Expression of Toll-like receptor 4 in dental pulp stem cells under the stimulation of lipopolysaccharide and Streptococcus mutans

doi:10.3969/j.issn.2095-4344.0668

Abstract

Abstract:

BACKGROUND: During the process of deep caries, dental pulp stem cells (DPSCs) are induced to differentiate into odontoblast-like cells after bacterial stimulation, forming reparative dentin to protect pulp tissues. It is considered to be an important part of innate immune response of dental pulp. However, the role of Toll-like receptor 4 (TLR4) as the most important receptor of innate immune response is not clear.
OBJECTIVE: To study the expression of TLR4 with the stimulation of lipopolysaccharide and Streptococcus mutans during deep caries.
METHODS: Primary cultured DPSCs were stimulated by lipopolysaccharide and Streptococcus mutans extract for 24 hours. RT-PCR was then performed to test the expression of TLR4 mRNA. Western blot and immunocytochemical staining were performed to test the expression of TLR4 protein in DPSCs.
RESULTS AND CONCLUSION: Stimulations with lipopolysaccharide and Streptococcus mutans extract significantly up-regulated the mRNA and protein expression of TLR4, indicating TLR4 is involved in the innate immune response of DPSCs in the case of deep caries.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Dental Pulp, Stem Cells, Lipopolysaccharides, Streptococcus mutans, Toll-Like Receptor 4, Tissue Engineering

CLC Number:

R394.2

Liu Ying, Gao Jie, Wu Bu-ling. Expression of Toll-like receptor 4 in dental pulp stem cells under the stimulation of lipopolysaccharide and Streptococcus mutans[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(33): 5333-5337.

Figures/Tables 1

References

[1] Liu Y, Gao Y, Zhan X, et al. TLR4 activation by lipopolysaccharide and Streptococcus mutans induces differential regulation of proliferation and migration in human dental pulp stem cells. J Endod. 2014;40(9):1375-1381.

[2] 麻丹丹,高杰,吴补领.改良组织块酶消化法培养人龋损牙髓干细胞的实验研究[J].牙体牙髓牙周病学杂志, 2011,21(7):371-374.

[3] Gronthos S, Mankani M, Brahim J, et al. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl AcadSci U S A. 2000;97(25):13625-13630.

[4] Arthur A, Rychkov G, Shi S, et al. Adult human dental pulp stem cells differentiate toward functionally active neurons under appropriate environmental cues. Stem Cells. 2008;26(7):1787-1795.

[5] Kim JK, Baker J, Nor JE, et al. mTor plays an important role in odontoblast differentiation. J Endod. 2011;37(8):1081-1085.

[6] 刘影,高杰,吴补领. 改良组织块酶消化法原代培养人牙髓干细胞的研究[J].口腔疾病防治,2018,26(3):166-170.

[7] Nam S, Won JE, Kim CH, et al. Odontogenic differentiation of human dental pulp stem cells stimulated by the calcium phosphate porous granules. J Tissue Eng. 2011;2011:812547.

[8] Ma D, Gao J, Yue J, et al. Changes in proliferation and osteogenic differentiation of stem cells from deep caries in vitro. J Endod. 2012;38(6):796-802.

[9] Gronthos S, Brahim J, Li W, et al. Stem cell properties of human dental pulp stem cells. J Dent Res. 2002;81(8):531-535.

[10] Wang CY, Tani-Ishii N, Stashenko P. Bone-resorptive cytokine gene expression in periapical lesions in the rat. Oral MicrobiolImmunol. 1997;12(2):65-71.

[11] 刘影,高岩,文军,徐帅妹.TLR4在健康和深龋人牙髓组织中的定位表达[J].口腔疾病防治,2017,25(3):158-163.

[12] Horst OV, Tompkins KA, Coats SR, et al. TGF-beta1 Inhibits TLR-mediated odontoblast responses to oral bacteria. J Dent Res. 2009;88(4):333-338.

[13] Okada M, Soda Y, Hayashi F, et al. PCR detection of Streptococcus mutans and S. sobrinus in dental plaque samples from Japanese pre-school children. J Med Microbiol. 2002;51(5): 443-447.

[14] Jiang HW, Zhang W, Ren BP, et al. Expression of toll like receptor 4 in normal human odontoblasts and dental pulp tissue. J Endod. 2006;32(8):747-751.

[15] Mutoh N, Tani-Ishii N, Tsukinoki K, et al. Expression of toll-like receptor 2 and 4 in dental pulp. J Endod. 2007;33(10):1183-1186.

