Aim: This study aimed to evaluate the effects of glycyrrhizin, as a potential therapeutic agent in endodontic surgery, on the proliferation and viability of diabetic human bone marrow-derived mesenchymal stem cells (hBM-MSCs).
Materials and methods: Diabetic human bone marrow-derived mesenchymal stem cells (hBM-MSCs) were isolated and characterized by flow cytometry. The cells were treated with different concentrations of Glycyrrhizin (Gly) (12.5, 25, 50, and 100 µg/mL) and 0.1% dimethyl sulfoxide (DMSO) as the control group. MTT assay was performed to evaluate the cell proliferation and viability after 24, 48, and 72 hours of the cell treatment with Gly. The optical density (OD) was measured at 570 nm. Each assay was repeated three times. The corrected OD and cell viability were determined. ANOVA followed by the Bonferroni post hoc test evaluated the statistical significance at p < 0.05.
Results: Flow cytometric analysis of the isolated cells showed positive expression of mesenchymal markers (CD105 and CD90) and negative expression of hematopoietic markers (CD34 and CD14). After 24 and 48 hours of cell treatment, Gly in 100 µg/mL concentration significantly decreased the diabetic hBM-MSC proliferation as compared with the control (p < 0.05). Gly in 12.5–50 µg/mL concentrations significantly increased the cell proliferation after 72 hours of treatment as compared with the control (p < 0.05). The diabetic hBM-MSC proliferation and viability at 12.5–50 µg/mL concentrations were significantly greater than that at 100 µg/mL concentration (p < 0.05).
Conclusion: Under the present study conditions, Gly (in 12.5–50 µg/mL concentrations) did not show cytotoxicity to diabetic hBM-MSCs and enhanced their proliferation. Gly may represent a potential therapeutic agent in endodontic surgery in diabetic patients.
Clinical significance: Preclinical assessment of Gly effects on diabetic hBM-MSCs is important for determining its effective concentration range, anticipating its therapeutic potential, and designing future in vivo studies.
Sreedevi P, Varghese N, Varugheese JM. Prognosis of periapical surgery using bonegrafts: A clinical study. J Conserv Dent 2011;14(1):68–72. DOI: 10.4103/0972-0707.80743.
von Arx T, Alsaeed M. The use of regenerative techniques in apical surgery: A literature review. Saudi Dent J 2011;23(3):113–127. DOI: 10.1016/j.sdentj.2011.02.004.
Setzer FC, Kratchman SI. Present status and future directions: Surgical endodontics. Int Endod J 2022;55(Suppl 4):1020–1058. DOI: 10.1111/iej.
Lin LM, Rosenberg PA. Repair and regeneration in endodontics. Int Endod J 2011;44(10):889–906. DOI: 10.1111/j.1365-2591.2011.01915.x.
Lin L, Chen MY, Ricucci D, et al. Guided tissue regeneration in periapical surgery. J Endod 2010;36(4):618–625. DOI: 10.1016/j.joen.2009.12.012.
Mao X, Liu Y, Chen C, et al. Mesenchymal stem cells and their role in dental medicine. Dent Clin North Am 2017;61(1):161–172. DOI: 10.1016/j.cden.2016.08.006.
Hwang NS, Zhang C, Hwang YS, et al. Mesenchymal stem cell differentiation and roles in regenerative medicine. Wiley Interdiscip Rev Syst Biol Med 2009;1(1):97–106. DOI: 10.1002/wsbm.26.
Andrzejewska A, Lukomska B, Janowski M. Concise review: Mesenchymal stem cells: From roots to boost. Stem Cells 2019;37(7):855–864. DOI: 10.1002/stem.3016.
Fouad AF. Diabetes mellitus as a modulating factor of endodontic infections. J Dent Educ 2003;67(4):459–467. PMID: 12749575.
Holland R, Gomes JE Filho, Cintra LTA, et al. Factors affecting the periapical healing process of endodontically treated teeth. J Appl Oral Sci. 2017;25(5):465–476. DOI: 10.1590/1678-7757-2016-0464.
Xu J, Zuo C. The fate status of stem cells in diabetes and its role in the occurrence of diabetic complications. Front Mol Biosci 2021;8:745035. DOI: 10.3389/fmolb.2021.745035.
Cassidy FC, Shortiss C, Murphy CG, et al. Impact of type 2 diabetes mellitus on human bone marrow stromal cell number and phenotypic characteristics. Int J Mol Sci 2020;21(7):2476. DOI: 10.3390/ijms21072476.
Saud B, Malla R, Shrestha K. A review on the effect of plant extract on mesenchymal stem cell proliferation and differentiation. Stem Cells Int 2019:7513404. DOI: 10.1155/2019/7513404.
Karobari MI, Adil AH, Assiry AA, et al. Herbal medications in endodontics and its application – A review of literature. Materials (Basel) 2022;15(9):3111. DOI: 10.3390/ma15093111.
Schmidt S, Heymann K, Melzig MF, et al. Glycyrrhizic acid decreases gentamicin-resistance in vancomycin-resistant Enterococci. Planta Med 2016;82(18):1540–1545. DOI: 10.1055/s-0042-114781.
Yang R, Wang LQ, Yuan BC, et al. The pharmacological activities of licorice. Planta Med 2015;81(18):1654–1669. DOI: 10.1055/s-0035-1557893.
Pastorino G, Cornara L, Soares S, et al. Liquorice (Glycyrrhiza glabra): A phytochemical and pharmacological review. Phytother Res 2018;32(12):2323–2339. DOI: 10.1002/ptr.6178.
