降钙素基因相关肽在成骨过程中表达的动物实验研究

目的:观察降钙素基因相关肽(calcitonin gene-related peptide,CGRP)在犬骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向成骨细胞分化过程中,及应用BMSCs构建组织工程骨提升犬上颌窦底后骨组织中的表达情况。方法:体外分离培养犬BMSCs,通过成骨诱导采用实时荧光定量PCR(real-time quantitative polymerase chain reaction,Q-PCR)和免疫荧光技术检测CGRP在不同时间点的表达情况。犬上颌窦底提升术后,分别在两侧上颌窦底植入BMSCs+Bio-Oss骨粉(实验组)和Bio-Oss骨粉(对照组)。12周后处死并获取动物标本,对窦底区骨组织行CGRP免疫组化染色。结果:体外实验示CGRP表达量随成骨诱导分化呈时间依赖性递增(P<0.05);上颌窦底提升实验组CGRP表达量明显高于对照组(P<0.05),且CGRP主要分布在骨再生代谢活跃区域。结论:CGRP表达量与骨再生过程密切相关,可考虑作为骨代谢活跃度的标志。

The Expression of Calcitonin Gene-related Peptide during Bone Formation

Objective： To observe the expressive levels of calcitonin gene-related peptide （CGRP） in bone marrow mesenchymal stem cells （BMSCs） during osteoinduction, and in the remodeled bone of maxillary sinus floor augmentation. Methods：In vitro, the BMSCs were isolated from Beagle dogs and were classified into osteogenic-induced group and non-induced group. Expression of the CGRP was detected by Q-PCR and immunofluorescence in the different time-points. In vivo, both right and left maxillary sinus of three healthy adult dogs were divided into experimental group and control group randomly. BMSCs complexed with Bio-Oss granules were applied for canine maxillary sinus augmentation models, while Bio-Oss granules alone served as controls. Immunohistochemical analysis were performed after 12 weeks on these bone substitutes. Results： Q-PCR and immunofluorescence examinations demonstrated the expression of CGRP in osteogenic-induced group was increased in a time-dependent manner. After maxillary sinus floor elevation, the expressive levels of CGRP in experimental group were superior than that of control group （P〈0.05）. CGRP was mainly distributed in the region where osteogenesis metabolism was active. Conclusion： The expression of CGRP was related to the progress of bone regeneration, and it can be regarded as an indicator of the activity of osteogenesis metabolism.