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Tunable viscoelastic polydimethylsiloxane substrates for cell mechanics and mechanobiology applications

机译：可调节的粘弹性聚二甲基硅氧烷基材，用于细胞力学和力学生物学应用

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Introduction: The physical and mechanical properties of the extracellular matrix (ECM) are known to influence and regulate cell fate and a wide variety of biological processes such as cell migration and differentiation. However, the underlying physical mechanisms behind the interactions between cells and their microenvironment is not well understood. Many studies have revealed profound effects of substrate elasticity on cellular responses. For instance, it has been shown that morphology, cytoskeletal structure, and cellular adhesion change in response to substrate elasticity. Substrate elasticity has also been shown to guide cell migration and to direct stem cell differentiation. In most of these studies, purely elastic materials were used to characterize cell-biomaterial interactions, however, most biological materials are viscoelastic and exhibit time-dependent deformation to the applied force, usually referred to as creep behavior. One clear consequence of the material's creep is energy dissipation. Unlike purely elastic materials where elastic energy is conserved, viscoelastic materials dissipate energy and constantly require input work to maintain a constant stress, commensurate with the rate of energy dissipation. The goal of this work is to establish a new platform to study mechanobiology and cell mechanics in response to viscoelastic properties of the ECM. Material and Method: Polydimethylsiloxane (PDMS) was purchased from Gelest. PDMS was functionalized for cell adhesion. A rheometer was used to characterize the viscoelastic properties of the substrates. Parallel plates with a diameter of 20 mm and a gap of 1000 μm were used. Frequency sweep and creep tests were performed. NIH 3T3 fibroblasts cells were cultured in a mixture of Dulbecco's Modified Eagle Medium, 10% fetal bovine serum, 1% Penicillin/Streptomycin at 37°C, in 5% CO2 humidified atmosphere. Cells were disassociated using 0.25% trypsin-EDTA when the cell confluency reached 70% and were cultured on the PDMS substrates' surface. Results: The fabricated PDMS substrates exhibit tunable viscoelastic properties. The shear and loss moduli were tunable between 4- 50 kPa and 0.2-7 kPa, respectively. Figure 1 shows the creep response of three PDMS samples. The shear stress was 2 kPa during creep test. This result indicates constant shear moduli in long time scale for these three samples but different time scale for their creep behavior. Cell culture results showed the spreading of NIH 3T3 cells on PDMS substrates. Discussion: Tunable viscoelastic PDMS substrates can provide us with a very strong tool to understand how cells sense and respond to the mechanical properties of their surrounding matrix. This viscoelastic platform can facilitate quantitative characterization of cell-ECM interactions to understand cell behavior during development and pathological conditions. Conclusions: PDMS-based tunable viscoelastic substrates with cell adhesion functionalities were fabricated and characterized for use in studying cell mechanics and mechanobiology.
机译：简介：已知细胞外基质（ECM）的物理和力学性能影响和调节细胞命运和各种生物过程，例如细胞迁移和分化。然而，细胞与其微环境之间的相互作用背后的潜在物理机制并不适合很好地理解。许多研究揭示了基材弹性对细胞反应的深刻影响。例如，已经显示出形态，细胞骨架结构和腹腔粘附响应于基底弹性而变化。还显示出基质弹性来引导细胞迁移并引导茎细胞分化。在这些研究中，使用纯弹性材料来表征细胞 - 生物材料相互作用，然而，大多数生物材料是粘弹性并且对施加的力表现出时效的变形，通常称为蠕变行为。物质蠕变的一个明确后果是能量耗散。与纯粹的弹性材料不同，弹性能量保守，粘弹性材料耗散能量并不断需要输入工作以保持恒定应力，以能量耗散的速度相称。这项工作的目标是建立一个新的平台，用于响应ECM的粘弹性的粘弹性性质研究机械学和细胞力学。材料和方法：从凝胶中购买聚二甲基硅氧烷（PDMS）。 PDMS官能化以用于细胞粘附。使用流变仪表征基材的粘弹性。使用直径为20mm和差距为1000μm的平行板。进行频率扫描和蠕变测试。 NIH 3T3成纤维细胞在Dulbecco的改性鹰培养基，10％胎牛血清，1％青霉素/链霉素的混合物中培养，在37℃下，在5％CO 2加湿气氛中。当细胞汇合达到70％时，使用0.25％胰蛋白酶-EDTA脱离细胞并在PDMS底物表面上培养。结果：制造的PDMS基材表现出可调粘弹性性能。分别在4-50kPa和0.2-7kPa之间进行剪切和损失模量。图1显示了三种PDMS样品的蠕变响应。蠕变试验期间剪切应力为2kPa。该结果表明这三个样本长时间刻度恒定的剪切模量，但是它们的蠕变行为的不同时间量表。细胞培养结果显示NIH 3T3细胞对PDMS基材的扩散。讨论：可调谐粘弹性PDMS基板可以为我们提供一个非常强大的工具，以了解细胞感觉如何感觉和响应其周围矩阵的机械性能。该粘弹性平台可以促进细胞-ECM相互作用的定量表征，以了解开发和病理条件期间的细胞行为。结论：基于PDMS的可调谐粘弹性基材，具有细胞粘附功能的粘弹性底物，并表征用于研究细胞力学和力学生物学。

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