Deuterium chloride

Poly(ethylene glycol) monomethyl ether MW 5000 kDa (mPEG), medium molecular weight chitosan (190–340 kDa), phthalic anhydride, anhydrous pyridine, 4-dimethylamino-pyridine (DMAP), succinic anhydride, anhydrous N,N-dimethylformamide (DMF), hydrazine monohydrate, hydroxybenzotrizole (HOBt), mucin from porcine stomach type III, egg yolk from chicken, and lactoferrin human were purchased from Sigma-Aldrich (St. Louis, MO, USA). Low molecular weight chitosan (oligosaccharide) (LMW OCs), 15 kDa, was obtained from Polysciences Inc., USA. Dulbecco’s Modified Eagle’s Medium (DMEM), Opti-MEM, and diethyl ether were obtained from Thermo Fisher Scientific Inc. (Waltham, MA, USA). Carbodiimide hydrochloride (EDC·HCl) and deuterium chloride were obtained from ACROS Organics (Morris Plains, NJ, USA). Deuterium oxide (D₂O) and dimethyl sulfoxide-d₆ were obtained from EMD Millipore Corporation (Billerica, MA, USA). pSpCas9-2A-GFP plasmid was purchased from Addgene (Plasmid #48138).

Medium molecular weight chitosan (MMW-CHT; ref.448877; 75% degree of deacetylation calculated by ¹H NMR – Figure S1 - Bruker DRX 300 Avance at 300.13 MHz; CHT was dissolved in a solution containing 98% D₂O and 2% DCl (v/v)), alginic acid sodium salt (ref. A3249), glacial acetic acid and phosphate buffer saline (PBS) were purchased from Sigma-Aldrich, and used as received. Sodium chloride (NaCl) was purchased from LabChem, and sodium hydroxide (NaOH) from AkzoNobel. Deuterium oxide and deuterium chloride (20% in Deuterium oxide) were purchased from Acros. Amicon® Ultra 4 mL centrifuge tubes with Ultracell 100 membrane (100 kDa cutoff), were purchased from Merck Millipore. Mouse fibroblast cell lines L929 (ref. 85011425) were purchased from Sigma-Aldrich, and human adipose-derived mesenchymal stem cells (hASCs; ref. ATCC® PCS-500-011™) from ATCC. Cell culture media Dubelcco’s Modified Eagle Medium (DMEM) was purchased from Sigma-Aldrich (ref. D2902). Minimum Essential Medium Eagle-Alpha Modification (αMEM; ref. 12000014), fetal bovine serum (FBS; ref. 10270106), Alamar Blue reagent (ref. DAL1025), Calcein AM (ref. C3099) and propidium iodide (ref. P1304MP), TrypLe™ Express Enzyme (ref. 12605010) and penicillin/streptomycin solution (ref. 15140122) were acquired from ThermoScientific.