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口腔软组织来源的愈合伤口成纤维细胞的基因表达谱:与临床结局、自噬激活和纤维化标志物表达的相关性。

Gene expression profiles of oral soft tissue-derived fibroblast from healing wounds: correlation with clinical outcome, autophagy activation and fibrotic markers expression.

机构信息

Department of Oral and Maxillofacial Sciences, Section of Periodontics, Sapienza University of Rome, Rome, Italy.

Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy.

出版信息

J Clin Periodontol. 2021 May;48(5):705-720. doi: 10.1111/jcpe.13439. Epub 2021 Feb 17.

Abstract

AIM

Our aim was to evaluate gene expression profiling of fibroblasts from human alveolar mucosa (M), buccal attached gingiva (G) and palatal (P) tissues during early wound healing, correlating it with clinical response.

MATERIALS AND METHODS

M, G and P biopsies were harvested from six patients at baseline and 24 hr after surgery. Clinical response was evaluated through Early wound Healing Score (EHS). Fibrotic markers expression and autophagy were assessed on fibroblasts isolated from those tissues by Western blot and qRT-PCR. Fibroblasts from two patients were subjected to RT profiler array, followed by network analysis of the differentially expressed genes. The expression of key genes was validated with qRT-PCR on all patients.

RESULTS

At 24 hr after surgery, EHS was higher in P and G than in M. In line with our clinical results, no autophagy and myofibroblast differentiation were observed in G and P. We observed significant variations in mRNA expression of key genes: RAC1, SERPINE1 and TIMP1, involved in scar formation; CDH1, ITGA4 and ITGB5, contributing to myofibroblast differentiation; and IL6 and CXCL1, involved in inflammation.

CONCLUSIONS

We identified some genes involved in periodontal soft tissue clinical outcome, providing novel insights into the molecular mechanisms of oral repair (ClinicalTrial.gov-NCT04202822).

摘要

目的

我们旨在评估人肺泡黏膜(M)、颊侧附着龈(G)和腭部(P)组织成纤维细胞在早期伤口愈合过程中的基因表达谱,并将其与临床反应相关联。

材料和方法

在基线和术后 24 小时,从 6 名患者中采集 M、G 和 P 活检组织。通过早期伤口愈合评分(EHS)评估临床反应。通过 Western blot 和 qRT-PCR 评估从这些组织中分离的成纤维细胞中的纤维化标志物表达和自噬情况。对两名患者的成纤维细胞进行 RT 基因芯片分析,然后对差异表达基因进行网络分析。通过 qRT-PCR 对所有患者验证关键基因的表达。

结果

术后 24 小时,EHS 在 P 和 G 中高于 M。与我们的临床结果一致,G 和 P 中未观察到自噬和肌成纤维细胞分化。我们观察到关键基因的 mRNA 表达存在显著差异:参与瘢痕形成的 RAC1、SERPINE1 和 TIMP1;参与肌成纤维细胞分化的 CDH1、ITGA4 和 ITGB5;以及参与炎症的 IL6 和 CXCL1。

结论

我们确定了一些与牙周软组织临床结果相关的基因,为口腔修复的分子机制提供了新的见解(ClinicalTrials.gov-NCT04202822)。

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