Toll-like receptor and C-type lectin receptor agonists attenuate osteogenic differentiation in human dental pulp stem cells.
Cell viability
Dental pulp cells
Osteogenic differentiation
Toll-like receptor
Journal
BMC oral health
ISSN: 1472-6831
Titre abrégé: BMC Oral Health
Pays: England
ID NLM: 101088684
Informations de publication
Date de publication:
31 Jan 2024
31 Jan 2024
Historique:
received:
12
11
2023
accepted:
14
01
2024
medline:
1
2
2024
pubmed:
1
2
2024
entrez:
31
1
2024
Statut:
epublish
Résumé
This study aimed to investigate the effects of various toll-like receptor (TLR) and C-type lectin receptor (CLR) ligands on osteogenic differentiation in human dental pulp stem cells (hDPSCs). hDPSCs were cultured and treated with various concentrations (0.01, 0.1, 1.0, and 10 µg/mL) of TLR or CLR agonists (PG-LPS, E.coli LPS, poly(I:C), Pam3CSK4, Furfurman, and Zymosan). Cell viability was determined by MTT assay. The effects of TLR and CLR agonists on osteogenic differentiation of hDPSCs were measured by alkaline phosphatase (ALP) activity, Alizarin Red S staining, and Von Kossa staining. In addition, the mRNA expression of osteogenesis-related genes (ALP, COL1A1, RUNX2, OSX, OCN and DMP1) was examined by RT-qPCR. A non-parametric analysis was employed for the statistical analyses. The statistically significant difference was considered when p < 0.05. Treatment with TLR and CLR agonists was associated with an increase in hDPSCs' colony-forming unit ability. Compared with the control group, TLR and CLR agonists significantly inhibited the osteogenic differentiation of hDPSCs by decreasing the ALP activity, mineralised nodule formation, and mRNA expression levels of osteogenesis-related genes (ALP, COL1A1, RUNX2, OSX, OCN and DMP1). The inhibition of TRIF but not Akt signalling rescued the effects of TLR and CLR agonist attenuating hDPSCs' mineralisation. The activation of TLRs or CLRs exhibited an inhibitory effect on osteogenic differentiation of hDPSCs via the TRIF-dependent signalling pathway.
Sections du résumé
BACKGROUND
BACKGROUND
This study aimed to investigate the effects of various toll-like receptor (TLR) and C-type lectin receptor (CLR) ligands on osteogenic differentiation in human dental pulp stem cells (hDPSCs).
METHODS
METHODS
hDPSCs were cultured and treated with various concentrations (0.01, 0.1, 1.0, and 10 µg/mL) of TLR or CLR agonists (PG-LPS, E.coli LPS, poly(I:C), Pam3CSK4, Furfurman, and Zymosan). Cell viability was determined by MTT assay. The effects of TLR and CLR agonists on osteogenic differentiation of hDPSCs were measured by alkaline phosphatase (ALP) activity, Alizarin Red S staining, and Von Kossa staining. In addition, the mRNA expression of osteogenesis-related genes (ALP, COL1A1, RUNX2, OSX, OCN and DMP1) was examined by RT-qPCR. A non-parametric analysis was employed for the statistical analyses. The statistically significant difference was considered when p < 0.05.
RESULTS
RESULTS
Treatment with TLR and CLR agonists was associated with an increase in hDPSCs' colony-forming unit ability. Compared with the control group, TLR and CLR agonists significantly inhibited the osteogenic differentiation of hDPSCs by decreasing the ALP activity, mineralised nodule formation, and mRNA expression levels of osteogenesis-related genes (ALP, COL1A1, RUNX2, OSX, OCN and DMP1). The inhibition of TRIF but not Akt signalling rescued the effects of TLR and CLR agonist attenuating hDPSCs' mineralisation.
CONCLUSIONS
CONCLUSIONS
The activation of TLRs or CLRs exhibited an inhibitory effect on osteogenic differentiation of hDPSCs via the TRIF-dependent signalling pathway.
Identifiants
pubmed: 38297241
doi: 10.1186/s12903-024-03894-7
pii: 10.1186/s12903-024-03894-7
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
148Subventions
Organisme : National Research Council of Thailand
ID : N41A640135
Organisme : Faculty Research Fund, Faculty of Dentistry, Chulalongkorn University
ID : N/A
Informations de copyright
© 2024. The Author(s).
