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Esmailpour, Z., Madadi, S., & Baazm, M. (2024). The antiapoptotic effects of conditioned medium from bone marrow-derived mesenchymal stromal stem cells on cyclophosphamide-induced testicular damage in rat: An experimental study. International Journal of Reproductive BioMedicine (IJRM), 22(2), 89–100. https://doi.org/10.18502/ijrm.v22i2.15706
https://doi.org/10.18502/ijrm.v22i2.15706
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authors
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Zeynab Esmailpour
Zeynab Esmailpour
Students Research Committee, Arak University of Medical Sciences, Arak, Iran.
-
Soheila Madadi
Soheila Madadi
Department of Anatomy, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran.
-
Maryam Baazm
Maryam Baazm
Department of Anatomy, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
Published Date
- Mar 25, 2024
The antiapoptotic effects of conditioned medium from bone marrow-derived mesenchymal stromal stem cells on cyclophosphamide-induced testicular damage in rat: An experimental study
Abstract
Background: Cyclophosphamide (CP) has some negative effects on the reproductive system. Stem cells and their metabolites are being utilized to enhance fertility after chemotherapy.
Objective: This study aimed to investigate the impact of conditioned medium (CM) derived from bone marrow mesenchymal stromal stem cells (BM-MSCs) on the toxic effects of CP on testicles.
Materials and Methods: BM-MSCs were isolated, a CM was collected and 25-fold concentrated. 24 male Wistar rats (8 wk, 200–250 gr) were randomly divided into following groups: control, CP, CP+DMEM, CP+CM. CP was given at a single dose of 100 mg/kg. 2 wk after the CP administration, CM was injected into the testicular efferent duct. Sperm parameters, testicular histopathology, and the level of testosterone were analyzed 2 months after treatment. The expression of B-cell lymphoma 2 (Bcl2) and Bcl2-associated X protein (Bax) genes were evaluated by real-time polymerase chain reaction.
Results: CP had a negative effect on testis histology (p < 0.001) and sperm quality (p < 0.001). It changed the expression of genes associated with apoptosis (p < 0.001). Treatment with CM reduced the expression of Bax (p < 0.001), while significantly increasing the expression of Bcl2 (p = 0.01). It improved sperm count (p = 0.03), viability (p < 0.001), motility (p < 0.001), spermatogonial count (p < 0.001), and epithelial thickness of testicular tubules (p = 0.02).
Conclusion: These findings suggest that CM produced from BM-MSCs may be valuable for therapeutic approaches in reproductive medicine and may lessen the side effects of CP.
Key words: Bone marrow mesenchymal stem cells, Cyclophosphamide, Conditioned medium, Apoptosis, Spermatogenesis.
References
[1] El Gharabawy GS, Abd Allah EE-DE, Amr IM, Elmitwalli M. Histological and immunohistochemical study of the effect of cyclophosphamide on testis of male adult albino rats and the possible protective role of vitamin E. Egypt J Hospital Med 2019; 77: 5930–5946.
[2] Fusco R, Salinaro AT, Siracusa R, D’Amico R, Impellizzeri D, Scuto M, et al. Hidrox® counteracts cyclophosphamideinduced male infertility through NRF2 pathways in a mouse model. Antioxidants 2021; 10: 778.
[3] Abram McBride J, Lipshultz LI. Male fertility preservation. Curr Urol Rep 2018; 19: 49.
[4] Yuan Q-L, Zhang Y-G, Chen Q. Mesenchymal stem cell (MSC)−derived extracellular vesicles: Potential therapeutics as MSC trophic mediators in regenerative medicine. Anat Rec 2020; 303: 1735–1742.
[5] Sagaradze G, Grigorieva O, Nimiritsky P, Basalova N, Kalinina N, Akopyan Z, et al. Conditioned medium from human mesenchymal stromal cells: Towards the clinical translation. Int J Mol Sci 2019; 20: 1656.
[6] Benavides-Castellanos MP, Garzón-Orjuela N, Linero I. Effectiveness of mesenchymal stem cell-conditioned medium in bone regeneration in animal and human models: A systematic review and meta-analysis. Cell Regen 2020; 9: 5.
[7] Veronesi F, Borsari V, Sartori M, Orciani M, Mattioli−Belmonte M, Fini M. The use of cell conditioned medium for musculoskeletal tissue regeneration. J Cell Physiol 2018; 233: 4423–4442.
[8] Ogata K, Katagiri W, Osugi M, Kawai T, Sugimura Y, Hibi H, et al. Evaluation of the therapeutic effects of conditioned media from mesenchymal stem cells in a rat bisphosphonate-related osteonecrosis of the jaw-like model. Bone 2015; 74: 95–105.
