组织工程骨修复兔股骨大段骨缺损,对降钙素基因相关肽Ⅰ型受体中长期表达的影响.方法:取36只新西兰大白兔,制作单侧股骨1.5 cm缺损模型,随机分3组,感觉神经组于骨缺损处植入成骨诱导7d的骨髓间充质干细胞-β-磷酸三钙组织工程骨,并在组织工程骨侧槽内植入自体感觉神经束;血管束组于骨缺损处植入成骨诱导7d的骨髓间充质干细胞-β-磷酸三钙组织工程骨,并在组织工程骨侧槽内植入自体股血管束;空白对照组于骨缺损处植入成骨诱导7d的骨髓间充质干细胞-β-磷酸三钙组织工程骨.植入后24,48周,进行X射线、免疫组织学及荧光定量PCR检测.结果与结论:①X射线检查:感觉神经组、血管束组X射线评分高于空白对照组(P<0.05),感觉神经组、血管束组X射线评分比较差异无显著性意义;②荧光定量PCR检测:血管束组降钙素基因相关肽Ⅰ型受体表达量高于感觉神经组、空白对照组(P<0.05),感觉神经组与空白对照组比较差异无显著性意义;③免疫组织化学观察:感觉神经组、血管束组降钙素基因相关肽Ⅰ型受体阳性表达强于空白对照组;④结果表明:植入含血管束的组织工程骨可促进降钙素基因相关肽Ⅰ型受体的表达.
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