Azizsoltani A, Piri K, Behzad S, et al. Ethyl acetate extract of licorice root (Glycyrrhiza glabra) enhances proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells. Iran J Pharm Res 2018;17(3):1057–1067. DOI: 10.22037/ijpr.2018.2246.
Misir S. Licorice root ethanol extract induces cell proliferation in human osteoblast cells. Turkish J Agric – Food Sci Technol 2021;9(4):803–806. DOI: 10.24925/turjaf.v9i4.803-806.4238.
do Nascimento MHM, de Araújo DR. Exploring the pharmacological potential of glycyrrhizic acid: From therapeutic applications to trends in nanomedicine. Futur Pharmacol 2022;2(1):1–15. DOI: 10.3390/futurepharmacol2010001.
Li Z, Chen C, Zhu X, et al. Glycyrrhizin suppresses RANKL-induced osteoclastogenesis and oxidative stress through inhibiting NF-κB and MAPK and activating AMPK/Nrf2. Calcif Tissue Int 2018;103(3): 324–337. DOI: 10.1007/s00223-018-0425-1.
Baghaei K, Hashemi SM, Tokhanbigli S, et al. Isolation, differentiation, and characterization of mesenchymal stem cells from human bone marrow. Gastroenterol Hepatol Bed Bench 2017;10(3):208–213. PMID: 29118937.
Kumar P, Nagarajan A, Uchil PD. Analysis of cell viability by the MTT assay. Cold Spring Harb Protoc 2018(6). DOI: 10.1101/pdb.prot095505.
Bergenholtz G, Wikesjö UM, Sorensen RG, et al. Observations on healing following endodontic surgery in nonhuman primates (Macaca fascicularis): Effects of rhBMP-2. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2006;101(1):116–125. DOI: 10.1016/j.tripleo.2005.02.085.
Su P, Tian Y, Yang C, et al. Mesenchymal stem cell migration during bone formation and bone diseases therapy. Int J Mol Sci 2018;19(8):2343. DOI: 10.3390/ijms19082343.
Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006;98(5):1076–1084. DOI: 10.1002/jcb.20886.
Kornicka K, Houston J, Marycz K. Dysfunction of mesenchymal stem cells isolated from metabolic syndrome and type 2 diabetic patients as result of oxidative stress and autophagy may limit their potential therapeutic use. Stem Cell Rev Rep 2018;14(3):337–345. DOI: 10.1007/s12015-018-9809-x.
Bai J, Xu J, Hang K, et al. Glycyrrhizic acid promotes osteogenic differentiation of human bone marrow stromal cells by activating the Wnt/β-catenin signaling pathway. Front Pharmacol 2021;12:607635. DOI: 10.3389/fphar.2021.607635.
Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 2006;8(4):315–317. DOI: 10.1080/14653240600855905.
Ghasemi M, Turnbull T, Sebastian S, et al. The MTT assay: Utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. Int J Mol Sci 2021;22(23):12827. DOI: 10.3390/ijms222312827.
Xu H, Fang L, Zeng Q, et al. Glycyrrhizic acid alters the hyperoxidative stress-induced differentiation commitment of MSCs by activating the Wnt/β-catenin pathway to prevent SONFH. Food Funct 2023;14(2):946–960. DOI: 10.1039/d2fo02337g.
Chrzanowski J, Chrzanowska A, Graboń W. Glycyrrhizin: An old weapon against a novel coronavirus. Phytother Res 2021;35(2): 629–636. DOI: 10.1002/ptr.6852.
Badr AE, Omar N, Badria FA. A laboratory evaluation of the antibacterial and cytotoxic effect of liquorice when used as root canal medicament. Int Endod J 2011;44(1):51–58. DOI: 10.1111/j.1365-2591.2010.01794.x.
Alghamdi F, Shakir M. The influence of Enterococcus faecalis as a dental root canal pathogen on endodontic treatment: A systematic review. Cureus 2020;12(3):e7257. DOI: 10.7759/cureus.7257.
Yasui S, Fujiwara K, Tawada A, et al. Efficacy of intravenous glycyrrhizin in the early stage of acute onset autoimmune hepatitis. Dig Dis Sci 2011;56(12):3638–3647. DOI: 10.1007/s10620-011-1789-5.
Hung CH, Kee KM, Chen CH, et al. A randomized controlled trial of glycyrrhizin plus tenofovir vs. tenofovir in chronic hepatitis B with severe acute exacerbation. Clin Transl Gastroenterol 2017;8(6):e104. DOI: 10.1038/ctg.2017.29.
Mou KH, Han D, Liu WL, et al. Combination therapy of orally administered glycyrrhizin and UVB improved active-stage generalized vitiligo. Braz J Med Biol Res 2016;49(8):e5354. DOI: 10.1590/1414-431X20165354.
Xu W, Li Y, Ju M, et al. A multicenter, randomized, double-blind, placebo-controlled study of compound glycyrrhizin capsules combined with a topical corticosteroid in adults with chronic eczema. Evid Based Complement Alternat Med 2020:6127327. DOI: 10.1155/2020/6127327.
Gomaa AA, Mohamed HS, Abd-Ellatief RB, et al Advancing combination treatment with glycyrrhizin and boswellic acids for hospitalized patients with moderate COVID-19 infection: A randomized clinical trial. Inflammopharmacology 2022;30(2): 477–486. DOI: 10.1007/s10787-022-00939-7.
