Stromal cell derived factor-1/chemokine receptor 4 signaling pathway is involved in bone morphogenetic protein 2-induced migration of bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2016.41.003

Abstract

Abstract:

BACKGROUND: Stromal cell derived factor-1 (SDF-1)/chemokine receptor 4 (CXCR-4) biological axis plays a chemotactic role in a variety of cells, making it possible to regulate the regeneration of a variety of tissues. Whether the bidogical is involved in bone morphogenetic protein-2 (BMP-2)-induced homing of bone marrow mesenchymal stem cells, however, is still unclear.
OBJECTIVE: To study the role of SDF-1/CXCR4 signaling pathway in BMP-2-induced migration of mouse bone marrow mesenchymal stem cells.
METHODS: Bone marrow mesenchymal stem cells in logarithmic growth were selected and intervened with SDF-1 (0, 50, 100 and 200 μg/L), BMP-2 (0, 50, 100 and 200 μg/L) and AMD3100 (50 μg/L) to induce cell migration detected by Transwell method.
RESULTS AND CONCLUSION: The migration of bone marrow mesenchymal stem cells was closely related to SDF-1 and BMP-2, and proportional to the concentration of both SDF-1 and BMP-2. When SDF-1 and BMP-2 were used jointly, the number of migrated cells was increased significantly, and highest number of migrated cells was obtained at 200 μg/L. Moreover, these migrated cells showed a nest-like distribution under microscopy. AMD3100 as an inhibitor markedly suppressed the migration of bone marrow mesencnymal stem cells induced by BMP-2, but the number of migrated cells was likely to increase with the increasing concentration of BMP-2 that exceeded a specific value. Overall, our findings show that SDF-1/CXCR4 signaling pathway is an important pathway in BMP-2-induced migration of bone marrow mesenchymal stem cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Morphogenetic Proteins, Receptors, CXCR4, Tissue Engineering

CLC Number:

R394.2

Yang Yi, Yi Lei, Yeerzhati Hajiaheman, Telieke Kanzhale, Jin Ge-le. Stromal cell derived factor-1/chemokine receptor 4 signaling pathway is involved in bone morphogenetic protein 2-induced migration of bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(41): 6098-6104.

Figures/Tables 2

2.1 不同质量浓度SDF-1诱导的骨髓间充质干细胞的迁移情况 SDF-1诱导下骨髓间充质干细胞迁移具有浓度依赖性，且不同质量浓度的SDF-1诱导的骨髓间充质干细胞的迁移数均高于0 μg/L SDF-1组，其中200 μg/L效果最为明显(图1)。 2.2 不同质量浓度骨形态发生蛋白2诱导的骨髓间充质干细胞的迁移情况加入骨形态发生蛋白2后，骨髓间充质干细胞迁移数明显增多，均高于0 μg/L骨形态发生蛋白2组，且细胞迁移数与骨形态发生蛋白2浓度成正比，各组骨髓间充质干细胞迁移数差异有显著性意义(F=446.93，P < 0.05)。当骨形态发生蛋白2浓度为200 μg/L时，细胞迁移数达到峰值(图2)。 2.3 SDF-1联合骨形态发生蛋白2对骨髓间充质干细胞迁移的影响同时加入SDF-1和骨形态发生蛋白2时，骨髓间充质干细胞迁移数量明显增多，各组骨髓间充质干细胞迁移数差异有显著性意义(F=553.54，P < 0.05)，并细胞迁移数和SDF-1与骨形态发生蛋白2浓度成正比，当200 μg/L SDF-1+200 μg/L骨形态发生蛋白2干预时，骨髓间充质干细胞迁移数量最多，并在镜下呈“巢穴”样分布(图3)。 2.4 阻断剂AMD3100对骨形态发生蛋白2诱导的骨髓间充质干细胞迁移的影响加入50 μg/L AMD3100时，骨髓间充质干细胞迁移数明显降低，50 μg/L AMD3100对50，100及200 μg/L骨形态发生蛋白2诱导的细胞迁移数的影响差异有显著性意义(F=142.53，P < 0.05)；即阻断剂AMD3100明显抑制了骨形态发生蛋白2诱导骨髓间充质干细胞迁移，但当增加骨形态发生蛋白2质量浓度达到一定量时，骨髓间充质干细胞迁移数随着骨形态发生蛋白2质量浓度的增加而增多(图4)。反向干预印证了SDF-1/CXCR4信号通路是参与了骨形态发生蛋白2诱导大鼠骨髓间充质干细胞迁移的重要信号通路。"

