Xu Xiaofeng, Lin Kaijin, Zhou Bowei, Peng Dongsheng, Wang Siyi, Zhao Wei, Zheng Minqian, Yang Jin, Guo Jianbin
Department of Stomatology, Affiliated Hospital of Putian University, Putian, China; Fujian Key Laboratory of Oral Diseases & Fujian Provincial Engineering Research Center of Oral Biomaterial & Stomatological Key Lab of Fujian College and University, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, China; Institute of Stomatology & Research Center of Dental and Craniofacial Implants, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, China.
Fujian Key Laboratory of Oral Diseases & Fujian Provincial Engineering Research Center of Oral Biomaterial & Stomatological Key Lab of Fujian College and University, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, China; Institute of Stomatology & Research Center of Dental and Craniofacial Implants, School and Hospital of Stomatology, Fujian Medical University, Fuzhou, China.
J Craniomaxillofac Surg. 2025 Oct 29. doi: 10.1016/j.jcms.2025.10.009.
Demineralized dentin matrix (DDM) is a bioactive tissue known to promote bone regeneration by enhancing osteoblast proliferation and osteogenic differentiation. Recent studies suggest that DDM may also stimulate fibroblast proliferation, adhesion, and migration. However, the inductive capacity of DDM on human gingival fibroblasts (HGFs) and its underlying mechanisms remain largely unexplored. This study aims to investigate the effects of DDM on HGFs and to elucidate the potential molecular mechanisms involved.
The effects of DDM on HGFs migration and adhesion were investigated using a multi-step approach. First, the optimal DDM concentration for co-culture with HGFs was determined using a cell migration assay. Next, Transwell migration assays and scanning electron microscopy were employed to evaluate the impact of DDM on HGFs migration and adhesion. Subsequently, high-throughput sequencing was performed on co-cultured HGFs and a control group to analyze and identify differentially expressed genes (DEGs). Finally, the expression of three DEGs was validated using RT-qPCR.
The optimal concentration of DDM for promoting HGFs migration was determined to be 10 mg/mL. The Transwell assay revealed a significantly higher number of migrating HGFs in the DDM-treated group compared to the control group. Scanning electron microscopy imaging showed that HGFs adhered to and spread on the DDM surface. Transcriptome analysis identified 64 DEGs between the HGFs monoculture group and the DDM co-culture group, with 6 genes upregulated and 57 downregulated. GO functional enrichment analysis and KEGG pathway enrichment analysis indicated that these DEGs were primarily enriched in biological processes related to immune and inflammatory responses. The expression levels of three key DEGs were further validated using qRT-PCR, and the results were consistent with the RNA-Seq data.
DDM exhibits biocompatibility and enhances HGFs migration. This suggests that DDM may play a crucial role in promoting gingival soft tissue healing by regulating biological processes and gene expression related to immune and inflammatory responses.
脱矿牙本质基质（DDM）是一种生物活性组织，已知其可通过增强成骨细胞增殖和成骨分化来促进骨再生。最近的研究表明，DDM还可能刺激成纤维细胞增殖、黏附和迁移。然而，DDM对人牙龈成纤维细胞（HGFs）的诱导能力及其潜在机制在很大程度上仍未得到探索。本研究旨在研究DDM对HGFs的影响，并阐明其中潜在的分子机制。
采用多步骤方法研究DDM对HGFs迁移和黏附的影响。首先，使用细胞迁移试验确定与HGFs共培养的最佳DDM浓度。接下来，采用Transwell迁移试验和扫描电子显微镜评估DDM对HGFs迁移和黏附的影响。随后，对共培养的HGFs和对照组进行高通量测序，以分析和鉴定差异表达基因（DEGs）。最后，使用RT-qPCR验证三个DEGs的表达。
确定促进HGFs迁移的DDM最佳浓度为10mg/mL。Transwell试验显示，与对照组相比，DDM处理组中迁移的HGFs数量显著更多。扫描电子显微镜成像显示，HGFs黏附并铺展在DDM表面。转录组分析确定了HGFs单培养组和DDM共培养组之间的64个DEGs，其中6个基因上调，57个基因下调。GO功能富集分析和KEGG通路富集分析表明，这些DEGs主要富集在与免疫和炎症反应相关的生物学过程中。使用qRT-PCR进一步验证了三个关键DEGs的表达水平，结果与RNA-Seq数据一致。
DDM具有生物相容性并增强HGFs迁移。这表明DDM可能通过调节与免疫和炎症反应相关的生物学过程和基因表达，在促进牙龈软组织愈合中发挥关键作用。
