Department of Orthodontics, Stomatological Hospital, Southern Medical University, No. 366 South Jiangnan Road, Haizhu District, Guangzhou, 510280, Guangdong, People's Republic of China.
Department of Orthodontics, Nanning Angel Stomatological Hospital, No. 20-1, Xinmin Road, Nanning, 530029, Guangxi, People's Republic of China.
Cell Tissue Bank. 2023 Mar;24(1):45-58. doi: 10.1007/s10561-022-10010-y. Epub 2022 May 28.
Human periodontal ligament stem cells (hPDLSCs) are vital in cellular regeneration and tissue repair due to their multilineage differentiation potential. Low intensity pulsed ultrasound (LIPUS) has been applied for treating bone and cartilage defects. This study explored the role of LIPUS in the immunomodulation and osteogenesis of hPDLSCs. hPDLSCs were cultured in vitro, and the effect of different intensities of LIPUS (30, 60, and 90 mW/cm) on hPDLSC viability was measured. hPDLSCs irradiated by LIPUS and stimulated by lipopolysaccharide (LPS) and LIPUS (90 mW/cm2) were co-cultured with peripheral blood mononuclear cells (PBMCs). Levels of immunomodulatory factors in hPDLSCs and inflammatory factors in PBMCs were estimated, along with determination of osteogenesis-related gene expression in LIPUS-irradiated hPDLSCs. The mineralized nodules and alkaline phosphatase (ALP) activity of hPDLSCs and levels of IκBα, p-IκBα, and p65 subunits of NF-κB were determined. hPDLSC viability was increased as LIPUS intensity increased. Immunomodulatory factors were elevated in LIPUS-irradiated hPDLSCs, and inflammatory factors were reduced in PBMCs. Osteogenesis-related genes, mineralized nodules, and ALP activity were promoted in LIPUS-irradiated hPDLSCs. The cytoplasm of hPDLSCs showed increased IκBα and p65 and decreased p-IκBα at increased LIPUS intensity. After LPS and LIPUS treatment, the inhibitory effect of LIPUS irradiation on the NF-κB pathway was partially reversed, and the immunoregulation and osteogenic differentiation of hPDLSCs were decreased. LIPUS irradiation enhanced immunomodulation and osteogenic differentiation abilities of hPDLSCs by inhibiting the NF-κB pathway, and the effect is dose-dependent. This study may offer novel insights relevant to periodontal tissue engineering.
人牙周韧带干细胞（hPDLSCs）具有多向分化潜能，在细胞再生和组织修复中至关重要。低强度脉冲超声（LIPUS）已应用于治疗骨和软骨缺损。本研究探讨了 LIPUS 在 hPDLSCs 免疫调节和成骨中的作用。体外培养 hPDLSCs，测量不同强度 LIPUS（30、60 和 90 mW/cm）对 hPDLSC 活力的影响。将 LIPUS 照射的 hPDLSCs 与脂多糖（LPS）和 LIPUS（90 mW/cm2）刺激的外周血单个核细胞（PBMCs）共培养。测定 hPDLSCs 中免疫调节因子和 PBMCs 中炎症因子的水平，以及 LIPUS 照射的 hPDLSCs 中成骨相关基因的表达。测定 hPDLSCs 矿化结节和碱性磷酸酶（ALP）活性以及 NF-κB 的 IκBα、p-IκBα 和 p65 亚基的水平。随着 LIPUS 强度的增加，hPDLSC 的活力增加。LIPUS 照射的 hPDLSCs 中免疫调节因子升高，PBMCs 中炎症因子减少。LIPUS 照射的 hPDLSCs 中成骨相关基因、矿化结节和 ALP 活性增加。随着 LIPUS 强度的增加，hPDLSCs 的细胞质中 IκBα 和 p65 增加，p-IκBα 减少。LPS 和 LIPUS 处理后，LIPUS 照射对 NF-κB 通路的抑制作用部分逆转，hPDLSCs 的免疫调节和成骨分化能力降低。LIPUS 照射通过抑制 NF-κB 通路增强 hPDLSCs 的免疫调节和成骨分化能力，且呈剂量依赖性。本研究可能为牙周组织工程提供新的见解。
