Osteogenic ability and autophagy level between normal and inflammatory periodontal ligament stem cells

doi:10.12307/2025.037

Chinese Journal of Tissue Engineering Research ›› 2025, Vol. 29 ›› Issue (1): 74-79.doi: 10.12307/2025.037

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Osteogenic ability and autophagy level between normal and inflammatory periodontal ligament stem cells

Mao Jiaqi1, Zhao Liru2, Yang Dongru1, Hu Yongqing1, Dai Bowen3, Li Shujuan1

1First Department of Periodontology, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China; 2Department of Orthodontics, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China; 3Hebei Medical University, Shijiazhuang 050017, Hebei Province, China

Received:2023-12-21 Accepted:2024-02-08 Online:2025-01-08 Published:2024-05-18
Contact: Li Shujuan, Master, Chief physician, Professor, First Department of Periodontology, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China
About author:Mao Jiaqi, Master, Attending physician, First Department of Periodontology, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China Zhao Liru, Master, Attending physician, Lecturer, Department of Orthodontics, Hebei Key Laboratory of Stomatology, Hebei Clinical Research Center for Oral Diseases, School and Hospital of Stomatology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China Mao Jiaqi and Zhao Liru contributed equally to this article.
Supported by:
Medical Science Research Project in Hebei Province, No. 20211078 (to ZLR); Medical Science Research Project in Hebei Province, No. 20230185 (to MJQ); 2019 Hebei Provincial Health Commission Government Funded Provincial Clinical Medicine Excellent Talent Training and Basic Research Project, No. 2019061441-2 (to LSJ); Elderly Disease Prevention and Control Project of Hebei Provincial Department of Finance in 2017, No. 361029 (to LSJ)

Abstract

Abstract: BACKGROUND: Inflammation affects the osteogenic differentiation of periodontal ligament stem cells, and the osteogenic ability and autophagy level of periodontal ligament stem cells are closely related. However, there are no relevant reports on whether inflammation affects the osteogenic ability and autophagy level of periodontal ligament stem cells at different stages of osteogenic differentiation.
OBJECTIVE: To explore alkaline phosphatase expression and autophagy periodontal ligament stem cells levels in periodontitis and normal conditions.
METHODS: Periodontal ligament stem cells from normal and periodontitis patients were isolated and cultured, and underwent Vimentin, pan-CK, and Stro-1 fluorescence staining. At 3, 7, and 14 days of osteogenic differentiation, western blot assay was used to detect the protein expression levels of alkaline phosphatase, LC3B, Beclin1, and ATG5 in normal and inflammatory periodontal ligament stem cells. The mRNA expression levels of alkaline phosphatase, bone sialoprotein, osteocalcin, Runx2, LC3B, Beclin1, and ATG5 were detected by real-time PCR.
RESULTS AND CONCLUSION: (1) Stro-1 was positive, Vimentin was positive, and pan CK was negative in periodontal ligament stem cells. (2) At 3, 7, and 14 days after osteogenic differentiation, compared with normal periodontal ligament stem cells, the mineralization nodules formed by periodontal ligament stem cells from inflammatory sources were significantly reduced (P < 0.01); the expression of alkaline phosphatase protein and mRNA was significantly lower (P < 0.05); the mRNA expression levels of bone sialoprotein, osteocalcin, and Runx2 were significantly decreased (P < 0.05). (3) At 7 and 14 days after osteogenic differentiation, compared with normal periodontal ligament stem cells, the expression levels of ATG5, LC3B, and Beclin1 proteins and mRNA of periodontal ligament stem cells were downregulated (P < 0.05). These findings suggest that inflammation reduces the activity of periodontal ligament stem cells in mineralizing nodule formation and the expression of alkaline phosphatase and weakens the autophagy potential of periodontal ligament stem cells at 7 and 14 days after osteogenic differentiation.

