正常人与慢性牙周炎伴有骨质疏松患者牙槽骨组织培养细胞的生物学特性  

Biological characteristics of cell lines derived from human dental alveolus

作　　者：陈世璋[1] 黄靖香[2] 孙明学[2] 赵斌[2] 余征[2] 

机构地区：[1]首都医科大学附属北京朝阳医院口腔科,北京100020 [2]解放军总医院骨科研究所

出　　处：《中国骨质疏松杂志》2004年第4期474-476,共3页Chinese Journal of Osteoporosis

摘　　要：目的 无菌取健康人和慢性牙周炎伴骨质疏松患者的牙槽骨组织,经体外培养,各建立一个能传代、冻存、复苏成活的种子细胞系。比较两种细胞系的生物学特性。为牙槽骨的缺损、修复、治疗提供理论和相关实验依据。方法从原代培养出来的4个细胞系(H-171、H-258、261、262)中,选出牙周炎伴骨质疏松患者组细胞系H-171,正常组H-258,用组化、免疫组化等染色,观察细胞形态。用细胞计数法计算出两种细胞倍增时间和分裂指数。经过多次传代、冻存、复苏,比较细胞的生长、老化规律。结果 1.异常牙槽骨组,原代培养1次成功,细胞形态为短梭形,3次冻存,3次复苏均存活,其倍增时间为53.4 h。平均分裂指数为46。细胞传代20次后,仍生长良好,以破骨细胞为主。2.健康牙槽骨组原代培养26例,因多种原因能传代的细胞系仅有3例,已传代10次,形态为长梭形,经2次冻存复苏,细胞存活、生长速度较H-171慢,倍增时间为65.9 h,平均分裂指数3.5‰。3.两种细胞均贴壁生长,具有成骨细胞的特点:AKP、甲苯胺兰、PAS、四环素标记矿化结节组化染色及Ⅰ型胶原、BMP-2免疫组化染色均为阳性。结论1.均具有成骨细胞特点。2.生长速度:H-171较H-258快,传代20次,生长活跃,H-258传到8代开始老化,生长速度减慢。Objective To investigate the biological characteristics of cell lines established from healthy and diseased human dental alveoli. Methods We first established primary cell lines from either healthy or diseased or human dental alveolus. Two cell lines, H-258 and H-171, respectively derived from healthy and diseased tissues, were selected for further studies on cell morphology, growth and aging by cell counting, histochemical and immunohistochemical staining. Results 1) Primary cell lines were successfully established from diseased human dental alveoli. After three cycles of freezing and thawing, cells still kept a good growth condition and no morphological alterations were observed. The doubling time was 53.4 hours and mean division index (MDI) was 4‰ . Cells could be maintained after twenty passages without obvious reduction of doubling time and MDI. 2) We could passage only three cell lines among twenty-six primary cell lines derived from healthy human dental alveoli. Cells are shown as long shuttles as compared to short shuttles of cells derived from healthy alveoli. After two cycles freezing and thawing, cells were still alive but with decreased growth speed. Cell doubling time was 85.9 hours and mean division index (MDI) was 3‰. 3) Both cell lines (H-171 and H-258) shared the characteristics with os-teoblast as they showed an attached growth type and had positive staining with AKP, PAS and BMP2 etc. Conclusions Primary cell lines established from diseased human dental alveoli showed greater growth potential than that from healthy dental alveoli. All cell lines shared characteristics with osteoblast. Therefore cell lines we developed may have clinical applications on the treatment of diseased dental alveoli.

关 键 词：牙槽骨 冻存 患者 骨质疏松 复苏 骨组织 细胞系 分裂指数 慢性牙周炎 原代培养 

分 类 号：R781[医药卫生—口腔医学] R681[医药卫生—临床医学]