Enhancement of dental pulp stem cell viability using Acmella oleraceae extract combined with bioactive glass
Keywords:
Bioactive glass, Sol-gel, Acmella oleraceae, DPSC, Cell ViabilityAbstract
This study aims to determine the effects of bioactive glass (BG) combined with Acmella oleracea (AO) extracts on dental pulp stem cells (DPSC) viability. DPSC were exposed to different combinations of BG-AO leave extract-conditioned medium. The BG 45S5 powder was synthesized using the sol-gel method. AO extract was prepared using ethanol extraction method. Gas Chromatography–Mass Spectrometry (GCMS) analysis of the AO ethanol extract was performed on a GCMS system consisting of an Agilent 6890 gas chromatograph coupled with an Agilent 5973 mass spectrometer. Sol-gel BG conditioned medium doped with AO extracts at various concentrations (25, 50, 100 and 250 µg/mL) with BG (1 mg/ mL) were prepared and exposed to DPSC. The DPSC was also treated using BG- and AO- only conditioned medium and non-treated cell as control. The DPSC cells’ responses were assessed using Alamar Blue (AB) assay. The results showed that GCMS analysis revealed the presence of amide, ester, terpenoid, fatty acid, alkene, terpene, carbohydrate, phenolic and alkane groups. Based on the AB assay, the BGAO- conditioned medium promoted DPSC viability. However, an increase in DPSC cell viability is clearly observed at Day 7 and 14 following exposure in BGAO-conditioned medium at the ratio of 1 mg/mL BG with 50 and 100 µg/mL of AO in comparison with AO alone. BGAO-conditioned medium at a dose of 25 µg/mL supported greater DPSC viability compared to other combination doses. The effect of combination of BG and AO towards DPSC at a certain dosage revealed continuous cell viability over the observation period and promoted cell growth that may be contributed by the combined effects of BG dissolution ions into the culture medium and also the presence of identified compound from the AO extracts namely phytol, linoleic acid, palmitic acid and 1, 4, 7,-Cycloundecatriene, 1, 5, 9, 9-tetramethyl, Z, Z, Z. Thus, it may have a significant potential to help in promoting dental and hard tissue regenerationReferences
Tomlin EM, Nelson SJ, Rossmann JA. Ridge Preservation for Implant Therapy: a Review of the Literature. The open dentistry journal. 2014;8:66-76.
Houreh AB, Labbaf S, Ting H-K, Ejeian F, Jones JR, Esfahani M-HN. Influence of calcium and phosphorus release from bioactive glasses on viability and differentiation of dental pulp stem cells. Journal of Materials Science. 2017;52(15):8928-41.
Hench LL. The story of Bioglass®. Journal of Materials Science: Materials in Medicine. 2006;17(11):967-78.
Rekhi A, Sarin S. Bioactive glass: A potential next generation biomaterial. SRM Journal of Research in Dental Sciences. 2016;7(1):27.
Mohd Zain NS, Tajudin SS, Mohd Noor SNF, Mohamad H. Biocompatibility Assessment of Newly Fabricated Melt Derived 45S5 Bioactive Glass on Dental Cells Proliferation. Journal of Biomedical and Clinical Sciences. 2017;1(1):7.
Sepulveda P, Jones JR, Hench LL. Characterization of melt-derived 45S5 and sol-gel-derived 58S bioactive glasses. Journal of biomedical materials research. 2001;58(6):734-40.
Faure J, Drevet R, Lemelle A, Ben Jaber N, Tara A, El Btaouri H, et al. A new sol-gel synthesis of 45S5 bioactive glass using an organic acid as catalyst. Materials science & engineering C, Materials for biological applications. 2015;47:407-12.
Cacciotti I, Lombardi M, Bianco A, Ravaglioli A, Montanaro L. Sol–gel derived 45S5 bioglass: synthesis, microstructural evolution and thermal behaviour. Journal of Materials Science: Materials in Medicine. 2012;23(8):1849-66.
Jones JR, Brauer DS, Hupa L, Greenspan DC. Bioglass and Bioactive Glasses and Their Impact on Healthcare. International Journal of Applied Glass Science. 2016;7(4):423-34.
Dubey S, Maity S, Singh M, Saraf SA, Saha S. Phytochemistry, Pharmacology and Toxicology of Spilanthes acmella: A Review. Advances in pharmacological sciences. 2013;2013:423750.
Nakatani N, Nagashima M. Pungent Alkamides from Spilanthes acmella L. var. oleracea Clarke. Bioscience, biotechnology, and biochemistry. 1992;56(5):759-62.
