中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (33): 5354-5359.doi: 10.3969/j.issn.2095-4344.2017.33.017

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

以牙髓干细胞构建组织工程化牙本质牙髓复合体修复牙髓损伤

王 璐，马 欣，韩耀伦，李亚东，骆书文   

  1. 河南省人民医院口腔科，河南省郑州市 450003
  • 修回日期:2017-10-10 出版日期:2017-11-28 发布日期:2017-12-01
  • 通讯作者: 骆书文。河南省人民医院口腔科，河南省郑州市 450003
  • 作者简介:王璐，1986年生，河南省人，汉族，2013年四川大学华西口腔医学院毕业，硕士，主治医师，主要从事口腔内科工作。
  • 基金资助:

    河南省卫生厅项目(201403153)

Dental pulp repair with dental pulp stem cells by construction of tissue-engineered dentin-pulp complex

Wang Lu, Ma Xin, Han Yao-lun, Li Ya-dong, Luo Shu-wen   

  1. Department of Stomatology, Henan Provincial People's Hospital, Zhengzhou 450003, Henan Province, China
  • Revised:2017-10-10 Online:2017-11-28 Published:2017-12-01
  • Contact: Luo Shu-wen, Department of Stomatology, Henan Provincial People's Hospital, Zhengzhou 450003, Henan Province, China
  • About author:Wang Lu, Master, Attending physician, Department of Stomatology, Henan Provincial People's Hospital, Zhengzhou 450003, Henan Province, China
  • Supported by:

    Henan Provincial Health Department Project, No. 201403153

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摘要：

文章快速阅读：

文题释义：
牙髓干细胞：牙髓干细胞属于成体干细胞，是存在于牙髓中具有多向分化潜能的成体干细胞，已成为全球基础及口腔医学界研究者研究的热点。
牙本质牙髓复合体：牙髓是牙体内惟一的软组织，具有自身修复的潜能，承担着向牙齿硬组织和形成牙本质提供营养的重要功能。性能良好的支架材料成为组织工程化牙本质牙髓复合体研究中的关键因素。

 

摘要
背景：研究证明，牙髓干细胞具有高度增殖、多向分化的干细胞特征，体外实验证明其在一定条件下可诱导分化成多种功能细胞或组织。目前，利用牙髓干细胞构建组织工程化牙本质牙髓复合体移植对牙体缺损进行修复有望成为可能。
目的：观察牙髓干细胞构建组织工程化牙本质牙髓复合体移植对大鼠牙体缺损的修复作用。
方法：建立去髓牙齿模型大鼠，随机分为模型组及牙髓复合体移植组。建模后，复合体移植组于模型大鼠髓腔中植入牙本质牙髓复合体，模型组大鼠不作处理。移植后3，5，7周采集标本，对大鼠牙齿组织切片苏木精-伊红和免疫荧光染色，采用图像软件系统测量大鼠牙本质厚度。
结果与结论：①牙髓细胞形态：牙髓细胞贴壁生长，大多数呈成长梭形、椭圆形，部分呈多角形；第3代细胞长梭形，呈纤维样生长，形态均一，免疫组织化学染色后，细胞核呈现为深蓝色，卵圆形，呈梭形着色深细胞为成纤维细胞样细胞，波形丝蛋白染色呈现为阳性；②大鼠牙齿组织苏木精-伊红染色：模型组大鼠牙齿牙本质细胞出现空泡状变性，髓腔内可见被破坏牙髓组织和碎屑，残留不规则的组织条梭，髓腔出现大量红细胞聚集和炎细胞，血管明显扩张。移植后7周，牙本质基质样组织层内可见成牙本质细胞成束装排列，原发性牙本质与再生牙本质界限明显；③免疫荧光染色：移植后3周，复合体移植组髓腔内细胞数量明显增多(P < 0.05)；移植后5周，髓腔内细胞数量明显大量增加，并分布于髓壁；④测定牙本质厚度：与模型组相比，牙本质牙髓复合体移植组在相同时间点大鼠牙本质厚度均明显较大(P < 0.05)，复合体移植组不同时间点牙本质厚度相比均差异有显著性意义(P < 0.05)；⑤结果证实，牙本质牙髓复合体能促进牙本质的再生和修复。

 

关键词: 干细胞, 动物模型, 牙髓干细胞, 牙本质牙髓复合体, 移植, SD大鼠, 去髓, 修复, 牙本质, 组织工程

Abstract:

BACKGROUND: Studies have shown that dental pulp stem cells have high proliferation and multi-directional differentiation abilities and can differentiate into a variety of cells under certain conditions. At present, the use of dental pulp stem cells to construct tissue-engineered dentin-pulp complex is expected to become a new strategy for human dental defect repair .
OBJECTIVE: To observe the effect of dental pulp stem cells on the repair of rat tooth defects by construction of tissue-engineered dentin-pulp complex.
METHODS: Twenty-four Sprague-Dawley rats were used to make animal models with dental pulp removal, and then model rats were randomly divided into model group and transplantation group. Rats in the transplantation group were subjected to tissue-engineered dentin-pulp complex transplantation, and those in the model group given no treatment. Tooth samples were collected at 3, 5, 7 weeks post transplantation and observed using hematoxylin-eosin staining and immunofluorescence staining. The dentin thickness of rats was measured by Image Pro Plus 6.0 image software system.
RESULTS AND CONCLUSION: (1) Dental pulp cells was mostly spindle/oval-shaped and partially polygonal. The third generation of cells with long spindle shape showed fibrous growth and uniform morphology. Findings from immunohistochemical staining showed spindle-shaped deep-colored cells with oval nuclei stained as dark blue were identified as fibroblast-like cells, and were positire for vimtin. (2) Findings from hematoxylin-eosin staining showed vacuolar degeneration of the cells, and hbdestroyed pulp tissue and debris, irregular cord-like tissue, and a large amount of red blood cells and inflammatory cells in the pulp cavity, accompanied by clearly visible vascular dilation. Seven weeks after transplantation, a bundle of odontoblasts were visible in the matrix-like tissues of the dentin, and there was a distinct boundary between the original dentin and regenerated dentin. (3) Findings from immunofluorescent staining showed that after dentin-pulp complex transplantation, the number of cells in the pulp cavity increased significantly at 3 weeks, and there was also a substantial increase in dental pulp cells at 5 weeks that were distributed on the wall of the pulp cavity. Compared with the model group, the dentin thickness in the transplantation group was significantly higher at each time after transplantation (P < 0.05), and in the transplantation group, there was also a significant difference in the dentin thickness at different time points (P < 0.05). To conclude, the tissue-engineered dentin-pulp complex can promote dentin regeneration and repair.   

 

 

Key words: Tissue Engineering, Stem Cells, Models, Animal

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