The regulation of PERK on osteogenic differentiation of human periodontal ligament cells

doi:10.19439/j.sjos.2025.06.002

Abstract

Abstract: PURPOSE: To explore whether RNA-activated protein kinase-like ER resident kinase(PERK) has a regulatory effect on the osteogenic differentiation of human periodontal ligament cells (PDLCs). METHODS: PDLCs were primarily cultured, and siRNA was used to silence the expression of PERK in PDLCs. PDLCs were cultured in osteoblast differentiation-induction medium. Subsequently, the osteogenic differentiation capacity was evaluated using alkaline phosphatase(ALP) staining and alizarin red staining. RESULTS: After osteogenic induction culture of PDLCs in vitro, ALP and alizarin red staining were enhanced, and the deposition of mineral salts increased. Meanwhile, the expression of phosphorylated PERK(p-PERK) in osteogenic PDLCs was higher than cells in the control group. Transfection of si-PERK significantly downregulated PERK protein expression in PDLCs (P<0.05). ALP and alizarin red staining showed that the PERK^-/- group had markedly lower ALP staining intensity and calcium salt deposition than the osteogenic induction group, along with decreased activating transcription factor 4(ATF4) protein expression. CONCLUSIONS: PERK regulates the osteogenic differentiation of PDLCs.

Key words: Human periodontal ligament cells, RNA-activated protein kinase-like ER-resident kinase, Osteogenic differentiation, ATF4

CLC Number:

R781.4

Li Lifen, Du Rong, Jiang Long. The regulation of PERK on osteogenic differentiation of human periodontal ligament cells[J]. Shanghai Journal of Stomatology, 2025, 34(6): 571-576.

