CAJ | 학술논문

目的探索应用基因转染的组织工程技术修复眶骨缺损的可行性.方法实验研究.体外分离、扩增兔骨髓基质干细胞(BMSC),将人骨形成蛋白2(BMP-2)基因通过腺病毒转染的方式导入BMSC,采用RT-PCR、免疫印迹法检测目的基因的表达情况,并观察基因增强后的BMSC体外增殖特性和成骨能力.将BMP-2基因修饰后的BMSC复合天然珊瑚材料构建组织工程骨.24只成年新西兰白兔制造双侧眼眶骨下缘12 mm长的节段性骨缺损,共计48侧.随机数字表法分为4组,每组12侧眶骨缺损,行组织工程骨回植修复.A组为以转染BMP-2的自体BMSC构建的组织工程骨,B组为未转染基因的自体BMSC构建的组织工程骨、C组为单纯珊瑚材料,D组为旷置组.分别在手术后4、8、16周取材进行大体观察、micro-CT骨密度分析、组织学观察和组织形态学检测,比较骨缺损修复效果.采用单因素3水平设计定量资料方差分析处理新生骨和正常骨密度之间的差异,并采用LSD法对3个平均值进行两两比较;采用两因素析因设计定量资料方差分析处理新生骨占骨缺损面积的百分比.结果 BMP-2基因修饰后的BMSC能够高表达目的基因,并且转染后的细胞体外成骨能力明显增强;构建的组织工程骨能够良好修复自体眶骨缺损,新生骨密度、成骨面积值均显著高于各对照组(F=11.46,F=7180.97;P<0.05).结论 BMP-2基因修饰的组织工程骨具有良好修复眶骨缺损的能力.(中华眼科杂志,2009,45:66-72)
목적 탐색응용기인전염적조직공정기술수복광골결손적가행성.방법 실험연구.체외분리、확증토골수기질간세포(BMSC),장인골형성단백2(BMP-2)기인통과선병독전염적방식도입BMSC,채용RT-PCR、면역인적법검측목적 기인적표체정황,병관찰기인증강후적BMSC체외증식특성화성골능력.장BMP-2기인수식후적BMSC복합천연산호재료구건조직공정골.24지성년신서란백토제조쌍측안광골하연12 mm장적절단성골결손,공계48측.수궤수자표법분위4조,매조12측광골결손,행조직공정골회식수복.A조위이전염BMP-2적자체BMSC구건적조직공정골,B조위미전염기인적자체BMSC구건적조직공정골、C조위단순산호재료,D조위광치조.분별재수술후4、8、16주취재진행대체관찰、micro-CT골밀도분석、조직학관찰화조직형태학검측,비교골결손수복효과.채용단인소3수평설계정량자료방차분석처리신생골화정상골밀도지간적차이,병채용LSD법대3개평균치진행량량비교;채용량인소석인설계정량자료방차분석처리신생골점골결손면적적백분비.결과 BMP-2기인수식후적BMSC능구고표체목적 기인,병차전염후적세포체외성골능력명현증강;구건적조직공정골능구량호수복자체광골결손,신생골밀도、성골면적치균현저고우각대조조(F=11.46,F=7180.97;P<0.05).결론 BMP-2기인수식적조직공정골구유량호수복광골결손적능력.(중화안과잡지,2009,45:66-72)
Objective To investigate the efficacy of tissue-engineered bone (human BMP-2 genetic modified BMSC combined with coral) in healing the segmental orbital defect in rabbits. Methods Rabbit BMSC were isolated and cultured in vitro, and cells of passage 2 were infected with adenovirus-mediated transfection of human BMP-2 gene (150 pfu/cell). After infection, the expression of BMP-2 was determined by RT-PCR and Western blot analysis, and cell proliferation and osteogenic differentiation were observed by flow cytometry, ALP and Alizarin red staining. A 12 mm bone defect in the infraorbital rim was induced by surgery in both orbits of 24 New Zealand white rabbits. The defects were repaired with modified tissue-engineered bone constructed with coral plus BMP-2 transfected BMSC (Group A, n=12), constructed by coral plus non-transfected BMSC (Group B, n=12) and grafts of coral alone (Group C, n=12), with untreated group (Group D, n=12) served as control. The osteogenesis of bone defect was monitored by gross observation, micro-CT measurement, histological and histomorphologic analysis at 4, 8, and 16 weeks after the implantation. Results After transfection, the BMP-2 expression was confirmed by RT-PCR and western blot, and the osteogenesis activity of BMSC could be obviously enhanced. The 12 mm segmental defect of rabbit orbit couldn't heal alone. Gross observation and micro-CT demonstrated well the bony-union in experimental group, with higher bone mineral density and more bone volume than other control groups (F=11.46,F=7180.97;P<0.05). Conclusion This study demonstrated that the rabbit orbital defect could be successfully repaired by tissue-engineered bone constructed with human BMP-2 gene modified BMSC and coral. (Chin J Ophthalmol, 2009,45:66-72)