Quercitrin for periodontal regeneration: effects on human gingival fibroblasts and mesenchymal stem cells

Manuel Gómez-Florit, Marta Monjo, Joana M Ramis, Manuel Gómez-Florit, Marta Monjo, Joana M Ramis

Abstract

Periodontal disease (PD) is the result of an infection and chronic inflammation of the gingiva that may lead to its destruction and, in severe cases, alveolar bone and tooth loss. The ultimate goal of periodontal treatment is to achieve periodontal soft and hard tissues regeneration. We previously selected quercitrin, a catechol-containing flavonoid, as a potential agent for periodontal applications. In this study, we tested the ability of quercitrin to alter biomarker production involved in periodontal regeneration on primary human gingival fibroblasts (hGF) and primary human mesenchymal stem cells (hMSC) cultured under basal and inflammatory conditions. To mimic PD inflammatory status, interleukin-1 beta (IL-1β) was used. The expression of different genes related to inflammation and extracellular matrix were evaluated and prostaglandin E2 (PGE2) production was quantified in hGFs; alkaline phosphatase (ALP) activity and calcium content were analysed in hMSCs. Quercitrin decreased the release of the inflammatory mediator PGE2 and partially re-established the impaired collagen metabolism induced by IL-1β treatment in hGFs. Quercitrin also increased ALP activity and mineralization in hMSCs, thus, it increased hMSCs differentiation towards the osteoblastic lineage. These findings suggest quercitrin as a novel bioactive molecule with application to enhance both soft and hard tissue regeneration of the periodontium.

Figures

Figure 1. Effect of quercitrin on hGF…
Figure 1. Effect of quercitrin on hGF stimulated with IL-1β.
(A) Experimental design: the effect of quercitrin (QUER) treatment on hGF was evaluated in four different inflammatory scenarios (stripped pattern); treatment for 1 day with interleukin-1 beta (IL-1β); treatment for 3 days with IL-1β; treatment with IL-1β during the first 3 days and culturing the cells for 14 days (therapeutic approach; T); and culturing the cells for 14 days plus IL-1β treatment during the last 3 days (preventive approach; P). (B) Cytotoxicity measured after 1 and 3 days of treatment. Dotted line represents high control (100% cytotoxicity). (C) Gene expression results: cells were treated with quercitrin (grey bars) or without (white bars), following the experimental design. Data were normalized to reference genes, expressed as percentage of control (vehicle), which was set to 100%. Values represent the mean ± SEM of two independent experiments. One, two and three symbols represent a significant difference between two groups with P ≤ 0.05, P < 0.01 and P < 0.001, respectively: (*) treatment versus control (vehicle); (#) quercitrin versus vehicle in the presence of IL-1β for each time point.
Figure 2. Quercitrin decreased PGE2 release on…
Figure 2. Quercitrin decreased PGE2 release on hGF stimulated with IL-1β.
Cells were treated with quercitrin (QUER; grey bars) or without (white bars) in the presence of interleukin-1 beta (IL-1β). Four different scenarios were set: treatment for 1 day with IL-1β and quercitrin; treatment for 3 days with IL-1β and quercitrin; treatment with quercitrin for 14 days plus IL-1β during the first 3 days (therapeutic approach; T); and treatment with quercitrin for 14 days plus IL-1β during the last 3 days (preventive approach; P). Values represent the mean ± SEM of two independent experiments. One, two and three symbols represent a significant difference between two groups with P ≤ 0.05, P versus vehicle in the presence of IL-1β for each time point.
Figure 3. Quercitrin did not show cytotoxic…
Figure 3. Quercitrin did not show cytotoxic effects on hMSCs.
Cells were grown under six different conditions: basal media; basal media with 200 μM quercitrin (QUER); osteogenic media (OS); osteogenic media with 200 μM quercitrin; osteogenic media supplemented with interleukin-1 beta (IL-1β); and osteogenic media supplemented with IL-1β and quercitrin. (A) Cytotoxicity measured after 2 days. Dotted line represents high control (100% cytotoxicity). (B) Metabolic activity measured after 19 days. Values represent the mean ± SEM of two independent experiments. One, two and three symbols represent a significant difference between two groups with P ≤ 0.05, P < 0.01 and P < 0.001, respectively: (*) treatment versus control (vehicle).
Figure 4. Quercitrin increased the osteoblastic differentiation…
Figure 4. Quercitrin increased the osteoblastic differentiation of hMSCs.
(A) ALP activity and (B) calcium content determined after 19 days of hMSCs growth under six different conditions: basal media; basal media with 200 μM quercitrin (QUER); osteogenic media (OS); osteogenic media with 200 μM quercitrin; osteogenic media supplemented with interleukin-1 beta (IL-1β); and osteogenic media supplemented with IL-1β and quercitrin. Values represent the mean ± SEM of two independent experiments. One, two and three symbols represent a significant difference between two groups with P ≤ 0.05, P < 0.01 and P < 0.001, respectively: (*) treatment versus control (vehicle); (#) quercitrin versus vehicle; (§) inflammation groups versus osteogenic groups.

