@2024 Afarand., IRAN
ISSN: 2252-0805 The Horizon of Medical Sciences 2018;24(4):270-276
ISSN: 2252-0805 The Horizon of Medical Sciences 2018;24(4):270-276
Analyzing the Effects of Adipogenic Potential in Hydro-Alcoholic Extract of Marijuana on Mesenchymal Bone Marrow Stem Cells of Adult Male Rats
ARTICLE INFO
Article Type
Original ResearchAuthors
Sazmand M. (1,2)Mehrbani D. (*)
Hosseini S.E. (2)
Amini M. (3)
(*) Burn and Wound Healing Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
(1,2) Department of Biology, College of Science, Fars Science and Research, Branch, Islamic Azad University, Fars, Iran
(2) Department of Biology, College of Science, Shiraz Branch, Islamic Azad University, Shiraz, Iran
(3) Laparascopy Research Center, Department of Surgery, Shiraz University of Medical Sciences, Shiraz, Iran
Correspondence
Address: Burn and Wound Healing Research Center, Shiraz University of Medical Sciences, Shiraz, IranPhone: +989171130337
Fax: +987136223418
davood_mehrabani@yahoo.com
Article History
Received: February 25, 2018Accepted: September 22, 2018
ePublished: October 10, 2018
ABSTRACT
Aims
Millions of people consume psychoactives such as marijuana. Mesenchyme
cells, which are derived from bone marrow stem cells, have multiple applications
in tissue engineering and regenerative medicine. The aim of this study was to
analyze the effects of marijuana on differentiation power of bone marrow stem
cells of adult male rats in adipose tissues.
Materials & Methods In this experimental study, the bone marrow stem cells of adult male rats were segregated with flushing method. Then, their mesenchymal identity was confirmed with a morphological study, PCR-RT method, and surface markers of CD45 and CD34 along with the related genes of CD90 and CD73. For analyzing the viability and differentiation power of the cells in adipose tissues in terms of marijuana usage, cell samples were exposed to different dosages of marijuana, such as 30 and 6000 ng/ml and cell survival was evaluated by MTT. The adipogenic differentiation potential of the treated cells were analyzed based on oil red staining. The results were analyzed by ANOVA and Schaffe tests.
Findings The results indicated that the extracted cells are non-adherent in flasks and, in moving from passage one to the next passage, their duplication speed increased. In passage three, they changed into fusiform cells. Based on RT-PCR, it was revealed that CD90 and CD73 were existent, but CD34 and CD45 are nonexistent. Marijuana treatment caused no change in the form of stem cells. 10days after exposure to adipogenic circumstances, in addition to increase of survival, their cytoplasm was repleted with adipose.
Conclusion The results of this study revealed that marijuana has positive effects on mesenchymal bone marrow stem cells in differentiating the adipocyte.
Materials & Methods In this experimental study, the bone marrow stem cells of adult male rats were segregated with flushing method. Then, their mesenchymal identity was confirmed with a morphological study, PCR-RT method, and surface markers of CD45 and CD34 along with the related genes of CD90 and CD73. For analyzing the viability and differentiation power of the cells in adipose tissues in terms of marijuana usage, cell samples were exposed to different dosages of marijuana, such as 30 and 6000 ng/ml and cell survival was evaluated by MTT. The adipogenic differentiation potential of the treated cells were analyzed based on oil red staining. The results were analyzed by ANOVA and Schaffe tests.
Findings The results indicated that the extracted cells are non-adherent in flasks and, in moving from passage one to the next passage, their duplication speed increased. In passage three, they changed into fusiform cells. Based on RT-PCR, it was revealed that CD90 and CD73 were existent, but CD34 and CD45 are nonexistent. Marijuana treatment caused no change in the form of stem cells. 10days after exposure to adipogenic circumstances, in addition to increase of survival, their cytoplasm was repleted with adipose.
Conclusion The results of this study revealed that marijuana has positive effects on mesenchymal bone marrow stem cells in differentiating the adipocyte.
CITATION LINKS
[1]Kidd PM. Multiple sclerosis, an autoimmune inflammatory disease: Prospects for its integrative management. Altern Med Rev. 2001;6(6):540-66.
