Human adipose-derived stromal cells (hASCs) are a promising cell source for bone tissue engineering. However, before the clinical application of hASCs for the treatment of bone defects, key questions require answers, including whether pre-osteoinduction (OI) and flow cytometric cell purification are indispensible steps for in vivo bone formation by hASCs. In this study, hASCs were purified by flow cytometric cell sorting (FCCS). The osteogenic capabilities of hASCs and purified hASCs with or without pre-osteoinduction were examined through in vitro and in vivo experiments. We found that pre-OI enhanced the in vitro osteogenic capacity of hASCs. However, 8 weeks after in vivo implantation, there were no significant differences between hASCs and hASCs that had undergone OI (hASCs+OI) or between purified hASCs and purified hASCs+OI (P>0.05). Interestingly, we also found that purified hASCs had an osteogenic potential similar to that of unpurified hASCs in vitro and in vivo. These results suggest that FCCS and in vitro pre-OI are not requirements for in vivo bone formation by hASCs.
References
[1]
Levi B, Longaker MT (2011) Concise review: adipose-derived stromal cells for skeletal regenerative medicine. Stem Cells 29 (4) 576–582.
[2]
Li X, Yao J, Wu L, Jing W, Tang W, et al. (2010) Osteogenic induction of adipose-derived stromal cells: not a requirement for bone formation in vivo. Artif Organs 34 (1) 46–54.
[3]
Liu Y, Zhou Y, Feng H, Ma GE, Ni Y (2008) Injectable tissue-engineered bone composed of human adipose-derived stromal cells and platelet-rich plasma. Biomaterials 29 (23) 3338–3345.
[4]
Zhou Y, Ni Y, Liu Y, Zeng B, Xu Y, et al. (2010) The role of simvastatin in the osteogenesis of injectable tissue-engineered bone based on human adipose-derived stromal cells and platelet-rich plasma. Biomaterials 31 (20) 5325–5335.
[5]
Levi B, James AW, Nelson ER, Vistnes D, Wu B, et al. (2010) Human adipose derived stromal cells heal critical size mouse calvarial defects. PLoS One 5 (6) e11177.
[6]
Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, et al. (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7 (2) 211–28.
[7]
Behr B, Tang C, Germann G, Longaker MT, Quarto N (2011) Locally applied vascular endothelial growth factor A increases the osteogenic healing capacity of human adipose-derived stem cells by promoting osteogenic and endothelial differentiation. Stem Cells 29 (2) 286–296.
[8]
Hong JM, Kim BJ, Shim JH, Kang KS, Kim KJ, et al. (2012) Enhancement of bone regeneration through facile surface functionalization of solid freeform fabrication-based three-dimensional scaffolds using mussel adhesive proteins. Acta Biomater 8 (7) 2578–2586.
[9]
Amos PJ, Bailey AM, Shang H, Katz AJ, Lawrence MB, et al. (2008) Functional binding of human adipose-derived stromal cells: effects of extraction method and hypoxia pretreatment. Ann Plast Surg 60 (4) 437–444.
[10]
Monaco E, Bionaz M, Hollister SJ, Wheeler MB (2011) Strategies for regeneration of the bone using porcine adult adipose-derived mesenchymal stem cells. Theriogenology 75 (8) 1381–1399.
[11]
Lu Z, Roohani-Esfahani SI, Wang G, Zreiqat H (2012) Bone biomimetic microenvironment induces osteogenic differentiation of adipose tissue-derived mesenchymal stem cells. Nanomedicine 8 (4) 507–515.
[12]
Correia C, Bhumiratana S, Yan LP, Oliveira AL, Gimble JM, et al. (2012) Development of silk-based scaffolds for tissue engineering of bone from human adipose-derived stem cells. Acta Biomater 8 (7) 2483–2492.
[13]
Ying X, Cheng S, Wang W, Lin Z, Chen Q, et al. (2012) Effect of lactoferrin on osteogenic differentiation of human adipose stem cells. Int Orthop 36 (3) 647–653.
