摘要
目的:应用AdBMP-2基因修饰体外培养的山羊耳软骨细胞,并与F127pluronic支架材料复合,进行裸鼠皮下异位植入研究,探索BMP-2基因修饰高等哺乳动物软骨细胞促进软骨组织重建的可行性。方法:体外培养山羊耳软骨细胞,转染AdBMP-2和AdLacZ基因,X-gal染色检测基因转染效率。将基因修饰的细胞与可吸收生物材料PluronicF127复合形成细胞—生物材料复合物,并注射到裸鼠皮下,分别在术后2周和4周取材,观察2周标本CollX染色,4周标本X-gal、阿利新蓝、BMP-2及VEGF免疫组化染色。结果:50pfu/细胞可以获得80%的转染效率,AdBMP-2组在2周时出现大量成熟的软骨组织,4周时出现较多的骨样组织,BMP-2和VEGF染色强阳性;而AdLacZ组2周时尚未形成明显的软骨样组织,4周出现小块的软骨组织,未见骨样组织及VEGF阳性表达。结论:实验条件下,BMP-2基因转染早期即促进了软骨的成熟和分化,但随后发生了骨化,提示可尝试将其应用于软骨原位缺损或筛选更为理想的生长因子,以促进软骨再生。
PURPOSE： To explore the feasibility of promoting cartilage regeneration by BMP-2 gene modified highmammal goat chondrocyte when combined with F127 matrix in a subcutaneous ectopic nude mice model. METHODS： Goat auricular chondrocytes were obtained and transduced with AdBMP-2 and AdLacZ over-expression viruses. Transfer efficiency was detected using X-gal examination.Gene transduced chondrocytes were combined with F127 pluronic scaffold and injected into nude mice subcutaneously.The in vivo effects were investigated via histology, CollX, X-gal, Alcian Blue,BMP-2 and VEGF immunohistochemistry（IHC） staining 2 and 4 weeks later. RESULTS： 50 pfu/cell had a transfer efficiency of 80%. AdBMP-2 group promoted mature chondrocytes formation at week 2 while ossification was detected at week 4 with BMP-2 and VEGF intensively expressed, while small amount of cartilage was detected until week 4 with BMP-2 and VEGF negative in AdLacZ group. CONCLUSIONS： In this experiment, BMP-2 gene transfer accelerates early cartilage maturation but also ossification at a early stage, which suggests that BMP-2 gene could only be applied at a more suitable place such as orthotropic cartilage defect model to promote cartilage regeneration or that alternative growth factors need to be explored to promote cartilage regeneration but not ossification.Supported by National Natural Science Foundation of China （Grant No.30400502,30772431）; Research Fund of Science and Technology Committee of Shanghai Municipality（Grant No.04dz05601, 05DJ14006, 055407034, 05QMX1426,07DZ22007）, Shanghai Leading Academic Discipline Project （Grant No.Y0203） and ＂Dawn＂ Program of Shanghai Municipal Education Commission（Grant No.07SG19）.
出处
《中国口腔颌面外科杂志》
CAS
2008年第1期38-42,共5页
China Journal of Oral and Maxillofacial Surgery
基金
国家自然科学基金(30400502
30772431)
上海市科学技术委员会科研基金(04dz05601
05DJ14006
055407034
05QMX1426
07DZ22007)
上海市重点(优势)学科建设项目(Y0203)
上海市教育委员会曙光计划(07SG19)~~
