Effects of tension on mitochondrial autophagy and osteogenic differentiation of periodontal ligament stem cells

Abstract

This study aimed to explore the osteogenic ability and mitochondrial autophagy of periodontal ligament stem cells (PDLSCs) under cyclic tensile stress (CTS). Primary PDLSCs were isolated from the periodontal membrane and cultured by passage. Alizarin red staining, alkaline phosphatase detection, reverse transcription polymerase chain reaction (RT-PCR), and Western blotting were used to detect the osteogenic differentiation level of PDLSCs. Mitochondrial autophagy in PDLSCs after CTS was measured using a mitochondrial autophagy detection kit, and the expression levels of autophagy-related proteins LC3B, LAMP1 and Beclin1 were measured using cellular immunofluorescence technology, RT-PCR and Western blot. After applying CTS, the osteogenic differentiation ability of PDLSCs was significantly improved, and the expression of alkaline phosphatase on the surface of the cell membrane and the formation of calcium nodules in PDLSCs were significantly increased respectively. We also studied the relevant mechanism of action and found that applying CTS can promote the osteogenic differentiation of PDLSCs and is related to the activation of mitochondrial autophagy. This study provides new insights into the mechanism of increased osteogenic differentiation on the tension side of orthodontic teeth and provides new experimental evidence for the involvement of mitochondrial autophagy in the regulation of osteogenic differentiation.

References

[1] Wise GE, King GJ. Mechanisms of tooth eruption and orthodontic tooth movement. J Dent Res. 2008;87:414-434.

[2] Cultrera G, Lo Giudice A, Santonocito S, et al. MicroRNA modulation during orthodontic tooth movement: a promising strategy for novel diagnostic and personalized therapeutic interventions. Int J Mol Sci. 2022;23:15501.

[3] Liu J, Li Q, Liu S, et al. Periodontal ligament stem cells in the periodontitis microenvironment are sensitive to static mechanical strain. Stem Cells Int. 2017;2017:1380851.

[4] Sen S, Erber R. Neuronal guidance molecules in bone remodeling and orthodontic tooth movement. Int J Mol Sci. 2022;23:10077.

[5] Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004;364:149-155.

[6] Liang Y, Shakya A, Liu X. Biomimetic tubular matrix induces periodontal ligament principal fiber formation and inhibits osteogenic differentiation of periodontal ligament stem cells. ACS Appl Mater Interfaces. 2022;14:36451-36461.

[7] Shao X, Hu Z, Zhan Y, Ma W, Quan L, Lin Y. MiR-26a-tetrahedral framework nucleic acids mediated osteogenesis of adipose-derived mesenchymal stem cells. Cell Prolifer. 2022;55:e13272.

[8] Forni MF, Peloggia J, Trudeau K, Shirihai O, Kowaltowski AJ. Murine mesenchymal stem cell commitment to differentiation is regulated by mitochondrial dynamics. Stem Cells. 2016;34:743-755.

[9] Shen T, Qiu L, Chang H, et al. Cyclic tension promotes osteogenic differentiation in human periodontal ligament stem cells. Int J Clin Exp Pathol. 2014;7:7872-7880.

[10] Jiang Z, Hua Y. Hydrogen sulfide promotes osteogenic differentiation of human periodontal ligament cells via p38-MAPK signaling pathway under proper tension stimulation. Arch Oral Biol. 2016;72:8-13.

[11] Chukkapalli SS, Lele TP. Periodontal cell mechanotransduction. Open Biol. 2018;8:180053.

[12] Shalehin N, Seki Y, Takebe H, et al. Gli1(+)-PDL cells contribute to alveolar bone homeostasis and regeneration. J Dent Res. 2022;101:1537-1543.

[13] Zhang T, Ma HS, Zhang XL, Shi S, Lin Y. Functionalized DNA nanomaterials targeting toll-like receptor 4 prevent bisphosphonate-related osteonecrosis of the jaw via regulating mitochondrial homeostasis in macrophages. Adv Funct Mater. 2023;33:2213401.

[14] Yan R, Cui WT, Ma WJ, Li J, Liu Z, Lin Y. Typhaneoside-tetrahedral framework nucleic acids system: mitochondrial recovery and antioxidation for acute kidney injury treatment. ACS Nano. 2023;17:8767-8781.

[15] Shi R, Meng X, Feng R, et al. Stress distribution and fracture resistance of repairing cracked tooth with fiber-reinforced composites and Onlay. Aust Endod J. 2022;48:458-464.

[16] Furhauser R, Florescu D, Benesch T, et al. Evaluation of soft tissue around single-tooth implant crowns: the pink esthetic score. Clin Oral Implants Res. 2005;16:639-644.

[17] Ruan JY, Gong ZL, Zhang RZ, et al. Evaluation of four different restorative Materials for restoration of the periodontal condition of wedge-shaped defect: a comparative study. Med Sci Monit. 2017;23:4462-4470.

[18] Peluso V, Rinaldi L, Russo T, et al. Impact of magnetic stimulation on periodontal ligament stem cells. Int J Mol Sci. 2021;23:188.