[2]Long JZ, Nomura DK, Vann RE, Walentiny DM, Booker L, Jin X, et al. Dual blockade of FAAH and MAGL identifies behavioral processes regulated by endocannabinoid crosstalk in vivo. Proc Natl Acad Sci U S A. 2009;106(48):20270-5.
[3]Alakbarov FU. Medicinal Properties of Cannabis According to Medieval Manuscripts of Azerbaijan. J Cannabis Ther. 2001;1(2):3-14.
[4]Farquhar-Smith WP. Pain and cannabinoids: Science and evidence. Pain Rev. 2002;9(1):41-67.
[5]Ruiz L, Miguel A, Dı́az-Laviada I. Δ9‐Tetrahydrocannabinol induces apoptosis in human prostate PC‐3 cells via a receptor‐independent mechanism. FEBS Lett.1999;458(3):400-4.
[6]Sánchez C, Galve-Roperh I, Canova C, Brachet Ph, Guzmán M. Δ9‐Tetrahydrocannabinol induces apoptosis in C6 glioma cells. FEBS Lett. 1998;436(1):6-10.
[7]Marselos M, Karamanakos P. Mutagenicity, developmental toxicity and carcinogenicity of cannabis. Addict Bio. 1999;4(1):5-12.
[8]Cavuoto P, Wittert GA. The Role of the Endocannabinoid System in the Regulation of Energy Expenditure. Best Pract Res Clin Endocrinol Metab. 2009;23(1):79-86.
[9]Borgelt LM, Franson KL, Nussbaum AM, Wang GS. The Pharmacologic and Clinical Effects of Medical Cannabis. Pharmacotherapy. 2013;33(2):195-209
[10]Svízenská I, Dubový P, Sulcová A. Cannabinoid Receptors 1 and 2 (CB1 and CB2), their distribution, ligands and functional involvement in nervous system structures--a short review. Pharmacol Biochem Behav. 2008;90(4):501-11.
[11]Biehl JK, Russel B. Introduction to stem cell therapy. J Cardiovasc Nurs. 2009;24(2):98-105.
[12]Poulos SP, Dodson MV, Hausman GJ. Cell line models for differentiation: Preadipocytes and adipocytes. Exp Bio Med. 2010;235(10):1185-93.
[13]Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem. 2006;98(5):1076-84.
[14]Pournassarkhakabaz B, Baharvand H. Human Mesenchymal Stem Cells and its therapeutic applications. Anat Sci J. 2007;5(19-20):167-215. [Persian]
[15]Taheri F, Haji Ghasem Kashani M, Ghorbanian MT, Hosseinpour L. Inductive effect of Deprenyl and Dimethyl sulfoxide on proliferation and survival of the mesenchymal stem cells. J Gorgan Univ Med Sci. 2012;14(3):10-8. [Persian]
[16]Ramezani T, Baharara J, Saghiri N. The Effect of Saffron Aqueous Extract (Crocus sativus L) on Osteogenic Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells. J Birjand Univ Med Sci. 2014;21(2):169-78.
[17]Polisetti N, Chaitanya VG, Babu PP, Vemuganti GK. Isolation, characterization and differentiation potential of rat bone marrow stromal cells. Neurol India. 2010;58(2):201-8.
[18]Karaoz E, Genç ZS, Demircan PÇ, Aksoy A, Duruksu G. Protection of rat pancreatic islet function and viability by coculture with rat bone marrow-derived mesenchymal stem cells. Cell Death Dis. 2010;1(4):36.
[19]Maxson S, Lopez EA, Yoo D, Danilkovitch-Miagkova A, LeRoux MA. Concise review: Role of mesenchymal stem cells in wound repair. Stem Cells Trans Med. 2012;1(2):142-9.
[20]Rahmanifar F, Tamadon A, Mehrabani D, Zare SH, Abasi S, Keshavarz S, et al. Histomorphometric evaluation of treatment of rat azoosper-mic seminiferous tubules by allotransplantation of bone marrow-derived mesenchymal stem cells. Iran J Basic Med Sci. 2016;19(6):653-61.
