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2 protocols using tyramine hydrochloride

1

Synthesis and Characterization of Phenol-Functionalized Biomaterials

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HA-Ph (3.5-Ph groups per 100 repeating units) and gelatin-Ph (4.1 × 10−4 mol-Ph/g) were prepared according to the previous protocols, and the phenol contents were determined based on the tyramine standard curve (Figure S1) [26 (link),27 (link)]. Sodium hyaluronate (average molecular weight: 550 kDa, HA-LQ) was purchased from Kewpie (Tokyo, Japan) (molecular weight was measured as per the protocol described in Section 2.4. Molecular Weight Measurement). Tyramine hydrochloride, 3-(4-hydroxyphenyl)propionic acid, gelatin type B from bovine skin, and phalloidin-iFluor 488 (ab176759) were purchased from Chem-Impex (Wood Dale, IL, USA), Tokyo Chemical Industry (Tokyo, Japan), Sigma-Aldrich (St. Louis, MO, USA), and Abcam (Cambridge, UK), respectively. N-Hydroxysuccinimide, water-soluble carbodiimide hydrochloride, hyaluronidase, catalase from ovine, HRP, 31 w/w% hydrogen peroxide (H2O2) aqueous solution, and phosphate-buffered saline (PBS) containing 4 w/w% paraformaldehyde were purchased from FUJIFILM Wako Pure Chemical (Osaka, Japan).
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2

Multifunctional Hydrogel Synthesis and Characterization

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PEG-OH (8-arm, 20 kDa, JenKem), carbic anhydride (Sigma-Aldrich), 5-norbornene-2-carboxylic acid (Sigma-Aldrich), 4-dimethylaminopyridine (DMAP, Alfa Aesar), dopamine hydrochloride (Sigma-Aldrich), tyramine hydrochloride (Chem-Impex), isopropylamine (TCI chemicals), N,N’-Diisopropylcarbodiimide (DIC, Chem-Impex), N,N’-Dicyclohexylcarbodiimide (DCC, Sigma-Aldrich), hydroxybenzotriazole (HOBt, ≥20 wt% water, Oakwood Chemical), N,N’-diisopropylethylamine (DIPEA, TCI chemicals), cysteine (Sigma-Aldrich), and lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP, ≥95%, Sigma-Aldrich), 4-arms thiolated PEG (PEG4SH, 10 kDa, Laysan Bio), 5,5’-dithiobis-(2-nitrobenzoic acid) (Ellman’s reagent, Life Technologies), pyridine (Fisher), anhydrous tetrahydrofuran (THF, Fisher), anhydrous dimethylformamide (DMF, amine-free, Fisher), anhydrous dichloromethane (DCM, Fisher), methanol (Sigma-Aldrich) and diethyl ether (Fisher) were used as received.
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