References

[1] Janicki P, Schmidmaier G. What should be the characteristics of the ideal bone graft substitute? Combining scaffolds with growth factors and/or stem cells. Injury. 2011;42 Suppl 2:S77-81.
[2] 段红永,管强,武欣,等.骨髓间充质干细胞诱导分化为组织工程血管用种子细胞的研究[J].中华实验外科杂志,2015, 32(4):748-752.
[3] 义勇,魏垒.基质细胞衍生因子-1与骨再生修复[J].国际骨科学杂志, 2016,37(3):190-193.
[4] 王鑫,许文静,彭江,等.间充质干细胞归巢在骨损伤修复中的作用及其影响[J].解放军医药杂志,2014,26(4):107- 110.
[5] Granero-Moltó F, Weis JA, Miga MI, et al. Regenerative effects of transplanted mesenchymal stem cells in fracture healing. Stem Cells. 2009;27(8): 1887-1898.
[6] 李忠伟,金格勒,唐周舟,等.外源性重组人骨形态发生蛋白-2应用于兔腰椎植骨融合中细胞增殖的实验研究[J].中华创伤骨科杂志,2009,11(5):464-468.
[7] 杨毅,金格勒,唐周舟,等.复合骨在兔腰椎融合过程中相关基因表达调控的影响[J].中华实验外科杂志,2010, 27(2): 245-247.
[8] Lee CH, Jin MU, Jung HM, et al. Effect of dual treatment with SDF-1 and BMP-2 on ectopic and orthotopic bone formation. PLoS One. 2015; 10(3): e0120051.
[9] 王新均,唐俊明,孔霞,等.不同迁移体系对SDF-1/CXCR4介导间充质干细胞迁移的影响[J].陕西医学杂志,2009, 38(3):262-266.
[10] 张伟,黄文志,张玉平.归巢因子基质细胞衍生因子1/CXCR4在角膜上的分布[J].中国组织工程研究,2015, 19(46):7466-7471.
[11] Schantz JT, Chim H, Whiteman M. Cell guidance in tissue engineering: SDF-1 mediates site-directed homing of mesenchymal stem cells within three-dimensional polycaprolactone scaffolds. Tissue Eng. 2007;13(11):2615-2624.
[12] Bobadilla M, Sainz N, Abizanda G, et al. The CXCR4/SDF1 axis improves muscle regeneration through MMP-10 activity. Stem Cells Dev. 2014; 23(12): 1417-1427.
[13] Brzoska E, Kowalski K, Markowska-Zagrajek A, et al. Sdf-1 (CXCL12) induces CD9 expression in stem cells engaged in muscle regeneration. Stem Cell Res Ther. 2015;6:46.
[14] Kucia M, Jankowski K, Reca R, et al. CXCR4-SDF-1 signalling, locomotion, chemotaxis and adhesion. J Mol Histol. 2004;35(3):233-245.
[15] Otsuru S, Tamai K, Yamazaki T, et al. Circulating bone marrow-derived osteoblast progenitor cells are recruited to the bone-forming site by the CXCR4/stromal cell-derived factor-1 pathway. Stem Cells. 2008;26(1):223-234.
[16] Saeed H, Ahsan M, Saleem Z, et al. Mesenchymal stem cells (MSCs) as skeletal therapeutics - an update. J Biomed Sci. 2016;23:41.
[17] 周永庆,胡图强,喻学洲,等.表皮生长因子对间充质干细胞迁移的影响及其信号机制[J].临床口腔医学杂志,2011, 27(10):587-590.
[18] 王鑫,苟文隆,徐小龙,等.骨髓间充质干细胞归巢促小鼠骨折愈合的实验研究[J].国际骨科学杂志,2015,36(3): 218-223.
[19] 王玉兰,何晓梅,唐薇,等.基质细胞衍生因子-1α/CXCR4/ CXCR7轴对骨髓间充质干细胞迁移的影响[J]. 解剖学报,2014,45(5):639-645.
[20] 陈鹏,张波,张连阳.间充质干细胞迁移的研究进展[J].中华临床医师杂志(电子版),2012,6(4):977-979.
[21] 徐秀萍,邱海波.间充质干细胞特异性归巢至损伤部位的研究进展[J].中华内科杂志,2016,55(6):483-485.
[22] 张博,刘娜,刘洪臣.SDF-1/CXCR-4 生物轴的研究进展[J].中华老年口腔医学杂志,2015,13(2):101-104.
[23] Liu X, Zhou C, Li Y, et al. SDF-1 promotes endochondral bone repair during fracture healing at the traumatic brain injury condition. PLoS One. 2013;8(1): e54077.
[24] 丁鹏,冯忠堂,杨智勇,等.趋化因子SDF-1体外趋化骨髓基质细胞迁移的实验研究[J].中华神经医学杂志,2007, 6(3): 25-227.
