total antioxidant capacity (teac) assay kits  (Millipore)

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: Schematic and illustrated technical overview of the main steps of the present study. The retained approach enabled a translational assessment of the cellular extracts under consideration, ranging from formulation options to ex vivo investigation of HA-containing combination product efficacy-related parameters. Steps 1 and 2 enabled the identification of optimally stable and sterilizable cellular derivatives. Step 3 enabled the characterization of the intrinsic properties and effects of the lyophilized cellular extracts. Step 4 enabled the study of the effects of the cellular extracts in a hydrogel environment with simulated oxidative stress within H 2 O 2 challenge assays (i.e., mimicking accelerated degradation conditions or a pathological in vivo environment). Steps 5 and 6 enabled the validation of the technical applicability of the extracts and combination products for potential therapeutic management of tendinopathies, based on formulation, preliminary safety, and efficacy-related parameters. FRAP, ferric reducing antioxidant power; HA, hyaluronic acid; 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: Formulation, Ex Vivo, In Vivo, Biomarker Discovery

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: Step-by-step presentation of the experimental plan of the main study, with reference to the questions addressed by the various assays and reference to the obtained experimental results (i.e., for each assay). A distinction is made between the functional attributes of the cellular extracts in lyophilizate form (i.e., intrinsic antioxidant activity) and the functional attributes of the cellular extracts reconstituted in HA (i.e., mainly the viscosity modulating functions, mediated by oxidative stress). References to the corresponding experimental results are provided (i.e., in blue font; figures, and tables). η*, complex viscosity; BSA, bovine serum albumin; FRAP, ferric reducing antioxidant power; HA, hyaluronic acid; LYO-WC, lyophilized whole-cell fraction; 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: Functional Assay, Antioxidant Activity Assay, Viscosity

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

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: TEAC values of various non-irradiated ( A ) and γ-irradiated (i.e., irradiation dose of 31 kGy, ( B )) progenitor tenocyte extracts, before and after 0.22 µm filtration, respectively. Results 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 )). Significant statistical differences (i.e., * or p value < 0.05), very significant statistical differences (i.e., ** or 0.001 < p value < 0.01), or extremely significant statistical differences (i.e., **** or p value < 0.0001) were found between the presented mean values. kGy, kiloGray; LYO-PLA, lyophilized placebo sample; LYO-LYS, lyophilized lysate fraction; LYO-MEM, lyophilized membrane fraction; LYO-SN, lyophilized soluble fraction; LYO-WC, lyophilized whole-cell fraction.

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: Irradiation, Filtration, Membrane

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: Experimental antioxidant capacities (i.e., TEAC values) of various stabilized whole-cell progenitor tenocyte samples (i.e., containing 7.5 × 10 6 cell equivalents/vial) or lyophilizate placebo samples containing two types of vial atmospheres (i.e., partial vacuum or atmospheric pressure air), respectively 1 . Values are presented as means assorted to the corresponding standard deviations. kGy, kiloGray; 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:

Journal: Antioxidants

Article Title: Aerobic Training Down-Regulates Pentraxin 3 and Pentraxin 3/Toll-Like Receptor 4 Ratio, Irrespective of Oxidative Stress Response, in Elderly Subjects

doi: 10.3390/antiox9020110

Figure Lengend Snippet: Effects of an 8-week aerobic training on plasma PTX3 ( a ) and oxidative biomarkers (GSH ( b ), TEAC ( c ), and total ROS/RNS ( d )) in both trained (TG) and control (CG) groups. Data are means ± SEM. GSH, reduced glutathione; PTX3, pentraxin 3; ROS/RNS, reactive oxygen/ nitrogen species; TEAC, trolox equivalent antioxidant capacity.

Article Snippet: The plasma pentraxin 3 (cat# ab214570, Abcam, Cambridge, UK), reduced glutathione (GSH, cat# E-BC-K030, Elabscience, Texas, USA) and total antioxidant capacity (TEAC) (ABTS assay, cat# AOX-1, Zenbio, Research Triangle Park, NC, USA) were measured through commercially available kits with the Epoch™ microplate spectrophotometer (BioTek Instruments, Winooski, VT, USA).

Techniques: