Cytotoxic and Osteoblast Differentiation Induction Properties of Crude Polar Extract of Piper sarmentosum leaves
Cytotoxicity and differentiation effect of P. sarmentosum
Keywords:Cytotoxicity, Differentiation, Ethyl acetate, Osteoblast, Piper sarmentosum
Piper sarmentosum or ‘kaduk’ is a well-known herb plant in Malaysia. Its extracts were found to exhibit a bone-protective effect against osteoporotic rats. Our study aim is to morphologically observe the effect of P. sarmentosum ethyl acetate extract on the differentiation of human peripheral blood stem cells (PBSCs) into osteoblasts. P. sarmentosum extracts (1-900 µg/mL) prepared using 1% dimethyl sulfoxide (DMSO) were used in the cytotoxicity assay. Then, the differentiation assay was performed using concentrations of 1-50 µg/mL. The untreated cells acted as the negative control, while the cells cultured in 50 µg/mL ascorbic acid and 10 mM β-glycerophosphate was a positive control. The cytotoxicity effect and proliferation capacity of the cells were analyzed using Trypan Blue exclusion method, while the differentiation of PBSCs was observed using von Kossa staining and ALP gene expression analysis. The result showed a decrease in cells’ viability in a dose-dependent manner during cytotoxicity assay. After 14 days of the differentiation, a constant rate of proliferation could be observed in the treated cells and positive control, while the untreated cells showed an increase in proliferation. The mineralization of extract-treated cells showed significant differences (p<0.05) as compared to the negative control. The expression of ALP was also upregulated on day 14 of differentiation compared to day 0. In conclusion, the extract was capable to induce osteoblast differentiation of human peripheral blood stem cell and potential to be used in tissue regeneration
Raman V, Galal AM, Khan IA (2012) An Investigation of the Vegetative Anatomy of Piper sarmentosum, and a Comparison with the Anatomy of Piper betle (Piperace-ae). American Journal of Plant Sciences 03 (08): 1135–1144. DOI: 10.4236/ajps.2012.38137.
Rukachaisirikul T, Siriwattanakit P, Sukcharoenphol K et al. (2004) Chemical constituents and bioactivity of Piper sarmentosum. Journal of Ethnopharmacology 93 (2–3): 173–176. DOI: 10.1016/j.jep.2004.01.022.
Subramaniam V, Ilham Adenan M, Rashih Ahmad A, Sahdan R (2003) Natural antioxidants: Piper sarmento-sum (Kadok) and Morinda elliptica (Mengkudu). Mal J Nutr 9 (1): 41–51.
Hafizah AH, Zaiton Z, Zulkhairi A et al. (2010) Piper sarmentosum as an antioxidant on oxidative stress in hu-man umbilical vein endothelial cells induced by hydrogen peroxide. Journal of Zheijiang University Science B 11 (5): 357–365. DOI: 10.1631/jzus.B0900397.
Chanprapai P, Chavasiri W (2017) Antimicrobial activity from Piper sarmentosum Roxb . against rice pathogenic bacteria and fungi. Journal of Integrative Agriculture 16 (11): 2513–2524. DOI: 10.1016/S2095-3119(17)61693-9.
Intan Zarina ZA, Fazry S, Jamar NH et al. (2020) The effects of Piper sarmentosum aqueous extracts on zebrafish ( Danio rerio ) embryos and caudal fin tissue regeneration. Scientific Reports 10 (1): 1–11. DOI: 10.1038/s41598-020-70962-7.
Shahrul Hisham ZA, Wan Omar WHH, Safian MF et al. (2009) Intrinsic anticarcinogenic effects of Piper sarmen-tosum ethanolic extract on a human hepatoma cell line. Cancer Cell International 9 (1): 6. DOI: 10.1186/1475-2867-9-6.
Estai MA, Suhaimi FH, Das S et al. (2011) Piper sarmen-tosum enhances fracture healing in ovariectomized osteo-porotic rats: A radiological study. Clinics 66 (5): 865–872. DOI: 10.1590/S1807-59322011000500025.
Peungvicha P, S. Thirawarapan S, Temsiririrkkul R et al. (1998) Hypoglycemic effect of the water extract of Piper sarmentosum in rats. Journal of Ethnopharmacology 60 (1): 27–32. DOI: 10.1016/S0378-8741(97)00127-X.
Ridtitid W, Ruangsang P, Reanmongkol W, Wongnawa M (2007) Studies of the anti-inflammatory and antipyretic activities of the methanolic extract of Piper sarmentosum Roxb. leaves in rats. Songklanakarin J. Sci. Technol. 29.
Weissman IL (2000) Translating Stem and Progenitor Cell Biology to the Clinic : Barriers and Opportunities. Science 287 (5457): 1442–1446.
Shahrul Hisham ZA, Rohaya MAW, Ismail I et al. (2005) Stem cells, cytokines and their receptors. Asia-Pacific Journal of Molecular Biology and Biotechnology 13 (1): 1–13.
Gussoni E, Soneoka Y, Strickland CD et al. (1999) Dys-trophin expression in the mdx mouse restored by stem cell transplantation. Nature 401 (6751): 390–394.
Lagasse E, Connors H, Al-Dhalimy M et al. (2000) Puri-fied hematopoietic stem cells can differentiate into hepato-cytes in vivo. Nature Medicine 6 (11): 1229–1234. DOI: 10.1038/81326.