[9] Cai Y-T, Xiong C-L, Shen S-L, Rao J-P, Liu T-S, Qiu F. Mesenchymal stem cell−secreted factors delayed spermatogenesis injuries induced by busulfan involving intercellular adhesion molecule regulation. Andrologia 2019; 51: e13285.
[10] Abdollahifar M-A, Azad N, Faraji Sani M, Raoofi A, Abdi S, Aliaghaei A, et al. Impaired spermatogenesis caused by busulfan is partially ameliorated by treatment with conditioned medium of adipose tissue derived mesenchymal stem cells. Biotech Histochem 2022; 97: 107–117.
[11] Mahiddine FY, Kim JW, Qamar AY, Ra JC, Kim SH, Jung EJ, et al. Conditioned medium from canine amniotic membrane-derived mesenchymal stem cells improved dog sperm post-thaw quality-related parameters. Animals 2020; 10: 1899.
[12] Shah SM, Saini N, Singh MK, Manik R, Singla SK, Palta P, et al. Testicular cell-conditioned medium supports embryonic stem cell differentiation toward germ lineage and to spermatocyte-and oocyte-like cells. Theriogenology 2016; 86: 715–729.
[13] Akyash F, Aflatoonian R, Farashahi Yazd E, Golzadeh J, Tahajjodi S, Moore H, et al. Testicular sperm extraction derived cells conditioned medium as an in vitro niche supports germ cells development from human embryonic stem cells. 35th Annual Meeting of the European Society for Human Reproduction and Embryology; 24–26 June 2019; Vienna, Austria. 2019; 34: i333.
[14] Khanmohammadi N, Sameni HR, Mohammadi M, Pakdel A, Mirmohammadkhani M, Parsaie H, et al. Effect of transplantation of bone marrow stromal cell-conditioned medium on ovarian function, morphology and cell death in cyclophosphamide-treated rats. Cell J 2018; 20: 10–18.
[15] Nihashi Y, Yamamoto M, Shimosato T, Takaya T. Myogenetic oligodeoxynucleotide restores differentiation and reverses inflammation of myoblasts aggravated by cancer-conditioned medium. Muscles 2022; 1: 111–120.
[16] Rezaei Sh, Hosseinimehr SJ, Zargari M, Karimpour Malekshah A, Mirzaei M, Talebpour Amiri F. Protective effects of sinapic acid against cyclophosphamide−induced testicular toxicity via inhibiting oxidative stress, caspase−3 and NF−kB activity in BALB/c mice. Andrologia 2021; 53: e14196.
[17] Aghaie S, Nikzad H, Amini Mahabadi J, Taghizadeh M, Azami-Tameh A, Taherian A, et al. Protective effect of combined pumpkin seed and ginger extracts on sperm characteristics, biochemical parameters and epididymal histology in adult male rats treated with cyclophosphamide. Anat Sci Int 2016; 91: 382–390.
[18] Gul M, Hildorf S, Dong L, Thorup J, Hoffmann ER, Jensen CFS, et al. Review of injection techniques for spermatogonial stem cell transplantation. Hum Reprod Update 2020; 26: 368–391.
[19] Mizoguchi T. [Characterization of bone marrow mesenchymal stem cells]. Clin Calcium 2017; 27: 779–787. (in Japanese)
[20] Zohour Soleimani M, Jalali Mashayekhi F, Mousavi Hasanzade M, Baazm M. Alteration in CatSper1 and 2 genes expression, sperm parameters and testis histology in varicocelized rats. Int J Reprod BioMed 2018; 16: 183– 190.
[21] Jalili C, Kamani M, Roshankhah S, Sadeghi H, Salahshoor MR. Effect of Falcaria vulgaris extracts on sperm parameters in diabetic rats. Andrologia 2018; 50: e13130.
[22] Hemati U, Moshajari M, Jalali Mashayekhi F, Bayat M, Moslemi A, Baazm M. The effect of curcumin on NRF2/Keap1 signalling pathway in the epididymis of mouse experimental cryptorchidism. Andrologia 2022; 54: e14532.
[23] Babaei A, Kheradmand N, Baazm M, Nejati N, Khalatbari M. Protective effect of vitamin E on sperm parameters in rats infected with Candida albicans. Andrologia 2020; 52: e13593.
[24] Gassei K, Orwig KE. Experimental methods to preserve male fertility and treat male factor infertility. Fertil Steril 2016; 105: 256–266.