References

[1] Abdulkareem AA, AI-Taweel FB, AI-Sharqi AJB, et al.Current concepts in the pathogenesis of periodontitis: from symbiosis to dysbiosis[J]. J Oral Microbiol, 2023, 15(1): 2197779.
[2] Kwon T, Lamster IB, Levin L.Current concepts in the management of periodontitis[J]. Int Dent J, 2021, 71(6): 462-476.
[3] Cheng L, Fan Y, Cheng J, et al.Long non-coding RNA ZYF-AS1 represses periodontitis tissue inflammation and oxidative damage via modulating microRNA-129-5p/DEAD-Box helicase 3 X-linked axis[J]. Bioengineered, 2022, 13(5): 12691-12705.
[4] Li C, Qi F, Zhou Q, et al.Betulinic acid promotes the osteogenic differentiation of human periodontal ligament cells by upregulating EGR1[J]. Acta Biochim Biophys Sin (Shanghai), 2021, 53(10): 1266-1276.
[5] Queiroz A, Albuquerque-Souza E, Gasparoni LM, et al.Therapeutic potential of periodontal ligament stem cells[J]. World J Stem Cells, 2021, 13(6): 605-618.
[6] Ranines LN, Zhao H, Wang Y, et al.PERK is a critical metabolic hub for immunosuppressive function in macrophages[J]. Nat Immunol, 2022, 23(3): 431-445.
[7] Guo J, Ren R, Sun K, et al.PERK controls bone homeostasis through the regulation of osteoclast differentiation and function[J]. Cell Death Dis, 2020, 11(10): 847-863.
[8] Vanhoutte D, Schips TG, Vo A, et al.Thbs induces lethal cardiac atrophy through PERK-ATF4 regulated autophagy[J]. Nat Commun, 2021, 12(1): 3928-3944.
[9] Wei J, Sheng X, Feng D, et al.PERK is essential for neonatal skeletal development to regulate osteoblast proliferation and differentiation[J]. J Cell physiol, 2008, 217(3): 693-707.
[10] Saito A, Ochiai K, Kondo S, et al.Endoplasmic reticulum stress response mediated by the PEK-eIF2(alpha)-ATF4 pathway is involved in osteoblast differentiation induced by BMP2[J]. J Biol Chem, 2011, 286(6): 4809-4818.
[11] Tanaka K, Kaji H, Yamaguchi T, et al.Involvement of the osteoinductive factors, Tmem119 and BMP-2, and the ER stress response PERK-eIF2α-ATF4 pathway in the commitment of myoblastic into osteoblastic cells[J]. Calcif Tissue Int, 2014, 94(4): 454-464.
[12] Wu T, Jiang Y, Shi W, et al.Endoplasmic reticulum stress: a novel targeted approach to repair bone defects by regulating osteogenesis and angiogenesis[J]. J Transl Med, 2023, 21(1): 480-496.
[13] Fraser D, Nguyen T, Benoit DSW.Matrix control of periodontal ligament cell activity via synthetic hydrogel scaffolds[J]. Tissue Eng Part A, 2021, 27(11-12): 733-747.
[14] Ying M, Zhang B.Daidzein promotes the proliferation and osteogenic differentiation of periodontal ligament stem cell[J]. Oral Dis, 2023, 29(3): 1226-1233.
[15] 文扬, 朱亚琴. PERK干扰慢病毒载体的构建及其在人牙髓细胞中的表达[J].上海口腔医学, 2017, 26(4): 363-367.
[16] Zhong M, Huang J, Wu Z, et al.Potential roles of selectins in periodontal diseases and associated systemic diseases: could they be targets for immunotherapy[J]. Int J Mol Sci, 2022, 23(22): 14280.
[17] Bian M, Wang W, Song C, et al.Autophagy-related genes predict the progression of periodontitis through the ceRNA network[J]. J Inflamm Res, 2022, 15: 1811-1824.
[18] Liu G, Zhang L, Zhou X, et al.Inducing the "re-development state" of periodontal ligament cells via tuning macrophage mediated immune microenvironment[J]. J Adv Res, 2024, 60: 233-248.
[19] Cao J, Zhang Q, Yang Q, et al.Epigenetic regulation of osteogenic differentiation of periodontal ligament stem cells in periodontitis[J]. Oral Dis, 2023, 29(7): 2529-2537.
[20] Costa CR, Amorim BR, de Magalhães P, et al. Effects of plants on osteogenic differentiation and mineralization of periodontal ligament cells: a systematic review[J]. Phytother Res, 2016, 30(4): 519-531.
[21] Wang YC, Li X, Shen Y, et al.PERK(protein kinase RNA-like ER kinase) branch of the unfolded protein response confers neuroprotection in ischemic stroke by supressing protein synthesis[J]. Stroke, 2020, 51(5): 1570-1577.
[22] Guo J, Ren R, Sun K, et al.PERK signaling pathway in bone metabolism: friend or foe?[J]. Cell Prolif, 2021, 54(4): e13011.
[23] Jin N, Jin N, Wang Z, et al.Osteopromotive carbon dots promote bone regeneration through the PERK-elF2α-ATF4 pathway[J]. Biomater Sci, 2020, 8(10): 2840-2852.
[24] Yang Y, Feng N, Liang R, et al.Porgranulin, a moderator of estrogen/estrogen receptor alpha binding, regulates bone homeostasis through PERK/p-eIF2 signaling pathway[J]. J Mol Med (Berl), 2022,100(8): 1191-1207.
[25] Qiu W, Sun Q, Li N, et al.Superoxide dismutase 2 scavenges ROS to promote osteogenic differentiation of human periodontal ligament stem cells by regulating Smad3 in alveolar bone-defective rats[J]. J Periodontal, 2024, 95(5): 469-482.
[26] Li J, Li C, Huang Z, et al.Empagliflozin alleviates atherosclerotic calcification by inhibiting osteogenic differentiation of vascular smooth muscle cells[J]. Front Pharmacol, 2023, 14: 1295463.
[27] Zhou X, Xiong H, Lu Y, et al.PKD2 deficiency suppresses amino acid biosynthesis in ADPKD by impairing the PERK-TBL2-eTF2-ATF4 pathway[J]. Biochem Biophys Res Commun, 2021, 561: 73-79.
[28] Feng Y, Han Z, Jiang W, et al.Promotion of osteogenesis in BMSC under hypoxia by ATF4 via the PERK-elF2alpha signaling pathway[J]. In Vitro Cell Dev Biol Anim, 2022, 58(10): 886-897.
[29] Zhu X, Yang L, Han X, et al.Oxidized phospholipids facilitate calcific aortic valve disease by elevating ATF4 through the PERK-eIF2alpha axis[J]. Aging (Albany NY), 2023, 15(14): 6834-6847.
[30] Vanhotte D, Schips TG, Vo A, et al.Thbs1 induces lethal cardiac atrophy through PERK-ATF4 regulated autophagy[J]. Nat Commun,2021,12(1): 3928-3944.
[31] Gu L, Wang Z, Gu H, et al.Atf4 regulates angiogenic differences between alveolar bone and long bone macrophages by regulating M1 polarization, based on single-cell RNA sequencing, RNA-seq and ATAC-seq analysis[J]. J Transl Med, 2023, 21(1): 193-220.
[32] Hao L, Zhong W, Dong H, et al.ATF4 activation promotes hepatic mitochondrial dysfuction by repressing NRF1-TFAM signaling in alcoholic steatohepatitis[J]. Gut, 2021, 70(10): 1933-1945.
[33] Li X, Zheng J, Chen S, et al.Oleandrin, a cardiac glycoside, induces immunogenic cell death via the PERK/elF2α/ATF4/CHOP pathway in breast cancer[J]. Cell Death Dis, 2021, 12(4): 314-329.
[34] Liu F, Wang X, Zheng B, et al.USF2 enhances the osteogenic differentiation of PDLCs by promoting ATF4 transcriptional activities[J]. J Periodontal Res, 2020, 55(1): 68-76.
[35] Verginadis II, Avgousti H, Monslow J, et al.A stromal integrated stress response activates perivascular cancer-associated fibroblasts to drive angiogenesis and tumor progresstion[J]. Nat Cell Biol, 2022, 24(6): 940-953.