Key words: periodontal ligament stem cell, periodontitis, alkaline phosphatase, autophagy, inflammation, osteogenic differentiation

CLC Number:

Mao Jiaqi, Zhao Liru, Yang Dongru, Hu Yongqing, Dai Bowen, Li Shujuan. Osteogenic ability and autophagy level between normal and inflammatory periodontal ligament stem cells[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(1): 74-79.

Figures/Tables 2

References

[1] CHEN FM, SUN HH, LU H, et al. Stem cell-delivery therapeutics for periodontal tissue regeneration. Biomaterials. 2012;33(27):6320-6344.
[2] QIU W, WU BL, FANG FC. Overview of noncoding RNAs involved in the osteogenic differentiation of periodontal ligament stem cells. World J Stem Cells. 2020;12(4):251-265.
[3] YU M, SUN L, BA P, et al. Progranulin promotes osteogenic differentiation of periodontal membrane stem cells in both inflammatory and non-inflammatory conditions. J Int Med Res. 2021; 49(8):3000605211032508.
[4] CHEN Q, LIU X, WANG D, et al. Periodontal Inflammation-Triggered by Periodontal Ligament Stem Cell Pyroptosis Exacerbates Periodontitis. Front Cell Dev Biol. 2021;9:663037.
[5] LU Y, ZHAO L, MAO J, et al. Rab27a-mediated extracellular vesicle secretion contributes to osteogenesis in periodontal ligament-bone niche communication. Sci Rep. 2023;13(1):8479.
[6] SHANG L, LIU Z, MA B, et al. Dimethyloxallyl glycine/nanosilicates-loaded osteogenic/angiogenic difunctional fibrous structure for functional periodontal tissue regeneration. Bioact Mater. 2020;6(4): 1175-1188.
[7] ZHANG X, YAN Q, LIU X, et al. LncRNA00638 promotes the osteogenic differentiation of periodontal mesenchymal stem cells from periodontitis patients under static mechanical strain. Stem Cell Res Ther. 2023;14(1):177.
[8] JIN S, JIANG H, SUN Y, et al. Osteogenic differentiation of periodontal membrane stem cells in inflammatory environments. Open Life Sci. 2022;17(1):1240-1248.
[9] ZHENG W, WANG S, WANG J, et al. Periodontitis promotes the proliferation and suppresses the differentiation potential of human periodontal ligament stem cells. Int J Mol Med. 2015;36(4):915-922.
[10] DEBNATH J, GAMMOH N, RYAN KM. Autophagy and autophagy-related pathways in cancer. Nat Rev Mol Cell Biol. 2023;24(8):560-575.
[11] ZACHARI M, LONGO M, GANLEY IG. Aberrant autophagosome formation occurs upon small molecule inhibition of ULK1 kinase activity. Life Sci Alliance. 2020;3(12):e202000815.
[12] GANLEY I. The Importance of Being Autophagic. N Engl J Med. 2021; 384(25):2449-2450.
[13] ZHANG Y, WANG P, WANG Y, et al. Gold Nanoparticles Promote the Bone Regeneration of Periodontal Ligament Stem Cell Sheets Through Activation of Autophagy. Int J Nanomedicine. 2021;16:61-73.
[14] MAO CY, WANG YG, ZHANG X, et al. Double-edged-sword effect of IL-1β on the osteogenesis of periodontal ligament stem cells via crosstalk between the NF-κB, MAPK and BMP/Smad signaling pathways. Cell Death Dis. 2016;7(7):e2296.
[15] SHAO X, HU Z, SU H, et al. Effects of tension on mitochondrial autophagy and osteogenic differentiation of periodontal ligament stem cells. Cell Prolif. 2023:e13561. doi: 10.1111/cpr.13561.
[16] LIU H, ZHENG J, ZHENG T, et al. Exendin-4 regulates Wnt and NF-κB signaling in lipopolysaccharide-induced human periodontal ligament stem cells to promote osteogenic differentiation. Int Immunopharmacol. 2019;75:105801.