[21]Mahmoudi Z, Soleimani M, Saiedi A, Iranshahi M, Azizsoltanli A. Effect of Ferula gummosa ethanolic extract on osteogenesis in human mesenchymal stem cells. J Med Plants. 2013;2(46):50-9. [Persian]
[22]Rosen ED, Hsu CH, Wang X, Sakai S, Freeman MW, Gonzalez FJ, et al. C/EBPα induces adipogenesis through PPARγ: a unified pathway. Genes Dev. 2002;16(1):22-6.
[23]O'sullivan SE. Cannabinoids go nuclear: Evidence for activation of peroxisome proliferator‐activated receptors. Br J Pharmacol. 2007;152(5):576-82.
[24]Frisher M, White S, Varbiro G, Voisey C, Perumal D, Crome I, et al. The role of cannabis and cannabinoids in diabetes. Br J Diabetes Vasc Dis. 2010;10(6):267-73.
[25]McPartland JM. The endocannabinoid System: An osteopathic perspective. J Am Osteopath Assoc. 2008;108(10):586-600.
[26]Idris AI, Sophocleous A, Landao-Bassonga E, Canals M, Milligan G, Baker D, et al. Cannabinoid receptor type 1 protects against age-related osteoporosis by regulating osteoblast and adipocyte differentiation in marrow stromal cells. Cell Metab. 2009;10(2):139-47.
[27]Nick Bakhte Dastjerdi M. The Effect of Dexamethasone on the Differentiation of Bone Marrow Scaffold Cells into Fatty Cells. Isfahan Med Sch J. 2005;23(77-76):17. [Persian]
[28]Lombard C, Hegde VL, Nagarkatti M, Nagarkatti PS. Perinatal exposure to Δ9-tetrahydrocannabinol triggers profound defects in T cell differentiation and function in fetal and postnatal stages of life, including decreased responsiveness to HIV antigens. J Pharmacol Exp Ther. 2011;339(2):607-17.
[29]Takada I, Kouzmenko AP, Kato S. Molecular switching of osteoblastogenesis versus adipogenesis: implications for targeted therapies. Expert Opin Ther Targets. 2009;13(5):593-603.
[30]Cao JJ. Effects of obesity on bone metabolism. J Orthop Surg Res. 2011;6:30.
[2]Long JZ, Nomura DK, Vann RE, Walentiny DM, Booker L, Jin X, et al. Dual blockade of FAAH and MAGL identifies behavioral processes regulated by endocannabinoid crosstalk in vivo. Proc Natl Acad Sci U S A. 2009;106(48):20270-5.
[3]Alakbarov FU. Medicinal Properties of Cannabis According to Medieval Manuscripts of Azerbaijan. J Cannabis Ther. 2001;1(2):3-14.
[4]Farquhar-Smith WP. Pain and cannabinoids: Science and evidence. Pain Rev. 2002;9(1):41-67.
[5]Ruiz L, Miguel A, Dı́az-Laviada I. Δ9‐Tetrahydrocannabinol induces apoptosis in human prostate PC‐3 cells via a receptor‐independent mechanism. FEBS Lett.1999;458(3):400-4.
[6]Sánchez C, Galve-Roperh I, Canova C, Brachet Ph, Guzmán M. Δ9‐Tetrahydrocannabinol induces apoptosis in C6 glioma cells. FEBS Lett. 1998;436(1):6-10.
[7]Marselos M, Karamanakos P. Mutagenicity, developmental toxicity and carcinogenicity of cannabis. Addict Bio. 1999;4(1):5-12.
[8]Cavuoto P, Wittert GA. The Role of the Endocannabinoid System in the Regulation of Energy Expenditure. Best Pract Res Clin Endocrinol Metab. 2009;23(1):79-86.
[9]Borgelt LM, Franson KL, Nussbaum AM, Wang GS. The Pharmacologic and Clinical Effects of Medical Cannabis. Pharmacotherapy. 2013;33(2):195-209
[10]Svízenská I, Dubový P, Sulcová A. Cannabinoid Receptors 1 and 2 (CB1 and CB2), their distribution, ligands and functional involvement in nervous system structures--a short review. Pharmacol Biochem Behav. 2008;90(4):501-11.
[11]Biehl JK, Russel B. Introduction to stem cell therapy. J Cardiovasc Nurs. 2009;24(2):98-105.