Références
Li D, Wu M. Pattern recognition receptors in health and diseases. Signal Transduct Target Ther. 2021;6(1):291.
doi: 10.1038/s41392-021-00687-0
pubmed: 34344870
pmcid: 8333067
Iwasaki A, Medzhitov R. Toll-like receptor control of the adaptive immune responses. Nat Immunol. 2004;5(10):987–95.
doi: 10.1038/ni1112
pubmed: 15454922
Rodriguez-Lozano FJ, Bueno C, Insausti CL, Meseguer L, Ramirez MC, Blanquer M, et al. Mesenchymal stem cells derived from dental tissues. Int Endod J. 2011;44(9):800–6.
doi: 10.1111/j.1365-2591.2011.01877.x
pubmed: 21477154
Huang GT, Gronthos S, Shi S. Mesenchymal stem cells derived from dental tissues vs. those from other sources: their biology and role in regenerative medicine. J Dent Res. 2009;88(9):792–806.
doi: 10.1177/0022034509340867
pubmed: 19767575
pmcid: 2830488
Fawzy El-Sayed KM, Klingebiel P, Dörfer CE. Toll-like receptor expression Profile of Human Dental Pulp Stem/Progenitor cells. J Endod. 2016;42(3):413–7.
doi: 10.1016/j.joen.2015.11.014
pubmed: 26769027
DelaRosa O, Dalemans W, Lombardo E. Toll-like receptors as modulators of mesenchymal stem cells. Front Immunol. 2012;3.
Lu YC, Yeh WC, Ohashi PS. LPS/TLR4 signal transduction pathway. Cytokine. 2008;42(2):145–51.
doi: 10.1016/j.cyto.2008.01.006
pubmed: 18304834
Kumar A, Zhang J, Yu FS. Toll-like receptor 3 agonist poly(I:C)-induced antiviral response in human corneal epithelial cells. Immunology. 2006;117(1):11–21.
doi: 10.1111/j.1365-2567.2005.02258.x
pubmed: 16423036
pmcid: 1782193
Funderburg NT, Jadlowsky JK, Lederman MM, Feng Z, Weinberg A, Sieg SF. The toll-like receptor 1/2 agonists Pam(3) CSK(4) and human β-defensin-3 differentially induce interleukin-10 and nuclear factor-κB signalling patterns in human monocytes. Immunology. 2011;134(2):151–60.
doi: 10.1111/j.1365-2567.2011.03475.x
pubmed: 21896010
pmcid: 3194223
Dillon S, Agrawal S, Banerjee K, Letterio J, Denning TL, Oswald-Richter K, et al. Yeast zymosan, a stimulus for TLR2 and dectin-1, induces regulatory antigen-presenting cells and immunological tolerance. J Clin Invest. 2006;116(4):916–28.
doi: 10.1172/JCI27203
pubmed: 16543948
pmcid: 1401484
Shayegan A, Zucchi A, De Swert K, Balau B, Truyens C, Nicaise C. Lipoteichoic acid stimulates the proliferation, migration and cytokine production of adult dental pulp stem cells without affecting osteogenic differentiation. Int EndodJ. 2021;54(4):585–600.
doi: 10.1111/iej.13448
Rothermund K, Calabrese TC, Syed-Picard FN. Differential effects of Escherichia coli– Versus Porphyromonas gingivalis–derived lipopolysaccharides on Dental Pulp Stem Cell differentiation in Scaffold-free Engineered tissues. J Endod. 2022;48(11):1378–86e2.
doi: 10.1016/j.joen.2022.08.010
pubmed: 36108879
pmcid: 9764159
He W, Wang Z, Zhou Z, Zhang Y, Zhu Q, Wei K, et al. Lipopolysaccharide enhances Wnt5a expression through toll-like receptor 4, myeloid differentiating factor 88, phosphatidylinositol 3-OH Kinase/AKT and nuclear factor Kappa B pathways in Human Dental Pulp Stem cells. J Endod. 2014;40(1):69–75.
doi: 10.1016/j.joen.2013.09.011
pubmed: 24331994
Ha Y-E, Ju So Y, Im J, Yun C-H, Park J-C, Hyun Han S. TLR3 recognition of viral double-stranded RNA in human dental pulp cells is important for the innate immunity. Int Immunopharmacol. 2023;119:110161.
doi: 10.1016/j.intimp.2023.110161
pubmed: 37060811
Sabatini C, Ayenew L, Khan T, Hall R, Lee T. Dental Pulp Cell Conditioning through Polyinosinic-Polycytidylic Acid activation of toll-like receptor 3 for amplification of trophic factors. J Endod. 2022;48(7):872–9.
doi: 10.1016/j.joen.2022.04.009
pubmed: 35447294
Tomic S, Djokic J, Vasilijic S, Vucevic D, Todorovic V, Supic G, Colic M. Immunomodulatory properties of mesenchymal stem cells derived from Dental Pulp and Dental follicle are susceptible to activation by toll-like receptor agonists. Stem Cells Dev. 2010;20(4):695–708.
doi: 10.1089/scd.2010.0145
Takegawa D, Nakanishi T, Hirao K, Yumoto H, Takahashi K, Matsuo T. Modulatory roles of Interferon-γ through Indoleamine 2, 3-dioxygenase induction in Innate Immune response of Dental Pulp cells. J Endod. 2014;40(9):1382–7.