[25] Picton HM, Wyns Ch, Anderson RA, Goossens E, Jahnukainen K, Kliesch S, et al. A European perspective on testicular tissue cryopreservation for fertility preservation in prepubertal and adolescent boys. Hum Reprod 2015; 30: 2463–2475.
[26] Anan HH, Wahba NS, Abdallah MA, Mohamed DA. Histological and immunohistochemical study of cyclophosphamide effect on adult rat testis. Int J Sci Rep 2017; 3: 39–48.
[27] Afkhami−Ardakani M, Hasanzadeh Sh, Shahrooz R, Delirezh N, Malekinejad H. Spirulina platensis (Arthrospira platensis) attenuates cyclophosphamide−induced reproductive toxicity in male Wistar rats: Evidence for sperm apoptosis and p53/Bcl−2 expression. J Food Biochem 2021; 45: e13854.
[28] Alkhalaf MI, Alansari WS, Alshubaily FA, Alnajeebi AM, Eskandrani AA, Tashkandi MA, et al. Chemoprotective effects of inositol hexaphosphate against cyclophosphamide-induced testicular damage in rats. Sci Rep 2020; 10: 12599.
[29] Shittu SA, Shittu Sh-T, Akindele OO, Kunle-Alabi OT, Raji Y. Protective action of N-acetylcysteine on sperm quality in cyclophosphamide-induced testicular toxicity in male Wistar rats. JBRA Assist Reprod 2019; 23: 83–90.
[30] Ghobadi E, Moloudizargari M, Asghari MH, Abdollahi M. The mechanisms of cyclophosphamide-induced testicular toxicity and the protective agents. Expert Opin Drug Metab Toxicol 2017; 13: 525–536.
[31] Al-Dhalemi DM, Al-Hedaby FM. Evaluation of protective effect of cardamomum extraction against cyclophosphamide induced testicular degeneration. Biochem Cell Arch 2021; 21: 4231.
[32] Sadeghzadeh F, Sadeghzadeh A, Changizi-Ashtiyani S, Bakhshi S, Mashayekhi FJ, Mashayekhi M, et al. The effect of hydro-alcoholic extract of Ceratonia Silique L. on spermatogenesis index in rats treated with cyclophosphamide: An experimental study. Int J Reprod BioMed 2020; 18: 295–306.
[33] Cetinkaya-Un B, Un B, Akpolat M, Andic F, Yazir Y. Human amnion membrane-derived mesenchymal stem cells and conditioned medium can ameliorate X-irradiation-induced testicular injury by reducing endoplasmic reticulum stress and apoptosis. Reprod Sci 2022; 29: 944–954.
[34] Liang X, Lin F, Ding Y, Zhang Y, Li M, Zhou X, et al. Conditioned medium from induced pluripotent stem cellderived mesenchymal stem cells accelerates cutaneous wound healing through enhanced angiogenesis. Stem Cell Res Ther 2021; 12: 295.
[35] Sharifian P, Yari S, Hasanein P, Manteghi Nezhad Y. Conditioned medium of bone marrow mesenchymal stem cells improves sperm parameters and reduces histological alteration in rat testicular ischaemia/reperfusion model. Andrologia 2022; 54: e14624.
[36] Saleem R, Mohamed-Ahmed S, Elnour R, Berggreen E, Mustafa K, Al-Sharabi N. Conditioned medium from bone marrow mesenchymal stem cells restored oxidative stressrelated impaired osteogenic differentiation. Int J Mol Sci 2021; 22: 13458.
[37] Qiao H, Zhang R, Gao L, Guo Y, Wang J, Zhang R, et al. Molecular imaging for comparison of different growth factors on bone marrow-derived mesenchymal stromal cells’ survival and proliferation in vivo. BioMed Res Int 2016; 2016: 1363902.
[2] Fusco R, Salinaro AT, Siracusa R, D’Amico R, Impellizzeri D, Scuto M, et al. Hidrox® counteracts cyclophosphamideinduced male infertility through NRF2 pathways in a mouse model. Antioxidants 2021; 10: 778.
[3] Abram McBride J, Lipshultz LI. Male fertility preservation. Curr Urol Rep 2018; 19: 49.
[4] Yuan Q-L, Zhang Y-G, Chen Q. Mesenchymal stem cell (MSC)−derived extracellular vesicles: Potential therapeutics as MSC trophic mediators in regenerative medicine. Anat Rec 2020; 303: 1735–1742.
[5] Sagaradze G, Grigorieva O, Nimiritsky P, Basalova N, Kalinina N, Akopyan Z, et al. Conditioned medium from human mesenchymal stromal cells: Towards the clinical translation. Int J Mol Sci 2019; 20: 1656.
