牙周膜干细胞BMP-2-PSH复合膜修复新西兰兔牙槽骨缺损

周静; 熊萍; 刘超峰; 陈丽琼; 张永辉; 史珂; 聂焱; 刘彦

doi:10.12259/j.issn.2095-610X.S20210503

牙周膜干细胞BMP-2-PSH复合膜修复新西兰兔牙槽骨缺损

doi: 10.12259/j.issn.2095-610X.S20210503

周静¹,
熊萍²,
刘超峰²,
陈丽琼²,
张永辉²,
史珂²,
聂焱²,
刘彦^2, ,

1.
昆明医科大学附属延安医院口腔科，云南昆明　650100
2.
昆明医科大学附属口腔医院第二门诊部，云南昆明　650100

基金项目: 云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目[2018FE001-（274）]

详细信息

作者简介:
周静（1983～），女，云南昆明人，医学硕士，主治医师，主要从事口腔牙周正畸临床研究工作

通讯作者:
刘彦，E-mail：lydyc@aliyun.com

中图分类号: R781.3
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出版历程
- 收稿日期: 2021-01-29
- 网络出版日期: 2021-06-02
- 刊出日期: 2021-05-20

Periodontal Ligament Stem Cell BMP-2-PSH Composite Membrane in Repairing Alveolar Bone Defect in New Zealand Rabbits

1.
Dept. of Stomatology，Yan’an Hospital Affiliated of Kunming Medical University，Kunming Yunnan 650100
2.
The 2nd clinic，The Affiliated Stomatology Hospital of Kunming Medical University，Kunming Yunnan 650100，China

摘要

摘要: 目的拟构建新西兰兔牙周缺损模型，确定hPDLSCs-BMP-2-PSH膜的生物安全性和其体内成骨修复作用。方法在确定BMP-2-PSH膜没有细胞毒性后将培养好的hPDLSCs细胞膜片复合至BMP-2-PSH膜上。分别在每只新西兰兔下颌左中切牙牙槽骨缺损内植入hPDLSCs/PSH复合膜作为实验组，右中切牙牙槽骨缺损内植入对照性多孔纤维膜材料作为对照组（或不植入任何材料作为对照），然后全瓣复位，缝合。术后即刻和每3周（连续观察12周）进行影像学（CT）检查，观察骨缺损愈合情况；同时，分别在术后第4、8、12周观察PDLSCs/BMP-2双膜材料的吸收度和各组样本新骨形成量和新骨形态。结果不同浓度PSH膜和BMP-2-PSH膜浸提液对hPDLSCs细胞不具有细胞毒性，hPDLSCs细胞能够成功复合到BMP-2-PSH膜上；CBCT及组织形态学分析显示，hPDLSCs/PSH复合膜能够有效促进牙槽骨缺损修复再生。结论 hPDLSCs/PSH复合膜具有良好生物安全性，有利于修复新西兰兔牙周缺损。
- 牙周膜干细胞 /
- BMP-2-PSH复合膜 /
- 牙周再生
Abstract: Objective It is planned to build a New Zealand rabbit periodontal defect model and determine the biological safety of hPDLSCs-BMP-2-PSH membrane and its in vivo osteogenic repair effect. Methods After confirming that the BMP-2-PSH membrane was not cytotoxic, the cultured hPDLSCs cell membrane was compounded to the BMP-2-PSH membrane. Respectively, HPDLSCs/PSH composite membrane was implanted into the alveolar bone defect of mandibular left incisor in each New Zealand rabbit as the experimental group, and the controlled porous fiber membrane material was implanted into alveolar bone defect of the right central incisor as the control group （or not implanted any material was used as a control）, then the whole flap was reset and sutured. Imaging （CT） examination was performed immediately after the surgery and every 3 weeks （observed continuously for 12 weeks） to observe the healing of bone defects; meanwhile, the absorbance of PDLSCs/BMP-2 double membrane materials, the amount of new bone formation and new bone morphology of samples in each group were observed at 4, 8 and 12 weeks after the surgery. Results Different concentrations of PSH membrane and BMP-2-PSH membrane extracts were not cytotoxic to hPDLSCs cells, hPDLSCs cells could be successfully compounded to BMP-2-PSH membranes; The analysis of CBCT and histomorphology showed that hPDLSCs/PSH composite membranes can effectively promote the repair and regeneration of alveolar bone defects. Conclusion The hPDLSCs/PSH composite membrane has the good biological safety and is beneficial to repair periodontal defects of New Zealand rabbits.
- Periodontal ligament stem cells /
- BMP-2-PSH composite membrane /
- Periodontal regeneration

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图 1 牙槽骨缺损的形成和PDLSCs/PSH复合膜的移植示意图

A：暴露的兔牙槽骨；B：使用高速外科手术囊制备的骨缺损（10 mm× 5 mm× 4 mm）；C：在牙槽骨缺损内移植PDLSCs/PSH复合膜材料；D：缝合缺损外皮瓣。

Figure 1. Schematic diagram of the formation of alveolar bone defect and the transplantation of PDLSCs/PSH composite membrane alveolar bone defect

下载: 全尺寸图片幻灯片

图 2 PSH纤维膜电镜形貌（×50000）

Figure 2. Electron microscopic morphology of PSH fiber membrane （×50000）

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图 3 hPDLSCs细胞增殖活力

Figure 3. Cell proliferation activity of hPDLSCs

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图 4 BMP-2-PSH膜及复合在其上的hPDLSCs细胞（红色箭头所示）（×2 000）

Figure 4. BMP-2-PSH membrane and hPDLSCs cells compounded on it （shown by the red arrow）（×2 000）

下载: 全尺寸图片幻灯片

图 5 大体标本、CT观察

Figure 5. Gross specimen，CT observation

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图 6 实验组和对照组家兔牙槽骨缺损处HE染色（×400）

A：无新生骨小梁；B新生骨小梁形成（蓝色箭头示）；C：胶原纤维较少；D大量胶原纤维（黑色箭头示）。

Figure 6. HE staining of alveolar bone defect in experimental and control rabbits（×400）

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