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
计量
- 文章访问数: 3048
- HTML全文浏览量: 2079
- PDF下载量: 26
- 被引次数: 0
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

HTML全文

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）

下载: 全尺寸图片幻灯片

图 3 hPDLSCs细胞增殖活力

Figure 3. Cell proliferation activity of hPDLSCs

下载: 全尺寸图片幻灯片

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

下载: 全尺寸图片幻灯片

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）

下载: 全尺寸图片幻灯片

参考文献(17)

[1]	Nazir M A. Prevalence of periodontal disease, its association with systemic diseases and prevention[J]. Int J Health Sci (Qassim),2017,11(2):72-80.
[2]	Kinane D F,Stathopoulou P G,Papapanou P N. Periodontal diseases[J]. Nat Rev Dis Primers,2017,3(1):17038. doi: 10.1038/nrdp.2017.38
[3]	Han J,Menicanin D,Gronthos S,et al. Stem cells,tissue engineering and periodontal regeneration[J]. Australian Dental Journal,2013,59(s1):117-30.
[4]	Lin N H,Gronthos S,Bartold P M. Stem cells and future periodontal regeneration[J]. Periodontology,2009,51(1):239-251. doi: 10.1111/j.1600-0757.2009.00303.x
[5]	Chen F M,Jin Y. Periodontal tissue engineering and regeneration:Current approaches and expanding opportunities[J]. Tissue Eng Part B Rev,2010,16(2):219-255.
[6]	Zhao L R,Mao J Q,Zhao B J,et al. Isolation and biological characteristics of exosomes derived from periodontal ligament stem cells[J]. Shanghai Kou Qiang Yi Xue,2019,28(4):343-348.
[7]	Liu J,Chen B,Bao J,et al. Macrophage polarization in periodontal ligament stem cells enhanced periodontal regeneration[J]. Stem Cell Res Ther,2019,10(1):320. doi: 10.1186/s13287-019-1409-4
[8]	黄涛,陈汉. 纳米羟基磷灰石牙体修复材料的生物性能[J]. 中国组织工程研究,2016,20(34):5045-5050.
[9]	Toth F,Gall JM,Tozser J,et al. Effect of inducible bone morphogenetic protein 2 expression on the osteogenic differentiation of dental pulp stem cells in vitro[J]. Bone,2019,132(12):115214.
[10]	Park S Y,Kim K H,Gwak E H,et al. Ex vivo bone morphogenetic protein 2 gene delivery using periodontal ligament stem cells for enhanced re‐osseointegration in the regenerative treatment of peri‐implantitis[J]. Journal of Biomedical Materials Research Part A,2015,103(1):38-47. doi: 10.1002/jbm.a.35145
[11]	Tan J,Zhang M,Hai Z,et al. Sustained release of two bioactive factors from supramolecular hydrogel promotes periodontal bone regeneration[J]. ACS Nano,2019,13(5):5616-5622. doi: 10.1021/acsnano.9b00788
[12]	Zheng DH,Wang XX,Ma D,et al. Erythropoietin enhances osteogenic differentiation of human periodontal ligament stem cells via Wnt/β-catenin signaling pathway[J]. Drug design,Development and Therapy,2019,13(6):2543-2552.
[13]	Liu H,Zheng J,Zheng T,et al. Exendin-4 regulates Wnt and NF-κB signaling in lipopolysaccharide-induced human periodontal ligament stem cells to promote osteogenic differentiation[J]. International Immunopharmacology,2019,75(9):105801.
[14]	Yan W,Cao Y,Yang H,et al. CB1 enhanced the osteo/dentinogenic differentiation ability of periodontal ligament stem cells via p38 MAPK and JNK in an inflammatory environment[J]. Cell Proliferation,2019,52(6):e12691.
[15]	Jia L,Xiong Y,Zhang W,et al. Metformin promotes osteogenic differentiation and protects against oxidative stress-induced damage in periodontal ligament stem cells via activation of the Akt/Nrf2 signaling pathway[J]. Experimental Cell Research,2020,386(2):111717. doi: 10.1016/j.yexcr.2019.111717
[16]	Liu Z,Guo L,Li R,et al. Transforming growth factor-β1 and hypoxia inducible factor-1α synergistically inhibit the osteogenesis of periodontal ligament stem cells[J]. International Immunopharmacology,2019,75(8):105834.
[17]	Xu Q,Liu Z,Guo L,et al. Hypoxia mediates runt-related transcription factor 2 expression via induction of vascular endothelial growth factor in periodontal ligament stem cells[J]. Molecules and Cells,2019,42(11):763-772.