[17] JIANG N, HE D, MA Y, et al. Force-Induced Autophagy in Periodontal Ligament Stem Cells Modulates M1 Macrophage Polarization via AKT Signaling. Front Cell Dev Biol. 2021;9:666631.
[18] CHENG M, ZHOU Q. Targeting EZH2 Ameliorates the LPS-Inhibited PDLSC Osteogenesis via Wnt/β-Catenin Pathway. Cells Tissues Organs. 2020;209(4-6):227-235.
[19] WANG P, TIAN H, ZHANG Z, et al. EZH2 Regulates Lipopolysaccharide-Induced Periodontal Ligament Stem Cell Proliferation and Osteogenesis through TLR4/MyD88/NF-κB Pathway. Stem Cells Int. 2021;2021:7625134.
[20] SU Y, AI S, SHEN Y, et al. Regulatory Effects of Three-Dimensional Cultured Lipopolysaccharide-Pretreated Periodontal Ligament Stem Cell-Derived Secretome on Macrophages. Int J Mol Sci. 2023; 24(8):6981.
[21] RAMENZONI LL, RUSSO G, MOCCIA MD, et al. Periodontal bacterial supernatants modify differentiation, migration and inflammatory cytokine expression in human periodontal ligament stem cells. PLoS One. 2019;14(7):e0219181.
[22] ZHAO X, SUN W, GUO B, et al. Circular RNA BIRC6 depletion promotes osteogenic differentiation of periodontal ligament stem cells via the miR-543/PTEN/PI3K/AKT/mTOR signaling pathway in the inflammatory microenvironment. Stem Cell Res Ther. 2022;13(1):417.
[23] LI L, LIU W, WANG H, et al. Mutual inhibition between HDAC9 and miR-17 regulates osteogenesis of human periodontal ligament stem cells in inflammatory conditions. Cell Death Dis. 2018;9(5):480.
[24] SUN J, DONG Z, ZHANG Y, et al. Osthole improves function of periodontitis periodontal ligament stem cells via epigenetic modification in cell sheets engineering. Sci Rep. 2017;7(1):5254.
[25] ZHANG R, LIANG Q, KANG W, et al. Metformin facilitates the proliferation, migration, and osteogenic differentiation of periodontal ligament stem cells in vitro. Cell Biol Int. 2020;44(1):70-79.
[26] MENG T, ZHOU Y, LI J, et al. Azithromycin Promotes the Osteogenic Differentiation of Human Periodontal Ligament Stem Cells after Stimulation with TNF-α. Stem Cells Int. 2018;2018:7961962.
[27] QIU X, YU Y, LIU H, et al. Remodeling the periodontitis microenvironment for osteogenesis by using a reactive oxygen species-cleavable nanoplatform. Acta Biomater. 2021;135:593-605.
[28] YANG XQ, YU SY, YU L, et al. Effects of tacrolimus on autophagy protein LC3 in puromycin-damaged mouse podocytes. J Int Med Res. 2020;48(12):300060520971422.
[29] KONG Z, YAO T. Role for autophagy-related markers Beclin-1 and LC3 in endometriosis. BMC Womens Health. 2022;22(1):264.
[30] ELBIALY A. In vivo autophagy quantification: Measuring LC3 and P62 puncta in 3D image system from zebrafish larvae. J Cell Biochem. 2021;122(10):1435-1444.
[31] SARKAR C, LIPINSKI MM. Autophagy in neuroinflammation after traumatic brain injury. Neural Regen Res. 2024;19(5):951-952.
[32] LI X, GAO L, ZHENG L, et al. BMP4-mediated autophagy is involved in the metastasis of hepatocellular carcinoma via JNK/Beclin1 signaling. Am J Transl Res. 2020;12(6):3068-3077.
[33] GUI X, YANG H, LI T, et al. Autophagy induction via STING trafficking is a primordial function of the cGAS pathway. Nature. 2019;567(7747): 262-266.
[34] LUO Y, GOU H, CHEN X, et al. Lactate inhibits osteogenic differentiation of human periodontal ligament stem cells via autophagy through the MCT1-mTOR signaling pathway. Bone. 2022;162:116444.

Osteogenic ability and autophagy level between normal and inflammatory periodontal ligament stem cells

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