[6] Benavides-Castellanos MP, Garzón-Orjuela N, Linero I. Effectiveness of mesenchymal stem cell-conditioned medium in bone regeneration in animal and human models: A systematic review and meta-analysis. Cell Regen 2020; 9: 5.
[7] Veronesi F, Borsari V, Sartori M, Orciani M, Mattioli−Belmonte M, Fini M. The use of cell conditioned medium for musculoskeletal tissue regeneration. J Cell Physiol 2018; 233: 4423–4442.
[8] Ogata K, Katagiri W, Osugi M, Kawai T, Sugimura Y, Hibi H, et al. Evaluation of the therapeutic effects of conditioned media from mesenchymal stem cells in a rat bisphosphonate-related osteonecrosis of the jaw-like model. Bone 2015; 74: 95–105.
[9] Cai Y-T, Xiong C-L, Shen S-L, Rao J-P, Liu T-S, Qiu F. Mesenchymal stem cell−secreted factors delayed spermatogenesis injuries induced by busulfan involving intercellular adhesion molecule regulation. Andrologia 2019; 51: e13285.
[10] Abdollahifar M-A, Azad N, Faraji Sani M, Raoofi A, Abdi S, Aliaghaei A, et al. Impaired spermatogenesis caused by busulfan is partially ameliorated by treatment with conditioned medium of adipose tissue derived mesenchymal stem cells. Biotech Histochem 2022; 97: 107–117.
[11] Mahiddine FY, Kim JW, Qamar AY, Ra JC, Kim SH, Jung EJ, et al. Conditioned medium from canine amniotic membrane-derived mesenchymal stem cells improved dog sperm post-thaw quality-related parameters. Animals 2020; 10: 1899.
[12] Shah SM, Saini N, Singh MK, Manik R, Singla SK, Palta P, et al. Testicular cell-conditioned medium supports embryonic stem cell differentiation toward germ lineage and to spermatocyte-and oocyte-like cells. Theriogenology 2016; 86: 715–729.
[13] Akyash F, Aflatoonian R, Farashahi Yazd E, Golzadeh J, Tahajjodi S, Moore H, et al. Testicular sperm extraction derived cells conditioned medium as an in vitro niche supports germ cells development from human embryonic stem cells. 35th Annual Meeting of the European Society for Human Reproduction and Embryology; 24–26 June 2019; Vienna, Austria. 2019; 34: i333.
[14] Khanmohammadi N, Sameni HR, Mohammadi M, Pakdel A, Mirmohammadkhani M, Parsaie H, et al. Effect of transplantation of bone marrow stromal cell-conditioned medium on ovarian function, morphology and cell death in cyclophosphamide-treated rats. Cell J 2018; 20: 10–18.
[15] Nihashi Y, Yamamoto M, Shimosato T, Takaya T. Myogenetic oligodeoxynucleotide restores differentiation and reverses inflammation of myoblasts aggravated by cancer-conditioned medium. Muscles 2022; 1: 111–120.
[16] Rezaei Sh, Hosseinimehr SJ, Zargari M, Karimpour Malekshah A, Mirzaei M, Talebpour Amiri F. Protective effects of sinapic acid against cyclophosphamide−induced testicular toxicity via inhibiting oxidative stress, caspase−3 and NF−kB activity in BALB/c mice. Andrologia 2021; 53: e14196.
[17] Aghaie S, Nikzad H, Amini Mahabadi J, Taghizadeh M, Azami-Tameh A, Taherian A, et al. Protective effect of combined pumpkin seed and ginger extracts on sperm characteristics, biochemical parameters and epididymal histology in adult male rats treated with cyclophosphamide. Anat Sci Int 2016; 91: 382–390.
[18] Gul M, Hildorf S, Dong L, Thorup J, Hoffmann ER, Jensen CFS, et al. Review of injection techniques for spermatogonial stem cell transplantation. Hum Reprod Update 2020; 26: 368–391.
[19] Mizoguchi T. [Characterization of bone marrow mesenchymal stem cells]. Clin Calcium 2017; 27: 779–787. (in Japanese)
[20] Zohour Soleimani M, Jalali Mashayekhi F, Mousavi Hasanzade M, Baazm M. Alteration in CatSper1 and 2 genes expression, sperm parameters and testis histology in varicocelized rats. Int J Reprod BioMed 2018; 16: 183– 190.