相关文章(20)

[1]	张晔琳, 马丽娅, 彭旭晖, 杨禾丰, 佘睿. hsa-let-7a-5p调控牙周膜干细胞增殖及凋亡, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20231028
[2]	马思佳, 彭艺, 吴剑花, 税艳青. 云南省耿马县佤族成人牙周健康状况调查及危险因素, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20220404
[3]	丁慧, 姚兰. 云南省农村65～74岁老年人牙周状况及其相关因素, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210407
[4]	马丽娅, 饶南荃, 杨禾丰. 间充质干细胞外泌体在口腔组织再生中的研究进展, 昆明医科大学学报. doi: 10.12259/j.issn.2095-610X.S20210527
[5]	吴芸菲, 张黎, 刘育崧, 符起亚. 牙周翻瓣术配合根管治疗牙髓-牙周综合征的临床效果, 昆明医科大学学报.
[6]	何红, 张婉丽, 李星星, 牛涛, 李红云. 固定式恒久性牙周夹板的临床应用, 昆明医科大学学报.
[7]	钱丽萍, 马灵芝, 余兵, 周治, 刘亚丽. miR-17对炎症微环境中PDLSCs骨向分化特性的影响, 昆明医科大学学报.
[8]	李艳, 胡江天, 彭松娟, 郑宇祥. 正畸治疗对不同牙周生物型牙槽骨密度变化的临床价值, 昆明医科大学学报.
[9]	徐林, 胡敏, 李钟辉, 范建楠. 骨髓间充质干细胞复合人工支架材料治疗骨缺损的研究进展, 昆明医科大学学报.
[10]	何丽明, 何永文. 口腔修复膜材料在牙种植中引导骨再生的临床效果, 昆明医科大学学报.
[11]	胡正雄. TGF-β2和geneX对BrdU标记骨髓间充质干细胞增殖与成骨分化的作用, 昆明医科大学学报.
[12]	王祥芸. 牙周基础治疗对2型糖尿病伴牙周炎与超敏C-反应蛋白的影响, 昆明医科大学学报.
[13]	姚寒曦. HtrA1 及其相关因子MGP、BMP-2 在人牙周膜细胞体外成骨分化和矿化过程中的动态表达, 昆明医科大学学报.
[14]	张艺凡. 派力奥、Vitapex治疗重度牙周-牙髓联合病变临床疗效观察, 昆明医科大学学报.
[15]	周妮. 昆明市育龄期妊娠妇女及非妊娠妇女牙周健康调查, 昆明医科大学学报.
[16]	仲维广. 联合培养法进行人牙周膜细胞的比较培养与鉴定, 昆明医科大学学报.
[17]	高嘉宏. 牙周基础治疗术后配合中草药牙得安牙粉控制牙周炎疗效观察, 昆明医科大学学报.
[18]	黄颖. 人脐带间充质干细胞对再生障碍性贫血患者T细胞相关因子调节的体外研究, 昆明医科大学学报.
[19]	吴幼生. 综合疗法治疗牙周牙髓联合病变96例临床报告, 昆明医科大学学报.
[20]	丁鹏. 新生大鼠海马神经干细胞表达趋化因子受体CCR2的体外研究, 昆明医科大学学报.