Bone marrow mesenchymal stem cells combined with calcium alginate gel modified by hTGF-β1 for the construction of tissue-engineered cartilage in three-dimensional conditions

Zhu,Shaobo; Zhang,Tao; Sun,Chen; Yu,Aixi; Qi,Baiwen; Cheng,Hao

doi:10.3892/etm.2012.765

Bone marrow mesenchymal stem cells combined with calcium alginate gel modified by hTGF-β1 for the construction of tissue-engineered cartilage in three-dimensional conditions

Authors:
- Shaobo Zhu
- Tao Zhang
- Chen Sun
- Aixi Yu
- Baiwen Qi
- Hao Cheng
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Affiliations: Department of Micro-Orthopedics, Zhongnan Hospital, Wuhan University, Wuhan 430071, P.R. China
Published online on: October 25, 2012 https://doi.org/10.3892/etm.2012.765
Pages: 95-101

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Abstract

The aim of this study was to investigate the feasibility of Ad-hTGF-β1-transfected bone marrow mesenchymal stem cells (BMMSCs) combined with calcium alginate gel for the construction of tissue-engineered cartilage under three-dimensional conditions. Rat BMMSCs were divided into three groups: the Ad-hTGF-β1 transfection group, the Ad-EGFP transfection group and the control group. The BMMSCs in the Ad-hTGF-β1 transfection group were continually grown. The compound of cell-calcium alginate gel was cultured in a constant temperature incubator. The morphology of cells was examined, and the proliferation of cells was detected by MTT assay. The results from real-time PCR showed that the average relative ratio of TGF-β1 and transcriptional coactivator with PDZ-binding motif (TAZ) in the Ad-hTGF-β1 group was comparable to that of the control group (P<0.05). Using western blotting and immunohistochemistry, strong expression of collagen II in the Ad-hTGF-β1 group was detected. The results from western blotting showed that the expression of TGF-β1 in the Ad-hTGF-β1 group was significantly increased compared with that of the other two groups. The differentiation of BMMSCs was induced by Ad-hTGF-β1 transfection into chondrocytes. TGF-β1 may promote the differentiation of BMMSCs into chondrocytes by TAZ. BMMSCs transfected by Ad-hTGF-β1 could be induced into chondrocytes. These three-dimensional conditions could preferably mimic cell growth patterns in vivo.

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1	Ghidoni I, Chlapanidas T, Bucco M, et al: Alginate cell encapsulation: new advances in reproduction and cartilage regenerative medicine. Cytotechnology. 58:49–56. 2008. View Article : Google Scholar : PubMed/NCBI
2	Lin YJ, Yen CN, Hu YC, et al: Chondrocytes culture in three-dimensional porous alginate scaffolds enhanced cell proliferation, matrix synthesis and gene expression. J Biomed Mater Res A. 88:23–33. 2009. View Article : Google Scholar : PubMed/NCBI
3	Kanda C, Barry WO and Marie-Paule I: Effects of growth factors on cell proliferation and matrix synthesis of low-density, primary bovine chondrocytes cultured in collagen I gels. J Orthop Res. 20:1070–1078. 2002. View Article : Google Scholar : PubMed/NCBI
4	Yao Xiawan, Wei Liu, Tianyi Liu, et al: Application of bone marrow stromal cells in vitro tubular cartilage tissue engineering experiments. Chin J Microsurg. 28:328–330. 2005.
5	Jian Zhao and Wei Xu: Bone marrow mesenchymal stem cells and embryonic stem cells into cartilage cells into possible molecular mechanisms [J]. Tissue Eng Reconstructive Surg. 2:53–57. 2006.PubMed/NCBI
6	Hong JH, Hwang ES, McManus MT, et al: TAZ, a transcriptional modulator of mesenchymal stem cell differentiation. Science. 309:1074–1078. 2005. View Article : Google Scholar : PubMed/NCBI
7	Zhao L, Jiang S and Hantash BM: TGF-beta1 induces osteogenic differentiation of murine bone marrow stromal cells. Tissue Eng Part A. 16:725–733. 2010. View Article : Google Scholar : PubMed/NCBI
8	Hong JH and Yaffe MB: TAZ: a beta-catenin-like molecule that regulates mesenchymal stem cell differentiation. Cell Cycle. 5:176–179. 2006. View Article : Google Scholar : PubMed/NCBI
9	Lund AW, Yener B, Stegemann JP and Plopper GE: The natural and engineered 3D microenvironment as a regulatory cue during stem cell fate determination. Tissue Eng Part B Rev. 15:371–380. 2009. View Article : Google Scholar : PubMed/NCBI
10	Purpura KA, Aubin JE and Zandstra PW: Sustained in vitro expansion of bone progenitors is cell density dependent. Stem Cells. 22:39–50. 2004. View Article : Google Scholar : PubMed/NCBI
11	McBeath R, Pirone DM, Nelson CM, et al: Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. Dev Cell. 6:483–495. 2004. View Article : Google Scholar : PubMed/NCBI
12	Liu WF and Chen CS: Cellular and multicellular form and function. Adv Drug Deliv Rev. 59:1319–1328. 2007. View Article : Google Scholar : PubMed/NCBI
13	Burg KJ, Porter S and Kellam JF: Biomaterial developments for bone tissue engineering. Biomaterials. 21:2347–2359. 2000. View Article : Google Scholar : PubMed/NCBI
14	Benya PD and Shaffer JD: Dediferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 30:215–224. 1982. View Article : Google Scholar : PubMed/NCBI
15	Vinatier C, Guicheux J, Daculsi G, et al: Cartilage and bone tissue engineering using hydrogels. Biomed Mater Eng. 16:S107–S113. 2006.PubMed/NCBI
16	Dobratz EJ, Kim SW, Voglewede A and Park SS: Injectable cartilage: using alginate and human chondrocytes. Arch Facial Plast Surg. 11:40–47. 2009. View Article : Google Scholar : PubMed/NCBI
17	Chang SC, Rowley JA, Tobias G, et al: Injection molding of chondrocyte / alginate constructs in the shape of facial implants. J Biomed Mater Res. 55:503–511. 2001. View Article : Google Scholar : PubMed/NCBI
18	Chang SC, Tobias G, Roy AK, Vacanti CA and Bonassar LJ: Tissue engineering of autologous cartilage for craniofacial reconstruction by injection molding. Plast Reconstr Surg. 112:793–799. 2003. View Article : Google Scholar : PubMed/NCBI
19	Yi Guoquan and Shibi Lu: Alginate properties and applications in cartilage tissue engineering. Chin J Orthopaedic Surg. 14:216–218. 2006.
20	Kamishina H, Miyabayashi T, Clemmons RM, et al: Three-dimensional culture of feline articular chondrocytes in alginate microspheres. J Vet Med Sci. 68:1239–1242. 2006. View Article : Google Scholar : PubMed/NCBI