摘要
目的:探讨体外培养和鉴定源于大鼠磨牙牙骨质的成牙骨质细胞(Cementoblast,CMB)的可行性,观察培养状态下CMB的形态与功能,就其是否能分泌碱性磷酸酶(Alkalinephoshatseactive-ity,ALP)、骨钙素的生物学特性进行初步研究,以期建立能排除牙周其他细胞干扰的CMB体外实验模型。方法:使用组织块结合酶消化的二步法,分别培养来自牙骨质的CMB,来自牙槽骨的成骨细胞和来自牙周韧带的成纤维细胞。然后对所培养的细胞分别进行骨钙素免疫组化染色和ALP活力测定,并利用茜素红S,检测在条件培养基下细胞形成矿化结节的情况。结果:3种牙周细胞都可从约12周大鼠第一磨牙根周获得进行原代培养。骨钙素在体外培养的CMB和成骨细胞中被强烈表达,在牙周韧带成纤维细胞中几乎不表达。成骨细胞ALP活性最强,在成纤维细胞中适中,在CMB中几乎无活性。CMB形成矿化结节茜素红S染色阳性。结论:CMB可以在体外成功培养,并可与牙槽骨成骨细胞、牙周韧带的成纤维细胞相区别。
AIM:To investigate the feasibility of culturing and identifying the cementoblasts(CMB) from molar cementum in vitro,and to observe the shape and function of CMB in the status of culture,and to study the biological characteristics of CMB secreting alkaline phoshatse(ALP) and osteocalcin of CMB so as to establish an experimental model of CMB in vitro which can eliminate the interaction from other periodontal cells . METHODS:By using a 2 step method of enzyme digestion/explant culture,osteoblasts from alveolar bone,periodontal ligament fibroblasts and CMBs from cementum were cultured respectively.Then immunohistochemical staining was used to assay the bone osteocalcin of the cultured cells,and the activities of ALP were measured.The formation of mineralized nodules of the cultured cells was assayed by alizarin red S under the conditional culture. RESULTS:Three types of periodontal cells were obtained from the periodontium surrounding the first molar in 12 week old rats for primary culture.Osteocalcin was strongly expressed in the osteoblasts and cementoblasts of the CMBs in vitro,but hardly expressed in the osteoblasts of periodontal ligament.The highest level of ALP activity was found in the osteoblasts,a moderate level in fibroblasts,and little level in CMB.CMB formed the mineralized nodules and were positively stained by alizarin red S. CONCLUSION:CMB can be successfully cultured in vitro which is different from periodontal ligament fibroblasts and osteoblasts from alveolar bone.
出处
《中国临床康复》
CSCD
2004年第14期2636-2638,F008,共4页
Chinese Journal of Clinical Rehabilitation
