Abstract
Osteoporosis due to estrogen deficiency is an increasing bone health issue worldwide: new strategies are being studied for regenerative medicine of bone pathologies in these patients. The most commonly used cells for tissue engineering therapy are the bone marrow mesenchymal stem cells (BMSCs), but they might be negatively affected by aging and estrogen deficiency. Besides the general advantages of adipose-derived mesenchymal stem cells (ADSCs) over BMSCs, ADSCs also seem to be less affected by aging than BMSCs, but in the literature, little is known about ADSCs in estrogen deficiency. The present study investigated the in vitro behavior of ADSCs, isolated from healthy (SHAM) and estrogen-deficient (OVX) rats. Phenotype, clonogenicity, viability, and osteogenic differentiation, at both cellular and molecular levels, were evaluated with or without osteogenic stimuli. Pro-inflammatory cytokines, growth factors, and adipogenic differentiation markers were also analyzed. There were no significant differences between OVX and SHAM ADSCs in some analyzed parameters. In addition, clonogenicity, osteopontin (Spp1) gene expression, alkaline phosphatase (ALP) activity at 2 weeks of culture, total collagen (COLL), osteocalcin (Bglap) gene expression and production, and matrix mineralization were significantly higher in OVX than in SHAM ADSCs. Besides the increase in some osteogenic markers, peroxisome proliferator-activated receptor gamma (Pparg) gene was also more expressed in OVX in osteogenic medium, with a concomitant estrogen receptor 1 (Esr1) gene expression decrease. These results underlined that ADSCs were not affected by estrogen deficiency in an osteogenic microenvironment.
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Aldahmash A, Zaher W, Al-Nbaheen M, Kassem M (2012) Human stromal (mesenchymal) stem cells: basic biology and current clinical use for tissue regeneration. Ann Saudi Med 32:68–77
Benisch P, Schilling T, Klein-Hitpass L, Frey SP, Seefried L, Raaijmakers N, Krug M, Regensburger M, Zeck S, Schinke T, Amling M, Ebert R, Jakob F (2012) The transcriptional profile of mesenchymal stem cell population in primary osteoporosis is distinct and shows overexpression of osteogenic inhibitors. Plos One 7:e45142. doi:10.1371/journal.pone.0045142
Bonofiglio D, Gabriele S, Aquila S, Catalano S, Gentile M, Middea E, Giordano F, Antò S (2005) Estrogen receptor alpha binds to peroxisome proliferator-activated receptor response element and negatively interferes with peroxisome proliferator-activated receptor gamma signaling in breast cancer cells. Clin Cancer Res 11:6139–6147
Boyce BF, Rosenberg E, de Papp AE, le Duong T (2012) The osteoclast, bone remodelling and treatment of metabolic bone disease. Eur J Clin Invest 42:1332–1341. doi:10.1111/j.1365-2362.2012.02717.x
Calleja-Agius J, Brincat MP (2009) Effects of hormone replacement therapy on connective tissue: why is this important? Best Pract Res Clin Obstet Gynaecol 23:121–127. doi:10.1016/j.bpobgyn.2008.10.003
Chen HT, Lee MJ, Chen CH, Chuang SC, Chang LF, Ho ML, Hung SH, Fu YC, Wang YH, Wang HI, Wang GJ, Kang L, Chang JK (2012) Proliferation and differentiation potential of human adipose-derived mesenchymal stem cells isolated from elderly patients with osteoporotic fractures. J Cell Mol Med 16:582–592. doi:10.1111/j.1582-4934.2011.01335.x
Cui G, Leng H, Wang K, Wang J, Zhu S, Jia J, Chen X, Zhang W, Qin L, Bai W (2013) Effects of remifemin treatment on bone integrity and remodeling in rats with ovariectomy-induced osteoporosis. PLoS One 8:e82815. doi:10.1371/journal.pone.0082815
