EFFECT OF ROLLING COMPRESSION LOADING BIOREACTOR ON CHONDROGENESIS OF RABBIT BONE MARROW MESENCHYMAL STEM CELLS WITH DIFFERENT LOADING PARAMETERS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

SUN Minglin ¹ , ZHU Lei ¹ , LUuml ¹ , Dan ²

1Department of Orthopedics, Affiliated Hospital of Chinese People’s Armed Police Force, Tianjin, 300162, P.R.China;;
2School of Mechanical Engineering, Tianjin University of Technology. Corresponding author: ZHU Lei, E-mail: zhu.lei619@gmail.com;

Corresponding author：

Keywords：

Bone marrow mesenchymal stem cells; Chondrocytes; Bioreactor; Rolling compression loading; Rabbit

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Objective To explore the effect of rolling compression loading bioreactor on chondrogenesis of rabbit bone marrow mesenchymal stem cells (BMSCs) with different loading parameters. Methods BMSCs were isolated from New Zealand rabbits, aged 2.5 months. BMSCs at passage 3 were used to prepare BMSCs-agarose gels (4 mm in diameter and height, respectively). Samples were divided into 8 groups: 10% (group A1), 20% (group A2), and 30% (group A3) compression groups (0.4 Hz, 3 h/ d) and 20 minutes (group B1), 3 hours (group B2), and 12 hours (group B3) rolling time groups and static culture (control groups). The living cell rate, the collagen type II and Aggrecan gene expressions, and glycosaminoglycan (GAG) content were determined, and histological staining was done at 24 hours, 7 days, 14 days, and 21 days after culture. Results At 14 and 21 days, the living cell rates of groups A1 and A2 were significantly higher than that of group A3 (P lt; 0.05), groups B1 and B2 were significantly higher than group B3 (P lt; 0.05). Collagen type II and Aggrecan gene expressions of the experimental groups at each time point were significantly higher than those of the control groups (P lt; 0.05); at 14 and 21 days, collagen type II and Aggrecan gene expressions of groups A1 and A2 were significantly higher than those of group A3, and groups B1 and B2 were also significantly higher than group B3 (P lt; 0.05). At 14 and 21 days, the GAG contents of groups A1 and A2 were significantly higher than those of group A3 (P lt; 0.05); groups B1 and B2 were also significantly higher than group B3 (P lt; 0.05). At 21 days, toluidine blue staining showed that obvious blue-staining and even cartilage lacunae were seen in groups A2 and B2, but light and quite rare blue-staining in groups A1, A3, B1, and B3. Conclusion The rolling compression loading bioreactor has great promotion effect on chondrogenesis of rabbit BMSCs with rolling parameters of 0.4 Hz, 3 hours, and 20% compression.

Citation： SUN Minglin,ZHU Lei,LUuml,Dan,ZHANG Chunqiu. EFFECT OF ROLLING COMPRESSION LOADING BIOREACTOR ON CHONDROGENESIS OF RABBIT BONE MARROW MESENCHYMAL STEM CELLS WITH DIFFERENT LOADING PARAMETERS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(1): 89-94. doi: Copy

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20.	Wang ZH, He XJ, Yang ZQ, et al. Cartilage tissue engineering with demineralized bone matrix gelatin and fibrin glue hybrid scaffold: an in vitro study. Artif Organs, 2010, 34(2): 161-166.