[14]
Cheng NC, Chang HH, Tu YK, Young TH (2012) Efficient transfer of human adipose-derived stem cells by chitosan/gelatin blend films. J Biomed Mater Res B Appl Biomater 100 (5) 1369–1377.
[15]
Santo VE, Duarte AR, Popa EG, Gomes ME, Mano JF, et al. (2012) Enhancement of osteogenic differentiation of human adipose derived stem cells by the controlled release of platelet lysates from hybrid scaffolds produced by supercritical fluid foaming. J Control Release 162 (1) 19–27.
[16]
Kang JM, Han M, Park IS, Jung Y, Kim SH (2012) Adhesion and differentiation of adipose-derived stem cells on a substrate with immobilized fibroblast growth factor. Acta Biomater 8 (5) 1759–1767.
[17]
Locke M, Feisst V, Dunbar PR (2011) Concise Review: Human Adipose-Derived Stem Cells: Separating Promise from Clinical Need. Stem Cells 29 (3) 404–411.
[18]
Rada T, Reis RL, Gomes ME (2011) Distinct stem cells subpopulations isolated from human adipose tissue exhibit different chondrogenic and osteogenic differentiation potential. Stem Cell Rev 7 (1) 64–76.
[19]
Al Battah F, De Kock J, Vanhaecke T, Rogiers V (2011) Current status of human adipose-derived stem cells: differentiation into hepatocyte-like cells. ScientificWorldJournal 11: 1568–1581.
[20]
Zhou Y, Yan Z, Zhang H, Lu W, Liu S, et al. (2011) Expansion and delivery of adipose-derived mesenchymal stem cells on three microcarriers for soft tissue regeneration. Tissue Eng Part A 17 (23–24) 2981–2997.
[21]
Schaffler A, Buchler C (2007) Concise review: adipose tissue-derived stromal cells–basic and clinical implications for novel cell-based therapies. Stem Cells 25 (4) 818–827.
[22]
Gimble JM, Katz AJ, Bunnell BA (2007) Adipose-derived stem cells for regenerative medicine. Circ Res 100 (9) 1249–1260.
[23]
McIntosh K, Zvonic S, Garrett S, Mitchell JB, Floyd ZE, et al. (2006) The immunogenicity of human adipose-derived cells: temporal changes in vitro. Stem Cells 24 (5) 1246–1253.
[24]
Yuan J, Cui L, Zhang WJ, Liu W, Cao Y (2007) Repair of canine mandibular bone defects with bone marrow stromal cells and porous β-tricalcium phosphate. Biomaterials 28 (6) 1005–1013.
[25]
Szpalski C, Nguyen PD, Cretiu Vasiliu CE, Chesnoiu-Matei I, Ricci JL, et al. (2012) Bony engineering using time-release porous scaffolds to provide sustained growth factor delivery. J Craniofac Surg 23 (3) 638–644.
[26]
Li X, Liu H, Niu X, Fan Y, Feng Q, et al. (2011) Osteogenic differentiation of human adipose-derived stem cells induced by osteoinductive calcium phosphate ceramics. J Biomed Mater Res B Appl Biomater 97 (1) 10–19.
[27]
Csaki C, Matis U, Mobasheri A, Ye H, Shakibaei M (2007) Chondrogenesis, osteogenesis and adipogenesis of canine mesenchymal stem cells: a biochemical, morphological and ultrastructural study. Histochem Cell Biol 128 (6) 507–520.
[28]
Niemeyer P, Kornacker M, Mehlhorn A, Seckinger A, Vohrer J, et al. (2007) Comparison of immunological properties of bone marrow stromal cells and adipose tissue-derived stem cells before and after osteogenic differentiation in vitro. Tissue Eng 13 (1) 111–121.
[29]
Hofmann S, Hagenmuller H, Koch AM, Muller R, Vunjak-Novakovic G, et al. (2007) Control of in vitro tissue-engineered bone-like structures using human mesenchymal stem cells and porous silk scaffolds. Biomaterials 28 (6) 1152–1162.
[30]
Endres M, Hutmacher DW, Salgado AJ, Kaps C, Ringe J, et al. (2003) Osteogenic induction of human bone marrow-derived mesenchymal progenitor cells in novel synthetic polymer-hydrogel matrices. Tissue Eng 9 (4) 689–702.
