BMP-2誘導人跟腱來源肌腱干細胞成軟骨分化的實驗研究_《中國修復重建外科雜志》

作者：

芮云峰 ^1,2 , 郭永剛 ³ , 林禹丞 ¹ , 馬良彧 ² , 成心錕 ² , 陳輝 ¹ , 王宸 ¹

1 東南大學附屬中大醫院骨科（南京，210009）;;
2 東南大學醫學院;;
3通遼市科爾沁第一人民醫院骨科;

關鍵詞：

肌腱干細胞成軟骨分化慢性腱病 BMP-2

DOI：

10.7507/1002-1892.20130328

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目的探討BMP-2體外誘導人跟腱來源肌腱干細胞（human Achilles tendon-derived stem cells，hATDSCs）成軟骨分化的作用。方法無菌條件下取急性跟腱斷裂患者自愿捐贈的跟腱組織，膠原酶消化獲得有核細胞；以500個/cm2細胞密度接種，細胞貼壁呈克隆樣生長，混合培養獲得原代hATDSCs；體外傳代培養至第3代，流式細胞儀檢測hATDSCs免疫表型；油紅O染色、茜素紅染色及番紅O/快綠染色分別鑒定hATDSCs成脂、成骨及成軟骨分化能力。取第3代細胞體外三維培養成微球后分為兩組，實驗組用重組人BMP-2（recombinant human BMP-2，rhBMP-2）干預，對照組不進行干預。3周后行微球大體觀察；組織學切片行HE染色、番紅O/快綠染色、Ⅱ型膠原免疫組織化學染色觀察；實時熒光定量PCR檢測成軟骨特異性基因SOX9、Ⅱ型膠原和軟骨聚集蛋白多糖（Aggrecan）表達。結果原代hATDSCs體外培養呈克隆樣集落生長；傳代后以紡錘形、成纖維樣細胞生長，形態均一。hATDSCs陽性表達MSCs特異性表面標志CD44、CD90、CD105，陰性表達CD34、 CD45和CD146。多向分化潛能鑒定示hATDSCs成脂誘導3周油紅O染色陽性，成骨誘導4周茜素紅染色陽性，成軟骨誘導3周番紅O/快綠染色陽性。rhBMP-2誘導hATDSCs 3周，實驗組微球形成，體積顯著大于對照組；HE染色、番紅O/快綠染色、Ⅱ型膠原免疫組織化學染色均呈陽性，實時熒光定量PCR檢測示SOX9、Ⅱ型膠原和Aggrecan mRNA表達顯著高于對照組（P lt; 0.05）。結論體外BMP-2可促進hATDSCs成軟骨分化和蛋白聚糖合成增加，為研究慢性腱病組織病理學變化提供細胞生物學依據。

引用本文： 芮云峰,郭永剛,林禹丞,馬良彧,成心錕,陳輝,王宸. BMP-2誘導人跟腱來源肌腱干細胞成軟骨分化的實驗研究. 中國修復重建外科雜志, 2013, 27(12): 1492-1498. doi: 10.7507/1002-1892.20130328 復制