Hadzir SN, Ibrahim SN, Abdul Wahab RM et al. (2014) Ascorbic acid induces osteoblast differentiation of human suspension mononuclear cells. Cytotherapy 16 (5): 674–682. DOI: 10.1016/j.jcyt.2013.07.013.
Manolagas SC (2000) Birth and Death of Bone Cells : Basic Regulatory Mechanisms and Implications for the Pathogenesis and Treatment of Osteoporosis. Endocr Rev 21 115–137.
Estai MA, Suhaimi F, Shuid AN et al. (2012) Biome-chanical evaluation of fracture healing following admin-istration of Piper sarmentosum in ovariectomised rats. Af-rican Journal of Pharmacy and Pharmacology 6 (3): 144–147. DOI: 10.5897/AJPP11.416.
Mohamad Asri SF, Soelaiman IN, Mohd Moklas MA et al. (2020) The Role of Piper sarmentosum Aqueous Ex-tract as a Bone Protective Agent, a Histomorphometric Study. International Journal of Molecular Sciences 21 (20): 7715.
Mohd Asri SF, Mohd Ramli ES, Soelaiman IN et al. (2016) Piper sarmentosum Effects on 11-Hydroxysteroid Dehydrogenase Type 1 Enzyme in Serum and Bone in Rat Model of Glucocorticoid-Induced Osteoporosis. Molecules 21 (11): 1523.
Cowan MM (1999) Plant Products as Antimicrobial Agents. Clinical microbiology reviews 12 (4): 564–582.
Houghton PJ, Raman A (1998) Laboratory handbook for the fractionation of natural extracts, 1st ed. New York, Ed Chapman and Hall.
Poojary MM, Vishnumurthy KA, Adhikari AV (2015) Extraction , characterization and biological studies of phy-tochemicals from Mammea suriga. Journal of Pharmaceu-tical Analysis 5 (3): 182–189. DOI: 10.1016/j.jpha.2015.01.002.
Bandoniene D, Murkovic M, Pfannhauser W et al. (2002) Detection and activity evaluation of radical scavenging compounds by using DPPH free radical and on-line HPLC-DPPH methods. European Food Reseaarch and Technology 214 (2): 143–147. DOI: 10.1007/s00217-001-0430-9.
Azahar NI, Mokhtar NM, Arifin MA (2020) Piper betle : a review on its bioactive compounds , pharmacological properties , and extraction process. In: IOP Conf. Ser. Mater. Sci. Eng. IOP Publishing, p 12044.
Opeña JM, Sotto RC (2022) Environmental Conditions, Phytochemical Constituents, and Antibacterial Activities of Two Philippine Medicinal Vitaceae Species. Journal of Tropical Life Science 12 (1): 11–19. DOI: 10.11594/jtls.12.01.02.
Akinsami A, Johnson O, Longdet I, Aguiyi J (2019) In Vitro Evaluation of Free Radical Scavenging, Fe2+ and SNP-Induced Lipid Peroxidation (Rat Brain) Activities of Methanolic Extracts from Three (3) Northern Nigerian Plants Leaf. Journal of Tropical Life Science 9 (1): 71–78. DOI: 10.11594/jtls.09.01.10.
Shahrul Hisham ZA, Kermani S, Intan Zarina ZA et al. (2013) Differentiation of Dental Pulp Stem Cells into Neuron-Like Cells in Serum-Free Medium. Stem Cells Int. 29 (6): 1519-1526.
Gorman DMO, Tierney CM, Brennan O, Brien FJO (2012) The Marine ‐ derived , Multi ‐ mineral formula , Aquamin , Enhances Mineralisation of Osteoblast Cells In Vitro. Phytotherapy Research 26 (3): 375–380.
Jeon J, Lee MS, Yang HS (2018) Differentiated osteo-blasts derived decellularized extracellular matrix to pro-mote osteogenic differentiation. Biomaterials Research 22 (1): 1–9.
Intan Zarina ZA, Shaipul Islam, Shazwana, Ling JP, Zainal Ariffin Z, Shahrul Hisham ZA (2021) The Effects Of Piper Sarmentosum Aqueous Extract On MC3T3-E1 Differentiation.pdf. NVEO-Natural Volatiles & Essential Oils Journal 4063–4979.
Wu T, Shu T, Le K et al. (2017) Icaritin, a novel plant-derived osteoinductive agent, enhances the osteogenic dif-ferentiation of human bone marrow- and human adipose tissue-derived mesenchymal stem cells. International Journal of Molecular Medicine 39 (4): 984–992. DOI: 10.3892/ijmm.2017.2906.
Hapidin H, Rozelan D, Abdullah H et al. (2015) Quercus infectoria gall extract enhanced the proliferation and ac-tivity of human fetal osteoblast cell line (hFOB 1.19). Malaysian Journal of Medical Sciences 22 (1): 12–22.
Wagner ER, Luther G, Zhu G et al. (2011) Defective Osteogenic Differentiation in the Development of Osteo-sarcoma. Sarcoma. DOI: 10.1155/2011/325238.
Choi M, Noh W, Park J et al. (2011) Gene expression pattern during osteogenic differentiation of human perio-dontal ligament cells in vitro. Journal of periodontal and implant science 41 (4): 167–175.
Copyright (c) 2022 Journal of Tropical Life Science
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
The work has not been published before (except in the form of an abstract or part of a published lecture or thesis) and it is not under consideration for publication elsewhere. When the manuscript is accepted for publication in this journal, the authors agree to automatic transfer of the copyright to the publisher.
Journal of Tropical Life Science is licensed under Creative Commons Attribution-NonCommercial 4.0 International License