[25] Ratanavaraporn J, Furuya H, Kohara H, et al. Synergistic effects of the dual release of stromal cell-derived factor-1 and bone morphogenetic protein-2 from hydrogels on bone regeneration. Biomaterials. 2011;32(11):2797-2811.
[26] Higashino K, Viggeswarapu M, Bargouti M, et al. Stromal cell-derived factor-1 potentiates bone morphogenetic protein-2 induced bone formation. Tissue Eng Part A. 2011;17(3-4):523-530.
[27] Shyu WC, Lin SZ, Yen PS, et al. Stromal cell-derived factor-1 alpha promotes neuroprotection, angiogenesis, and mobilization/homing of bone marrow-derived cells in stroke rats. J Pharmacol Exp Ther. 2008;324(2): 834-849.
[28] Inai K, Burnside JL, Hoffman S, et al. BMP-2 induces versican and hyaluronan that contribute to post-EMT AV cushion cell migration. PLoS One. 2013;8(10): e77593.
[29] 姚琳,林江.骨髓间质干细胞归巢至损伤组织的研究进展[J].国际口腔医学杂志, 2016,43(2):177-180.
[30] 但洋,刘鹏,刘琦斌,等.骨形态发生蛋白-2、-4和-6在脂肪间充质干细胞向软骨细胞诱导中的调节作用[J].中华试验外科杂志,2014,31(4):808-811.
[31] Thieme S, Ryser M, Gentsch M, et al. Stromal cell-derived factor-1alpha-directed chemoattraction of transiently CXCR4-overexpressing bone marrow stromal cells into functionalized three-dimensional biomimetic scaffolds. Tissue Eng Part C Methods. 2009;15(4):687-696.
[32] Zhu W, Boachie-Adjei O, Rawlins BA, et al. A novel regulatory role for stromal-derived factor-1 signaling in bone morphogenic protein-2 osteogenic differentiation of mesenchymal C2C12 cells. J Biol Chem. 2007; 282(26):18676-18685.
[33] 曹鑫,陈慧锦,金格勒,等.人重组骨形态发生蛋白-2通过p38信号通路调节人脂肪间充质干细胞血管内皮生长因子表达机制的研究[J].中华创伤骨科杂志,2014,16(1): 67-70.
[34] Zhu Q, Liu C, Cai C. A novel robot visual homing method based on SIFT features. Sensors (Basel). 2015;15(10):26063-26084.
[35] La WG, Jin M, Park S, et al. Delivery of bone morphogenetic protein-2 and substance P using graphene oxide for bone regeneration. Int J Nanomedicine. 2014;9 Suppl 1:107-116.
[36] Kadekar D, Kale V, Limaye L. Differential ability of MSCs isolated from placenta and cord as feeders for supporting ex vivo expansion of umbilical cord blood derived CD34(+) cells. Stem Cell Res Ther. 2015;6: 201.
[37] 王俊文,程文俊,张青松,等.同种异体骨混合自体骨复合骨形态发生蛋白-2修复胫骨骨缺损的研究[J].中华实验外科杂志,2013,30(2):394-396.
[38] Hettiaratchi MH, Miller T, Temenoff JS, et al. Heparin microparticle effects on presentation and bioactivity of bone morphogenetic protein-2. Biomaterials. 2014; 35(25):7228-7238.
[39] 刘源,于江,白洁,等.海螵蛸-骨形态发生蛋白复合材料对骨缺损大鼠成骨及再血管化的影响[J].中华创伤杂志, 2013,29(6):548-553.
[40] 张悦.CXCR4 基因修饰骨髓间充质干细胞体外迁移实验[D].天津:天津科技大学,2008.
[41] De Becker A, Riet IV. Homing and migration of mesenchymal stromal cells: How to improve the efficacy of cell therapy? World J Stem Cells. 2016;8(3): 73-87.
[42] 张金池,关霖,刘翔,等.碱性成纤维生长因子基因修饰骨髓间充质干细胞治疗鼠后肢缺血[J].中华实验外科杂志, 2016,33(5):1197-1200.