1.	Riley G. Tendinopathy—from basic science to treatment. Nat Clin Pract Rheumatol, 2008, 4(2): 82-89.
2.	Rees JD, Maffulli N, Cook J. Management of tendinopathy. Am J Sports Med, 2009, 37(9): 1855-1867.
3.	Xu Y, Murrell GA. The basic science of tendinopathy. Clin Orthop Relat Res, 2008, 466(7): 1528-1538.
4.	Rui YF, Lui PP, Li G, et al. Isolation and characterization of multipotent rat tendon-derived stem cells. Tissue Eng Part A, 2010, 16(5): 1549-1558.
5.	Rui YF, Lui PP, Chan LS, et al. Does erroneous differentiation of tendon-derived stem cells contribute to the pathogenesis of calcifying tendinopathy? Chin Med J (Engl), 2011, 124(4): 606-610.
6.	Rui YF, Lui PP, Wong YM, et al. Altered fate of tendon-derived stem cells isolated from a failed tendon-healing animal model of tendinopathy. Stem Cells Dev, 2013, 22(7): 1076-1085.
7.	Yee Lui PP, Wong YM, Rui YF, et al. Expression of chondro-osteogenic BMPs in ossified failed tendon healing model of tendinopathy. J Orthop Res, 2011, 29(6): 816-821.
8.	Rui YF, Lui PP, Rolf CG, et al. Expression of chondro-osteogenic BMPs in clinical samples of patellar tendinopathy. Knee Surg Sports Traumatol Arthrosc, 2012, 20(7): 1409-1417.
9.	Hashimoto Y, Yoshida G, Toyoda H, et al. Generation of tendon-to-bone interface “enthesis” with use of recombinant BMP-2 in a rabbit model. J Orthop Res, 2007, 25(11): 1415-1424.
10.	Rui YF, Lui PP, Wong YM, et al. BMP-2 stimulated non-tenogenic differentiation and promoted proteoglycan deposition of tendon-derived stem cells (TDSCs) in vitro. J Orthop Res, 2013, 31(5): 746-753.
11.	Lee WY, Lui PP, Rui YF. Hypoxia-mediated efficient expansion of human tendon-derived stem cells in vitro. Tissue Eng Part A, 2012, 18(5-6): 484-498.
12.	Bi Y, Ehirchiou D, Kilts TM, et al. Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche. Nat Med, 2007, 13(10): 1219-1227.
13.	Zhang J, Wang JH. Characterization of differential properties of rabbit tendon stem cells and tenocytes. BMC Musculoskelet Disord, 2010, 11: 10.
14.	Mienaltowski MJ, Adams SM, Birk DE. Regional differences in stem cell/progenitor cell populations from the mouse achilles tendon. Tissue Eng Part A, 2013, 19(1-2): 199-210.
15.	Yin Z, Chen X, Chen JL, et al. The regulation of tendon stem cell differentiation by the alignment of nanofibers. Biomaterials, 2010, 31(8): 2163-2175.
16.	Tang Y, Tang W, Lin Y, et al. Combination of bone tissue engineering and BMP-2 gene transfection promotes bone healing in osteoporotic rats. Cell Biol Int, 2008, 32(9): 1150-1157.
17.	Chen CH, Liu HW, Tsai CL, et al. Photoencapsulation of bone morphogenetic protein-2 and periosteal progenitor cells improve tendon graft healing in a bone tunnel. Am J Sports Med, 2008, 36(3): 461-473.
18.	Knippenberg M, Helder MN, Zandieh Doulabi B, et al. Osteogenesis versus chondrogenesis by BMP-2 and BMP-7 in adipose stem cells. Biochem Biophys Res Commun, 2006, 342(3): 902-908.
19.	Asahina I, Sampath TK, Hauschka PV. Human osteogenic protein-1 induces chondroblastic, osteoblastic, and/or adipocytic differentiation of clonal murine target cells. Exp Cell Res, 1996, 222(1): 38-47.
20.	Date T, Doiguchi Y, Nobuta M, et al. Bone morphogenetic protein-2 induces differentiation of multipotent C3H10T1/2 cells into osteoblasts, chondrocytes, and adipocytes in vivo and in vitro. J Orthop Sci, 2004, 9(5): 503-508.
21.	Zeiter S, Lezuo P, Ito K. Effect of TGF beta1, BMP-2 and hydraulic pressure on chondrogenic differentiation of bovine bone marrow mesenchymal stromal cells. Biorheology, 2009, 46(1): 45-55.
22.	段智霞, 鄭啟新, 郭曉東, 等. 骨形成蛋白 2 活性多肽體外定向誘導骨髓間充質干細胞向成骨方向分化的劑量依賴性研究. 中國修復重建外科雜志, 2007, 21(10): 1118-1122.
23.	An C, Cheng Y, Yuan Q, et al. IGF-1 and BMP-2 induces differentiation of adipose-derived mesenchymal stem cells into chondrocytes-like cells. Ann Biomed Eng, 2010, 38(4): 1647-1654.
24.	Qing W, Guang-Xing C, Lin G, et al. The osteogenic study of tissue engineering bone with BMP2 and BMP7 gene-modified rat adipose-derived stem cell. J Biomed Biotechnol, 2012, 2012: 410879.
25.	Rui YF, Lui PP, Lee YW, et al. Higher BMP receptor expression and BMP-2-induced osteogenic differentiation in tendon-derived stem cells compared with bone-marrow-derived mesenchymal stem cells. Int Orthop, 2012, 36(5): 1099-1107.

