The effect of Toll-like receptor 4 in nicotine suppressing the osteogenic potential of periodontal ligament stem cells

doi:10.7518/hxkq.2017.04.005

Abstract

Abstract:

Objective To explore the impact of nicotine on proliferation and osteogenic capability of periodontal ligament stem cells (PDLSCs), and the role of Toll-like receptor 4 (TLR4) in nicotine, suppressing the osteogenic capability of PDLSCs. Methods PDLSCs were cultured in vitro, and the flow cytometer was used to identify the surface antigen markers of PDLSCs. WST-1 was used to detect the proliferation ability of PDLSCs, which were stimulated by different concentrations of nicotine. Alizarin red staining was used to observe the formation of mineralized nodules after PDLSCs stimulation with different con-centrations of nicotine. Real-time polymerase chain reaction (RT-PCR) and Western blot were used to detect the change in osteogenic potential of PDLSCs stimulated by nicotine, after TAK-242, and with the inhibitor of TLR4. Results PDLSCs expressed mesenchymal stem cell-associated mar-kers CD90 and CD105. When the concentration of nicotine was 10^-4mol·L^-1, the PDLSC proliferation could be suppressed after 3 d compared with the control group (P<0.05). The amount of mineralized nodules reduced after osteogenic differentiation at 21 d by alizarin red staining. RT-PCR and Western blot showed the expression levels of alkaline phosphatase (ALP), and osteocalcin (OCN), and the Runt-related transcription factor-2 (Runx-2) were lower than in the control group when nicotine suppressed the PDLSCs (P<0.05). This effect was atte-nuated after TAK-242 was added. Conclusion Nicotine suppresses the proliferation and osteogenic capability of PDLSCs, which may be regulated by TLR4.

Key words: nicotine, periodontal ligament stem cells, Toll-like receptors 4, osteogenic differentiation

CLC Number:

R781.4

Luan Yan, Deqin Yang. The effect of Toll-like receptor 4 in nicotine suppressing the osteogenic potential of periodontal ligament stem cells[J]. West China Journal of Stomatology, 2017, 35(4): 368-372.

Figures/Tables 7

Fig 1

Tab 1

Fig 2

Fig 3

Fig 4

Fig 5

Fig 6

References 13

[1]	Bergström J.Tobacco smoking and chronic destructive perio-dontal disease[J]. Odontology, 2004, 92(1):1-8.
[2]	Ferraiolo DM.Predicting periodontitis progression[J]. Evid Based Dent, 2016, 17(1):19-20.
[3]	Chambrone L, Chambrone D, Lima LA, et al.Predictors of tooth loss during long-term periodontal maintenance: a systematic review of observational studies[J]. J Clin Perio-dontol, 2010, 37(7):675-684.
[4]	Lei M, Li K, Li B, et al.Mesenchymal stem cell characteris-tics of dental pulp and periodontal ligament stem cells after in vivo transplantation[J]. Biomaterials, 2014, 35(24):6332-6343.
[5]	Barua RS, Sharma M, Dileepan KN. Cigarette smoke amp-lifies inflammatory response and atherosclerosis progression through activation of the H1R-TLR2/4-COX2 Axis[J]. Front Immunol, 2015(6):572.
[6]	Lee CC, Avalos AM, Ploegh HL.Accessory molecules for Toll-like receptors and their function[J]. Nat Rev Immunol, 2012, 12(3):168-179.
[7]	Zhou Z, Li B, Dong Z, et al.Nicotine deteriorates the osteoge-nic differentiation of periodontal ligament stem cells through α7 nicotinic acetylcholine receptor regulating Wnt pathway[J]. PLoS ONE, 2013, 8(12):e83102.
[8]	Duane B.Further evidence that periodontal bone loss increases with smoking and age[J]. Evid Based Dent, 2014, 15(3):72-73.
[9]	Matta SG, Balfour DJ, Benowitz NL, et al.Guidelines on nicotine dose selection for in vivo research[J]. Psychophar-macology, 2007, 190(3):269-319.
[10]	Jia Q, Jiang W, Ni L.Down-regulated non-coding RNA (lncRNA-ANCR) promotes osteogenic differentiation of periodontal ligament stem cells[J]. Arch Oral Biol, 2015, 60(2):234-241.
[11]	Tilp C, Bucher H, Haas H, et al.Effects of conventional tobacco smoke and nicotine-free cigarette smoke on airway inflammation, airway remodelling and lung function in a triple allergen model of severe asthma[J]. Clin Exp Allergy, 2016, 46(7):957-972.
[12]	Mahanonda R, Sa-Ard-Iam N, Eksomtramate M, et al. Ciga-rette smoke extract modulates human beta-defensin-2 and interleukin-8 expression in human gingival epithelial cells[J]. J Periodont Res, 2009, 44(4):557-564.
[13]	Kim TH, Kim SJ, Lee SM.Stimulation of the α7 nicotinic acetylcholine receptor protects against sepsis by inhibiting Toll-like receptor via phosphoinositide 3-kinase activation[J]. J Infect Dis, 2014, 209(10):1668-1677.