Dalle Carbonare L, Valenti MT, Zanatta M, Donatelli L, Lo Cascio V (2009) Circulating mesenchymal stem cells with abnormal osteogenic differentiation in patients with osteoporosis. Arthritis Rheum 60:3356–3365. doi:10.1002/art.24884
de Girolamo L, Lopa S, Arrigoni E, Sartori MF, Baruffaldi Preis FW, Brini AT (2009) Human adipose-derived stem cells isolated from young and elderly women: their differentiation potential and scaffold interaction during in vitro osteoblastic differentiation. Cytotherapy 11:793–803. doi:10.3109/14653240903079393
Fei J, Tamski H, Cook C, Santanam N (2013) MicroRNA regulation of adipose derived stem cells in aging rats. Plos One 8:1–11. doi:10.1371/journal.pone.0059238
Fraser JK, Wulur I, Alfonso Z, Hedrick MH (2006) Fat tissue: an underappreciated source of stem cells for biotechnology. Trends Biotechnol 24:150–154
Giavaresi G, Fini M, Giardino R, Salamanna F, Sartori M, Borsari V, Spriano S, Bellini CM, Brayda-Bruno M (2011) In vivo preclinical evaluation of the influence of osteoporosis on the anchorage of different pedicle screw designs. Eur Spine J 20:1289–1296. doi:10.1007/s00586-011-1831-5
Gimble JM, Nuttall ME (2012) The relationship between adipose tissue and bone metabolism. Clin Biochem 45:874–879. doi:10.1016/j.clinbiochem.2012.03.006
Hoch AI, Binder BY, Genetos DC, Leach JK (2012) Differentiation-dependent secretion of proangiogenic factors by mesenchymal stem cells. PLoS One 7:e35579. doi:10.1371/journal.pone.0035579
Hoogendoorn RJ, Lu ZF, Kroeze RJ, Bank RA, Wuisman PI, Helder MN (2008) Adipose stem cells for intervertebral disc regeneration: current status and concepts for the future. J Cell Mol Med 12:2205–2216. doi:10.1111/j.1582-4934.2008.00291.x
Hoshiba T, Kawazoe N, Chen G (2012) The balance of osteogenic and adipogenic differentiation in human mesenchymal stem cells by matrices that mimic stepwise tissue development. Biomaterials 33:2015–2031. doi:10.1016/j.biomaterials.2011.11.061
Khan WS, Adesida AB, Tew SR, Andrew JG, Hardingham TE (2009) The epitope characterization and the osteogenic differentiation potential of human fat pad-derived stem cells is maintained with ageing in later life. Injury 40:150–157. doi:10.1016/j.injury.2008.05.029
Kim D, Monaco E, Maki A, de Lima AS, Kong HJ, Hurley WL, Wheeler MB (2010) Morphologic and transcriptomic comparison of adipose- and bone-marrow-derived porcine stem cells cultured in alginate hydrogels. Cell Tissue Res 341:359–370. doi:10.1007/s00441-010-1015-3
Kim H, Lee K, Ko CY, Kim HS, Shin HI, Kim T, Lee SH, Jeong D (2012) The role of nacreous factors in preventing osteoporotic bone loss through both osteoblast activation and osteoclast inactivation. Biomaterials 33:7489–7496. doi:10.1016/j.biomaterials.2012.06.098
Li Y, Zou S, Wang D, Feng G, Bao C, Hu J (2010) The effect of hydrofluoric acid treatment on titanium implant osseointegration in ovariectomized rats. Biomaterials 31:3266–3273. doi:10.1016/j.biomaterials.2010.01.028
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408
Mirsaidi A, Kleinhans KN, Rimann M, Tiaden AN, Stauber M, Rudolph KL, Richards PJ (2012) Telomere length, telomerase activity and osteogenic differentiation are maintained in adipose-derived stromal cells from senile osteoporotic SAMP6 mice. J Tissue Eng Regen Med 6:378–390. doi:10.1002/term.440
Mojallal A, Lequeux C, Shipkov C, Duclos A, Braye F, Rohrich R, Brown S, Damour O (2011) Influence of age and body mass index on the yield and proliferation capacity of adipose-derived stem cells. Aesthetic Plast Surg 35:1097–1105. doi:10.1007/s00266-011-9743-7
