部分去蛋白脱钙骨复合成骨细胞体内植入形成软骨或骨的可能性实验

背景采用理化技术制备的天然生物骨衍生材料具有天然网状孔隙系统,具有良好的细胞相容性,有利于成骨细胞的附着及生长,可用于成骨细胞的支架材料.目的观察部分去蛋白脱钙骨作为成骨细胞支架材料体内植入的成骨作用.设计随机对照实验.单位昆明医学院第一附属医院中心实验室.材料部分去蛋白脱钙骨;人胎骨膜成骨细胞;4周龄BALB/c裸小鼠20只,体质量25～28 g,雌雄不限.方法实验于2003-01/06在昆明医学院第一附属医院中心实验室完成.将部分去蛋白脱钙骨与人胎骨膜成骨细胞体外复合培养1周后植入裸鼠体内,每只裸鼠共植4块材料,脊柱左侧植入材料与细胞的复合物,作为实验侧,右侧单纯植入材料,作为空白对照侧.于术后4周及8周取出材料行碱性磷酸酶活性及常规组织学检查.主要观察指标裸鼠植入材料取出后行一般观察、碱性磷酸酶活性及常规组织学检查.结果20只裸鼠均进入结果分析.①裸鼠植入材料一般观察植入材料的周围组织未见坏死、化脓、积液等迹象,材料内有软组织长入、包裹.②碱性磷酸酶活性测定结果部分去蛋白脱钙骨复合成骨细胞体内植入8周时碱性磷酸酶活性比4周时强[(22.854±6.018),(11.286±4.268)nkat/g],并比单纯植入的材料强得多[(1.217±0.083),(2.717±0.583)nkat/g].③常规组织学检查结果实验侧4周见有软骨形成,8周时部分软骨形成骨组织,并见有髓腔,且软骨或新骨形成增多,而对照侧未见软骨或骨形成.结论部分去蛋白脱钙骨复合成骨细胞体内植入后能形成软骨或骨,并随植入时间的延长,软骨或新骨形成增多;部分去蛋白脱钙骨可用于成骨细胞的支架材料.

Possibility of osteogenesis of partially deproteinised decalcified bone composited with osteoblast in vivo

BACKGROUND: Natural bilogical bone-derived materials processed with physical and chemical methods possess natural network pore system. They have good cellular compatibility, and help osteoblasts attach and grow on them, and can be used as the scaffolds of osteoblasts.OBJECTIVE: To observe the osteogenetic effect of partially deproteinised decalcified bone (PDDB) as scaffolds of osteoblasts in vivo.DESIGN: A randomized and controlled trial.SETTING: Central Laboratory of the First Hospital of Kunming Medical College.MATERIALS: Partially deproteinised decalcified bone; human embryonic periosteum-derived osteoblasts; twenty 4-week-old BALB/c nude mice,with the body mass of 25 to 28 g, of either gender.METHODS: This experiment was conducted at the Central Laboratory of the First Affiliated Hospital of Kuming Medical College from January to June 2003. PDDB composited with human embryonic periostea-induced osteoblasts were implanted into the nude mice after cultured for 1 week in vivo, 4 scaffolds in each nude mouse. Composite of scalfolds and cells implanted on the left side of the spine was set as experimental side and simple implanted material on the right side of the spine was set as blank control side. Then alkaline phosphatase activity and routine histological examination were performed 4 and 8 weeks after operation.MAIN OUTCOME MEASURES: General observation, alkaline phosphatase activity and routine histological examination were performed after the materials were taken out.RESULTS: Twenty nude mice entered the stage of result analysis. ① General observation of the implanted materials: No necrosis, ecpyesis, or fluidifying was found around the implanted materials, but ingrowth and enwrapption of soft tissues were found. ② Measurement of alkaline phosphatase activity: alkaline phosphatase activity after PDDB composited osteoblasts in vivo was stronger at week 8 than at week 4 [(22.854±6.018) nkat/g vs(11.286±4.268) nkat/g], and was much stronger than that of the simple implanted materials [(1.217±0.083) nkat/g vs (2.717±0.583) nkat/g]. ③ Results of routine histological examination: Cartilage formed at week 4 and part of cartilage formed new bone and marrow cavity at week 8 at the experimental side, cartilage and new bone formed much more as time went by, but there was no any cartilage or bone formation at the control side.CONCLUSION: Cartilage and bone form after PDDB composited with osteoblasts are implanted, and more cartilage and new bone form as time passes. PDDB can be used as the scaffolds of osteoblasts.