Chemical reagents

All tissue culture and molecular biology reagents were purchased from Life Technologies (Paisley, UK) unless stated otherwise. Chemical reagents were purchased from Sigma Aldrich (Poole, UK).

All tissue culture and molecular biology reagents were purchased from Life Technologies (Paisley, UK) unless stated otherwise. Chemical reagents were purchased from Sigma Aldrich (Poole, Dorset, UK).

Chemical reagents were obtained from Sigma Chemical Company (St. Louis, MO, USA) and HiMedia Labs (Mumbai, India). Nimbolide was purchased from Asthagiri Herbal Research Foundation, Chennai. Antibodies for GAPDH, TIMP-2, RECK, MMP-2, MMP-9, VEGF, VEGFR2 and HIF1α were purchased from Santa Cruz Biotechnology, USA. p-VEGFR2^Tyr1175, Notch-1, NICD, ADAM-10 and histone (H2B) antibodies were from Cell Signaling Technology, USA. Fetal bovine serum was from GIBCO, Invitrogen, NY, USA. Power SYBR Green PCR master mix was obtained from Applied Biosystems, California, USA. FuGENE transfection reagent was procured from Promega. Oligonucleotide primers were procured from Sigma Genosys, San Ramon, USA. All other reagents used were of analytical grade.

Chemical reagents were obtained from Sigma–Aldrich (St. Louis, MO, USA) unless otherwise specified. RPMI1640, fetal bovine serum (FBS), and penicillin/streptomycin were purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). Trypsin/EDTA was purchased from GE Healthcare Life Sciences (Logan, UT, USA). E-Cadherin rabbit polyclonal antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). AMPK-α rabbit polyclonal antibody, rabbit monoclonal antibodies specific to p-AMPK-α (Thr172), JAK2, p-JAK2 (Tyr1008), and occludin, and mouse monoclonal antibodies detecting STAT3, p-STAT3 (Tyr705), cleaved caspase 1, and IL-1β were purchased from Cell Signaling Technology Inc. (Boston, MA, USA). NOX2 and ICAM-1 rabbit monoclonal antibodies, and TGF-β, TNF-α, NOX1, p-p47phox, pro-caspase-1, and claudin 2 rabbit polyclonal antibodies were purchased from Abcam (Cambridge, MA, USA). Rabbit polyclonal antibodies specific for IL-10 or IL-6 were from Abbiotec (San Diego, CA, USA), and NLRP3 rabbit polyclonal antibody was purchased from Novus Biologicals (Centennial, CO, USA). Tofacitinib citrate salt was obtained from L.C Laboratories (Woburn, MA, USA). VAS2870 was purchased from Sigma-Aldrich (St. Louis, MO, USA). BJ-3105 was synthesized by Byeong-Seon Jeong as reported [47 ].

The chemical reagents were obtained from Sigma (St. Louis, MO, USA). The antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA) or Abcam (Cambridge, MA, USA). Polyvinylidene difluoride (PVDF) membranes were obtained from Millipore Corporation (Bedford, MA, USA). Dokdo-MARK^TM protein size marker was obtained from ElpisBiotech (Daejeon, Korea). (±)-Sodium 3-hydroxybutyrate was purchased from Sigma-Aldrich (St. Louis, MO, USA). All other materials obtained were of the highest available grade. The PKA inhibitor was obtained from Cayman Chemical (MI, USA).

Four collagen cross-linkers—grape seed extract (GSE), green tea extract (GTE), cranberry juice extract (CJE), and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide in combination with N-hydroxysuccinimide (EDC/NHS)—were used in this study. All treatment solutions, including the cross-linking solutions and control solution, were prepared with 0.96% phosphate buffered saline (PBS) solution according to our previous studies [9 (link)]. The concentrations of the polyphenols were kept the same among the natural cross-linking solutions based on the polyphenol percentage reported by manufacturers (Table 1). Because one of the major components in green tea extract is insoluble cellulose, the GTE solution was centrifuged and only the supernatant was used. The pH of the CJE solution was adjusted to near neutrality with NaOH to minimize any pH effects. The chemical cross-linker 0.3 M EDC/0.12 M NHS solution was prepared according to the literature [26 (link)].
Chemical reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). Formulations of reagents were based on those of previous studies [9 (link),10 (link),11 (link)]. Collagenase (from Clostridium histolyticum, type I, ≥125 U/mg) solution was made at 0.1% (w/v) in TESCA buffer (pH = 7.4, 0.36 mM CaCl₂, 50 mM N-tris (hydroxymethyl) methyl-2-aminoethanesulfonic acid).