Roato Ilaria, Baima Giacomo, Orrico Clarissa, Mosca Balma Alessandro, Alotto Daniela, Romano Federica, Ferracini Riccardo, Aimetti Mario, Mussano Federico
Department of Surgical Sciences, University of Turin, 10126 Turin, Italy.
Dipartimento di Scienze Chirurgiche, Politecnico di Torino, 10129 Turin, Italy.
Biomedicines. 2023 Sep 14;11(9):2535. doi: 10.3390/biomedicines11092535.
Periodontal ligament (PDL) has become an elective source of mesenchymal stem cells (PDLSCs) in dentistry. This research aimed to compare healthy PDLSCs (hPDLSCs) and periodontitis PDLSCs (pPDLSCs) to ascertain any possible functional differences owing to their milieux of origin. Cells were tested in terms of colony-forming unit efficiency; multi differentiating capacity; immunophenotype, stemness, and senescent state were studied by flow cytometry, immunofluorescence, and β-galactosidase staining; gene expression using RT-PCR. Both hPDLSCs and pPDLSCs were comparable in terms of their immunophenotype and multilineage differentiation capabilities, but pPDLSCs showed a senescent phenotype more frequently. Thus, a selective small molecule inhibitor of DNA methyltransferase (DNMT), RG108, known for its effect on senescence, was used to possibly reverse this phenotype. RG108 did not affect the proliferation and apoptosis of PDLSCs, and it showed little effect on hPDLSCs, while a significant reduction of both p16 and p21 was detected along with an increase of SOX2 and OCT4 in pPDLSCs after treatment at 100 μM RG108. Moreover, the subset of PDLSCs co-expressing OCT4 and p21 decreased, and adipogenic potential increased in pPDLSCs after treatment. pPDLSCs displayed a senescent phenotype that could be reversed, opening new perspectives for the treatment of periodontitis.
牙周韧带（PDL）已成为牙科领域间充质干细胞（PDLSCs）的一个理想来源。本研究旨在比较健康牙周韧带干细胞（hPDLSCs）和牙周炎牙周韧带干细胞（pPDLSCs），以确定由于其起源环境可能存在的任何功能差异。对细胞进行了集落形成单位效率测试；研究了多向分化能力；通过流式细胞术、免疫荧光和β-半乳糖苷酶染色研究了免疫表型、干性和衰老状态；使用逆转录聚合酶链反应（RT-PCR）检测基因表达。hPDLSCs和pPDLSCs在免疫表型和多谱系分化能力方面具有可比性，但pPDLSCs更频繁地表现出衰老表型。因此，一种已知对衰老有影响的DNA甲基转移酶（DNMT）选择性小分子抑制剂RG108被用于可能逆转这种表型。RG108不影响PDLSCs的增殖和凋亡，对hPDLSCs影响不大，而在100μM RG108处理后的pPDLSCs中，检测到p16和p21均显著降低，同时SOX2和OCT4增加。此外，处理后的pPDLSCs中，共表达OCT4和p21的PDLSCs亚群减少，成脂潜能增加。pPDLSCs表现出一种可逆转的衰老表型，为牙周炎的治疗开辟了新的前景。
