Sun Mingxia, Tan Wanye, Wang Ketao, Dong Zuoqing, Peng Huahai, Wei Fengcai
Attending Doctor, Department of Stomatology, QiLu Hospital of Shandong University, Jinan, Shandong, China.
J Oral Maxillofac Surg. 2013 Oct;71(10):1789-99. doi: 10.1016/j.joms.2013.03.007. Epub 2013 May 12.
This report describes the effect of periosteal-derived cells transfected with adenovirus-mediated bone morphogenetic protein-2 (BMP-2) on the repair of mandibular defects in rabbits.
Periosteal-derived cells were transfected with a replication-defective adenoviral vector encoding BMP-2, and the expression of BMP-2 was examined in transfected cells using in situ hybridization and enzyme-linked immunosorbent assay. In addition, the proliferation ability and activity of alkaline phosphatase of transfected cells were examined using the 3-[4,5-dimethylthiazol-2-Yl]-2,5-diphenyltetrazolium bromide method and enzymology, respectively. In vitro critical-size defects (about 10 × 6 mm) were made bilaterally in each rabbit mandible, and individual sites were implanted with tissue-engineered bone modified with an adenovirus construct encoding the recombinant human BMP-2 gene (Ad-BMP-2), tissue-engineered bone without modification, single bioactive glass ceramic, or no implants (control). New bone formation was evaluated by histochemical stain.
BMP-2 expression in the supernate of infected cells was detected from the first day after Ad-BMP-2 transfection and remained at a high level for at least 2 weeks. Alkaline phosphatase expression in transfected cells was significantly greater than in uninfected cells. The group of Ad-BMP-2-modified periosteal-derived cells formed more new bone than the other group at any time point.
Gene-modified tissue-engineered bone grafts have greater osteogenic potential than single tissue-engineered bone and single bioactive glass ceramic graft. Ex vivo Ad-BMP-2 transfer to periosteal-derived cells can increase bone formation in critical-size bone defects. Further studies are needed to determine if modified engineered cells can be developed for safe and effective clinical applications.
本报告描述了用腺病毒介导的骨形态发生蛋白-2（BMP-2）转染的骨膜来源细胞对兔下颌骨缺损修复的影响。
用编码BMP-2的复制缺陷型腺病毒载体转染骨膜来源细胞，采用原位杂交和酶联免疫吸附测定法检测转染细胞中BMP-2的表达。此外，分别采用3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐法和酶学方法检测转染细胞的增殖能力和碱性磷酸酶活性。在每只兔下颌骨两侧制作体外临界尺寸缺损（约10×6mm），各个部位分别植入用编码重组人BMP-2基因的腺病毒构建体修饰的组织工程骨（Ad-BMP-2）、未修饰的组织工程骨、单一生物活性玻璃陶瓷或不植入任何物质（对照）。通过组织化学染色评估新骨形成情况。
Ad-BMP-2转染后第1天即检测到感染细胞上清液中有BMP-2表达，且至少2周内维持在高水平。转染细胞中碱性磷酸酶的表达明显高于未感染细胞。在任何时间点，Ad-BMP-2修饰的骨膜来源细胞组形成的新骨均多于其他组。
基因修饰的组织工程骨移植物比单一的组织工程骨和单一生物活性玻璃陶瓷移植物具有更大的成骨潜力。体外将Ad-BMP-2转移至骨膜来源细胞可增加临界尺寸骨缺损处的骨形成。需要进一步研究以确定是否能开发出经修饰的工程细胞用于安全有效的临床应用。
