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4 protocols using pcl mw 80 kda

1

Bioactive Hydrogel Scaffold Synthesis

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PCL (Mw=80 kDa), pluronic-F-127, gelatin, sodium borohydride, and Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dichloromethane (DCM) and N, N-dimethylformamide (DMF) were purchased from BDH Chemicals (Dawsonville, GA, USA). Dulbecco’s modified eagle medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin were obtained from Invitrogen (Carlsbad, CA, USA). CD 31, Ki67, TNF-α, IL-4, IL-6, IL-10 primary antibodies and Goat Anti-Rabbit IgG H&L (FITC), Goat Anti-Mouse IgG H&L (Alexa Fluor® 647), Goat Anti-Rat IgG H&L (Cy5®), Donkey Anti-Goat IgG H&L (Cy5®) secondary antibodies, phalloidin-iFluor 488 reagent (ab176753) were purchased from Abcam (Cambridge, MA, USA). CD206 and CCR7 primary antibodies were purchased from NovusBio (Centennial, CO, USA). The Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies, Inc (Rockville, MD, USA).
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2

Cytotoxicity Evaluation of Poly(ε-Caprolactone) on ARPE-19 Cells

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PCL (Mw, 80 kDa) was purchased from Sigma–Aldrich Co. (St. Louis, MO, USA). Formic acid (FA) and acetic acid (AA) were also bought from Merck Co. (Darmstadt, Germany). Human placentas were taken from the cesarean section of Milad hospital (Isfahan, Iran). ARPE-19 cells were purchased from the Isfahan Royan Institute (Isfahan, Iran). Dulbecco’s modified Eagle medium (DMEM (+) Glutamax. High glucose), fetal bovine serum (FBS), trypsin/EDTA, phosphate-buffered saline (PBS), penicillin–streptomycin, and MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) were obtained from Bio-Idea (Tehran, Iran).
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3

Neural Stem Cell Differentiation Protocol

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PCL (Mw= 80 kDa), pluronic-F-127, gelatin, sodium borohydride, Triton X-100 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dichloromethane (DCM) and N, N-dimethylformamide (DMF) were purchased from BDH Chemicals (Dawsonville, GA, USA). Dulbecco's modified eagle medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin were obtained from Invitrogen (Carlsbad, CA, USA). Basic fibroblast growth factor (FGF-2), epidermal growth factor (EGF), brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF) were obtained from Peptrotech (Rocky Hill, NJ, USA). The ReNcell Neural Stem Cell Medium was purchased from Millipore (Billerica, MA, USA). Tuj1 primary antibody and laminin were obtained from Millipore Sigma (Burlington, MA, USA). Goat anti-mouse IgG H&L (Alexa Fluor® 546) secondary antibody, Alexa Fluor™ 546 Phalloidin, DAPI were ordered from Abcam (Cambridge, MA, USA).
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4

3D Printed PCL Scaffolds for Tissue Engineering

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PCL scaffolds (l × w × h:10 mm × 10 mm × 10 mm; volume: 1,000 mm3) were printed using a 3DPLN2 bioprinter (3DPL, Tehran, Iran) with a Thermo polymer extruder equipped with a needle (inner diameter: 700 µm). PCL (Mw 80 kDa; Sigma-Aldrich®, St. Louis, MO, United States) was heated to 100°C before being extruded at 2 bar through a pre-heated needle. PCL strands were plotted layer-by-layer with an alternating 0°/90° lay-down pattern. In total, 30 scaffolds were printed for this study.
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