Effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture

doi:10.3969/j.issn.2095-4344.2014.47.009

Abstract

Abstract:

BACKGROUND: Previous researches have focused on the effect of phosphorus compounds on stem cells from animals or from human. But there is no study on the effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture.
OBJECTIVE: To explore the effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture.
METHODS: There were six groups in the experiment. Human bone marrow mesenchymal stem cells were inoculated in three-dimensional polystyrene scaffolds and then subjected to serum-free growth medium (group 3-GM) or serum-free growth medium containing 4 mmol/L (group 3-4P), 8 mmol/L (group 3-8P) phosphorus ions for 21 days, respectively. Cells cultured on the two-dimensional polystyrene scaffolds were used as control groups (groups 2-GM, 2-4P, 2-8P). Cellular proliferation was examined by cell counting kit-8; the mRNA expressions of osteogenic marker genes were assessed by RT-PCR; the formation of mineralized nodules for the osteogenic differentiation of human bone marrow mesenchymal stem cells was examined by Alizarin red staining.
RESULTS AND CONCLUSION: Compared with the two-dimensional culture, the growth of human bone marrow mesenchymal stem cells induced by phosphorus ions were more obvious in the three-dimensional polystyrenes scaffolds at days 4, 7, 14 and 21 (P < 0.05). Compared with the group 3-GM, the mRNA expression of collagen type I in groups 3-4P and 3-8P increased more significantly at days 7 and 14 (P < 0.05); the mRNA expression of alkaline phosphatase in groups 3-4P and 3-8P increased more significantly at day 14 (P < 0.05); the mRNA expression of osteocalcin in groups 3-4P and 3-8P increased more significantly at days 14 and 21 (P < 0.05). Mineralized nodules were formed in groups 3-4P and 3-8P but not in group 3-GM at day 21. So we concluded that phosphorus ions can promote proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells in three-dimensional polystyrenes scaffolds. Compared with the two-dimensional cell culture, the promoting growth effect of phosphorus ions on human bone marrow mesenchymal stem cells in three dimensional polystyrenes scaffolds are more obvious.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: tissue engineering, phosphorus, stem cells

CLC Number:

R318

Lei Qun, Chen Jiang, Huang Wen-xiu, Wu Dong, Lin Dong. Effect of phosphorus ions on human bone marrow mesenchymal stem cells under three-dimensional culture [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(47): 7591-7596.