References

    1. Tonetti M. S. et al. Principles in prevention of periodontal diseases. J. Clin. Periodontol. 42, S5–S11 (2015).
    1. Shin S. Y., Rios H. F., Giannobile W. V & Oh T. In Stem Cell Biology and Tissue Engineering in Dental Sciences (eds Vishwakarma A. et al.) Ch. 36, 459–469 (Elsevier Inc., 2015).
    1. Dentino A., Lee S., Mailhot J. & Hefti A. F. Principles of periodontology. Periodontol. 2000 61, 16–53 (2013).
    1. Heitz-Mayfield L. J. A. & Lang N. P. Surgical and nonsurgical periodontal therapy. Learned and unlearned concepts. Periodontol. 2000 62, 218–31 (2013).
    1. Han J., Menicanin D., Gronthos S. & Bartold P. Stem cells, tissue engineering and periodontal regeneration. Aust. Dent. J. 59, 1–14 (2013).
    1. Esposito M., Grusovin M. G., Papanikolaou N., Coulthard P. & Worthington H. V. Enamel matrix derivative (Emdogain(R)) for periodontal tissue regeneration in intrabony defects. Cochrane database Syst. Rev. CD003875, 10.1002/14651858.CD003875.pub3 (2009).
    1. Ramseier C. A, Rasperini G., Batia S. & Giannobile W. V. Advanced reconstructive technologies for periodontal tissue repair. Periodontol. 2000 59, 185–202 (2012).
    1. Preshaw P. M. Host response modulation in periodontics. Periodontol. 2000 48, 92–110 (2008).
    1. Pietta P. G. Flavonoids as antioxidants. J. Nat. Prod. 63, 1035–1042 (2000).
    1. Izzi V. et al. The effects of dietary flavonoids on the regulation of redox inflammatory networks. Front. Biosci. Landmark Ed. 17, 2396–418 (2012).
    1. An J., Zuo G. Y., Hao X. Y., Wang G. C. & Li Z. S. Antibacterial and synergy of a flavanonol rhamnoside with antibiotics against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA). Phytomedicine 18, 990–993 (2011).
    1. Cushnie T. P. T. & Lamb A. J. Recent advances in understanding the antibacterial properties of flavonoids. Int. J. Antimicrob. Agents 38, 99–107 (2011).
    1. Gómez-Florit M., Monjo M. & Ramis J. M. Identification of Quercitrin as Potential Therapeutic Agent for Periodontal Applications. J. Periodontol. 85, 966–974 (2014).
    1. Satué M., Arriero M. D. M., Monjo M. & Ramis J. M. Quercitrin and taxifolin stimulate osteoblast differentiation in MC3T3-E1 cells and inhibit osteoclastogenesis in RAW 264.7 cells. Biochem. Pharmacol. 86, 1476–86 (2013).
    1. Luo D., Or T. C. T., Yang C. L. H. & Lau A. S. Y. Anti-inflammatory activity of iridoid and catechol derivatives from Eucommia ulmoides oliver. ACS Chem. Neurosci. 5, 855–866 (2015).
    1. Heneka M. T. et al. Locus ceruleus controls Alzheimer’s disease pathology by modulating microglial functions through norepinephrine. Proc. Natl. Acad. Sci. USA 107, 6058–63 (2010).
    1. Zheng L. T., Ryu G.-M., Kwon B.-M., Lee W.-H. & Suk K. Anti-inflammatory effects of catechols in lipopolysaccharide-stimulated microglia cells: inhibition of microglial neurotoxicity. Eur. J. Pharmacol. 588, 106–13 (2008).
    1. Furuhashi I., Iwata S., Shibata S., Sato T. & Inoue H. Inhibition by licochalcone A, a novel flavonoid isolated from liquorice root, of IL-1beta-induced PGE2 production in human skin fibroblasts. J. Pharm. Pharmacol. 57, 1661–6 (2005).
    1. Kida Y. et al. Interleukin-1 stimulates cytokines, prostaglandin E2 and matrix metalloproteinase-1 production via activation of MAPK/AP-1 and NF-kappaB in human gingival fibroblasts. Cytokine 29, 159–68 (2005).
    1. Ono M. et al. Quest for anti-inflammatory substances using IL-1β-stimulated gingival fibroblasts. In Vivo (Brooklyn). 25, 763–768 (2011).
    1. Yucel-Lindberg T. & Båge T. Inflammatory mediators in the pathogenesis of periodontitis. Expert Rev. Mol. Med. 15, e7 (2013).
    1. Nokhbehsaim M. et al. Effects of enamel matrix derivative on periodontal wound healing in an inflammatory environment in vitro. J. Clin. Periodontol. 38, 479–90 (2011).
    1. Palmqvist P., Lundberg P., Lundgren I., Hänström L. & Lerner U. H. IL-1beta and TNF-alpha regulate IL-6-type cytokines in gingival fibroblasts. J. Dent. Res. 87, 558–563 (2008).
    1. Reed D. A. & Diekwisch T. G. H. In Stem Cell Biology and Tissue Engineering in Dental Sciences (eds Vishwakarma A. et al.) Ch. 35, 445–458 (Elsevier Inc., 2015).