1. Riley G. Tendinopathy—from basic science to treatment. Nat Clin Pract Rheumatol, 2008, 4(2): 82-89.
2. Rees JD, Maffulli N, Cook J. Management of tendinopathy. Am J Sports Med, 2009, 37(9): 1855-1867.
3. Xu Y, Murrell GA. The basic science of tendinopathy. Clin Orthop Relat Res, 2008, 466(7): 1528-1538.
4. Rui YF, Lui PP, Li G, et al. Isolation and characterization of multipotent rat tendon-derived stem cells. Tissue Eng Part A, 2010, 16(5): 1549-1558.
5. Rui YF, Lui PP, Chan LS, et al. Does erroneous differentiation of tendon-derived stem cells contribute to the pathogenesis of calcifying tendinopathy? Chin Med J (Engl), 2011, 124(4): 606-610.
6. Rui YF, Lui PP, Wong YM, et al. Altered fate of tendon-derived stem cells isolated from a failed tendon-healing animal model of tendinopathy. Stem Cells Dev, 2013, 22(7): 1076-1085.
7. Yee Lui PP, Wong YM, Rui YF, et al. Expression of chondro-osteogenic BMPs in ossified failed tendon healing model of tendinopathy. J Orthop Res, 2011, 29(6): 816-821.
8. Rui YF, Lui PP, Rolf CG, et al. Expression of chondro-osteogenic BMPs in clinical samples of patellar tendinopathy. Knee Surg Sports Traumatol Arthrosc, 2012, 20(7): 1409-1417.
9. Hashimoto Y, Yoshida G, Toyoda H, et al. Generation of tendon-to-bone interface “enthesis” with use of recombinant BMP-2 in a rabbit model. J Orthop Res, 2007, 25(11): 1415-1424.
10. Rui YF, Lui PP, Wong YM, et al. BMP-2 stimulated non-tenogenic differentiation and promoted proteoglycan deposition of tendon-derived stem cells (TDSCs) in vitro. J Orthop Res, 2013, 31(5): 746-753.
11. Lee WY, Lui PP, Rui YF. Hypoxia-mediated efficient expansion of human tendon-derived stem cells in vitro. Tissue Eng Part A, 2012, 18(5-6): 484-498.
12. Bi Y, Ehirchiou D, Kilts TM, et al. Identification of tendon stem/progenitor cells and the role of the extracellular matrix in their niche. Nat Med, 2007, 13(10): 1219-1227.
13. Zhang J, Wang JH. Characterization of differential properties of rabbit tendon stem cells and tenocytes. BMC Musculoskelet Disord, 2010, 11: 10.
14. Mienaltowski MJ, Adams SM, Birk DE. Regional differences in stem cell/progenitor cell populations from the mouse achilles tendon. Tissue Eng Part A, 2013, 19(1-2): 199-210.
15. Yin Z, Chen X, Chen JL, et al. The regulation of tendon stem cell differentiation by the alignment of nanofibers. Biomaterials, 2010, 31(8): 2163-2175.
16. Tang Y, Tang W, Lin Y, et al. Combination of bone tissue engineering and BMP-2 gene transfection promotes bone healing in osteoporotic rats. Cell Biol Int, 2008, 32(9): 1150-1157.
17. Chen CH, Liu HW, Tsai CL, et al. Photoencapsulation of bone morphogenetic protein-2 and periosteal progenitor cells improve tendon graft healing in a bone tunnel. Am J Sports Med, 2008, 36(3): 461-473.
18. Knippenberg M, Helder MN, Zandieh Doulabi B, et al. Osteogenesis versus chondrogenesis by BMP-2 and BMP-7 in adipose stem cells. Biochem Biophys Res Commun, 2006, 342(3): 902-908.
19. Asahina I, Sampath TK, Hauschka PV. Human osteogenic protein-1 induces chondroblastic, osteoblastic, and/or adipocytic differentiation of clonal murine target cells. Exp Cell Res, 1996, 222(1): 38-47.
20. Date T, Doiguchi Y, Nobuta M, et al. Bone morphogenetic protein-2 induces differentiation of multipotent C3H10T1/2 cells into osteoblasts, chondrocytes, and adipocytes in vivo and in vitro. J Orthop Sci, 2004, 9(5): 503-508.
21. Zeiter S, Lezuo P, Ito K. Effect of TGF beta1, BMP-2 and hydraulic pressure on chondrogenic differentiation of bovine bone marrow mesenchymal stromal cells. Biorheology, 2009, 46(1): 45-55.
22. 段智霞, 鄭啟新, 郭曉東, 等. 骨形成蛋白 2 活性多肽體外定向誘導骨髓間充質干細胞向成骨方向分化的劑量依賴性研究. 中國修復重建外科雜志, 2007, 21(10): 1118-1122.
23. An C, Cheng Y, Yuan Q, et al. IGF-1 and BMP-2 induces differentiation of adipose-derived mesenchymal stem cells into chondrocytes-like cells. Ann Biomed Eng, 2010, 38(4): 1647-1654.
24. Qing W, Guang-Xing C, Lin G, et al. The osteogenic study of tissue engineering bone with BMP2 and BMP7 gene-modified rat adipose-derived stem cell. J Biomed Biotechnol, 2012, 2012: 410879.
25. Rui YF, Lui PP, Lee YW, et al. Higher BMP receptor expression and BMP-2-induced osteogenic differentiation in tendon-derived stem cells compared with bone-marrow-derived mesenchymal stem cells. Int Orthop, 2012, 36(5): 1099-1107.

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BMP-2誘導人跟腱來源肌腱干細胞成軟骨分化的實驗研究

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BMP-2誘導人跟腱來源肌腱干細胞成軟骨分化的實驗研究

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