Moon SJ, Ahn IE, Jung H, Yi H, Kim J, Kim Y, Kwok SK, Park KS, Min JK, Park SH, Kim HY, Ju JH (2013) Temporal differential effects of proinflammatory cytokines on osteoclastogenesis. Int J Mol Med 31:769–777. doi:10.3892/ijmm.2013.1269
Pachon-Pena G, Yu G, Tucker A, Wu X, Vendrell J, Bunnell BA, Gimble JM (2010) Stromal stem cells from adipose tissue and bone marrow of age-matched female donors display distinct immunophenotypic profiles. J Cell Physiol 226:843–851. doi:10.1002/jcp.22408
Pino AM, Rosen CJ, Rodriguez JP (2012) In osteoporosis, differentiation of mesenchymal stem cells (MSCs) improves bone marrow adipogenesis. Biol Res 45:279–287. doi:10.4067/S0716-97602012000300009
Ray R, Novotny NM, Crisostomo PR, Lahm T, Abarbanell A, Meldrum DR (2008) Sex steroids and stem cell function. Mol Med 14:493–501. doi:10.2119/2008-00004.Ray
Rodriguez JP, Astudillo P, Rios S, Pino AM (2008) Involvement of adipogenic potential of human bone marrow mesenchymal stem cells (MSCs) in osteoporosis. Curr Stem Cell Res Ther 3:208–218
Rodriguez JP, Rios S, Fernandez M, Santibanez F (2004) Differential activation of ERK1,2 MAP kinase signaling pathway in mesenchymal stem cells from control and osteoporotic postmenopausal women. J Cell Biochem 92:745–754
Shi YY, Nacamuli RP, Salim A, Longaker MT (2005) The osteogenic potential of adipose-derived mesenchymal stem cells is maintained with aging. Plast Reconstruct Surg 116:1686–1696
Smith BJ, Bu SY, Wang Y, Rendina E, Lim YF, Marlow D, Clarke SL, Cullen DM, Lucas EA (2014) A comparative study of the bone metabolic response to dried plum supplementation and PTH treatment in adult, osteopenic ovariectomized rat. Bone 58:151–159. doi:10.1016/j.bone.2013.10.005
Tobita M, Orbay H, Mizuno H (2011) Adipose-derived stem cells: current findings and future perspectives. Discov Med 11:160–170
Torricelli P, Veronesi F, Pagani S, Maffulli N, Masiero S, Frizziero A, Fini M (2012) In vitro tenocyte metabolism in aging and oestrogen deficiency. Age 35:2125–2136. doi:10.1007/s11357-012-9500-0
Valenti MT, Garbin U, Pasini A, Zanatta M, Stranieri C, Manfro S, Zucal C, Dalle Carbonare L (2011) Role of Ox-PPACs in the differentiation of mesenchymal stem cells (MSCs) and Runx2 and PPARγ2 expression in MSCs-like of osteoporotic patients. Plos One 6:e20363. doi:10.1371/journal.pone.0020363
Veronesi F, Torricelli P, Borsari V, Tschon M, Rimondini L, Fini M (2011) Mesenchymal stem cells in aging and osteoporotic population. Crit Rev Eukaryot Gene Expr 21:363–377
Wan Y (2010) PPARγ in bone homeostasis. Trends Endocrinol Metab 21:722–728. doi:10.1016/j.tem.2010.08.006
Wu W, Niklason L, Steinbacher DM (2013) The effect of age on human adipose-derived stem cells. Plast Reconstr Surg 131:27–37. doi:10.1097/PRS.0b013e3182729cfc
Zhu M, Kohan E, Bradley J, Hedrick M, Benhaim P, Zuk P (2009) The effect of age on osteogenic, adipogenic and proliferative potential of female adipose-derived stem cells. J Tissue Eng Reg Med 3:290–301. doi:10.1002/term.165
Acknowledgments
This paper was partially supported by Rizzoli Orthopaedic Institute, “5 PER MILLE Project” (CUPD31J09000260001). The authors wish to thank Dr. Luca Cattini (Laboratory of Immuno-rheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopedic Research Institute) for the valuable contribution regarding flow cytometry characterization.
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Veronesi, F., Pagani, S., Della Bella, E. et al. Estrogen deficiency does not decrease the in vitro osteogenic potential of rat adipose-derived mesenchymal stem cells. AGE 36, 9647 (2014). https://doi.org/10.1007/s11357-014-9647-y
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DOI: https://doi.org/10.1007/s11357-014-9647-y