组别	1 d	2 d	3 d	4 d	5 d
10^-4 mol·L^-1	0.37±0.27	0.37±0.71	0.37±0.02*	0.20±0.02*	0.17±0.01*
10^-5 mol·L^-1	0.35±0.01	0.43±0.03	1.01±0.07	1.25±0.06	1.50±0.05*
10^-6 mol·L^-1	0.34±0.01	0.57±0.02	1.38±0.03	1.78±0.03	2.13±0.12
对照组	0.34±0.01	0.48±0.01	1.29±0.02	1.97±0.03	2.25±0.06

[1]	Tang Xiaoxue, Zhou Zheng, Li Qiqi, Jiang Dandan. Effects of sitagliptin activation of the stromal cell-derived factor-1/CXC chemokine receptor 4 signaling pathway on the proliferation, apoptosis, inflammation, and osteogenic differentiation of human periodontal ligament stem cells induced by lipopolysaccharide [J]. West China Journal of Stomatology, 2024, 42(1): 37-45.
[2]	Li Shenghong, Peng Shiyuan, Luo Xiaoling, Wang Yipei, Xu Xiaomei. Effect of naringenin on the anti-inflammatory, vascularization, and osteogenesis differentiation of human periodontal ligament stem cells via the stromal cell-derived factor 1/C-X-C motif chemokine receptor 4 signaling axis stimulated by lipopolysaccharide [J]. West China Journal of Stomatology, 2023, 41(2): 175-184.
[3]	Dai Zhenning, Zheng Weihan, Li Shiyu. Receptor activator of nuclear factor-κB ligand and tumor necrosis factor-α promotes osteoclast differentiation through the exosomes of inflammatory periodontal ligament stem cells [J]. West China Journal of Stomatology, 2022, 40(4): 377-385.
[4]	Zhang Peng, Zuo Dongchuan, Mou Siyu, Zhong Yutong, Yuan Xiaoping, Zeng Jin. Effect of inward rectifier potassium 2.1 channel on the osteogenic differentiation of human dental follicle cells and its mechanism [J]. West China Journal of Stomatology, 2022, 40(2): 139-147.
[5]	Pang Ming, Wei Hongxia, Chen Xi. Long non-coding RNA potassium voltage-gated channel subfamily Q member 1 overlapping transcript 1 regulates the proliferation and osteogenic differentiation of human periodontal ligament stem cells by targeting miR-24-3p [J]. West China Journal of Stomatology, 2021, 39(5): 547-554.
[6]	Wang Lin, Wang Xi, Ji Nan, Li Haimei, Cai Shixin. Mechanisms of the mechanically activated ion channel Piezo1 protein in mediating osteogenic differentiation of periodontal ligament stem cells via the Notch signaling pathway [J]. West China Journal of Stomatology, 2020, 38(6): 628-636.
[7]	Wen Jiahui, Wu Yanmin, Chen Lili. Functions of non-coding RNAs in the osteogenic differentiation of human periodontal ligament-derived cells [J]. West China Journal of Stomatology, 2020, 38(3): 330-337.
[8]	Lin Yongsheng,Wang Fengzhi,Lei Xiaojing,He Jianmin. Comparative study with the effect of stromal cell derived factor-1 on osteogenic differentiation of human healthy and inflammatory periodontal ligament stem cells [J]. West China Journal of Stomatology, 2019, 37(5): 469-475.
[9]	Jiehang Li,Zhifei Su,Xuan Bai,He Yuan,Jiyao Li. Effect of zoladronate on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells [J]. West China Journal of Stomatology, 2019, 37(3): 242-247.
[10]	Feng Liao,Yao Liu,Hanghang Liu,Jian Hu,Shuang Zhao,Shimao Yang. Effect of Angelica sinensis polysaccharide on the osteogenic differentiation of bone marrow mesenchymal stem cells of rats with high glucose levels [J]. West China Journal of Stomatology, 2019, 37(2): 193-199.
[11]	Jin Sun,Yun Liu,Qian Qu,Juan Qu,Wei Luo,Feng Zhang,Min. Wu. Effect of histone acetylation on osteogenic differentiation of periodontal ligament stem cells derived from periodontitis tissue [J]. West China Journal of Stomatology, 2019, 37(1): 102-105.
[12]	Ting Hui, Guangcan Zhang, Dandan Feng, Ping Ji. Role of neuropeptide substance P and the bone morphogenetic protein signaling pathway in osteogenic differentiation of ST2 cells [J]. West China Journal of Stomatology, 2018, 36(4): 378-383.
[13]	Yu Ma, Shuhui Li, Xinxin Ding, Peiling. Wu. Effects of tumor necrosis factor-α on osteogenic differentiation and Notch signaling pathway in human periodontal ligament stem cells [J]. West China Journal of Stomatology, 2018, 36(2): 184-189.
[14]	Wei Lei, Yue Ma, Aishu Ren. Neurotrophin 3 promotes osteogenic differentiation of human dental follicle cells [J]. West China Journal of Stomatology, 2018, 36(1): 23-28.
[15]	Wucheng Guo, Jieli Cheng, Zhengyi Yang, Yi Zhang, Enliang He, Jun Qian, Jingjing Song, Jin Sun, Lin Yuan. K (lysine) acetyltransferase 2A affects the osteogenic differentiation of periodontal ligament stem cells through the canonical Wnt pathway [J]. West China Journal of Stomatology, 2018, 36(1): 39-45.