[19] Xie Y, He JJ, Li SH, et al. A transdermal drug delivery system based on nucleic acid nanomaterials for skin photodamage treatment. Adv Funct Mater. 2023;2303580.

[20] Chen R, Wen D, Fu W, et al. The treatment effect of DNA framework nucleic acids on diffuse microvascular endothelial cell injury after subarachnoid hemorrhage. Cell Prolif. 2022;55:e13206.

[21] Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2006;8:315-317.

[22] Zhang B, Qin X, Zhou M, et al. Tetrahedral DNA nanostructure improves transport efficiency and anti-fungal effect of histatin 5 against Candida albicans. Cell Prolif. 2021;54:e13105.

[23] Shi SR, Chen TY, Lu WT, Chen Y, Xiao D, Lin Y. Amelioration of osteoarthritis via tetrahedral framework nucleic acids delivering Microrna-124 for cartilage regeneration. Adv Funct Mater. 2023;2305558.

[24] Yang Y, Wang BK, Chang ML, Wan ZQ, Han GL. Cyclic stretch enhances osteogenic differentiation of human periodontal ligament cells via YAP activation. Biomed Res Int. 2018;2018:2174824.

[25] Podesta MA, Remuzzi G, Casiraghi F. Mesenchymal stromal cells for transplant tolerance. Front Immunol. 2019;10:1287.

[26] Harper JW, Ordureau A, Heo JM. Building and decoding ubiquitin chains for mitophagy. Nat Rev Mol Cell Biol. 2018;19:93-108.

[27] Rambold AS, Kostelecky B, Elia N, Lippincott-Schwartz J. Tubular network formation protects mitochondria from autophagosomal degradation during nutrient starvation. Proc Natl Acad Sci U S A. 2011;108:10190-10195.

[28] Gomes LC, Di Benedetto G, Scorrano L. During autophagy mitochondria elongate, are spared from degradation and sustain cell viability. Nat Cell Biol. 2011;13:589-598.

[29] Chiricosta L, Gugliandolo A, Diomede F, et al. Moringin pretreatment inhibits the expression of genes involved in mitophagy in the stem cell of the human periodontal ligament. Molecules. 2019;24:3217.

[30] Nollet M, Santucci-Darmanin S, Breuil V, et al. Autophagy in osteoblasts is involved in mineralization and bone homeostasis. Autophagy. 2014;10:1965-1977.

[31] Ma S, Ding R, Cao J, Liu Z, Li A, Pei D. Mitochondria transfer reverses the inhibitory effects of low stiffness on osteogenic differentiation of human mesenchymal stem cells. Eur J Cell Biol. 2023;102:151297.

[32] Chen T, Xiao D, Li Y, et al. Tetrahedral framework nucleic acids regulate osteogenic differentiation potential of osteoporotic adipose-derived stem cells. Chin Chem Lett. 2022;33:2517-2521.

[33] Wen Y, Zhang M, Yao Y, et al. Biological regulation on synovial fibroblast and the treatment of rheumatoid arthritis by nobiletin-loaded tetrahedral framework nucleic acids cargo tank. Chin Chem Lett. 2023;34:107549.

[34] Zhao YX, Li SH, Feng MG, et al. Effects of puerarin-loaded tetrahedral framework nucleic acids on osteonecrosis of the femoral head. Small. 2023;e2302326.

[35] Mauthe M, Orhon I, Rocchi C, et al. Chloroquine inhibits autophagic flux by decreasing autophagosome-lysosome fusion. Autophagy. 2018;14:1435-1455.

[36] Gurney MA, Huang C, Ramil JM, et al. Measuring cardiac autophagic flux in vitro and in vivo. Methods Mol Biol. 2015;1219:187-197.

[37] Xu LX, Tang XJ, Yang YY, et al. Neuroprotective effects of autophagy inhibition on hippocampal glutamate receptor subunits after hypoxia-ischemia-induced brain damage in newborn rats. Neural Regen Res. 2017;12:417-424.

[38] Rabinovich-Nikitin I, Rasouli M, Reitz CJ, et al. Mitochondrial autophagy and cell survival is regulated by the circadian clock gene in cardiac myocytes during ischemic stress. Autophagy. 2021;17:3794-3812.

[39] Singh AK, Singh S, Tripathi VK, Bissoyi A, Garg G, Rizvi SI. Rapamycin confers neuroprotection against aging-induced oxidative stress, mitochondrial dysfunction, and neurodegeneration in old rats through activation of autophagy. Rejuvenation Res. 2019;22:60-70.

[40] Jeske R, Liu C, Duke L, et al. Upscaling human mesenchymal stromal cell production in a novel vertical-wheel bioreactor enhances extracellular vesicle secretion and cargo profile. Bioact Mater. 2023;25:732-747.

[41] Mizushima N, Yoshimori T. How to interpret LC3 immunoblotting. Autophagy. 2007;3:542-545.

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2023 2023 The Authors. Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.