Osteogenic induction of menstrual blood mesenchymal stem cell by different Ferula species extracts

Document Type : Original Research Article


1 Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

2 Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran


Objective: Ferula spp. have many applications in complementary medicine and are recognized as the most important sources of natural products for bone health and regeneration especially in postmenopausal women. Therefore, the aim of this study was to investigate the effects of the extracts from three Ferula species on proliferation and osteogenesis potential of human menstrual blood-derived mesenchymal stem cells (MenSCs).
Materials and Methods: The possible cytotoxic activity of three members of Ferula spp. (at concentrations of 5 to 100 μg/ml) was determined using MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay. Alkaline phosphatase (ALP) activity assay, Alizarin Red-S staining, and the expression analysis of an osteoblastic gene were performed to evaluate osteogenic differentiation potential.
Results: The extracts of F. flabelliloba and F. szowitsiana decreased the viability and growth of MenSCs while F. foetida increased the proliferation of cells after 72 hr incubation. Treatment of MenSCs with selected plant extracts revealed that F. foetida and F. szowitsiana could enhance the osteogenic potential of MenSCs in terms of ALP activity. The Runx-2 expression in the presence of F. foetida was significantly greater than observed following treatment with 17β-estradiol (as positive control).
Conclusion: The results of this study suggest that F. foetida and F. szowitsiana may have therapeutic values as a nutraceutical with respect to their considerable influence on osteogenic potential of mesenchymal stem cells.


