Chondrogenic differentiation of adipose-derived stem cells induced by growth differentiation factor-5 cultured on the type I collagen scaffold

doi:10.3969/j.issn.2095-4344.2015.19.008

Abstract

Abstract:

BACKGROUND: Growth differentiation factor-5 can induce adipose-derived stem cells into chondrocytes in our previous studies, but it has not been reported that the adipose-derived stem cells induced by growth differentiation factor-5 can differentiate into chondrocytes on the type I collagen scaffold.
OBJECTIVE: To investigate the chondrogenic differentiation ability of adipose-derived stem cells induced by growth differentiation factor-5 cultured on the type I collagen scaffold.
METHODS: Adipose derived stem cells were isolated from rabbit adipose tissue, the cells morphology was observed using inverted phase contrast microscope and the phenotypes were identified using immunofluorescence. The exogenous growth differentiation factor-5 was added to the cultural media with the type I collagen scaffold so as to induce the chondrogenic differentiation. The cells morphology was observed using hematoxylin-eosin staining and scanning electron microscope after the induction by growth differentiation factor-5 for 14 days. Meanwhile, the type II collagen and aggrecan mRNA expressions of the induced cells were measured using RT-PCR after the induction by growth differentiation factor-5 for 7, 14, and 21 days.
RESULTS AND CONCLUSION: The primary cultured adipose-derived stem cells proliferated adherently with the fusiform and polygonal distribution under inverted phase contrast microscope. The positive CD44, CD49d and negative CD106 were detected by immunofluorescence. The adipose-derived stem cells induced by growth differentiation factor-5 were well adhered to the type I collagen scaffold and strongly proliferated. The large amounts of extracellular matrix existed on the surface of the induced cells under scanning electron microscope. RT-PCR agarose gel electrophoresis indicated that the type II collagen and aggrecan mRNA expressions of the adipose-derived stem cells induced by growth differentiation factor-5 with the type I collagen scaffold were significantly increased. Growth differentiation factor-5 can successfully induce the chondrogenic differentiation of adipose-derived stem cells cultured on the type I collagen scaffold.

Key words: Collagen Type I, Stents, Growth Differentiation Factors, Chondrocytes

CLC Number:

R394.2

Liu Zhen-ning, Han Chang-xu, Zhao Min. Chondrogenic differentiation of adipose-derived stem cells induced by growth differentiation factor-5 cultured on the type I collagen scaffold [J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.2015.19.008.

Figures/Tables 2

References

[1] Kon E, Roffi A, Filardo G, et al. Scaffold-Based Cartilage Treatments: With or Without Cells? A Systematic Review of Preclinical and Clinical Evidence. Arthroscopy. 2015;31(4): 767-775.

[2] Filardo G, Kon E, Roffi A, et al. Scaffold-based repair for cartilage healing: a systematic review and technical note. Arthroscopy. 2013;29(1):174-186.

[3] Solheim E, Hegna J, Inderhaug E, et al. Results at 10-14 years after microfracture treatment of articular cartilage defects in the knee. Knee Surg Sports Traumatol Arthrosc. 2014. in press.

[4] Ortved KF, Begum L, Mohammed HO, et al. Implantation of rAAV5-IGF-I transduced autologous chondrocytes improves cartilage repair in full-thickness defects in the equine model. Mol Ther. 2015;23(2):363-373.

[5] Makris EA, Gomoll AH, Malizos KN, et al. Repair and tissue engineering techniques for articular cartilage. Nat Rev Rheumatol. 2015;11(1):21-34.

[6] Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7(2):211-228.

[7] Faroni A, Smith RJ,Reid AJ.Adipose derived stem cells and nerve regeneration. Neural Regen Res. 2014;9(14): 1341-1346.

[8] Hötten G, Neidhardt H, Jacobowsky B, et al. Cloning and expression of recombinant human growth/differentiation factor 5. Biochem Biophys Res Commun. 1994;204(2): 646-652.

[9] 刘振宁,贾长青,韩长旭,等.生长分化因子5诱导兔脂肪干细胞成软骨细胞分化的实验研究[J].中国修复重建外科杂志,2009, 23(4):483-489.

[10] Im GI, Jung NH, Tae SK. Chondrogenic differentiation of mesenchymal stem cells isolated from patients in late adulthood: the optimal conditions of growth factors. Tissue Eng. 2006;12(3):527-536.

[11] Huang X, Yang D, Yan W, et al. Osteochondral repair using the combination of fibroblast growth factor and amorphous calcium phosphate/poly(L-lactic acid) hybrid materials. Biomaterials. 2007;28(20):3091-3100.

[12] 赵明璨,刘畅.脂肪干细胞在软骨组织工程中的研究进展[J].中国生物医学工程学报.2014,33(4):475-481.

[13] Nickel J, Kotzsch A, Sebald W, et al. A single residue of GDF-5 defines binding specificity to BMP receptor IB. J Mol Biol. 2005;349(5):933-947.

[14] Yoon HH, Bhang SH, Shin JY, et al. Enhanced cartilage formation via three-dimensional cell engineering of human adipose-derived stem cells. Tissue Eng Part A. 2012;18 (19-20):1949-1956.

[15] Lehmann M, Martin F, Mannigel K, et al. Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes. Eur J Histochem. 2013;57(4):e31.

[16] Wang PY, Tsai WB. Modulation of the proliferation and matrix synthesis of chondrocytes by dynamic compression on genipin-crosslinked chitosan/collagen scaffolds. J Biomater Sci Polym Ed. 2013;24(5):507-519.

[17] Huselstein C, Li Y, He X. Mesenchymal stem cells for cartilage engineering. Biomed Mater Eng. 2012;22(1-3): 69-80.

[18] 张文元,杨亚冬,李颖,等.蚕丝/聚乳酸-羟基乙酸共聚物支架降解液与骨髓间充质干细胞的增殖[J].中国组织工程研究,2013, 17(25):4676-4683.

[19] Heidemann W, Jeschkeit-Schubbert S, Ruffieux K, et al. pH-stabilization of predegraded PDLLA by an admixture of water-soluble sodiumhydrogenphosphate--results of an in vitro- and in vivo-study. Biomaterials. 2002;23(17):3567-3574.

[20] Wang Y, Kim UJ, Blasioli DJ, et al. In vitro cartilage tissue engineering with 3D porous aqueous-derived silk scaffolds and mesenchymal stem cells. Biomaterials. 2005;26(34): 7082-7094.

[21] Ragetly GR, Griffon DJ, Lee HB, et al. Effect of chitosan scaffold microstructure on mesenchymal stem cell chondrogenesis. Acta Biomater. 2010;6(4):1430-1436.

[22] Buma P, Pieper JS, van Tienen T, et al. Cross-linked type I and type II collagenous matrices for the repair of full-thickness articular cartilage defects--a study in rabbits. Biomaterials. 2003; 24(19):3255-3263.

[23] 张军鹏.运动性软骨损伤与组织工程化软骨及生物支架材料的修复[J].中国组织工程研究与临床康复,2011,15 (8):1491-1494.

[24] Wakitani S, Goto T, Pineda SJ, et al. Mesenchymal cell-based repair of large, full-thickness defects of articular cartilage. J Bone Joint Surg Am. 1994;76(4):579-592.

[25] Schlegel W, Nürnberger S, Hombauer M, et al. Scaffold-dependent differentiation of human articular chondrocytes. Int J Mol Med. 2008;22(5):691-699.