Gold nanoparticles enhance proliferation and osteogenic differentiation of periodontal ligament stem cells by PINK1-mediated mitophagy

Abstract

Objective: 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.

Methods: 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.

Results: 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.

Conclusion: AuNPs enhance proliferation and osteogenic differentiation of PDLSCs through PINK1-mediated mitophagy.

Keywords: Gold nanoparticles; Mitophagy; Osteogenic differentiation; PINK1; Periodontal ligament stem cells; Proliferation.