(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (MTT), rhodamine 123 (Rho123), 5-fluorouracil, Pluronic F-68, and dimethylsulfoxide (DMSO) were from Sigma–Aldrich Chemie GmbH (Paris, France). PLGA Resomer® RG 503 H (was obtained from Evonik Industries, Essen, Germany. Phosphate buffer saline (PBS) was from Merck (Darmstadt, Germany). Fetal bovine serum (FBS) was from Gibco, Alfagene, Carcavelos, Portugal. RPMI-1640 medium (Roswell Park Memorial Institut), DMEM (Dulbecco’s Modified Eagle Medium), penicillin–streptomycin solution, antibiotic–antimycotic solution, L-glutamine, and trypsin/EDTA were from PAA (Vienna, Austria). Saccharomyces cerevisiae (ATCC 2601) cell culture, yeast extract peptone (YPD), HCT-116 (lung), MCF-7 (breast), and H460 (colon) human cancer cell lines were from the National Cancer Institute (Frederick, MD, USA). All other chemicals used in this study were of analytical grade and were purchased locally.
Plga resomer rg 503 h
Manufactured by Evonik
Sourced in Germany
PLGA Resomer® RG 503 H is a biodegradable and biocompatible copolymer composed of lactic acid and glycolic acid. It is a white to off-white powder.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Dimethyl sulfoxide (dmso), by Chempur (2 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Thiazolyl blue tetrazolium bromide, by Merck Group (2 mentions)
Pluronic f68, by Merck Group (1 mentions)
Phosphate buffer saline pbs, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Rpmi 1640 cell culture medium, by Lonza (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Merck Group (1 mentions)
Nile red, by Merck Group (1 mentions)
Dspe peg2000, by Lipoid (1 mentions)
Phospholipon 90g, by Lipoid (1 mentions)
7 hydroxyflavone, by Merck Group (1 mentions)
Tween 80, by Thermo Fisher Scientific (1 mentions)
Pd98059, by Selleck Chemicals (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Uranyl acetate, by Agar Scientific (1 mentions)
Pluronic f 127, by Merck Group (1 mentions)
Rpmi 1640 cell culture media, by Lonza (1 mentions)
7 protocols using plga resomer rg 503 h
1
Evaluation of Anticancer Compounds
2
Formulation and Characterization of PLGA-Based Nanocarriers
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PLGA Resomer RG 503 H, poly(D,L-lactide-co-glycolide), 50:50, Mw 24,000–38,000 was acquired from Evonik, Essen, Germany. SPC 90G (soy phosphatidylcholine, Phospholipon 90G), DDAB (dimethyldioctadecylammonium (bromide salt), and DSPE-PEG 2000 (N-(carbonyl-methoxypolyethylenglycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine 2000) were purchased from Lipoid, Ludwigshafen, Germany. Ursolic acid was purchased from Wuxi Cima, China. Thiazolyl Blue Tetrazolium Bromide, 4’,6-diamidino −2-phenylindole dihydrochloride (DAPI) and Nile Red were purchased from Merck, Darmstadt, Germany. RPMI-1640 cell culture medium was purchased from Lonza, Verviers, Belgium, Fetal bovine serum, GlutaMAX (L-alanyl-L-glutamine dipeptide in 0.85% NaCl), and 100× antibiotic-antimycotic were purchased from Life Technologies (Gibco/Life Technologies, Warszawa, Poland). Dimethylsulfoxide (DMSO) was purchased from ChemPur, Piekary Slaskie, Poland. Uranyl acetate and copper mesh (400 Mesh) with formware filters and carbon shells were purchased from Agar Scientific, Essex, UK.
3
Synthesis and Characterization of PLGA Nanoparticles
4
E. coli L-asparaginase Purification and PLGA Formulation
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L-asparaginase from E. coli was purified and provided ex gratia by PDC II Laboratory, National Institute of Immunology (NII) (New Delhi, India). Poly (lactic-co-glycolic acid) (PLGA) Resomer RG 503H, 50:50, 0.4 dL/g; MW 45,000, was provided as a gift by Evonik Industries AG (Essen, Germany). All the required chemicals were obtained from Sigma Aldrich (Mumbai, India). All were of high quality and HPLC grade. Double-distilled water was used throughout the experimentation. For evaluation of the particle size and polydispersity index, Zetasizer Nano ZS, Malvern Instruments (Malvern, United Kingdom) was used. For microscopic evaluation and observation, scanning electron microscopy, SEM; Hitachi, S-3400N (Tokyo, Japan) and transmission electron microscopy, TEM; CM-10; Philips, (Amsterdam, Netherlands) were carried out.