Mauth C, Huwig A, Graf-Hausner U, Roulet JF. Mauth C, Huwig A, Graf-Hausner U, Roulet J-F: Restorative Applications for Dental Pulp Therapy. In: Ashammakhi N, Reis R, Chiellini E (Eds.): Topics in Tissue Engineering. Vol. 3, 20072007. 1-32 p.
Cazzola M, Corazzari I, Prenesti E, Bertone E, Vernè E, Ferraris S. Bioactive glass coupling with natural polyphenols: Surface modification, bioactivity and anti-oxidant ability. Applied Surface Science. 2016;367:237-48.
Prachayasittikul S, Suphapong S, Worachartcheewan A, Lawung R, Ruchirawat S, Prachayasittikul V. Bioactive metabolites from Spilanthes acmella Murr. Molecules. 2009;14(2):850-67.
Savadi R, Yadav R, Yadav N. Study on immunomodulatory activity of ethanolic extract of Spillunthes acmella Murr. Leaves. Indian Journal of Natural Products and Resources.2010;1.
Volponi AA, Pang Y, Sharpe PT. Stem cell-based biological tooth repair and regeneration. Trends in cell biology. 2010;20(12):715-22.
Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proceedings of the National Academy of Sciences of the United States of America. 2000;97(25):13625-30.
Bronckaers A, Hilkens P, Fanton Y, Struys T, Gervois P, Politis C, et al. Angiogenic properties of human dental pulp stem cells. Plos One. 2013;8(8):e71104.
Noor SNFM, Zain NSM, Wei PY, Azizan NS, Mohamad H. Development of sol-gel bioactive glass for hard tissue regeneration. American Institute of Physics. 2016;1791:020017.
Bae SS, Ehrmann BM, Ettefagh KA, Cech NB. A validated liquid chromatography-electrospray ionization-mass spectrometry method for quantification of spilanthol in Spilanthes acmella (L.) Murr. Phytochemical analysis : PCA. 2010;21(5):438-43.
Franca JV, Queiroz MSR, do Amaral BP, Simas NK, da Silva NCB, Leal ICR. Distinct growth and extractive methods of Acmellaoleracea (L.) R.K. Jansen rising different concentrations of spilanthol: An important bioactive compound in human dietary. Food research international (Ottawa, Ont). 2016;89(Pt 1):781-9.
Anholeto LA, Oliveira PR, Rodrigues RAF, Spindola CDS, Labruna MB, Pizano MA, et al. Potential action of extract of Acmella oleracea (L.) R.K. Jansen to control Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) ticks. Ticks and tick-borne diseases. 2017;8(1):65-72.
Gupta N, Patel AR, Ravindra RP. Design of Akkalkara (Spilanthes acmella) formulations for antimicrobial and topical anti-inflammatory activities. International Journal of Pharma and Bio Sciences. 2012. 161-70 p.
Barman S, Sahu N, Deka S, Dutta S, Das S. Anti-inflammatory and analgesic activity of leaves of Spilanthes acmella (ELSA) in experimental animal models. Pharmacologyonline. 2009;1:1027-34.
Sudevan S, Sundar S, Ranganayaki P, Guptha A, Shafina J, Ramasamy V. Studies on in-vitro Anti-inflammatory activity of Acmella oleracea metabolic compounds. Journal of Chemical and Pharmaceutical Sciences. 2015. 227-32 p.
Lin X-H, Wu Y-B, Lin S, Zeng J-W, Zeng P-Y, Wu J-Z. Effects of Volatile Components and Ethanolic Extract from Eclipta prostrata on Proliferation and Differentiation of Primary Osteoblasts. Molecules. 2010;15(1):241.
Platt ID, El-Sohemy A. Regulation of osteoblast and adipocyte differentiation from human mesenchymal stem cells by conjugated linoleic acid. The Journal of nutritional biochemistry. 2009;20(12):956-64.
Maria-Ferreira D, da Silva LM, Mendes DAGB, Cabrini DdA, Nascimento AM, Iacomini M, et al. Rhamnogalacturonan from Acmella oleracea (L.) R.K. Jansen: Gastroprotective and Ulcer Healing Properties in Rats. Plos one. 2014;9(1):e84762.
Yamane LT, de Paula E, Jorge MP, de Freitas-Blanco V, #xf4, Santana n, et al. Acmella oleracea and Achyrocline satureioides as Sources of Natural Products in Topical Wound Care. Evidence-Based Complementary and Alternative Medicine. 2016;2016:9.
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