Defining the role of matrix compliance and proteolysis in three-dimensional cell spreading and remodeling
Defining the role of matrix compliance and proteolysis in three-dimensional cell spreading and remodeling
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DOI：
10.1529/biophysj.107.105841
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发表时间：
2008-04-01
影响因子：
3.4
通讯作者：
Seliktar, Dror
中科院分区：
文献类型：
--
作者：
Dikovsky, Daniel;Bianco-Peled, Havazelet;Seliktar, Dror
最近的研究已经确定细胞外基质（ECM）的顺应性作为一个有影响力的因素，在决定的锚定依赖性细胞的命运。我们探索了一种方法来检查ECM顺应性对三维培养中细胞形态和重塑的影响。为此，开发了生物ECM类似物材料以伪独立地改变其生物化学和物理性质。制备了一组18种材料变体，其剪切模量范围为10至700 Pa。平滑肌细胞被封装在这些材料和延时视频显微镜被用来显示基质模量，蛋白水解生物降解，细胞扩散，和细胞压实的矩阵之间的关系。对每种材料变体的基质的蛋白水解敏感性、基质压实程度和细胞形态进行定量，以与模量数据相关。最初的细胞扩散到水凝胶基质中取决于材料的蛋白水解敏感性，而细胞压实的程度被证明与材料的模量更相关。抑制基质金属蛋白酶深刻地影响了最初的细胞扩散和重塑，即使在最顺应性的材料。我们的结论是，平滑肌细胞利用蛋白水解形成板状伪足和牵引力收缩和重塑其周围的微环境。因此，基质模量可用于控制细胞重塑和压实的程度。这项研究进一步表明，ECM中基质模量和蛋白水解抗性之间的相互联系可能部分解偶联，以深入了解细胞如何解释其物理三维微环境。
Recent studies have identified extracellular matrix (ECM) compliance as an influential factor in determining the fate of anchorage-dependent cells. We explore a method of examining the influence of ECM compliance on cell morphology and remodeling in three-dimensional culture. For this purpose, a biological ECM analog material was developed to pseudo-independently alter its biochemical and physical properties. A set of 18 material variants were prepared with shear modulus ranging from 10 to 700 Pa. Smooth muscle cells were encapsulated in these materials and time-lapse video microscopy was used to show a relationship between matrix modulus, proteolytic biodegradation, cell spreading, and cell compaction of the matrix. The proteolytic susceptibility of the matrix, the degree of matrix compaction, and the cell morphology were quantified for each of the material variants to correlate with the modulus data. The initial cell spreading into the hydrogel matrix was dependent on the proteolytic susceptibility of the materials, whereas the extent of cell compaction proved to be more correlated to the modulus of the material. Inhibition of matrix metalloproteinases profoundly affected initial cell spreading and remodeling even in the most compliant materials. We concluded that smooth muscle cells use proteolysis to form lamellipodia and tractional forces to contract and remodel their surrounding microenvironment. Matrix modulus can therefore be used to control the extent of cellular remodeling and compaction. This study further shows that the interconnection between matrix modulus and proteolytic resistance in the ECM may be partly uncoupled to provide insight into how cells interpret their physical three-dimensional microenvironment.