State Key Laboratory of Oral Disease & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Oral Regenerative Medicine, West China School of Stomatology, Sichuan University, Chengdu, 610041, People's Republic of China.
Engineering Research Center of Oral Translational Medicine, Ministry of Education, West China School of Stomatology, Sichuan University, Chengdu, 610041, People's Republic of China.
Int J Nanomedicine. 2022 Feb 22;17:799-819. doi: 10.2147/IJN.S350869. eCollection 2022.
Lipopolysaccharide (LPS) pretreatment can enhance the therapeutic effect of dental follicle stem cells-derived small extracellular vesicles (DFC-sEV) for periodontitis, and this study aimed to investigate the underlying mechanisms and clinical application Of LPS-preconditioned DFC-sEV in periodontitis.
The protein spectrum of DFC-sEV before and after LPS pretreatment was determined by liquid chromatography-tandem mass spectrometry and bioinformatic analysis. Their effects on inflammatory periodontal ligament stem cells (PDLSCs) and macrophages were investigated for cell proliferation, migration, type 2 macrophage (M2) polarization, and intracellular reactive oxygen species (ROS) levels separately. In addition, the regulation of ROS/Jun amino-terminal kinases (JNK) and ROS/extracellular signal-related kinases (ERK) signaling by LPS-preconditioned DFC-sEV was also studied to reveal the antioxidant mechanism. In vivo, two kinds of DFC-sEV loaded with 0.2% hyaluronic acid (HA) gel were applied for canine periodontitis to evaluate the therapeutic potential.
The proteomic analysis showed that thirty-eight proteins were differentially expressed in LPS-preconditioned DFC-sEV, and interestingly, the highly expressed proteins were mainly involved in antioxidant and enzyme-regulating activities. In addition to promoting PDLSCs and macrophage proliferation, LPS-preconditioned DFC-sEV inhibited intracellular ROS as an antioxidant. It reduced the RANKL/OPG ratio of PDLSCs by inhibiting ROS/JNK signaling under inflammatory conditions and promoted macrophages to polarize toward the M2 phenotype via ROS/ERK signaling. Furthermore, LPS-preconditioned DFC-sEV loaded with the HA injectable system could sustainably release sEV and enhance the therapeutic efficacy for periodontitis in canines.
LPS-preconditioned DFC-sEV could be effectively used as an auxiliary method for periodontitis treatment via antioxidant effects in a subgingival environment, and loading it with HA is feasible and effective for clinical applications.
脂多糖（LPS）预处理可以增强牙髓干细胞来源的小细胞外囊泡（DFC-sEV）治疗牙周炎的疗效，本研究旨在探讨 LPS 预处理的 DFC-sEV 在牙周炎中的潜在机制和临床应用。
通过液相色谱-串联质谱和生物信息学分析，确定 LPS 预处理前后 DFC-sEV 的蛋白质谱。分别研究它们对炎性牙周膜干细胞（PDLSCs）和巨噬细胞的影响，包括细胞增殖、迁移、2 型巨噬细胞（M2）极化和细胞内活性氧（ROS）水平。此外，还研究了 LPS 预处理的 DFC-sEV 对 ROS/Jun 氨基末端激酶（JNK）和 ROS/细胞外信号调节激酶（ERK）信号的调节作用，以揭示其抗氧化机制。在体内，将两种负载 0.2%透明质酸（HA）凝胶的 DFC-sEV 用于犬牙周炎，以评估其治疗潜力。
蛋白质组学分析显示，LPS 预处理的 DFC-sEV 中有 38 种蛋白质表达差异，有趣的是，高表达的蛋白质主要参与抗氧化和酶调节活性。除了促进 PDLSCs 和巨噬细胞增殖外，LPS 预处理的 DFC-sEV 还作为抗氧化剂抑制细胞内 ROS。在炎症条件下，它通过抑制 ROS/JNK 信号通路降低 PDLSCs 中的 RANKL/OPG 比值，并通过 ROS/ERK 信号通路促进巨噬细胞向 M2 表型极化。此外，负载 HA 可注射系统的 LPS 预处理的 DFC-sEV 可以持续释放 sEV，并增强犬牙周炎的治疗效果。
LPS 预处理的 DFC-sEV 可通过龈下环境中的抗氧化作用有效用作牙周炎治疗的辅助方法，负载 HA 是可行且有效的临床应用方法。
