中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2007年
42期
8594-8597
,共4页
付崇建%郁冰冰%杨连甲%曹新生%张立藩
付崇建%鬱冰冰%楊連甲%曹新生%張立藩
부숭건%욱빙빙%양련갑%조신생%장립번
失重模拟%细胞培养%骨髓基质细胞%成骨细胞
失重模擬%細胞培養%骨髓基質細胞%成骨細胞
실중모의%세포배양%골수기질세포%성골세포
背景:骨组织在特殊物理环境如失重环境下,代谢活动会发生显著的变化,而成骨细胞是骨代谢和骨形成的核心部分,其对重力环境的变化敏感.目的:观察模拟失重条件对大鼠股骨骨髓基质细胞数量体外成骨能力的影响,揭示骨丢失的机制.设计:随机配对,对照实验.单位:解放军第四军医大学航空航天医学系和口腔医学院病理科.材料:选用20只成年健康雄性SD大鼠.实验开始当日按体质量随机分为对照组和悬吊组,每组10只.碱性磷酸酶试剂盒由北京中生生物工程高技术公司生产.方法:实验于1999-11/2000-07在解放军第四军医大学口腔医学院病理科完成.将SD大鼠随机配对分为鼠尾悬吊组和对照组,每组10只.悬吊组大鼠做尾部悬吊28 d,大鼠始终保持30°头低位及后肢自由悬垂不负重状态.对照组正常饲养.实验期满,取股骨,将股骨骨髓基质细胞进行原代和传代细胞培养.主要观察指标:采用细胞计数法和噻唑蓝法绘制原代和传代培养细胞的生长曲线,进行碱性磷酸酶活性及体外矿化小结形成量的检测.结果:①碱性磷酸酶活性:原代和传代培养悬吊组低于对照组,差异有显著性意义(P＜0.05).②钙化小结形成数:悬吊组少于对照组,差异有显著性意义(P＜0.05).③细胞生长:原代和传代股骨间充质细胞的生长曲线呈"S"形,悬吊组和对照组细胞倍增时间相近.④股骨骨髓基质细胞数:原代细胞培养系中,悬吊组比对照组约少50%(P＜0.05).结论:模拟失重条件下,大鼠骨髓基质细胞数明显减少,后肢承重骨成骨细胞数减少,体外成骨能力降低.
揹景:骨組織在特殊物理環境如失重環境下,代謝活動會髮生顯著的變化,而成骨細胞是骨代謝和骨形成的覈心部分,其對重力環境的變化敏感.目的:觀察模擬失重條件對大鼠股骨骨髓基質細胞數量體外成骨能力的影響,揭示骨丟失的機製.設計:隨機配對,對照實驗.單位:解放軍第四軍醫大學航空航天醫學繫和口腔醫學院病理科.材料:選用20隻成年健康雄性SD大鼠.實驗開始噹日按體質量隨機分為對照組和懸弔組,每組10隻.堿性燐痠酶試劑盒由北京中生生物工程高技術公司生產.方法:實驗于1999-11/2000-07在解放軍第四軍醫大學口腔醫學院病理科完成.將SD大鼠隨機配對分為鼠尾懸弔組和對照組,每組10隻.懸弔組大鼠做尾部懸弔28 d,大鼠始終保持30°頭低位及後肢自由懸垂不負重狀態.對照組正常飼養.實驗期滿,取股骨,將股骨骨髓基質細胞進行原代和傳代細胞培養.主要觀察指標:採用細胞計數法和噻唑藍法繪製原代和傳代培養細胞的生長麯線,進行堿性燐痠酶活性及體外礦化小結形成量的檢測.結果:①堿性燐痠酶活性:原代和傳代培養懸弔組低于對照組,差異有顯著性意義(P＜0.05).②鈣化小結形成數:懸弔組少于對照組,差異有顯著性意義(P＜0.05).③細胞生長:原代和傳代股骨間充質細胞的生長麯線呈"S"形,懸弔組和對照組細胞倍增時間相近.④股骨骨髓基質細胞數:原代細胞培養繫中,懸弔組比對照組約少50%(P＜0.05).結論:模擬失重條件下,大鼠骨髓基質細胞數明顯減少,後肢承重骨成骨細胞數減少,體外成骨能力降低.