5
PLGA-PEG Nanoparticles for Ursolic Acid Delivery
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PLGA Resomer® RG 503 H, Poly(d ,l -lactide-co-glycolide), 50:50, Mw 24,000–38,000 was acquired from Evonik, Essen, Germany. PLGA-PEG 2000 (PEG average Mn 2000, PLGA average Mn 11,500, lactide:glycolide 50:50) and PLGA-PEG 5000 (PEG average Mn 5000, PLGA Mn 20,000, lactide:glycolide 50:50) were purchased from Merck, Darmstadt, Germany. Ursolic acid was purchased from Wuxi Cima, China. Pluronic F-127 and Thiazolyl Blue Tetrazolium Bromide were purchased from Merck, Germany. RPMI-1640 cell culture media was purchased from Lonza, Basel, Belgium., Fetal bovine serum, GlutaMAX™ (L-alanyl-L-glutamine dipeptide in 0.85% NaCl) and 100× antibiotic-antimycotic were purchased from Life Technologies (Gibco/Life Technologies, Warsaw, Poland). Dimethylsulfoxide (DMSO) was purchased from ChemPur, Piekary Śląskie Poland. Uranyl acetate and copper mesh (400 Mesh) with formware filter and carbon shell, were purchased from Agar Scientific, Essex, UK.
6
Curcumin-Loaded PLGA Nanoparticles with Chitosan Coating
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PLGA (Resomer RG503H) was purchased from Evonik Industries AG (Essen, Germany). CUR was obtained from Sigma Aldrich (Steinheim, Germany). Chitosan hydrochloride was supplied by Heppe Medical Chitosan GmbH (Halle/Saale, Germany). Polyvinyl alcohol (PVA, Mowiol 4-88) was provided by Kuraray (Hattersheim, Germany). Acetonitrile (ACN, HPLC gradient grade) was purchased from Fisher Scientific UK Ltd (Loughborough, UK). Water used was Millipore Q-Gard 2 by the water purification system of Merck Millipore (MA, USA).
Nanoparticles were prepared according to a previously reported modified solvent-displacement method [Citation23]. Batch sizes of 500 mg PLGA were dissolved in acetonitrile under vigorous stirring at room temperature for 10 min. Then, 500 μl of a 1% (m/V) solution of CUR in acetonitrile was added. The total prepared amount of 30-ml polymer-drug solution was injected into 200 ml stirring aqueous phase containing 2% PVA using a syringe pump with a flow rate of 0.5 ml/min. The dispersion was stirred overnight for solvent evaporation and then purified three times by centrifugation at 10,000 RCF for 20 min. For positively charged chitosan-coated nanoparticles, chitosan hydrochloride was dissolved in the aqueous phase in addition to PVA.
Nanoparticles were prepared according to a previously reported modified solvent-displacement method [Citation23]. Batch sizes of 500 mg PLGA were dissolved in acetonitrile under vigorous stirring at room temperature for 10 min. Then, 500 μl of a 1% (m/V) solution of CUR in acetonitrile was added. The total prepared amount of 30-ml polymer-drug solution was injected into 200 ml stirring aqueous phase containing 2% PVA using a syringe pump with a flow rate of 0.5 ml/min. The dispersion was stirred overnight for solvent evaporation and then purified three times by centrifugation at 10,000 RCF for 20 min. For positively charged chitosan-coated nanoparticles, chitosan hydrochloride was dissolved in the aqueous phase in addition to PVA.
7
Cytotoxicity and Genotoxicity Evaluation
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RPMI 1640, horse serum, fetal calf serum (FCS), non-essential amino acids and phosphate buffered saline (PBS) were purchased from GIBCO Invitrogen (Cergy-Pontoise, France). PolyLactic coGlycolic Acid copolymer (PLGA, Resomer® RG503H) was from Evonik (Essen, Deutschland). Benzo(a)pyrene, naphthalene, di-ethyl-hexyl phthalate, referred as ‘pollutant’ and H2-DCFDA, pyocyanine, giemsa reagent, penicillin, streptomycin, amphotericin B, cyclophosphamide (CPA, CAS No. 6055-19-2), mitomycin C (MitoC, CAS No. 50-07-7), methyl methane sulfonate (MMS, CAS No. 66-27-3), Triton X-100, EDTA, HEPES, trizma base, propidium iodide, KCl, NaCl, sodium bicarbonate, sodium pyruvate, pluronic F68 solution, Trypan Blue, bovine serum albumin (BSA) were obtained from Sigma-Aldrich (Saint-Quentin Fallavier, France). Normal melting point agarose and low melting point agarose were purchased from Bio-rad (Marnes-la-Coquette, France). Acetic acid was purchased from VWR (Fontenay sous-bois, France). Dimethyl sulfoxide (DMSO) from Acros Organics (Noisy le Grand, France). Trypsin was obtained from Biochrom AG (Berlin, Germany), while NaOH, L-glutamine and absolute ethanol were from Merck (Darmstadt, Germany). The repair endonuclease hOgg1 was purchased from New England Bio-Labs via Ozyme (Saint Quentin en Yvelines, France).
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