[16] Wang CY, Tani-Ishii N, Stashenko P. Bone-resorptive cytokine gene expression in periapical lesions in the rat. Oral MicrobiolImmunol. 1997;12(2):65-71.

[17] Rutherford RB, Gu K. Treatment of inflamed ferret dental pulps with recombinant bone morphogenetic protein-7. Eur J Oral Sci. 2000;108(3):202-206.

[18] Hu H, Li Z, Zhu X, et al. Gua Lou GuiZhi decoction suppresses LPS-induced activation of the TLR4/NF-κB pathway in BV-2 murine microglial cells. Int J Mol Med. 2013;31(6):1327-1332.

[19] He W, Qu T, Yu Q, et al. LPS induces IL-8 expression through TLR4, MyD88, NF-kappaB and MAPK pathways in human dental pulp stem cells. IntEndod J. 2013;46(2):128-136.

[20] Zhang J, Zhang Y, Lv H, et al. Human stem cells from the apical papilla response to bacterial lipopolysaccharide exposure and anti-inflammatory effects of nuclear factor I C. J Endod. 2013; 39(11):1416-1422.

[21] Medvedev AE, Kopydlowski KM, Vogel SN. Inhibition of lipopolysaccharide-induced signal transduction in endotoxin- tolerized mouse macrophages: dysregulation of cytokine, chemokine, and toll-like receptor 2 and 4 gene expression. J Immunol. 2000;164(11):5564-5574.

[22] Papp T, Hollo K, Meszar-Katona E, et al. TLR signalling can modify the mineralization of tooth germ. ActaOdontol Scand. 2016; 74(4):307-314.

[23] Li JG, Lin JJ, Wang ZL, et al. Melatonin attenuates inflammation of acute pulpitis subjected to dental pulp injury. Am J Transl Res. 2015;7(1):66-78.

[24] Rachmawati D, Peferoen LA, Vogel DY, et al. Metal ions potentiate microglia responsiveness to endotoxin. J Neuroimmunol. 2016; 291:89-95.

[25] 岳静,高杰,徐树军,等.变形链球菌超声提取物对牙髓细胞增殖和天然免疫受体基因 mRNA表达的影响[J].实用口腔医学杂志, 2013, 29(1):16-19.

[26] Kesavalu L, Lucas AR, Verma RK, et al. Increased atherogenesis during Streptococcus mutans infection in ApoE-null mice. J Dent Res. 2012;91(3):255-260.

[27] Hirao K, Yumoto H, Takahashi K, et al. Roles of TLR2, TLR4, NOD2, and NOD1 in pulp fibroblasts. J Dent Res. 2009;88(8): 762-767.

[28] He W, Wang Z, Luo Z, et al. LPS promote the odontoblastic differentiation of human dental pulp stem cells via MAPK signaling pathway. J Cell Physiol. 2015;230(3):554-561.

[29] Levin LG, Rudd A, Bletsa A, et al. Expression of IL-8 by cells of the odontoblast layer in vitro. Eur J Oral Sci.1999;107(2): 131-137.

[30] Nagaoka S, Tokuda M, Sakuta T, et al. Interleukin-8 gene expression by human dental pulp fibroblast in cultures stimulated with Prevotella intermedia lipopolysaccharide. J Endod. 1996; 22(1):9-12.

[31] Otsuka M, Okinaga T, Ariyoshi W, et al. Ameloblastin Upregulates Inflammatory Response Through Induction of IL-1β in Human Macrophages. J Cell Biochem. 2017;118(10):3308-3317.

[32] Botero TM, Son JS, Vodopyanov D, et al. MAPK signaling is required for LPS-induced VEGF in pulp stem cells. J Dent Res. 2010;89(3):264-269.

[33] Sun DG, Xin BC, Wu D, et al. miR-140-5p-mediated regulation of the proliferation and differentiation of human dental pulp stem cells occurs through the lipopolysaccharide/toll-like receptor 4 signaling pathway. Eur J Oral Sci. 2017;125(6):419-425.

[34] Lawrence H, Mawdesley AE, Holland JP, et al. Targeting Toll-like receptor 4 prevents cobalt-mediated inflammation. Oncotarget. 2016;7(7):7578-7585.

[35] Papp T, Hollo K, Meszar-Katona E, et al. TLR signalling can modify the mineralization of tooth germ. ActaOdontol Scand. 2016;74(4):307-314.