doi: 10.1016/j.joen.2014.03.018
pubmed: 25146019
Bulanawichit W, Nguyen TNY, Ritprajak P, Nowwarote N, Osathanon T. Cell Wall Mannan of Candida attenuates osteogenic differentiation by Human Dental Pulp cells. J Endod. 2023;49(2):190–7.
doi: 10.1016/j.joen.2022.11.010
pubmed: 36586575
Veerayutthwilai O, Byers MR, Pham TTT, Darveau RP, Dale BA. Differential regulation of immune responses by odontoblasts. Oral Microbiol Immunol. 2007;22(1):5–13.
doi: 10.1111/j.1399-302X.2007.00310.x
pubmed: 17241164
Li D, Fu L, Zhang Y, Yu Q, Ma F, Wang Z, et al. The effects of LPS on adhesion and migration of human dental pulp stem cells in vitro. J Dent. 2014;42(10):1327–34.
doi: 10.1016/j.jdent.2014.07.007
pubmed: 25093548
Kato H, Taguchi Y, Tominaga K, Umeda M, Tanaka A. Porphyromonas gingivalis LPS inhibits osteoblastic differentiation and promotes pro-inflammatory cytokine production in human periodontal ligament stem cells. Arch Oral Biol. 2014;59(2):167–75.
doi: 10.1016/j.archoralbio.2013.11.008
pubmed: 24370188
Yu S, Cho HH, Joo HJ, Bae YC, Jung JS. Role of MyD88 in TLR agonist-induced functional alterations of human adipose tissue-derived mesenchymal stem cells. Mol Cell Biochem. 2008;317(1):143–50.
doi: 10.1007/s11010-008-9842-1
pubmed: 18581201
Liu Y, Gao Y, Zhan X, Cui L, Xu S, Ma D, 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):1375–81.
doi: 10.1016/j.joen.2014.03.015
pubmed: 25146018
He W, Wang Z, Luo Z, Yu Q, Jiang Y, Zhang Y, et al. LPS promote the Odontoblastic differentiation of Human Dental Pulp Stem cells via MAPK signaling pathway. J Cell Physiol. 2015;230(3):554–61.
doi: 10.1002/jcp.24732
pubmed: 25104580
Hwa Cho H, Bae YC, Jung JS. Role of toll-like receptors on human adipose‐derived stromal cells. Stem Cells. 2006;24(12):2744–52.
doi: 10.1634/stemcells.2006-0189
pubmed: 16902195
Kukolj T, Trivanović D, Djordjević IO, Mojsilović S, Krstić J, Obradović H, et al. Lipopolysaccharide can modify differentiation and immunomodulatory potential of periodontal ligament stem cells via ERK1,2 signaling. J Cell Physiol. 2018;233(1):447–62.
doi: 10.1002/jcp.25904
pubmed: 28295277
Wang Z, Pan J, Wright JT, Bencharit S, Zhang S, Everett ET, et al. Putative stem cells in Human Dental Pulp with Irreversible Pulpitis: an exploratory study. J Endod. 2010;36(5):820–5.
doi: 10.1016/j.joen.2010.02.003
pubmed: 20416426
pmcid: 2907071
Arora S, Cooper PR, Friedlander LT, Seo B, Rizwan SB, Rich AM, Hussaini HM. Potentiality and inflammatory marker expression are maintained in Dental Pulp cell cultures from Carious Teeth. Int J Mol Sci [Internet]. 2022; 23(16).
Wang W, Yuan C, Geng T, Liu Y, Zhu S, Zhang C, et al. Lipopolysaccharide inhibits osteogenic differentiation of periodontal ligament stem cells partially through toll-like receptor 4-mediated ephrinB2 downregulation. Clin Oral Investig. 2020;24(10):3407–16.
doi: 10.1007/s00784-020-03211-w
pubmed: 31974644
Nomiyama K, Kitamura C, Tsujisawa T, Nagayoshi M, Morotomi T, Terashita M, Nishihara T. Effects of Lipopolysaccharide on newly established Rat Dental pulp–derived cell line with Odontoblastic Properties. J Endod. 2007;33(10):1187–91.
doi: 10.1016/j.joen.2007.05.015
pubmed: 17889687
Akira S, Takeda K. Toll-like receptor signalling. Nat Rev Immunol. 2004;4(7):499–511.
doi: 10.1038/nri1391
pubmed: 15229469
Liu J, Chen L, Tao X, Tang K. Phosphoinositide 3-kinase/Akt signaling is essential for prostaglandin E2-induced osteogenic differentiation of rat tendon stem cells. BBRC. 2013;435(4):514–9.
pubmed: 23206708