Journal: Antioxidants
Article Title: Lyophilized Progenitor Tenocyte Extracts: Sterilizable Cytotherapeutic Derivatives with Antioxidant Properties and Hyaluronan Hydrogel Functionalization Effects
doi: 10.3390/antiox12010163
Figure Lengend Snippet: Comparative assessment of the TEAC values and hydrogel viscosity modulating properties of various doses of progenitor tenocyte whole cell samples (i.e., lyophilizates reconstituted in aqueous solvent), before and after submicron filtration and γ-irradiation, respectively. TEAC dose-response of reconstituted non-irradiated ( A ) or γ-irradiated (31 kGy, ( B )) whole cell samples containing 1.5 to 7.5 million cell equivalents/vial before and after 0.22 µm filtration, with the corresponding placebo controls. Results notably outlined a strong response (i.e., relative increase) of the γ-irradiated samples in TEAC measurements compared respectively to the same non-irradiated samples (( B ) vs. ( A )). Complex viscosity η* of reconstituted (i.e., in a hydrogel of HA 2.2–2.4 MDa MW at 1% in H 2 O:PBS 1:1) non-irradiated ( C ) or γ-irradiated (31 kGy, ( D )) whole cell samples containing 1.5 to 7.5 million cell equivalents/vial, with the corresponding placebo controls. Each sample was analyzed following the addition of H 2 O 2 (i.e., challenge item) or PBS (i.e., internal non-challenged controls) and incubation for 1 h at 37 °C. Very significant statistical differences (i.e., ** or 0.001 < p value < 0.01) or extremely significant statistical differences (i.e., *** or 0.0001 < p value < 0.001; **** or p value < 0.0001) were found between the presented mean values. HA, hyaluronic acid; kGy, kiloGray; LYO-WC, lyophilized whole cell fraction; MW, molecular weight; PBS, phosphate buffered saline; TEAC, Trolox equivalent antioxidant capacity.
Article Snippet: Eur.-grade dextran 40,000 (Pharmacosmos, Wiesbaden, Germany); D(+)-glucose, D(+)-mannose, and bovine serum albumin (BSA, Sigma-Aldrich, Buchs, Switzerland); mannitol, saccharose, lactose, and sorbitol (Hänseler, Herisau, Switzerland); D(+)-galactose, D(+)-mannose, and D(–)-fructose (Acros Organics, Geel, Belgium); xilitol (Alfa Aesar, Kandel, Germany); D(+)-trehalose (Apollo Scientific, Stockport, UK); Total Antioxidant Capacity (TEAC) Assay Kits, Ferric Reducing Antioxidant Power (FRAP) Assay Kits, 2-hydroxyethyl cellulose, and hydrogen peroxide at 30% w / w (Sigma-Aldrich, Buchs, Switzerland); DMEM cell culture medium (Life Technologies, Carlsbad, CA, USA); Penicillin-streptomycin (Thermo Fisher Scientific, Waltham, MA, USA); Millipore Stericup and Millex GS filter-sterilizing membranes with 0.22 μm pores (Merck, Darmstadt, Germany); nested 2R tubular glass lyophilization vials and bulk 6R clear glass vials (Schott, Mainz, Germany); laminated lyophilization stoppers (Adelphi Healthcare Packaging, Haywards Heath, UK and Flaver, Reinach, Switzerland); and lyophilization bags (Teclen, Oberpframmern, Germany).
Techniques: Viscosity, Solvent, Filtration, Irradiation, Incubation, Molecular Weight, Saline