低强度脉冲超声通过调节整合素α11（ITGA11）和粘着斑通路促进大鼠骨髓间充质干细胞的成骨分化和牙种植体的骨整合。

LIPUS promotes osteogenic differentiation of rat BMSCs and osseointegration of dental implants by regulating ITGA11 and focal adhesion pathway.

作者信息

Liang Chao, Zhang Yuqing, Yan Yuwei, Geng Wei, Li Jun, Liu Xiu

机构信息

Department of Dental Implant Center, Beijing Stomatological Hospital, School of Stomatology, Capital Medical University, Beijing, China.

Beijing Institute of Dental Research, Beijing Stomatological Hospital, School of Stomatology, Capital Medical University, Beijing, China.

出版信息

BMC Oral Health. 2025 Jan 4;25(1):22. doi: 10.1186/s12903-024-05411-2.

DOI:10.1186/s12903-024-05411-2

PMID:39755586

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11700459/

Abstract

BACKGROUND

Low-intensity pulsed ultrasound (LIPUS) has been used as an effective noninvasive method for treating fractures and osteoarthrosis, but the application in the field of oral implantation is in its infancy. This study aimed to clarify the effect and mechanism of LIPUS on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and implant osseointegration, and to provide an experimental basis for future clinical applications.

METHODS

Dental implants were inserted into Wistar rat femurs, and LIPUS was performed for 4 weeks. Micro-CT and toluidine blue staining were used to assess implant osseointegration. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to identify enriched functional terms and signalling pathways for differentially expressed genes from LIPUS-treated rat BMSC RNAseq data obtained from the GEO database. The random forest method was used to identify key risk genes according to the mean decrease Gini (MDG) coefficient. Then, LIPUS was applied to treat rat BMSCs, and alkaline phosphatase (ALP) staining, alizarin red staining, RT-PCR and western blotting were used to determine whether LIPUS could promote BMSC osteogenic differentiation via integrin α11 (ITGA11) and the focal adhesion pathway.

RESULTS

Our in vivo experimentations verified that LIPUS significantly increased new bone formation and osseointegration around the implant in rats. Bioinformatics analysis of RNA-seq data revealed that the upregulated genes in BMSCs after LIPUS treatment were significantly enriched in osteoblast differentiation-related functions and focal adhesion-related pathways. Random forest analysis revealed that ITGA11 was the most significant factor affecting BMSC osteogenic differentiation among the differentially expressed genes. In addition, LIPUS significantly increased ALP expression and mineralized nodule formation in rat BMSCs by upregulating ITGA11 and increasing the activity of FAK/PI3K/AKT/GSK3β/β-catenin pathway.

CONCLUSIONS

LIPUS can effectively promote implant osseointegration in rats and improve rat BMSC osteogenic differentiation by upregulating ITGA11 and increasing the activity of the downstream focal adhesion pathway.

摘要

背景

低强度脉冲超声（LIPUS）已被用作治疗骨折和骨关节炎的一种有效的非侵入性方法，但在口腔种植领域的应用尚处于起步阶段。本研究旨在阐明LIPUS对骨髓间充质干细胞（BMSCs）成骨分化及种植体骨整合的影响和机制，为未来临床应用提供实验依据。

方法

将牙种植体植入Wistar大鼠股骨，进行4周的LIPUS治疗。采用显微CT和甲苯胺蓝染色评估种植体骨整合情况。利用基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析，从GEO数据库获得的LIPUS处理的大鼠BMSC RNAseq数据中鉴定差异表达基因的富集功能术语和信号通路。采用随机森林法根据平均减少基尼系数（MDG）确定关键风险基因。然后，应用LIPUS处理大鼠BMSCs，采用碱性磷酸酶（ALP）染色、茜素红染色、RT-PCR和蛋白质免疫印迹法，以确定LIPUS是否能通过整合素α11（ITGA11）和粘着斑途径促进BMSCs成骨分化。

结果

我们的体内实验证实，LIPUS显著增加了大鼠种植体周围的新骨形成和骨整合。RNA-seq数据的生物信息学分析表明，LIPUS处理后BMSCs中上调的基因在成骨细胞分化相关功能和粘着斑相关途径中显著富集。随机森林分析显示，ITGA11是差异表达基因中影响BMSCs成骨分化的最显著因素。此外，LIPUS通过上调ITGA11并增加FAK/PI3K/AKT/GSK3β/β-连环蛋白通路的活性，显著增加了大鼠BMSCs中ALP的表达和矿化结节的形成。