1. Bi L, Cao Z, Hu Y, et al. Effects of different cross-linking conditions on the properties of genipin-cross-linked chitosan/collagen scaffolds for cartilage tissue engineering. J Mater Sci Mater Med, 2010, 22(1): 51-62.
2. Chen JP, Li SF, Chiang YP. Bioactive collagen-grafted poly-L-lactic acid nanofibrous membrane for cartilage tissue engineering. J Nanosci Nanotechnol, 2010, 10(8): 5393-5398.
3. Cinbiz MN, Tigli RS, Beskardes IG, et al. Computational fluid dynamics modeling of momentum transport in rotating wall perfused bioreactor for cartilage tissue engineering. J Biotechnol, 2010, 150(3): 389-395.
4. Gong Y, Su K, Lau TT, et al. Microcavitary hydrogel-mediating phase transfer cell culture for cartilage tissue engineering. Tissue Eng Part A, 2010, 16(12): 3611-3622.
5. Havla JB, Lotz AS, Richter E, et al. Cartilage tissue engineering for auricular reconstruction: In vitro evaluation of potential genotoxic and cytotoxic effects of scaffold materials. Toxicol In Vitro, 2010, 24(3): 849-853.
6. Jin CZ, Choi BH, Park SR, et al. Cartilage engineering using cell-derived extracellular matrix scaffold in vitro. J Biomed Mater Res A, 2010, 92(4): 1567-1577.
7. Williams GM, Lin JW, Sah RL. Cartilage reshaping via in vitro mechanical loading. Tissue Eng, 2007, 13(12): 2903-2911.
8. Mithoefer K, Williams RJ 3rd, Warren RF, et al. The microfracture technique for the treatment of articular cartilage lesions in the knee. A prospective cohort study. J Bone Joint Surg (Am), 2005, 87(9): 1911-1920.
9. Oliveira JT, Santos TC, Martins L, et al. Gellan gum injectable hydrogels for cartilage tissue engineering applications: In vitro studies and preliminary in vivo evaluation. Tissue Eng Part A, 2010, 16(1): 343-353.
10. Vavken P, Arrich F, Pilz M, et al. An in vitro model of biomaterial-augmented microfracture including chondrocyte-progenitor cell interaction. Arch Orthop Trauma Surg, 2010, 130(5): 711-716.
11. Tran SC, Cooley AJ, Elder SH. Effect of a mechanical stimulation bioreactor on tissue engineered, scaffold-free cartilage. Biotechnol Bioeng, 2011, 108(6): 1421-1429.
12. Anderer U, Libera J. In vitro engineering of human autogenous cartilage. J Bone Miner Res, 2002, 17(8): 1420-1429.
13. Mauck RL, Soltz MA, Wang CC, et al. Functional tissue engineering of articular cartilage through dynamic loading of chondrocyte-seeded agarose gels. J Biomech Eng, 2000, 122(3): 252-260.
14. Huang AH, Farrell MJ, Mauck RL. Mechanics and mechanobiology of mesenchymal stem cell-based engineered cartilage. J Biomech, 2010, 43(1): 128-136.
15. Hwang NS, Zhang C, Hwang YS, et al. Mesenchymal stem cell differentiation and roles in regenerative medicine. Wiley Interdiscip Rev Syst Biol Med, 2009, 1(1): 97-106.
16. Richardson SM, Hoyland JA, Mobasheri R, et al. Mesenchymal stem cells in regenerative medicine: Opportunities and challenges for articular cartilage and intervertebral disc tissue engineering. J Cell Physiol, 2010, 222(1): 23-32.
17. Hendriks J, Riesle J, van Blitterswijk CA. Co-culture in cartilage tissue engineering. J Tissue Eng Regen Med, 2007, 1(3): 170-178.
18. Alsalameh S, Amin R, Gemba T, et al. Identification of mesenchymal progenitor cells in normal and osteoarthritic human articular cartilage. Arthritis Rheum, 2004, 50(5): 1522-1532.
19. 赵斌, 王玉, 黄靖香, 等. 关节软骨源性多孔支架复合自体软骨细胞修复兔软骨缺损的长期疗效观察. 中国矫形外科杂志, 2010, 18(11): 937-940.
20. Wang ZH, He XJ, Yang ZQ, et al. Cartilage tissue engineering with demineralized bone matrix gelatin and fibrin glue hybrid scaffold: an in vitro study. Artif Organs, 2010, 34(2): 161-166.

Chinese Journal of Reparative and Reconstructive Surgery

EFFECT OF ROLLING COMPRESSION LOADING BIOREACTOR ON CHONDROGENESIS OF RABBIT BONE MARROW MESENCHYMAL STEM CELLS WITH DIFFERENT LOADING PARAMETERS

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