Akaberi M, Iranshahy M, Iranshahi M. 2015.
Review of the traditional uses,
phytochemistry, pharmacology and
toxicology of giant fennel (Ferula
communis L. subsp. communis). Iran J
Basic Med Sci, 18: 1050-1062.
Beral V. 2003. Breast cancer and hormonereplacement therapy in the Million Women
Study. Lancet, 362: 419-427.
Brunk CF, Jones KC, James TW. 1979. Assay
for nanogram quantities of DNA in cellular
homogenates. Anal Biochem, 92: 497-500.
Cavani F, Ferretti M, Carnevale G, Bertoni L,
Zavatti M, Palumbo C. 2012. Effects of
different doses of ferutinin on bone
formation/resorption in ovariectomized
rats. J Bone Miner Metab, 30: 619-629.
Cole ZA, Dennison EM, Cooper C. 2008.
Osteoporosis epidemiology update. Curr
Rheumatol Rep, 10: 92-96.
Collaborative Group on Epidemiological
Studies of Ovarian. 2015. Menopausal
hormone use and ovarian cancer risk:
individual participant meta-analysis of 52
epidemiological studies. Lancet, 385:
Dalirfardouei R, JamialahmadiK, Mahdipour E.
2018. A feasible method for the isolation of
mesenchymal stem cells from menstrual
blood and their exosomes. Tissue Cell, 55:
Dalirfardouei R, Jamialahmadi K, Jafarian AH,
Mahdipour E. 2019. Promising effects of
exosomes isolated from menstrual bloodderived mesenchymal stem cell on woundhealing process in diabetic mouse model. J
Tissue Eng Regen Med, 13: 555-568.
Ducy P, Zhang R, Geoffroy V, Ridall
AL,KarsentyG. 1997. Osf2/Cbfa1: A
transcriptional activator of osteoblast
differentiation. Cell, 89: 747-754.
Ferretti M, Bertoni L, Cavani F, Zavatti M,
Resca E, Carnevale G, Benelli A, Zanoli P,
Palumbo C. 2010. Influence of ferutinin on
bone metabolism in ovariectomized rats. II:
Role in recovering osteoporosis. J Anat,
217: 48-56.
Franceschi RT, XiaoG, Jiang D,
Gopalakrishnan R, Yang S, Reith E. 2003.
Multiple signaling pathways converge on
the Cbfa1/Runx2 transcription factor to
regulate osteoblast differentiation. Connect
Tissue Res, 44: 109-116.
Geoffroy V, Kneissel M, Fournier B, Boyde A,
Matthias P. 2002. High bone resorption in
adult aging transgenic mice overexpressing
cbfa1/runx2 in cells of the osteoblastic
lineage. Mol Cell Biol, 22: 6222-6233.
Golub EE, Boesze-Battaglia K. 2007. The role
of alkaline phosphatase in mineralization.
Curr Opin Orthop, 18: 444-448.
Iranshahy M, Iranshahi M. 2011. Traditional
uses, phytochemistry and pharmacology of
asafoetida (Ferula assa-foetida oleo-gumresin)—A review. JEthnopharmacol, 134:
Kim YJ, Lee MH, Wozney JM, ChoJY, Ryoo
HM. 2004. Bone morphogenetic protein-2-
induced alkaline phosphatase expression is
stimulated by Dlx5 and repressed by Msx2.
J Biol Chem, 279: 50773-50780.
Komori T, Yagi H, Nomura S, Yamaguchi A,
Sasaki K, Deguchi K, Shimizu Y, Bronson
R, Gao Y-H, Inada M. 1997. Targeted
disruption of Cbfa1results in a complete
lack of bone formation owing to
maturational arrest of osteoblasts. Cell, 89:
Liu W, Toyosawa S, Furuichi T, Kanatani N,
Yoshida C, Liu Y, Himeno M, Narai S,
Yamaguchi A, Komori T. 2001.
Osteogenic activity of Ferula species
AJP, Vol. 11, No. 3, May-Jun 2021 291
Overexpression of Cbfa1 in osteoblasts
inhibits osteoblast maturation and causes
osteopenia with multiple fractures. J Cell
Biol, 155: 157-166.
Mahmoudi Z, Soleimani M, Saidi A, Iranshahi
M, Azizsoltanli A. 2013. Effect of Ferula
gummosa ethanolic extract on osteogenesis
in human mesenchymal stem cells. J Med
Plants, 2: 50-59.
Mahmoudi Z, Soleimani M, Saidi A,
Khamisipour G, & Azizsoltani A. 2013.
Effects of Foeniculum vulgare ethanol
extract on osteogenesis in human
mecenchymal stem cells. Avicenna J
Phytomed, 3: 135-142.
Matin M, Nakhaeizadeh H, Bahrami A,
Iranshahi M, Arghiani N, Behnam Rassouli
F. 2014. Ferutinin, an apoptosis inducing
terpenoid from Ferula ovina. Asian Pac J
Cancer Prev, 15: 2123-2128.
Mirhashemi S, Kalantar Motamedi MH,
Mirhashemi AH, Mehrvarz S, Danial Z.
2017. Osteoporosis in Iran. Hosp Pract Res,
2: 57-57.
Moazzami Farida SH, Ghorbani A, Ajani Y,
Sadr M, Mozaffarian V. 2018.
Ethnobotanical applications and their
correspondence with phylogeny in
Apiaceae-Apioideae. Res J
Pharmacognosy, 5: 79-97.
Palumbo C, Cavani F, Bertoni L,Ferretti M.
2012. Role of phytoestrogen ferutinin in
preventing/recovering bone loss: results
from experimental ovariectomized rat
models. In: DionyssiotisY(Ed.),
Osteoporesis, pp. 711-732, London,
Palumbo C, Ferretti M, Bertoni L, Cavani F,
Resca E, Casolari B, Carnevale G, Zavatti
M, Montanari C, Benelli A, Zanoli P. 2009.
Influence of ferutinin on bone metabolism
in ovariectomized rats. I: role in preventing
osteoporosis. J Bone Miner Metab, 27: 538-
Pfaffl MW, Horgan GW, Dempfle L. 2002.
Relative expression software tool (REST)
for group-wise comparison and statistical
analysis of relative expression results in
real-time PCR. Nucleic Acids Res, 30: e36.
Riss TL, Moravec RA, Niles AL, Duellman S,
Benink HA, Worzella TJ, Minor L. (2013).
Cell viability assay. Eli Lilly & Company
and the National Center for Advancing
Translational Sciences.
Saeed IA, Ali L, Jabeen A, Khasawneh M,
Rizvi TA, AshrafSS. 2012. Estrogenic
activities of ten medicinal herbs from the
middle east.J Chromatogr Sci, 51: 33-39.
Safaeian L, Ghannadi A, Javanmard SH,
Vahidian MH. 2015. The effect of
hydroalcoholic extract of Ferula foetida
stems on blood pressure and oxidative
stress in dexamethasone-induced
hypertensive rats. Res Pharm Sci, 10: 326-
Safi R, Hamade A, Bteich N, El Saghir J, Assaf
MD, El-Sabban M et al. 2018. A ferutinin
analogue with enhanced potency and
selectivity against ER-positive breast
cancer cells in vitro. Biomed Pharmacother,
105: 267-273.
Salaminia S, Mohsenzadeh Y, Motedayen M,
Sayehmiri F, Dousti M. 2019. Hormone
replacement therapy and postmenopausal
cardiovascular events: a meta-analysis. Iran
Red Crescent Med J, 21: e82298.
Zainabadi K, Liu CJ, Guarente L. 2017. SIRT1
is a positive regulator of the master
osteoblast transcription factor, RUNX2.
PLOS ONE, 12: e0178520.
Zavatti M, Resca E, Bertoni L, Maraldi T,
Guida M, Carnevale G, Ferrari A, De Pol
A. 2013. Ferutinin promotes proliferation
and osteoblastic differentiation in human
amniotic fluid and dental pulp stem cells.
Life Sci, 92: 993-1003.
Zhao Z, Zhao M, Xiao G, Franceschi RT. 2005.
Gene transfer of the Runx2 transcription
factor enhances osteogenic activity of bone
marrow stromal cells in vitro and in vivo.
Mol Ther, 12: 247-253.