CAJ | 학술논문

背景:利用组织工程学技术将各种凝胶系统作为支架材料对骨缺损进行修复日益受到重视.所选择的支架材料必须具有无毒、无致畸的特点.目的:观察海藻酸钠-明胶共混体系/成骨细胞凝胶修复兔颅骨缺损的愈合过程中对染色体畸变的影响.设计、时间及地点:材料学体内动物实验,于2007-10/2008-03在北京世纪坛医院和北京大学医学部组织胚胎学教研室完成.材料:清洁级2月龄新西兰纯种大白兔12只,随机分为2组:实验组8只,雌性4只,雄性4只,对照组4只均为雌性.海藻酸钠干粉为美国Sigma公司产品,明胶干粉为河北绿岛公司产品.方法:将12只兔编号后抽取骨髓,密度梯度离心法分离骨髓基质干细胞,加入成骨细胞诱导液进行体外培养,诱导分化的成骨细胞经传代增殖为10~7数量级.制备海藻酸钠与明胶质量比为2:3的透明粉红色胶状液体,引入兔成骨细胞,细胞终浓度为5×10~9L~(-1),与CaCl_2溶液混合,形成果冻样海藻酸钠-明胶共混体系/成骨细胞凝胶.12只兔颅骨均制备直径1.5 cm的极限缺损,1周后实验组植入凝胶复合体0.5 mL进行修复,对照组不植入任何材料.主要观察指标:缺损修复后3个月取心脏血观察细胞遗传学的变化.结果:①每只实验动物观察100个中期分裂细胞,均未见染色体型畸变.对照兔在400个中期分裂相中见4个,单体畸变率为1%.实验兔在800个中期分裂相中见12个,单体畸变率为1.5%.两组均在正常范围内.②每只实验动物做2个染色体核型分析,染色体数目2n=44,对照兔核型为44,XX,正常雌性;雄性实验兔为44,XY,未见异常;雌性实验兔为44,XX,未见到异常.结论:海藻酸钠-明胶共混体系/成骨细胞凝胶在细胞遗传学角度上是安全的.
배경:이용조직공정학기술장각충응효계통작위지가재료대골결손진행수복일익수도중시.소선택적지가재료필수구유무독、무치기적특점.목적:관찰해조산납-명효공혼체계/성골세포응효수복토로골결손적유합과정중대염색체기변적영향.설계、시간급지점:재료학체내동물실험,우2007-10/2008-03재북경세기단의원화북경대학의학부조직배태학교연실완성.재료:청길급2월령신서란순충대백토12지,수궤분위2조:실험조8지,자성4지,웅성4지,대조조4지균위자성.해조산납간분위미국Sigma공사산품,명효간분위하북록도공사산품.방법:장12지토편호후추취골수,밀도제도리심법분리골수기질간세포,가입성골세포유도액진행체외배양,유도분화적성골세포경전대증식위10~7수량급.제비해조산납여명효질량비위2:3적투명분홍색효상액체,인입토성골세포,세포종농도위5×10~9L~(-1),여CaCl_2용액혼합,형성과동양해조산납-명효공혼체계/성골세포응효.12지토로골균제비직경1.5 cm적겁한결손,1주후실험조식입응효복합체0.5 mL진행수복,대조조불식입임하재료.주요관찰지표:결손수복후3개월취심장혈관찰세포유전학적변화.결과:①매지실험동물관찰100개중기분렬세포,균미견염색체형기변.대조토재400개중기분렬상중견4개,단체기변솔위1%.실험토재800개중기분렬상중견12개,단체기변솔위1.5%.량조균재정상범위내.②매지실험동물주2개염색체핵형분석,염색체수목2n=44,대조토핵형위44,XX,정상자성;웅성실험토위44,XY,미견이상;자성실험토위44,XX,미견도이상.결론:해조산납-명효공혼체계/성골세포응효재세포유전학각도상시안전적.
BACKGROUND: Utilizing tissue engineering technique, various gel systems are served as scaffolds to repair bone defect. The scaffolds should have features of nontoxic and no teratological effects to the body. OBJECTIVE: To observe the effect of sodium alginate-gelatin/osteoblast gel on chromosomal pattern aberration in rabbits. DESIGN, TIME AND SETTING: The in vivo material animal experiments were conducted at the Beijing Shijitan Hospital and Department of Histology and Embryology, Peking University Health Science Center from October 2007 to March 2008. MATERIALS: A total of 12 New Zealand rabbits, aged 2 months, with clean grade, were randomly divided into 2 groups. The experimental group contains 4 female and 4 male rabbits, and the remaining 4 females were served as the control group. Sodium alginate dried powder were purchased from Sigma, USA, and the gelatin dried powder were supplied by Liidao Company, Hebei, China. METHODS: Following numbering, bone marrow was collected from 12 rabbits. Bone marrow stromal stem cells (BMSCs) were isolated by the density gradient centrifugation, and then in vitro cultured with osteoblast inductor. Osteoblasts following passage were an order of magnitude of 10~7. Bright pink gelatiniform liquid with mass ratio of sodium alginate and gelatin at ratio of 2:3 was prepared. Rabbit osteoblasts with final concentration of 5×10~9/L were mixed with CaCb solution to form fruit jelly-shaped sodium alginate-gelatin/osteoblast gel. Critical-sized calvarial defects were created in diameter of 1.5 cm in 12 rabbits. After 1 week, cell/scaffold complex (0.5 mL) was implanted to repair the bone defect in the experimental group. There was no treatment in the control group. MAIN OUTCOME MEASURES: The change of chromosomal pattern was observed at 3 months following reparation. RESULTS: No Chromosome somatotype aberration was found in 100 metaphases in the experimental group. From 400 metaphases of the control group, 4 abnormal cells were found, with 1% chromatid-type aberration ratio. Meantime, 12 abnormal cells in 800 metaphases of the control group were found, with 1.5% chromatid-type aberration ratio. The numerical value was within the normal range. Chromosome karyotype analysis: the chromosome number of each experimental rabbit was 2n=44, karyotype of the control rabbit was 44, XX, which was normal female; or 44, XY, normal male, no abnormal was found. The female rabbit in the experiment group was 44, XX, no abnormal was seen. CONCLUSION: From the cytogenetoxicity point of view, sodium alginate-gelatin/osteoblast gel is safe in repairing bone defects.