口腔医学研究 ›› 2016, Vol. 32 ›› Issue (3): 244-248.DOI: 10.13701/j.cnki.kqyxyj.2016.03.008

• 基础研究论著 • 上一篇    下一篇

大鼠牙源性细胞复合nHAC/PLA支架体内构建牙体组织-骨复合体样结构的研究

赵征1*, 刘洪臣2, 黄征难1, 鄂玲玲2, 杨海青1   

  1. 1. 解放军第401医院口腔科 山东 青岛 266071;
    2. 解放军总医院口腔科 北京 100853
  • 收稿日期:2015-11-19 出版日期:2016-03-28 发布日期:2016-03-29
  • 通讯作者: 赵征,电话:(0532)51870540
  • 作者简介:赵征(1973~ ),女,山东青岛人,博士后,副主任医师,主要从事口腔内科学临床及基础的研究工作。
  • 基金资助:
    中国博士后科学基金特别资助金(编号:201003774)

Study on Tooth Tissue-Bone Complex-Type Structure Formed in Vivo by Rat Odontogenic Cells Combined with nHAC/PLA Scaffold.

ZHAO Zheng1, LIU Hong-chen2, HUANG Zheng-nan1, E Ling-ling2, YANG Hai-qing1   

  1. 1. Department of Stomatology, The 401st Hospital of Chinese PLA, Qingdao 266071, China;
    2. Department of Stomatology, PLA General Hospital, Beijing 100853, China
  • Received:2015-11-19 Online:2016-03-28 Published:2016-03-29

摘要: 目的: 探讨不同组合生长因子诱导的大鼠牙源性上皮细胞(rDECs)、间充质细胞(rDMCs)复合nHAC/PLA支架体内形成牙体组织-骨复合体样结构的能力。方法: 应用酶消化法和差速消化法分离培养rDECs和rDMCs,免疫荧光染色鉴定其来源。将各组细胞-支架复合物移植裸鼠皮下3月、5月后分别取材,检测移植物形成牙体组织-骨复合体样结构能力。结果: 成功分离培养、鉴定大鼠牙源性上皮细胞和间充质细胞。细胞-支架复合物移植后3月,组①、②、③在细胞-支架接触部位均发现类骨质、新生成熟骨和血管形成,并有OCN、OPN的阳性表达。组②促成骨作用最强。对照组④没有新生骨形成,仅有少量胶原纤维和血管。移植后5月,组①、③均形成没有髓腔的牙根样结构,部分区域有DSPP、DMP1、CAP表达。组②形成周边有新生骨、中央有单根牙体样组织,中心为含有牙髓样组织和血管的髓腔样结构,并有DSPP、DMP1、OPN、OCN在髓腔内壁牙本质基质、部分牙髓样组织和血管内的阳性表达,ameloblastin、CAP分别在釉质样、牙骨质样组织中表达阳性。对照组④形成新生骨,OCN强阳性表达,未形成牙体样结构。结论: 经TGF-β1+BMP4诱导的rDECs + rDMCs-nHAC/PLA体内移植物促成骨和成牙作用最强,移植后5月形成牙体组织-骨复合体样结构。

关键词: 大鼠牙源性上皮细胞, 大鼠牙源性间充质细胞, 纳米羟基磷灰石-胶原/聚乳酸

Abstract: Objective: To investigate the capability of tooth tissue-bone complex-like structure in vivo formed by rDECs+rDMCs-nHAC/PLA compounds induced by different growth factors. 〖WT5”HZ〗Methods: Rat dental epithelial cells (rDECs)and rat dental mesenchymal cells (rDMCs) were isolated, cultured and identified.The cell-scaffold compounds were transplanted into athymic mice subcutaneously and taken out after 3 or 5 months. The capability of tooth tissue-bone complex-like structure formed by the graftswas detected. Results: BothrDECs and rDMCscould be isolated, cultured and identified.After 3 months, the osteoid, new bone and blood vessel were found in group 1 to 3. The osteogenesis in group2 (TGF-β1+BMP4)was most powerful. There were only a few collagen fibers and blood vessels in the control (group 4). After 5 months, tooth root-like structure without pulp cavity was seen in group 1 and 3. In group 2, new bone and tooth-type tissueswere formed. Ameloblastin and CAP were expressed positively in emailloid and cementoid tissues respectively. In group 4, there was new bone but no tooth-like structure. Conclusion: The most powerful way that affects theosteogenesis and tooth-forming forrDECs+rDMCs-nHAC/PLA grafts is TGF-β1+BMP4.

Key words: rDECs , rDMCs , nHAC/PLA

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