CAJ | 학술논문

目的评价BMP2基因修饰的犬脂肪源性基质细胞(ADSCs)与β-磷酸三钙(β-TCP)复合修复自体大段骨缺损的疗效.方法从比格犬背部脂肪组织中提取基质细胞,转染腺病毒介导的人BMP2基因(Adv-hBMP2),通过ELISA和裸鼠体内异位成骨实验鉴定BMP2的表达及异位成骨活性;取比格犬11只,制作双侧尺骨2.5 cm骨缺损模型,缺损处旷置(4侧)或随机填充:单纯TCP(6侧),AD-SCs+TCP(6侧),Adv-hBMP2-ADSCs+TCP(6侧).所有动物16周后处死.结果 ELISA显示犬ADSCs被腺病毒转染后可高表达具有成骨活性的BMP2;裸鼠体内异位成骨实验证实所分泌的BMP2具有骨诱导活性.骨缺损修复的X线片显示,单纯TCP和ADSCs+TCP组至16周时,骨缺损均未愈合;BMP2基因修饰ADSCs+TCP组6例中2例愈合,4例部分愈合.显微摄片和组织学观察示:单纯TCP和AD-SCs+TCP组仅在骨缺损断端形成编织骨,缺损中央被纤维组织填充,而Adv-hBMP2-ADSCs+TCP组骨.缺损处皮质连续,新生骨组织主要为编织骨,部分改建形成板层骨.组织形态学分析示BMP2基因修饰ADSCs明显促进了新骨形成.结论 BMP2基因修饰ADSCs可以修复犬尺骨大段骨缺损.
목적 평개BMP2기인수식적견지방원성기질세포(ADSCs)여β-린산삼개(β-TCP)복합수복자체대단골결손적료효.방법 종비격견배부지방조직중제취기질세포,전염선병독개도적인BMP2기인(Adv-hBMP2),통과ELISA화라서체내이위성골실험감정BMP2적표체급이위성골활성;취비격견11지,제작쌍측척골2.5 cm골결손모형,결손처광치(4측)혹수궤전충:단순TCP(6측),AD-SCs+TCP(6측),Adv-hBMP2-ADSCs+TCP(6측).소유동물16주후처사.결과 ELISA현시견ADSCs피선병독전염후가고표체구유성골활성적BMP2;라서체내이위성골실험증실소분비적BMP2구유골유도활성.골결손수복적X선편현시,단순TCP화ADSCs+TCP조지16주시,골결손균미유합;BMP2기인수식ADSCs+TCP조6례중2례유합,4례부분유합.현미섭편화조직학관찰시:단순TCP화AD-SCs+TCP조부재골결손단단형성편직골,결손중앙피섬유조직전충,이Adv-hBMP2-ADSCs+TCP조골.결손처피질련속,신생골조직주요위편직골,부분개건형성판층골.조직형태학분석시BMP2기인수식ADSCs명현촉진료신골형성.결론 BMP2기인수식ADSCs가이수복견척골대단골결손.
Objective To evaluate osteogenetic effectiveness of porous β-tricalcium phosphate(β-TCP) ceramic mixed with human bone morphogenetic protein2 gene(Adv-hBMP2)modified adipose derived stromal cells (ADSCs) in the repair of critical-sized bone defects..Methods The ADSCs taken from the back of beagle dogs were modified by the BMP2 gene.The expression and bone-induction ability of BMP2 was identified by ELISA and ectopic bone formation in nude mice.The cells were applied to a β-tricalcium phosphate (TGP)carrier and implanted into ulnar bone defects in the canine model.18 ulnar bone defects were divided into three groups randomly and filled with granular TCP alone,granular TCP and ADSCs,or TCP and ADSCs transduced with Adv-hBMP2 respectively.All dogs were followed clinically and roentgenographically for 16 weeks and then sacrificed.Results ELISA and ectopic bone formation in nude mice showed the recombinant ADSCs could express BMP2 highly and stably.No bone defects healed after implanting granular TCP alone or granular TCP and ADSCs.In the TCP and ADSCs transduced with AdvhBMP2 group,two defects healed,four partly healed.Histological examination showed woven bone at the both end of the cortices but entirelv fibrous tissue in the middle in which defects filled with TCP alone or TCP and ADSCs.Defects filled with TCP and transduced ADSCs showed substatial new bone formation.Histomorphometry showed TCP combined with ADSCs did not significantly increase new bone area compared with TCP alone.TCP and recombinant ADSCs produced a significant increase in newly formed bone area.Conclusion ADSCs tansduced with BMP2 gene in a TCP carrier can enhance bone regeneratmn to repair the critically-sized bone defect.