A custom-built SES apparatus is employed for the fabrication of two-dimensional (2D) non-woven PCL microfibrous meshes. A volume of 5 mL PCL working solution with 12% [w/v] final concentration is prepared by mixing PCL pellets with HFIP by gentle overnight magnetic stirring at 1000 RPM. A stainless-steel needle tip (21 gauge) with blunt end is attached on a plastic Luer-lock syringe (3 mL). 1 mL of final PCL working solution is slowly loaded in the syringe barrel without introducing any bubbles. The loaded syringe is clamped on a programmable syringe pump (Harvard Apparatus). The positive lead of a high DC voltage power supply is attached on the needle tip and the ground lead on an aluminum collector vertically placed at a distance equal to 12–13 cm. PLL coated glass coverslips are taped on the grounded aluminum collector. Non-woven fibrous meshes are collected on the glass coverslips at a volumetric flow (Q) rate equal to 10 μL/min (Q = 10 μL/min) and voltage potential (Vp) equal to 15 (Vp = 15 kV) for different spinning times. Samples obtained for spinning time equal to 1 min and 3 min are designated as “SES-1 min” and “SES-3 min”, respectively.
Programmable syringe pump
Manufactured by Harvard Apparatus
The Programmable Syringe Pump is a laboratory device used to precisely control the flow rate and volume of liquids dispensed from a syringe. It is capable of accurately delivering small, controlled amounts of fluids for various experimental and analytical applications.
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6 protocols using programmable syringe pump
1
Fabrication of 2D PCL Microfibrous Meshes
2
Simulating Interstitial Flow in HUVEC Cultures
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Fluid flow was controlled with a programmable syringe pump (Harvard Apparatus). The flow medium was the same as the growth medium (EGM). Before initiation of flow-based experiments, clear polypropylene barbed elbow fittings (1/16 inch, Cole-Parmer) connected to silicone tubing (Saint-Gobain) were inserted into the 1.5-mm diameter inlet/outlet ports of the HUVEC channels. The opposite inlet/outlet ports of the HUVEC channels were connected to luer adapters (Cole-Parmer) which served as fluid reservoirs. To subject endothelial cells solely to interstitial flow (transverse convection), we pulled media through the collagen matrix from the reservoirs connected to the other endothelial-lined channel, while exposing both channels to negligible levels of tangential shear. A pressure gradient across the collagen gel is also generated. In this configuration, fluid extravasates from one HUVEC channel, convects through the collagen and intravasates into the other channel. The flow rate was 7.3 μl/h corresponding to a flow velocity of 60 μm/sec. Devices without imposed flow served as controls (“static”). In this case, the media was changed every day.
3
Perfusion Culture of Endothelial Cells
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Cells in the macroscale device were subjected to flow for 1 h using a programmable syringe pump (Harvard Apparatus) (Fig. 1d ). The flow rate for both configurations was 0.7 ml/h corresponding to a bulk flow velocity of 0.5 μm/sec. The flow medium was the same as the growth medium (i.e. EGM). Exogenous VEGF (50 ng/ml) was added to the media as per experimental requirements. Gels not exposed to flow served as control (“static”).
4
Electrospinning of PVP/MEH-PPV Composite Fibers
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Solutions are prepared
by dissolving 30 mg of MEH-PPV (MW = 150–250
kDa, Sigma Aldrich) and 30 mg of PVP (MW = 1.3 × 106 Da, Alfa Aesar) in 1 mL of a mixture
of chloroform (CHCl3, Sigma Aldrich) and dimethyl sulfoxide
(DMSO, Sigma Aldrich) with a volumetric ratio of 9:1 (v/v). The solutions
are stirred for 24 h. Next, WO3 NWs (Sigma Aldrich) are
added to the solution to obtain a weight ratio of 20% with respect
to the dry weight of the PVP/MEH-PPV mixture. The solutions are sonicated
for 2 h and then injected through a 21G needle with a flow rate of
2 mL/h by using a programmable syringe pump (Harvard Apparatus). A
7 kV voltage bias (EL60R0.6-22, Glassman High Voltage) is applied
between the needle and a drum with a grounded disk (8 cm diameter
and 1 cm thickness) rotating at 2000 rpm and positioned at a distance
of 5 cm from the needle. For optical characterization, fibers are
deposited on quartz substrates (1 × 1 cm2, thickness
1 mm).
by dissolving 30 mg of MEH-PPV (MW = 150–250
kDa, Sigma Aldrich) and 30 mg of PVP (MW = 1.3 × 106 Da, Alfa Aesar) in 1 mL of a mixture
of chloroform (CHCl3, Sigma Aldrich) and dimethyl sulfoxide
(DMSO, Sigma Aldrich) with a volumetric ratio of 9:1 (v/v). The solutions
are stirred for 24 h. Next, WO3 NWs (Sigma Aldrich) are
added to the solution to obtain a weight ratio of 20% with respect
to the dry weight of the PVP/MEH-PPV mixture. The solutions are sonicated
for 2 h and then injected through a 21G needle with a flow rate of
2 mL/h by using a programmable syringe pump (Harvard Apparatus). A
7 kV voltage bias (EL60R0.6-22, Glassman High Voltage) is applied
between the needle and a drum with a grounded disk (8 cm diameter
and 1 cm thickness) rotating at 2000 rpm and positioned at a distance
of 5 cm from the needle. For optical characterization, fibers are
deposited on quartz substrates (1 × 1 cm2, thickness
1 mm).
5
Interstitial Fluid Flow in Cell Constructs
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During imaging, interstitial fluid flow was applied to the cell/gel construct within the flow chamber with phenol red-free and serum-free DMEM. Here, fluid flow was applied through pores within the gel in which cells were embedded. An environmental chamber (Ibidi) was used to maintain temperature (37 °C) and CO2 level (5%). A programmable syringe pump (Harvard Apparatus) was used to apply interstitial fluid flow to the cells by controlling the flow rate. A pulsatile (0.2 Hz) flow at four different flow rates (2, 5, 10, and 20 μl/min) was used.
6
Alginate Encapsulation of Porcine Islets
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