不同參數滾壓模式對兔BMSCs向軟骨細胞分化促進作用的研究_《中國修復重建外科雜志》

作者：

孫明林 ¹ , 朱雷 ¹ , 呂丹 ¹ , 張春秋 ²

1 武警后勤學院附屬醫院骨科（天津，300162）;;
2 天津理工大學機械工程學院;

關鍵詞：

BMSCs 軟骨細胞生物反應器滾壓載荷兔

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摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

目的觀察在不同滾壓參數下，滾壓載荷生物反應器對BMSCs-瓊脂糖復合體中兔BMSCs向軟骨細胞誘導分化的作用。方法分離2.5月齡新西蘭兔BMSCs，培養至第3代后與低熔點瓊脂糖復合成凝膠塊，并在自制模具中形成直徑4 mm、高4 mm的圓柱形BMSCs-瓊脂糖復合體。將樣本分別在0.4 Hz、3 h/d基本參數下比較壓縮量10%、20%、30%（分別為A1、A2、A3組），以及0.4 Hz、20%壓縮量基本參數下比較20 min、3 h、12 h滾壓時間（分別為B1、B2、B3組）對BMSCs向軟骨細胞分化的作用，以靜態培養作為對照組。在培養24 h及7、14、21 d時取各組標本進行死活細胞檢測、實時定量PCR檢測、糖胺聚糖（glycosaminoglycan，GAG）含量檢測及組織學染色觀察。結果死活細胞檢測：14、21 d時，A1、A2組活細胞比率均顯著高于A3組，B1、B2組活細胞比率均顯著高于B3組，差異均有統計學意義（P lt; 0.05）。實時定量PCR檢測：各實驗組7、14、21 dⅡ型膠原及蛋白聚糖（Aggrecan）mRNA表達均高于對照組，差異有統計學意義（P lt; 0.05）；14、 21 d時，A1、A2組Ⅱ型膠原、Aggrecan mRNA表達均高于A3組，B1、B2組均高于B3組，差異有統計學意義（P lt; 0.05）。以上指標A1、A2組間及B1、B2組間比較，差異均無統計學意義（P gt; 0.05）。GAG含量檢測：14、21 d時，A1、A2組GAG含量均顯著高于對照組和A3組，B1、B2組GAG含量也顯著高于對照組和B3組，差異均有統計學意義（P lt; 0.05）；21 d時A1、A2組間差異均有統計學意義（t=2.830，P=0.027）。組織學觀察：21 d時，A2組和B2組出現明顯藍染，可見軟骨陷窩樣結構；而A1、A3組和B1、B3組藍染效果較差，分布稀少。結論滾壓載荷生物反應器在0.4 Hz、20%壓縮量、3 h滾壓時間參數下，能更有效地促進BMSCs-瓊脂糖復合體中兔BMSCs向軟骨細胞誘導分化。

引用本文： 孫明林,朱雷,呂丹,張春秋. 不同參數滾壓模式對兔BMSCs向軟骨細胞分化促進作用的研究. 中國修復重建外科雜志, 2013, 27(1): 89-94. doi: 復制

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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.
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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. 趙斌, 王玉, 黃靖香, 等. 關節軟骨源性多孔支架復合自體軟骨細胞修復兔軟骨缺損的長期療效觀察. 中國矯形外科雜志, 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.

《中國修復重建外科雜志》

不同參數滾壓模式對兔BMSCs向軟骨細胞分化促進作用的研究

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《中國修復重建外科雜志》

不同參數滾壓模式對兔BMSCs向軟骨細胞分化促進作用的研究

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

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