Rizhao Hospital of Traditional Chinese Medicine, China.
Disease Control and Prevention Center of Rizhao, China.
Arch Oral Biol. 2023 Jun;150:105692. doi: 10.1016/j.archoralbio.2023.105692. Epub 2023 Mar 29.
Evidence suggests that gold nanoparticles (AuNPs) improve osteogenic differentiation of periodontal ligament stem cells (PDLSCs), PTEN-induced putative kinase 1 (PINK1) dependent mitophagy modulates inter-clonal communication among PDLSCs with osteogenic heterogeneity, but the mechanism remains vague. Therefore, the current research assessed the influence of AuNPs on proliferation, osteogenic differentiation and mitophagy of PDLSCs and the potential mechanism was analyzed.
Gold nanospheres with a diameter of 5, 10, 20, 40, and 80 nm were synthesized and characterized through transmission electron microscopy, and rat PDLSCs were isolated using flow sorting. Next, PDLSCs were treated with AuNPs or PINK1 lentivirus to obtain its overexpression or suppression. Proliferation and osteogenic differentiation were evaluated by CCK-8, ALP staining, ARS staining, and immunoblotting of OCN, OPN, RUNX2, ALP, BMP2, and COL1. Mitochondrial quality, homeostasis and quantity were assessed though JC-1 staining, immunoblotting of Tom20, Tim23 and HSP60 and mitochondrial ROS detection. PINK1, Parkin, Beclin1 and LC3 expression was quantified to investigate mitophagy, using RT-qPCR and immunoblotting and the formation of RFP-GFP-LC3-labeled autophagosomes were also measured.
The proliferation ability of PDLSCs almost reached the maximum under 20 nm AuNPs for 24 h. AuNPs enhanced the proliferation and osteogenic differentiation of PDLSCs, improved mitochondrial quality and homeostasis as well as attenuated mitochondrial quantity. Additionally, mitophagy was enhanced by PDLSCs. Activation of PINK1 synergistically enhanced AuNPs-mediated mitophagy, mitochondrial quality, homeostasis and osteogenic differentiation in PDLSCs, obtaining opposite effects when PINK1 was suppressed.
AuNPs enhance proliferation and osteogenic differentiation of PDLSCs through PINK1-mediated mitophagy.
有证据表明，金纳米粒子（AuNPs）可促进牙周膜干细胞（PDLSCs）的成骨分化，PTEN 诱导的假定激酶 1（PINK1）依赖性线粒体自噬调节具有成骨异质性的 PDLSCs 间的克隆间通讯，但机制尚不清楚。因此，本研究评估了 AuNPs 对 PDLSCs 增殖、成骨分化和线粒体自噬的影响，并分析了潜在的机制。
通过透射电子显微镜合成并表征了直径为 5、10、20、40 和 80nm 的金纳米球，并用流式分选法分离大鼠 PDLSCs。然后，用 AuNPs 或 PINK1 慢病毒处理 PDLSCs，以获得其过表达或抑制。通过 CCK-8、ALP 染色、ARS 染色和 OCN、OPN、RUNX2、ALP、BMP2 和 COL1 的免疫印迹评估增殖和成骨分化。通过 JC-1 染色、Tom20、Tim23 和 HSP60 的免疫印迹和线粒体 ROS 检测评估线粒体质量、平衡和数量。通过 RT-qPCR 和免疫印迹定量测定 PINK1、Parkin、Beclin1 和 LC3 的表达，以研究线粒体自噬，并测量 RFP-GFP-LC3 标记的自噬体的形成。
在 24h 内，20nm AuNPs 下 PDLSCs 的增殖能力几乎达到最大值。AuNPs 增强了 PDLSCs 的增殖和成骨分化，改善了线粒体质量和平衡，减少了线粒体数量。此外，还增强了线粒体自噬。PINK1 的激活协同增强了 PDLSCs 中 AuNPs 介导的线粒体自噬、线粒体质量、平衡和成骨分化，而当 PINK1 被抑制时则产生相反的效果。
AuNPs 通过 PINK1 介导的线粒体自噬增强 PDLSCs 的增殖和成骨分化。
