Dulbecco s modified eagle s medium nutrient mixture f 12 dmem f12

The activating reagents for amide bond formation, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), and N-hydroxysuccinimide (NHS) were supplied by Acros Organics (Geel, Belgium). N-isopropylacrylamide (NIPAM) and 2,2′-azobis(isobutyronitrile) (AIBN) obtained from Showa Chemical (Tokyo, Japan) were recrystallized from n-hexane and methanol, respectively. Hyaluronic acid (HA) (average molecular weight = 1.3 × 10⁶ Da) was purchased from Bloomage Freda Biopharm Co. (Jinan, China). Alcian blue, Safranin O, 2-morpholinoethane sulfonic acid (MES), 2-aminoethanethiol (AET), 2,4,6-trinitrobenzene sulfonic acid (TNBSA), and Dulbecco’s modified Eagle’s medium/nutrient mixture F-12 (DMEM-F12) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serums (FBS) and Live/Dead cell viability/cytotoxicity kit were obtained from Thermo Fisher Scientific (Waltham, MA, USA). For methyl tetrazolium salt (MTS) assays, the CellTiter96 AQueous One Solution Cell Proliferation Assay kit was obtained from Promega (Madison, WI, USA).

PLGA with a lactide-to-glycolide ratio of 85:15 (intrinsic viscosity 0.96 dL/g) was purchased from Green Chemical Inc., Taipei, Taiwan, while calcium hydrogen phosphate (Ca₂HPO₄·2H₂O) was purchased from Showa Co., Gyoda, Japan. Calcium carbonate (CaCO₃), sodium hydroxide (NaOH), cetylpyridinium chloride and poly(vinyl alcohol) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Dichloromethane (CH₂Cl₂) was procured from Alfa Aesar, Ward Hill, MA, USA, while Dulbecco’s Modified Eagle’s Medium (DMEM) and fetal bovine serum (FBS) were purchased from Invitrogen (Carlsbad, CA, USA) and Thermo Fisher Scientific (Waltham, MA, USA), respectively. Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 (DMEM/F-12) was purchased from Sigma-Aldrich (St. Louis, MO, USA). All chemicals were used as received without further purification.