Two commercially available PDMS, Sylgard 527 gel and Sylgard 184 elastomer (Dow Corning), were used to create PDMS substrates with variable mechanical properties. Sylgard 527 was selected in this work, as a soft substrate, due to its very low elastic modulus (<10 kPa) within the physiological range of elastic moduli of the in vivo brain tissue (30 (link)). Once mixed (5:4), the PDMS 527 was poured into 35 mm diameter petri dishes to create ∼1-2 mm thick films. For the Stiff substrates a 50:1 mixture of Sylgard 184 was spin coated onto a slab of 1mm thick of PDMS 10:1 (Sylgard 184) to create ∼10 µm thick film. PDMS substrates were cured at 65°C overnight (12–24 hours) before all experiments. Substrates were washed with 70% ethanol and DI water and sterilized with UVB before seeding.
Sylgard 527 gel
Manufactured by Dow
Sourced in United States
Sylgard 527 gel is a two-part silicone-based potting and encapsulating compound. It is designed for applications requiring low viscosity, high elasticity, and electrical insulation. The product offers good thermal stability and chemical resistance.
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Lab products found in correlation
5 protocols using sylgard 527 gel
1
Engineered PDMS Substrates for Cell Culture
2
3D Printed Macropore-Mimetic Hemispheres for Tissue Engineering
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Macropore-mimetic hemispheres (MMH) were designed in Tinker CAD (https://www.tinkercad.com/ , accessed on 15 December 2021) with pore sizes that matched the average pore sizes determined by SEM image analysis (Figure 1 B) and were exported as an STL file (Figure S3A –C). MMH tools were 3D printed at the Duderstadt Center Fabrication Studio at the University of Michigan using a Stratasys J750 Polyjet 3D Printer (Rehovot, Israel) with 27 μm laser resolution and rigid opaque photopolymer.
Commercially available polydimethylsiloxane (PDMS), Sylgard 527 gel and Sylgard 184 elastomer (Dow Corning, Midland, MI, USA) were blended to match the mechanical properties of our PLLA scaffold materials. The ratio of Sylgard 184 to Sylgard 527 was 1:8, by weight [83 (link)]. The mixture was degassed in a desiccator and cast at room temperature, then cured for 24 h at 65 °C.
Prior to cell seeding, constructs were sterilized with ethylene oxide and 70% ethanol solution. Normal cell culture protocols were followed otherwise.
Commercially available polydimethylsiloxane (PDMS), Sylgard 527 gel and Sylgard 184 elastomer (Dow Corning, Midland, MI, USA) were blended to match the mechanical properties of our PLLA scaffold materials. The ratio of Sylgard 184 to Sylgard 527 was 1:8, by weight [83 (link)]. The mixture was degassed in a desiccator and cast at room temperature, then cured for 24 h at 65 °C.
Prior to cell seeding, constructs were sterilized with ethylene oxide and 70% ethanol solution. Normal cell culture protocols were followed otherwise.
3
Tunable PDMS Substrates for Cell Studies
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Polydimethylsiloxane (PDMS) substrates with tunable mechanical properties were created by blending two types of commercially available PDMS types, Sylgard 184 elastomer and Sylgard 527 gel (Dow Corning). Briefly, Sylgard 184 was made by mixing 10 parts base to 1 part curing agent by weight. Sylgard 527 was prepared by mixing equal weights of part A and part B. After both types of Sylgard 184 and 527 were prepared individually, they were then combined in different mass ratios and mixed for 5 min with a glass stir rod, as previously reported (Napiwocki et al., 2020 (link); Palchesko et al., 2012 (link)). For this study, PDMS substrates with a Young's modulus of 10 kPa were utilized, which corresponds to 1:50 ratio of Sylgard 184:527. After the PDMS blends were mixed together, they were then poured into a 100‐mm diameter Petri dish and cured overnight at 60℃. To overcome challenges associated with handling low‐modulus PDMS substrates, the PDMS was cured on top of a foundation layer of Sylgard 184 with a final thickness of 2 mm. Once cured, the PDMS substrates were removed from the Petri dish and cut into 1 cm squares with a razor blade and UV sterilized prior to the transfer of ECM.
4
Fabrication of Soft PDMS Substrates
5
Fabrication of PDMS Substrates with Tunable Stiffness
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PDMS substrates with varying stiffness were fabricated through blending sylgard 184 gel and sylgard 527 gel (Dow Corning, USA) as previously described (Palchesko, Zhang, Sun, & Feinberg, 2012) (link).
Briefly, base liquid and curing agent of sylgard 184 gel were mixed at a mass ratio of 10:1, while the part A and B of sylgard 527 gel were mixed equally. Subsequently, these two gels were blended with varying mass percentage of sylgard 184 from 0 to 100. Once defoamed in a Thinky-Conditioning mixer (Thinky Corporation, Japan), PDMS was poured into tissue culture plates to create ~2-mm thick films. All substrates were cured at 100 o C for 4 h, followed by treatment with UV-Ozone cleaner (Novascan Technologies, Ames, IA, USA) for 30 min and immediately coating with 25 μg/ml fibronectin (FN) (Corning) for 2 h before use for cell culture. FN coating was confirmed by fluorescence microscopic imaging of Alexa Fluor® 647 mouse anti-FN (BD Biosciences, San Jose, CA). The Young's modulus of PDMS substrates was determined by uniaxial tensile testing using a Shimadzu universal tester (Shimadzu, Japan) as previously described (Zhang et al., 2014) (link). Substrate topography was examined using an atomic force microscope (AFM) of Scanning Near-field Optical Microscopy (SNOM, NTEGRA Solaris, NT-MDT, Russia) in tapping mode with a scan point size of 256 × 256 over an area of 5 μm × 5 μm.
Briefly, base liquid and curing agent of sylgard 184 gel were mixed at a mass ratio of 10:1, while the part A and B of sylgard 527 gel were mixed equally. Subsequently, these two gels were blended with varying mass percentage of sylgard 184 from 0 to 100. Once defoamed in a Thinky-Conditioning mixer (Thinky Corporation, Japan), PDMS was poured into tissue culture plates to create ~2-mm thick films. All substrates were cured at 100 o C for 4 h, followed by treatment with UV-Ozone cleaner (Novascan Technologies, Ames, IA, USA) for 30 min and immediately coating with 25 μg/ml fibronectin (FN) (Corning) for 2 h before use for cell culture. FN coating was confirmed by fluorescence microscopic imaging of Alexa Fluor® 647 mouse anti-FN (BD Biosciences, San Jose, CA). The Young's modulus of PDMS substrates was determined by uniaxial tensile testing using a Shimadzu universal tester (Shimadzu, Japan) as previously described (Zhang et al., 2014) (link). Substrate topography was examined using an atomic force microscope (AFM) of Scanning Near-field Optical Microscopy (SNOM, NTEGRA Solaris, NT-MDT, Russia) in tapping mode with a scan point size of 256 × 256 over an area of 5 μm × 5 μm.
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