    1. Noguchi K. & Ishikawa I. The roles of cyclooxygenase-2 and prostaglandin E2 in periodontal disease. Periodontol. 2000 43, 85–101 (2007).
    1. Gutiérrez-Venegas G. & Contreras-Sánchez A. Luteolin and fisetin inhibit the effects of lipopolysaccharide obtained from Porphyromonas gingivalis in human gingival fibroblasts. Mol. Biol. Rep. 40, 477–85 (2013).
    1. Comalada M. et al. In vivo quercitrin anti-inflammatory effect involves release of quercetin, which inhibits inflammation through down-regulation of the NF-κB pathway. Eur. J. Immunol. 35, 584–592 (2005).
    1. Dai X., Ding Y., Zhang Z., Cai X. & Li Y. Quercetin and quercitrin protect against cytokine-induced injuries in RINm5F β-cells via the mitochondrial pathway and NF-κB signaling. Int. J. Mol. Med. 31, 265–271 (2013).
    1. Fang S.-H., Rao Y. K. & Tzeng Y.-M. Anti-oxidant and inflammatory mediator’s growth inhibitory effects of compounds isolated from Phyllanthus urinaria. J. Ethnopharmacol. 116, 333–40 (2008).
    1. Ghosh S. & Hayden M. S. New regulators of NF-kappaB in inflammation. Nat. Rev. Immunol. 8, 837–48 (2008).
    1. Hoesel B. & Schmid J. A. The complexity of NF-κB signaling in inflammation and cancer. Mol. Cancer 12, 86 (2013).
    1. Kim H. P., Son K. H., Chang H. W. & Kang S. S. Anti-inflammatory plant flavonoids and cellular action mechanisms. J. Pharmacol. Sci. 96, 229–245 (2004).
    1. Trzeciakiewicz A., Habauzit V. & Horcajada M.-N. When nutrition interacts with osteoblast function: molecular mechanisms of polyphenols. Nutr. Res. Rev. 22, 68–81 (2009).
    1. Ding M., Zhao J., Bowman L., Lu Y. & Shi X. Inhibition of AP-1 and MAPK signaling and activation of Nrf2/ARE pathway by quercitrin. Int. J. Oncol. 36, 59–67 (2010).
    1. Soell M., Elkaim R. & Tenenbaum H. Cathepsin C, Matrix metalloproteinases, and their tissue inhibitors in gingiva and gingival crevicular fluid from periodontitis-affected patients. J. Dent. Res. 81, 174–178 (2002).
    1. Kawaguchi H. et al. Enhancement of periodontal tissue regeneration by transplantation of bone marrow mesenchymal stem cells. J. Periodontol. 75, 1281–1287 (2004).
    1. Li B. & Jin Y. In Stem Cell Biology and Tissue Engineering in Dental Sciences (eds Vishwakarma A. et al.) Ch. 37, 471–482 (Elsevier Inc., 2015).
    1. Chen F. M., Wu L. A., Zhang M., Zhang R. & Sun H. H. Homing of endogenous stem/progenitor cells for in situ tissue regeneration: Promises, strategies, and translational perspectives. Biomaterials 32, 3189–3209 (2011).
    1. Monsarrat P. et al. Concise Review: Mesenchymal stromal cells used for periodontal regeneration: a systematic review. Stem Cells Transl. Med. 3, 768–774 (2014).
    1. Sonomoto K. et al. Interleukin-1β induces differentiation of human mesenchymal stem cells into osteoblasts via the Wnt-5a/receptor tyrosine kinase-like orphan receptor 2 pathway. Arthritis Rheum. 64, 3355–63 (2012).
    1. Loebel C. et al. The calcification potential of human MSCs can be enhanced by interleukin-1β in osteogenic medium. J. Tissue Eng. Regen. Med. n/a–n/a, 10.1002/term.1950 (2014).
    1. Ferreira E. et al. Inflammatory cytokines induce a unique mineralizing phenotype in mesenchymal stem cells derived from human bone marrow. J. Biol. Chem. 288, 29494–505 (2013).
    1. Hughes F. J., Ghuman M. & Talal A. Periodontal regeneration: a challenge for the tissue engineer? Proc. Inst. Mech. Eng. H. 224, 1345–1358 (2010).
    1. Carnes D. L., Maeder C. L. & Graves D. T. Cells with osteoblastic phenotypes can be explanted from human gingiva and periodontal ligament. J. Periodontol. 68, 701–707 (1997).
    1. Córdoba A. et al. Flavonoid-modified surfaces: multifunctional bioactive biomaterials with osteopromotive, anti-inflammatory, and anti-fibrotic potential. Adv. Healthc. Mater. 4, 540–549 (2015).
    1. Battino M., Ferreiro M. S., Fattorini D. & Bullon P. In vitro antioxidant activities of mouthrinses and their components. J. Clin. Periodontol. 29, 462–7 (2002).
    1. Palaska I., Papathanasiou E. & Theoharides T. C. Use of polyphenols in periodontal inflammation. Eur. J. Pharmacol. 720, 77–83 (2013).

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