[31]
Kim HP, Ji YH, Rhee SC, Dhong ES, Park SH, et al. (2012) Enhancement of bone regeneration using osteogenic-induced adipose-derived stem cells combined with demineralized bone matrix in a rat critically-sized calvarial defect model. Curr Stem Cell Res Ther 7 (3) 165–172.
[32]
Dudas JR, Marra KG, Cooper GM, Penascino VM, Mooney MP, et al. (2006) The osteogenic potential of adipose-derived stem cells for the repair of rabbit calvarial defects. Ann Plast Surg 56 (5) 543–548.
[33]
Cowan CM, Shi Y-Y, Aalami OO, Chou Y-F, Mari C, et al. (2004) Adipose-derived adult stromal cells heal critical-size mouse calvarial defects. Nat Biotechnol 22 (5) 560–567.
[34]
Miyahara Y, Nagaya N, Kataoka M, Yanagawa B, Tanaka K, et al. (2006) Monolayered mesenchymal stem cells repair scarred myocardium after myocardial infarction. Nat Med 12 (4) 459–465.
[35]
Phinney DG (2007) Biochemical heterogeneity of mesenchymal stem cell populations: clues to their therapeutic efficacy. Cell Cycle 6 (23) 2884–2889.
[36]
Cooper MS, Hewison M, Stewart PM (1999) Glucocorticoid activity, inactivity and the osteoblast. J Endocrinol 163 (2) 159–164.
[37]
Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, et al. (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13 (12) 4279–4295.
[38]
Niemeyer P, Vohrer J, Schmal H, Kasten P, Fellenberg J, et al. (2008) Survival of human mesenchymal stromal cells from bone marrow and adipose tissue after xenogenic transplantation in immunocompetent mice. Cytotherapy 10 (8) 784–795.
[39]
Coelho PG, Coimbra ME, Ribeiro C, Fancio E, Higa O, et al. (2009) Physico/chemical characterization and preliminary human histology assessment of a β-TCP particulate material for bone augmentation. Mater Sci Eng C Mater Biol Appl: C 29 (7) 2085–2091.
[40]
Morelhao SL, Coelho PG, Honnicke MG (2010) Synchrotron X-ray imaging via ultra-small-angle scattering: principles of quantitative analysis and application in studying bone integration to synthetic grafting materials. Eur Biophys J 39 (5) 861–865.
[41]
Xu J, Zheng Z, Fang D, Gao R, Liu Y, et al. (2012) Mesenchymal stromal cell-based treatment of jaw osteoradionecrosis in Swine. Cell Transplant 21 (8) 1679–1686.
[42]
Liu Y, Wang L, Kikuiri T, Akiyama K, Chen C, et al. (2011) Mesenchymal stem cell-based tissue regeneration is governed by recipient T lymphocytes via IFN-gamma and TNF-alpha. Nat Med 17 (12) 1594–1601.
[43]
Sakaguchi Y, Sekiya I, Yagishita K, Muneta T (2005) Comparison of human stem cells derived from various mesenchymal tissues: superiority of synovium as a cell source. Arthritis Rheum 52 (8) 2521–2529.
[44]
Zhou YS, Liu YS, Tan JG (2006) Is 1, 25-dihydroxyvitamin D3 an ideal substitute for dexamethasone for inducing osteogenic differentiation of human adipose tissue-derived stromal cells in vitro? Chin Med J (Engl) 119 (15) 1278–1286.
[45]
Ge W, Shi L, Zhou Y, Liu Y, Ma GE, et al. (2011) Inhibition of osteogenic differentiation of human adipose-derived stromal cells by retinoblastoma binding protein 2 repression of RUNX2-activated transcription. Stem Cells 29 (7) 1112–1125.
[46]
Fan Z, Yamaza T, Lee JS, Yu J, Wang S, et al. (2009) BCOR regulates mesenchymal stem cell function by epigenetic mechanisms. Nat Cell Biol 11 (8) 1002–1009.
