微生物群衍生的后生元对人牙周膜间充质基质细胞的抗炎、抗氧化和再生作用的评估。

Evaluation of the anti-inflammatory, antioxidant and regenerative effects of microbiota-derived postbiotics in human periodontal ligament mesenchymal stromal cells.

作者信息

Demirhan Hazal Kibar, Omer Oglou Emine, Aksoy Zeynep Busra, Kiran Fadime

机构信息

Pharmabiotic Technologies Research Laboratory, Department of Biology, Faculty of Science, Ankara University, Ankara, 06100, Turkey.

Graduate School of Natural and Applied Sciences, Ankara University, Ankara, 06110, Turkey.

出版信息

Clin Oral Investig. 2025 Apr 23;29(5):262. doi: 10.1007/s00784-025-06341-1.

Abstract

OBJECTIVE

This study investigates the regenerative and protective effects of postbiotics (cell-free supernatant) derived from the Lactiplantibacillus plantarum EIR/IF-1 strain on human periodontal ligament mesenchymal stromal cells (hPDL-MSCs).

MATERIALS AND METHODS

hPDL-MSCs were isolated from periodontal ligament tissues (PDL) of wisdom teeth using enzymatic digestion and subsequently characterized through immunophenotyping. The effect of postbiotics on the viability of hPDL-MSCs was assessed using the MTT assay and flow cytometry, while their impact on cell migration was evaluated via the scratch assay. Anti-inflammatory effects of postbiotics were investigated on lipopolysaccharide (LPS, derived from Porphyromonas gingivalis)-stimulated hPDL-MSCs through Enzyme-Linked Immunosorbent Assay (ELISA). Additionally, the antioxidant effects of postbiotics were analyzed in hydrogen peroxide (H₂O₂)-induced hPDL-MSCs by measuring reactive oxygen species (ROS) levels using flow cytometry. The expression of collagen type I (COL1A1) gene was further assessed by quantitative reverse transcription PCR and immunofluorescence staining.

RESULTS

Treatment with postbiotics (250 µg/mL) significantly increased the viability and migration capability of hPDL-MSCs, while enhancing collagen production for PDL repair. Treatment with postbiotics for 24 h resulted in a 54.53 ± 2.01% reduction in intracellular ROS levels compared to untreated HO-induced hPDL-MSCs. Furthermore, postbiotics significantly decreased the production of pro-inflammatory cytokines (IL-8, IL-6, and IL-1β), and increased the anti-inflammatory cytokine IL-10 (2.67-fold) compared to untreated LPS-stimulated hPDL-MSCs.

CONCLUSION

Our findings indicate that postbiotics exhibit biological activity throughout all stages of the healing process, beginning with the modulation of the inflammatory response to LPS stimulation, followed by the promotion of cell migration, proliferation, and collagen synthesis. Given the unmet need for safe and adjuvant therapeutic approaches that promote comprehensive periodontal regeneration in periodontal diseases, this study presents postbiotics as a promising candidate.

CLINICAL RELEVANCE

Postbiotics could be integrated into regenerative therapies as a novel bioactive material to improve the healing and regenerative outcomes in periodontal defects by both controlling inflammation and stimulating tissue repair processes.

摘要

目的

本研究调查植物乳杆菌EIR/IF-1菌株产生的后生元(无细胞上清液)对人牙周膜间充质基质细胞(hPDL-MSCs)的再生和保护作用。

材料与方法

采用酶消化法从智齿的牙周膜组织(PDL)中分离出hPDL-MSCs,随后通过免疫表型分析对其进行表征。使用MTT法和流式细胞术评估后生元对hPDL-MSCs活力的影响,同时通过划痕试验评估其对细胞迁移的影响。通过酶联免疫吸附测定(ELISA)研究后生元对脂多糖(LPS,牙龈卟啉单胞菌来源)刺激的hPDL-MSCs的抗炎作用。此外,通过流式细胞术测量活性氧(ROS)水平,分析后生元在过氧化氢(H₂O₂)诱导的hPDL-MSCs中的抗氧化作用。通过定量逆转录PCR和免疫荧光染色进一步评估I型胶原(COL1A1)基因的表达。

结果

用后生元(250 µg/mL)处理可显著提高hPDL-MSCs的活力和迁移能力,同时增强用于牙周膜修复的胶原蛋白生成。与未处理的H₂O₂诱导的hPDL-MSCs相比,用后生元处理24小时可使细胞内ROS水平降低54.53±2.01%。此外,与未处理的LPS刺激的hPDL-MSCs相比,后生元显著降低促炎细胞因子(IL-8、IL-6和IL-1β)的产生,并使抗炎细胞因子IL-10增加(2.67倍)。

结论

我们的研究结果表明,后生元在愈合过程的所有阶段均表现出生物活性,首先是调节对LPS刺激的炎症反应,随后促进细胞迁移、增殖和胶原蛋白合成。鉴于牙周疾病中促进全面牙周再生的安全辅助治疗方法尚未得到满足,本研究提出后生元是一个有前景的候选物。

临床意义

后生元可作为一种新型生物活性材料整合到再生疗法中,通过控制炎症和刺激组织修复过程来改善牙周缺损的愈合和再生效果。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75f9/12014813/a68fce625c87/784_2025_6341_Fig1_HTML.jpg

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