Comparative effect of physicomechanical and biomolecular cues on zone-specific chondrogenic differentiation of mesenchymal stem cells.
Comparative effect of physicomechanical and biomolecular cues on zone-specific chondrogenic differentiation of mesenchymal stem cells.
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DOI：
10.1016/j.biomaterials.2016.03.034
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发表时间：
2016-06
期刊：
影响因子：
14
通讯作者：
Jabbari E
中科院分区：
文献类型：
--
作者：
Moeinzadeh S;Pajoum Shariati SR;Jabbari E
目前用于关节软骨再生的组织工程方法很少将组织恢复到正常状态，因为生成的组织缺乏天然软骨的复杂的带状组织。由于缺乏对物理、机械和生物分子线索与祖细胞的区域特异性软骨分化之间关系的了解，关节软骨的区域再生受到阻碍。这项工作以 3D 方式研究了 TGF-β1、区域特异性生长因子、最佳基质刚度和添加纳米纤维对分化的人间充质干细胞 (hMSC) 表达关节软骨浅层、中间和钙化区域特异的软骨形成标记物的影响。生长因子包括浅层、中层和钙化区的 BMP-7、IGF-1 和羟基磷灰石 (HA)；最佳基体刚度为80 kPa、2.1 MPa、320 MPa；纳米纤维水平、随机和垂直于凝胶表面排列。将具有区域特异性细胞密度的 hMSC 封装在工程水凝胶中，并在有或没有 TGF-β1、区域特异性生长因子、最佳基质模量和纤维添加的情况下培养，并在碱性软骨形成培养基中培养。通过 mRNA、蛋白质和生化分析来测量封装细胞的表达。结果表明，区域特异性基质硬度对 hMSC 向浅表和钙化区表型的软骨形成分化具有主导作用。添加平行于凝胶表面方向的排列纳米纤维显着增强 hMSCs 浅层软骨分化中 Col II 的表达。相反，生物分子因子IGF-1与TGF-β1联合对hMSCs的中区成软骨分化具有主导作用。这项工作的结果可能会导致模仿关节软骨的带状组织的多层移植物的开发。
Current tissue engineering approaches to regeneration of articular cartilage rarely restore the tissue to its normal state because the generated tissue lacks the intricate zonal organization of the native cartilage. Zonal regeneration of articular cartilage is hampered by the lack of knowledge for the relation between physical, mechanical, and biomolecular cues and zone-specific chondrogenic differentiation of progenitor cells. This work investigated in 3D the effect of TGF-β1, zone-specific growth factors, optimum matrix stiffness, and adding nanofibers on the expression of chondrogenic markers specific to the superficial, middle, and calcified zones of articular cartilage by the differentiating human mesenchymal stem cells (hMSCs). Growth factors included BMP-7, IGF-1, and hydroxyapatite (HA) for the superficial, middle, and calcified zones, respectively; optimum matrix stiffness was 80 kPa, 2.1 MPa, and 320 MPa; and nanofibers were aligned horizontal, random, and perpendicular to the gel surface. hMSCs with zone-specific cell densities were encapsulated in engineered hydrogels and cultured with or without TGF-β1, zone-specific growth factor, optimum matrix modulus, and fiber addition and cultured in basic chondrogenic medium. The expression of encapsulated cells was measured by mRNA, protein, and biochemical analysis. Results indicated that zone-specific matrix stiffness had a dominating effect on chondrogenic differentiation of hMSCs to the superficial and calcified zone phenotypes. Addition of aligned nanofibers parallel to the direction of gel surface significantly enhanced expression of Col II in the superficial zone chondrogenic differentiation of hMSCs. Conversely, biomolecular factor IGF-1 in combination with TGF-β1 had a dominating effect on the middle zone chondrogenic differentiation of hMSCs. Results of this work could potentially lead to the development of multilayer grafts mimicking the zonal organization of articular cartilage.