[21] Jalili C, Kamani M, Roshankhah S, Sadeghi H, Salahshoor MR. Effect of Falcaria vulgaris extracts on sperm parameters in diabetic rats. Andrologia 2018; 50: e13130.
[22] Hemati U, Moshajari M, Jalali Mashayekhi F, Bayat M, Moslemi A, Baazm M. The effect of curcumin on NRF2/Keap1 signalling pathway in the epididymis of mouse experimental cryptorchidism. Andrologia 2022; 54: e14532.
[23] Babaei A, Kheradmand N, Baazm M, Nejati N, Khalatbari M. Protective effect of vitamin E on sperm parameters in rats infected with Candida albicans. Andrologia 2020; 52: e13593.
[24] Gassei K, Orwig KE. Experimental methods to preserve male fertility and treat male factor infertility. Fertil Steril 2016; 105: 256–266.
[25] Picton HM, Wyns Ch, Anderson RA, Goossens E, Jahnukainen K, Kliesch S, et al. A European perspective on testicular tissue cryopreservation for fertility preservation in prepubertal and adolescent boys. Hum Reprod 2015; 30: 2463–2475.
[26] Anan HH, Wahba NS, Abdallah MA, Mohamed DA. Histological and immunohistochemical study of cyclophosphamide effect on adult rat testis. Int J Sci Rep 2017; 3: 39–48.
[27] Afkhami−Ardakani M, Hasanzadeh Sh, Shahrooz R, Delirezh N, Malekinejad H. Spirulina platensis (Arthrospira platensis) attenuates cyclophosphamide−induced reproductive toxicity in male Wistar rats: Evidence for sperm apoptosis and p53/Bcl−2 expression. J Food Biochem 2021; 45: e13854.
[28] Alkhalaf MI, Alansari WS, Alshubaily FA, Alnajeebi AM, Eskandrani AA, Tashkandi MA, et al. Chemoprotective effects of inositol hexaphosphate against cyclophosphamide-induced testicular damage in rats. Sci Rep 2020; 10: 12599.
[29] Shittu SA, Shittu Sh-T, Akindele OO, Kunle-Alabi OT, Raji Y. Protective action of N-acetylcysteine on sperm quality in cyclophosphamide-induced testicular toxicity in male Wistar rats. JBRA Assist Reprod 2019; 23: 83–90.
[30] Ghobadi E, Moloudizargari M, Asghari MH, Abdollahi M. The mechanisms of cyclophosphamide-induced testicular toxicity and the protective agents. Expert Opin Drug Metab Toxicol 2017; 13: 525–536.
[31] Al-Dhalemi DM, Al-Hedaby FM. Evaluation of protective effect of cardamomum extraction against cyclophosphamide induced testicular degeneration. Biochem Cell Arch 2021; 21: 4231.
[32] Sadeghzadeh F, Sadeghzadeh A, Changizi-Ashtiyani S, Bakhshi S, Mashayekhi FJ, Mashayekhi M, et al. The effect of hydro-alcoholic extract of Ceratonia Silique L. on spermatogenesis index in rats treated with cyclophosphamide: An experimental study. Int J Reprod BioMed 2020; 18: 295–306.
[33] Cetinkaya-Un B, Un B, Akpolat M, Andic F, Yazir Y. Human amnion membrane-derived mesenchymal stem cells and conditioned medium can ameliorate X-irradiation-induced testicular injury by reducing endoplasmic reticulum stress and apoptosis. Reprod Sci 2022; 29: 944–954.
[34] Liang X, Lin F, Ding Y, Zhang Y, Li M, Zhou X, et al. Conditioned medium from induced pluripotent stem cellderived mesenchymal stem cells accelerates cutaneous wound healing through enhanced angiogenesis. Stem Cell Res Ther 2021; 12: 295.
[35] Sharifian P, Yari S, Hasanein P, Manteghi Nezhad Y. Conditioned medium of bone marrow mesenchymal stem cells improves sperm parameters and reduces histological alteration in rat testicular ischaemia/reperfusion model. Andrologia 2022; 54: e14624.
[36] Saleem R, Mohamed-Ahmed S, Elnour R, Berggreen E, Mustafa K, Al-Sharabi N. Conditioned medium from bone marrow mesenchymal stem cells restored oxidative stressrelated impaired osteogenic differentiation. Int J Mol Sci 2021; 22: 13458.
[37] Qiao H, Zhang R, Gao L, Guo Y, Wang J, Zhang R, et al. Molecular imaging for comparison of different growth factors on bone marrow-derived mesenchymal stromal cells’ survival and proliferation in vivo. BioMed Res Int 2016; 2016: 1363902.