摘要
目的构建重组人BMP-2(recombinant human BMP-2,rhBMP-2)修饰的β磷酸三钙(βtricalciumphosphate,β-TCP)/胶原生物支架材料,初步探讨其在牙组织工程中发挥的作用,评价其作为牙组织工程支架材料的可行性。方法将纳米级β-TCP粉末与胶原在强碱性水溶液中复合,制备β-TCP/胶原生物支架材料,并与rhBMP-2复合,制备牙组织工程支架材料。SPF级8～10周龄SD大鼠46只,雄性34只,雌性12只,体重250～300 g。体视显微镜下分离新生SD大鼠的下颌骨,取牙胚消化成细胞悬液,与支架材料复合培养制备组织工程牙胚。通过扫描电镜观察、细胞黏附率测定和MTT测定细胞增殖情况评价支架材料对牙胚细胞体外生长的影响。将组织工程牙胚植入SD大鼠肾被膜下作为实验组(n=12),另分别植入单独牙胚细胞团块(细胞对照组,n=12)及单独支架材料(材料对照组,n=4)作为对照,4、8周时取出标本行大体及组织学观察。结果扫描电镜示β-TCP/胶原生物支架材料呈疏松多孔状,质地柔软,亲水性良好;复合培养3 d后牙胚细胞可紧密贴附于支架材料,生长状态良好。牙胚细胞接种至支架材料上经4、8、12 h孵育后,细胞黏附率分别为27.20%±2.37%、44.52%±1.87%、73.81%±4.15%。MTT检测示牙胚细胞在β-TCP/胶原生物支架材料上的增殖情况与未放置支架材料相似,差异无统计学意义(P>0.05)。各组植入物移植于大鼠肾被膜下4、8周后大体观察可见白色钙化物形成;植入后4周,实验组镜下可见明显牙样形态及典型的釉质和牙本质样结构形成,细胞对照组可见釉质和牙本质样结构,但排列相对紊乱;植入后8周,实验组釉质及牙本质样结构较4周成熟,层次也更加清晰,细胞对照组也可见较为成熟的釉质和牙本质样结构;植入后4、8周,材料对照组均未见牙样结构形成。结论 rh BMP-2修饰的β-TCP/胶原生物支架材料与牙胚细胞生物相容性良好,可作为牙组织工程
Objective To explore a novel nanometer biomaterial which could induce the regeneration of tooth tissues intelligently,and to evaluate the feasibility of using this kind of biomaterial as the scaffold for tooth tissue engineering by investigating the role it plays in tooth tissue engineering.Methods The scaffold for tooth tissue engineering containing recombinant human bone morphogenetic protein 2（rhBMP-2） was prepared by mixing nanoscale β tricalcium phosphate（β-TCP）/collagen particles.Forty-six 8-10 weeks old specific pathogen free Sprague Dawley（SD） rats,including 34 females and 12 males,weighing 250-300 g,were involved in this study.Tooth germs were removed under a stereomicroscope from the mandible of newborn SD rat,then digested and suspended.Scanning electronic microscope（SEM）,adhesion rate of cells,and MTT assay were used to evaluate the effects of the scaffold on the tooth germ cells cultured in vitro.The tissue engineered tooth germ which was constructed by tooth germ cells and scaffold was transplanted under SD rat s kidney capsule as the experimental group（n=12）;the tooth germ cells（cell-control group,n=12） or scaffold without cells（material-control group,n=4） were transplanted separately as control groups.Specimens were harvested to perform general and histological observations at 4 and 8 weeks after transplantation.Results β-TCP/collagen showed a loose and porous appearance with soft texture and excellent hydrophilicity.Tooth germ cells grew well and could attach to the scaffold tightly 3 days after coculture.The adhesion rates of tooth germ cells were 27.20% ± 2.37%,44.52% ± 1.87%,and 73.81% ± 4.15% when cocultured with scaffold for 4,8,and 12 hours,respectively.MTT assay showed that the cell proliferation status of experimental group was similar to that of the control group,showing no significant difference（P 〉 0.05）.Some white calcified specimens could be harvested at 4-8 weeks after transplantation.At 4 weeks after transplantation,some typical structures of
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2011年第2期149-154,共6页
Chinese Journal of Reparative and Reconstructive Surgery
基金
浙江省自然科学基金资助项目(Y206114)
浙江省科技计划资助项目(2008C33063)~~
关键词
牙组织工程
支架材料
β磷酸三钙
胶原
重组人BMP-2
大鼠
Tooth tissue engineering Scaffold material βtricalcium phosphate Collagen Recombinant human bone morphogenetic protein 2 Rat