[12]Poulos SP, Dodson MV, Hausman GJ. Cell line models for differentiation: Preadipocytes and adipocytes. Exp Bio Med. 2010;235(10):1185-93.
[13]Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem. 2006;98(5):1076-84.
[14]Pournassarkhakabaz B, Baharvand H. Human Mesenchymal Stem Cells and its therapeutic applications. Anat Sci J. 2007;5(19-20):167-215. [Persian]
[15]Taheri F, Haji Ghasem Kashani M, Ghorbanian MT, Hosseinpour L. Inductive effect of Deprenyl and Dimethyl sulfoxide on proliferation and survival of the mesenchymal stem cells. J Gorgan Univ Med Sci. 2012;14(3):10-8. [Persian]
[16]Ramezani T, Baharara J, Saghiri N. The Effect of Saffron Aqueous Extract (Crocus sativus L) on Osteogenic Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells. J Birjand Univ Med Sci. 2014;21(2):169-78.
[17]Polisetti N, Chaitanya VG, Babu PP, Vemuganti GK. Isolation, characterization and differentiation potential of rat bone marrow stromal cells. Neurol India. 2010;58(2):201-8.
[18]Karaoz E, Genç ZS, Demircan PÇ, Aksoy A, Duruksu G. Protection of rat pancreatic islet function and viability by coculture with rat bone marrow-derived mesenchymal stem cells. Cell Death Dis. 2010;1(4):36.
[19]Maxson S, Lopez EA, Yoo D, Danilkovitch-Miagkova A, LeRoux MA. Concise review: Role of mesenchymal stem cells in wound repair. Stem Cells Trans Med. 2012;1(2):142-9.
[20]Rahmanifar F, Tamadon A, Mehrabani D, Zare SH, Abasi S, Keshavarz S, et al. Histomorphometric evaluation of treatment of rat azoosper-mic seminiferous tubules by allotransplantation of bone marrow-derived mesenchymal stem cells. Iran J Basic Med Sci. 2016;19(6):653-61.
[21]Mahmoudi Z, Soleimani M, Saiedi A, Iranshahi M, Azizsoltanli A. Effect of Ferula gummosa ethanolic extract on osteogenesis in human mesenchymal stem cells. J Med Plants. 2013;2(46):50-9. [Persian]
[22]Rosen ED, Hsu CH, Wang X, Sakai S, Freeman MW, Gonzalez FJ, et al. C/EBPα induces adipogenesis through PPARγ: a unified pathway. Genes Dev. 2002;16(1):22-6.
[23]O'sullivan SE. Cannabinoids go nuclear: Evidence for activation of peroxisome proliferator‐activated receptors. Br J Pharmacol. 2007;152(5):576-82.
[24]Frisher M, White S, Varbiro G, Voisey C, Perumal D, Crome I, et al. The role of cannabis and cannabinoids in diabetes. Br J Diabetes Vasc Dis. 2010;10(6):267-73.
[25]McPartland JM. The endocannabinoid System: An osteopathic perspective. J Am Osteopath Assoc. 2008;108(10):586-600.
[26]Idris AI, Sophocleous A, Landao-Bassonga E, Canals M, Milligan G, Baker D, et al. Cannabinoid receptor type 1 protects against age-related osteoporosis by regulating osteoblast and adipocyte differentiation in marrow stromal cells. Cell Metab. 2009;10(2):139-47.
[27]Nick Bakhte Dastjerdi M. The Effect of Dexamethasone on the Differentiation of Bone Marrow Scaffold Cells into Fatty Cells. Isfahan Med Sch J. 2005;23(77-76):17. [Persian]
[28]Lombard C, Hegde VL, Nagarkatti M, Nagarkatti PS. Perinatal exposure to Δ9-tetrahydrocannabinol triggers profound defects in T cell differentiation and function in fetal and postnatal stages of life, including decreased responsiveness to HIV antigens. J Pharmacol Exp Ther. 2011;339(2):607-17.
[29]Takada I, Kouzmenko AP, Kato S. Molecular switching of osteoblastogenesis versus adipogenesis: implications for targeted therapies. Expert Opin Ther Targets. 2009;13(5):593-603.
[30]Cao JJ. Effects of obesity on bone metabolism. J Orthop Surg Res. 2011;6:30.