中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (32): 5102-5106.doi: 10.12307/2022.906

• 口腔组织构建 oral tissue construction • 上一篇    下一篇

抑制miR-203-3p表达对糖尿病小鼠牙种植体周围骨形成的影响

李孔梅，罗诒财，李  昊   

  1. 广西医科大学口腔医学院口腔修复科，广西壮族自治区南宁市  530021
  • 收稿日期:2021-10-18 接受日期:2021-11-27 出版日期:2022-11-18 发布日期:2022-05-12
  • 通讯作者: 李昊，博士，副主任医师，广西医科大学口腔医学院口腔修复科，广西壮族自治区南宁市 530021
  • 作者简介:李孔梅，女，1997年生，广西壮族自治区玉林市人，汉族，广西医科大学在读硕士，主要从事口腔种植修复方面的研究。
  • 基金资助:
    国家自然科学基金(82060195，81600833)，项目负责人：李昊

Effect of inhibiting miR-203-3p expression on bone formation around dental implants in diabetic mice

Li Kongmei, Luo Yicai, Li Hao   

  1. Department of Prosthodontics, College of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Received:2021-10-18 Accepted:2021-11-27 Online:2022-11-18 Published:2022-05-12
  • Contact: Li Hao, MD, Associate chief physician, Department of Prosthodontics, College of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • About author:Li Kongmei, Master candidate, Department of Prosthodontics, College of Stomatology, Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, Nos. 82060195 and 81600833 (to LH)

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摘要：

文题释义：
miR-203-3p：是miRNA家族的一员，在多种细胞的增殖、分化以及炎症反应等过程中具有调节作用。既往研究表明，miR-203-3p可调节多种疾病状态下的骨形成，通过调控miR-203-3p表达治疗多种炎症、肿瘤等疾病是近年来研究的热点。
种植体周围骨形成：牙种植修复缺失牙可以达到很好的功能和美学效果，是治疗牙缺失的重要方式之一。牙种植成功的关键是种植体周围有良好的骨形成，而糖尿病骨状态易导致牙槽骨缺损难以愈合、牙种植体周围骨形成不良，从而导致种植体易松动、牙种植失败，因此，如何改善糖尿病状态种植体周围骨形成是口腔医学中的难点。

背景：糖尿病状态下牙种植体周围骨形成不佳，机制不明确。近年研究表明，miR-203-3p可调节多种疾病状态下的骨形成，在糖尿病颌骨组织中高表达，然而是否对牙种植体周围骨形成也有影响，尚不清楚。
目的：探讨抑制miR-203-3p表达对糖尿病小鼠牙种植体周围骨形成的影响。
方法：C57BL/6J小鼠48只，随机取血糖正常小鼠12只作为正常组，其余小鼠构建糖尿病模型后，随机分为糖尿病组、糖尿病+antago miR-NC组、糖尿病+antago miR-203-3p组，每组12只。所有小鼠构建左侧上颌牙种植模型，糖尿病+antago miR-NC组注射阴性对照antago miR-NC(0.25 mmol/L)，糖尿病+ antago miR-203-3p组注射antago miR-203-3p(0.25 mmol/L)，其余两组注射等体积生理盐水。3个月后取小鼠左侧上颌骨组织，Micro-CT、苏木精-伊红染色观察种植体周围骨形成情况；qRT-PCR检测种植体周围骨组织中miR-203-3p、Smad1、碱性磷酸酶、Runx2、Osterix、Ⅰ型胶原基因表达。
结果与结论：①Micro-CT检测显示，与糖尿病组、糖尿病+antago miR-NC组比较，糖尿病+antago miR-203-3p组种植体周围牙槽骨组织形成情况较好，骨体积分数、骨小梁厚度较高(P < 0.05)；②苏木精-伊红染色显示，糖尿病+antago miR-203-3p组种植体周围骨形成Lane-Sandhu评分高于糖尿病组、糖尿病+antago miR-NC组(P < 0.05)；③qRT-PCR结果显示，糖尿病+antago miR-203-3p组种植体周围骨组织miR-203-3p基因表达水平低于糖尿病组、糖尿病+antago miR-NC组(P < 0.05)，Smad1、碱性磷酸酶、Runx2、Osterix、Ⅰ型胶原基因表达水平高于糖尿病组、糖尿病+antago miR-NC组(P < 0.05)；④结果表明，在高糖状态下，抑制miR-203-3p表达有利于种植体周围骨形成，这些作用可能与调节Smad信号通路及碱性磷酸酶、Runx2、Osterix、Ⅰ型胶原等成骨相关因子表达有关。

https://orcid.org/0000-0002-4536-0929 (李孔梅)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 糖尿病, miR-203-3p, 种植体, 骨形成, 小鼠, Smad1, Runx2, Osterix, 碱性磷酸酶, Ⅰ型胶原

Abstract: BACKGROUND: Up to now, dental implant has become an important restoration for patients with tooth loss, and bone formation around dental implants is the key to the success of dental implantation. There is often a poor bone formation around dental implants under diabetic conditions and the mechanism remains unclear. Recent studies have shown that miR-203-3p regulates bone formation in different diseases, and strongly expressed in jaw bone tissues under diabetic conditions. However, it is unclear whether miR-203-3p can influence bone formation around dental implants.
OBJECTIVE: To investigate the effect of inhibiting miR-203-3p expression on bone formation around dental implants in diabetic mice.
METHODS: Twelve mice with normal blood glucose were randomly selected as normal group from forty-eight C57BL/6J mice. The other 36 mice were selected to make diabetic models and then randomly divided into diabetic group, diabetes+antago miR-NC group, and diabetes+antago miR-203-3p group (n=12 per group). Single implant was inserted into the left maxilla of each mouse. Diabetic+antago miR-NC group was injected with antago miR-203-3p negative control, antago miR-NC (0.25 mmol/L). Diabetes+antago miR-203-3p group was injected with antago miR-203-3p (0.25 mmol/L). The other two diabetic groups were injected with the equal volume of normal saline. After 3 months, the left maxillary bone tissue of mice was collected, and the bone formation around dental implants was detected using micro-CT and hematoxylin-eosin staining. qRT-PCR was used to detect the gene expression of miR-203-3p, Smad1, alkaline phosphatase, Runx2, Osterix, and type I collagen in bone tissue around dental implants.  
RESULTS AND CONCLUSION: Micro-CT results showed that compared with the diabetic group and diabetes+antago miR-NC group, the diabetes+antago miR-203-3p group showed better alveolar bone formation around dental implants and higher bone volume fraction and bone trabecular thickness (P < 0.05). Hematoxylin-eosin staining results showed that the Lane-Sandhu score of bone formation around dental implants was higher in the diabetes+antago miR-203-3p group than the diabetic group and the diabetes+antago miR-NC group (P < 0.05). qRT-PCR results showed that the expression level of miR-203-3p gene in bone tissue around dental implants was lower (P < 0.05), while the expression levels of Smad1, alkaline phosphatase, Runx2, Osterix, and type I collagen genes were higher in the diabetes+antago miR-203-3p group than the diabetic group and the diabetes+antago miR-NC group (P < 0.05). All these findings indicate that the inhibition of miR-203-3p expression is beneficial to bone formation around dental implants in diabetic hyperglycemia, and these effects may be related to the regulation of Smad signaling pathway and the expression of osteogenesis-related factors such as alkaline phosphatase, Runx2, Osterix, and type I collagen.

Key words: diabetes mellitus, miR -203-3p, implant, bone formation, mouse, Smad1, Runx2, Osterix, alkaline phosphatase, type I collagen

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