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1

Fabrication and Characterization of BETA Scaffold

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The BETA scaffold was manufactured, as previously described [21 (link)]. Briefly, the copolymer emulsion of poly(ε-caprolactone) (PCL: Sigma-Aldrich, Saint Louis, MO, USA, Mn 80,000), and gelatin (Type A from porcine skin, Sigma) dissolved in TFE ((2,2,2-trifluoroethanol; Roth) were spin-coated (2000 rpm) and were dried under vacuum (300 mbar). The optimum mixing ratio of 9.35% PCL and 6.34% gelatin [w/v solvent] was used for the co-polymer of PCL/gelatin as previously described [22 (link)]. Membranes were sterilized before cell culture experiments with PBS, ethanol 80%, and UV exposure. Scanning electron microscopy (SEM, Zeiss Crossbeam 340, Carl Zeiss AG, Oberkochen, Germany) was used to study the morphology of the BETA scaffolds. The cells were fixed in 2% v/v glutaraldehyde (GA; Sigma-Aldrich) and then dehydrated in gradient acetone solutions (Sigma-Aldrich) followed by hexamethyldisilazane (HDMS, Sigma-Aldrich) for 15 min. The samples were subsequently sputter-coated with platinum and analyzed at an operating voltage of 2 kV.
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2

Gelatin-Chitosan Hydrogel Fabrication

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Gelatin (type A, from porcine skin) and chitosan (low molecular weight) was purchased from Sigma-Aldrich (Shanghai, China). Methacrylic anhydride was purchased from Aladdin (Shanghai, China). 3,4-Dihydroxyhydrocinnamic acid, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), ethanol (AR, 99.7%), DL-dithiothreitol (DTT) and zinc chloride (ZnCl2) were purchased from Macklin (Shanghai, China). CCK-8 proliferation assay kit was purchased from Beyotime (Shanghai, China). Dulbecco’s modified Eagle’s medium/Ham’s F-12 medium (DMEM/F-12), induction media (osteogenic, chondrogenic and adipogenic) and fetal bovine serum (FBS) were ordered from Invitrogen. Cell culture plate was ordered from Corning.
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3

Photopolymerizable Hydrogel Fabrication

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Triethylamine (TEA), Diphenyl (2,4,6-trimethylbenzoyl) phosphine oxide (TPO), anhydrous dichloromethane (DCM), D-chloroform (CDCl3) polyethylene glycol di-acrylate (PEGDA) with molecular weight of 700 Da and gelatin Type A from porcine skin were purchased from Millipore Sigma Aldrich (St. Louis, MO). Sebacic acid (SA), glycerol, 4-Dimethylaminopyridine (DMAP), Ethyl Acetate (EA) were purchased from Fisher Scientific (Waltham, MA). Acryloyl chloride (AC) was purchased from Alfa Aesar (Haverhill, MA). For cell culture and viability tests, Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS) and penicillin/streptomycin (P/S) Calcein AM were purchased from Thermo Fisher Scientific (Waltham, MA). Ethidium Homodimer was purchased from Biotium (Hayward, CA).
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