Figures/Tables 3

References

[1] Bokhari M, Carnachan RJ, Cameron NR, et al. Novel cell culture device enabling three-dimensional cell growth and improved cell function. Biochem Biophys Res Commun. 2007;354(4):1095-1100.
[2] Inanc B, Elcin AE, Elcin YM. Osteogenic induction of human periodontal ligament fibroblasts under two-and three- dimensional culture conditions. Tissue Eng. 2006;12(2):257-266.
[3] Parekkadan B, Milwid JM. Mesenchymal stem cells as therapeutics. Annu Rev Biomed Eng. 2010;15(12):87-117.
[4] Asari S, Itakura S, Ferreri K, et al. Mesenchymal stem cells suppress B-cell terminal differentiation. Exp Hematol. 2009; 37(5):604-615.
[5] Ramasamy R, Tong CK, Seow HF, et al. The immunosuppressive effects of human bone marrow derived mesenchymal stem cells target T cell prolifferation but not its effector function. Cell Immunol. 2008;251(2):131-136.
[6] Honda Y, Anada T, Kamakura S, et al. Elevated extracellular calcium stimulates secretion of bone morphogenetic protein 2 by a macrophage cell line. Biochem Biophys Res Commun. 2006;345(3):1155-1160.
[7] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25(4):402-408.
[8] Joe AW, Gregory-Evans K. Mesenchymal stem cells and potential applications in treating ocular disease. Curr Eye Res. 2010;35(11):941-952.
[9] Giordano R, Canesi M, Isalberti M, et al. Autologous mesenchymal stem cell therapy for progressive supranuclear palsy: translation into a phase I controlled, randomized clinical study. J Transl Med. 2014;12:14.
[10] Gopal K, Amirhamed HA, Kamarul T. Advances of human bone marrow-derived mesenchymal stem cells in the treatment of cartilage defects: a systematic review. Exp Biol Med (Maywood). 2014;239(6):663-669.
[11] Moghadasali R, Azarnia M, Hajinasrollah M, et al. Intra-renal arterial injection of autologous bone marrow mesenchymal stromal cells ameliorates cisplatin-induced acute kidney injury in a rhesus Macaque mulatta monkey model. Cytotherapy. 2014;16(6):734-749.
[12] Pan XH, Yang XY, Yao X, et al. Bone-marrow mesenchymal stem cell transplantation to treat diabetic nephropathy in tree shrews. Cell Biochem Funct. 2014;32(5):453-463.
[13] Huang NF, Li S. Mesenchymal stem cells for vascular regeneration. Regen Med. 2008;3(6):877-892.
[14] Bruder SP, Fink DJ, Caplan AL. Mesenchymal stem cells in bonedevelopment, bone repair and skeletal regeneration therapy. J Cell Biochem. 1994;56(3):283-294.
[15] Badet L, Benhamou PY, Wojtusciszyn A, et al. Expectations and strategies regardingislet transplantation: metabolic data from the GRAGIL 2 trial. Transplantation. 2007;84(1):89-96.
[16] Janssens S, Dubois C, Bogaert J, et al. Autologous bone marrow-derived stem-cell transfer in patients with ST-segment elevation myocardial infarction: doubleblind, randomized controlled trials. Lancet. 2006;367(9505):113-121.
[17] Wang C, Wang S, Li K, et al. Preparation of Laponite Bioceramics for Potential Bone Tissue Engineering Applications. PLoS One. 2014;9(6):e99585.
[18] Nazemi K, Moztarzadeh F, Jalali N, et al. Synthesis and Characterization of Poly(lactic-co-glycolic) Acid Nanoparticles-Loaded Chitosan/Bioactive Glass Scaffolds as a Localized Delivery System in the Bone Defects. Biomed Res Int. 2014;2014:898930.
[19] Salehi S, Grünert AK, Bahners T, et al. New nanofibrous scaffold for corneal tissue engineering. Klin Monbl Augenheilkd. 2014;231(6):626-630.
[20] Stevanato L, Sinden JD. The effects of microRNAs on human neural stem cell differentiation in two- and three-dimensional cultures. Stem Cell Res Ther. 2014;5(2):49.
[21] Ülker HE, Ülker M, Gümü? et al. Cytotoxicity testing of temporary luting cements with two- and three-dimensional cultures of bovine dental pulp-derived cells. Biomed Res Int. 2013;2013:910459.
[22] Saltzman WM. Weaving cartilage at zero g: the reality of tissue engineering in space. Proc Natl Acad Sci U S A. 1997;94(25):13380-13382.
[23] Komlev VS, Barinov SM, Bozo II, et al. Bioceramics composed of octacalcium phosphate demonstrate enhanced biological behavior. ACS Appl Mater Interfaces. 2014;6(19): 16610-16620 .
[24] Zorin VL, Komlev VS, Zorina AI, et al. Octacalcium phosphate ceramics combined with gingiva-derived stromal cells for engineered functional bone grafts. Biomed Mater. 2014;9(5): 055005.
[25] Ling LE, Feng L, Liu HC, et al. The effect of calcium phosphate composite scaffolds on the osteogenic differentiation of rabbit dental pulp stem cells. J Biomed Mater Res A. 2014 [Epub ahead of print].
[26] Bléry P, Corre P, Malard O, et al. Evaluation of new bone formation in irradiated areas using association of mesenchymal stem cells and total fresh bone marrow mixed with calcium phosphate scaffold. J Mater Sci Mater Med. 2014 [Epub ahead of print].
[27] Baykan E, Koc A, Eser Elcin A, et al. Evaluation of a biomimetic poly(ε-caprolactone)/β-tricalcium phosphate multispiral scaffold for bone tissue engineering: in vitro and in vivo studies. Biointerphases. 2014;9(2):029011.
[28] Qiao PY, Li FF, Dong LM, et al. Delivering MC3T3-E1 cells into injectable calcium phosphate cement through alginate-chitosan microcapsules for bone tissue engineering. J Zhejiang Univ Sci B. 2014;15(4):382-392.
[29] Rendenbach C, Yorgan TA, Heckt T, et al. Effects of extracellular phosphate on gene expression in murine osteoblasts. Calcif Tissue Int. 2014;94(5):474-483.
[30] LeGeros RZ. Properties of Osteoconductive Biomaterials: Calcium Phosphates. Clin Orthop Related Res. 2002;395: 81-89.
[31] Song IH, Dennis JE. Simple evaluation method for osteoinductive capacity of cells or scaffolds using ceramic cubes. Tissue Cell. 2014;46(5):372-378.
[32] Perez RA, Riccardi K, Altankov G, et al. Dynamic cell culture on calcium phosphate microcarriers for bone tissue engineering applications. J Tissue Eng. 2014;5: 2041731414543965.
[33] Brennan MA, Renaud A, Amiaud J, et al. Pre-clinical studies of bone regeneration with human bone marrow stromal cells and biphasic calcium phosphate. Stem Cell Res Ther. 2014; 5(5):114.
[34] Zhou R, Xu W, Chen F, et al. Amorphous calcium phosphate nanospheres/polylactide composite coated tantalum scaffold: Facile preparation, fast biomineralization and subchondral bone defect repair application. Colloids Surf B Biointerfaces. 2014[Epub ahead of print].
[35] Rajzer I, Menaszek E, Kwiatkowski R, et al. Electrospun gelatin/poly(ε-caprolactone) fibrous scaffold modified with calcium phosphate for bone tissue engineering. Mater Sci Eng C Mater Biol Appl. 2014;44:183-190.
[36] Sanda M, Shiota M, Fujii M, et al. Capability of new bone formation with a mixture of hydroxyapatite and beta-tricalcium phosphate granules. Clin Oral Implants Res. 2014[Epub ahead of print].