배경:골조직재특수물리배경여실중배경하,대사활동회발생현저적변화,이성골세포시골대사화골형성적핵심부분,기대중력배경적변화민감.목적:관찰모의실중조건대대서고골골수기질세포수량체외성골능력적영향,게시골주실적궤제.설계:수궤배대,대조실험.단위:해방군제사군의대학항공항천의학계화구강의학원병이과.재료:선용20지성년건강웅성SD대서.실험개시당일안체질량수궤분위대조조화현조조,매조10지.감성린산매시제합유북경중생생물공정고기술공사생산.방법:실험우1999-11/2000-07재해방군제사군의대학구강의학원병이과완성.장SD대서수궤배대분위서미현조조화대조조,매조10지.현조조대서주미부현조28 d,대서시종보지30°두저위급후지자유현수불부중상태.대조조정상사양.실험기만,취고골,장고골골수기질세포진행원대화전대세포배양.주요관찰지표:채용세포계수법화새서람법회제원대화전대배양세포적생장곡선,진행감성린산매활성급체외광화소결형성량적검측.결과:①감성린산매활성:원대화전대배양현조조저우대조조,차이유현저성의의(P＜0.05).②개화소결형성수:현조조소우대조조,차이유현저성의의(P＜0.05).③세포생장:원대화전대고골간충질세포적생장곡선정"S"형,현조조화대조조세포배증시간상근.④고골골수기질세포수:원대세포배양계중,현조조비대조조약소50%(P＜0.05).결론:모의실중조건하,대서골수기질세포수명현감소,후지승중골성골세포수감소,체외성골능력강저.
BACKGROUND: Under specially physical environment, for example weightlessness, metabolism of bone tissue may have remarkable changes; however, osteoblast is a core of bone metabolism and bone formation, so it is very sensitive to changes of gravity environment.OBJECTIVE: To observe the effects of simulated weightlessness on bone marrow stromal cell count and osteogentic capacity of weight bearing bone in rats so as to reveal the mechanism of bone loss.DESIGN : Randomized pairing and controlled study.SETTING: College of Aerospace Medicine and Department of Pathology of Stomatology College, the Fourth Military Medical University of Chinese PLA.MATERIALS: A total of 20 adult healthy male SD rats were selected in this study. At the beginning of experiment, rats based on their body mass were randomly divided into control group and suspension group with 10 in each group. Alkaline phosphatase kit was provided by Beijing Zhongsheng Bioengineering High-technological Company.METHODS: The experiment was carried out in the Department of Pathology, Collage of Stomatology, the Fourth Military Medical University of Chinese PLA from November 1999 to July 2000. Rats were randomly divided into tail suspension group and control group with 10 in each group. Rats in the tail suspension group were given tail suspension for 28 days. Their heads maintained 30° low position, and their hindlimbs freely suspended and were not given weight loading. While, rats in the control group were fed normally. At the end of experiment, bone marrow stromal cells were obtained from femur for primary and transferring cultures.MAIN OUTCOME MEASURES: Cell counting and methylthianolyldiphenyl-tetrazolium bromide (MTT) assay were used to draw growth curve of cells in primary and transferring cultures and to measure activity of alkaline phosphatase and forming quantity of mineralized nodules in vitro.RESULTS : ① Activity of alkaline phosphatase: Activity of alkaline phosphatase of cells in the primary and transferring cultures in the suspension group was lower than that in the control group, and there was significant difference between them (P＜0.05). ② Forming quantity of mineralized nodules: Forming quantity of mineralized nodules in the suspension group was less than that in the control group, and there was significant difference between them (P＜0.05). ③ Cell growth: Growth curve of femoral stromal cells in primary and transferring culture manifested as S. Doubling time of cells in the suspension group was similar to that in the control group. ④ Amount of bone marrow stromal cells in femur: Amount of bone marrow stromal cells in primary culture in the suspension group was decreased as 50% as that in the control group (P＜0.05).CONCLUSION: Under simulated weightlessness, amount of bone marrow stromal cells decreases obviously; in addition, amount of osteoblast also decreases in weight bearing bone of hindlimbs and osteogentic capacity in vitro decreases simultaneously.
