Pluronics® is a member of the PEO-PPO-PEO triblock co-polymers. Pluronics® F-127 (PF-127, Sigma St Louis, MO) is a commercially available thermosensitive hydrogel. The aqueous solution of PF-127 at a concentration between 20-30% (wt.%) reversibly turns into a gel at room temperature [41 ]. The PF-127 (Sigma, St. Louis, MO) aqueous solutions were prepared by the cold method [42 (link)] by adding PF-127 slowly to cold (4°C) distilled water with gentle mixing until complete dissolution of the polymer. Stock concentration (28%) of PF-127 was stored at 4°C and cell suspension and compounds (liposomal nanoparticles, described above) were added to the stock to prepare the final concentration of PF-127 at 18-20% for this study.
Pf 127
Manufactured by Merck Group
Sourced in United States, China
PF-127 is a thermoreversible, water-soluble copolymer developed by Merck Group for use in various laboratory applications. It exhibits a reversible thermal gelation behavior, forming a semi-solid gel at physiological temperatures. The core function of PF-127 is to serve as a medium for the controlled release and delivery of various substances in laboratory settings.
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Lab products found in correlation
Axio observer z1, by Zeiss (3 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (2 mentions)
Ag490, by Merck Group (1 mentions)
Alamar blue solution, by Thermo Fisher Scientific (1 mentions)
D luciferin, by GoldBio (1 mentions)
Triethylamine, by J&K Scientific (1 mentions)
Hyaluronic acid, by Merck Group (1 mentions)
Hydroxypropyl methylcellulose hpmc k4m, by Merck Group (1 mentions)
Potassium dihydrogen phosphate, by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Pluronics f 127, by Merck Group (1 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Triton 100x, by Merck Group (1 mentions)
Penicillin streptomycin, by Biosera (1 mentions)
Alizarin red, by Merck Group (1 mentions)
Triethanolamine, by Fagron (1 mentions)
Ethanol, by Tedia (1 mentions)
Resazurine sodium salt, by Merck Group (1 mentions)
23 protocols using pf 127
1
Thermosensitive Pluronics F-127 Hydrogel
2
Nematode Attraction to Plant Root Tips
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Attraction to seedling root tips was assessed as described6 (link). Freshly hatched RKN J2 were suspended at approximately 200 per ml in 23% (wt/vol) PF-127 (Sigma-Aldrich, USA) augmented with a buffer composed of equimolar Tris and morpholinoethanesulfonic acid (MES) (one mM each or as indicated in text), unadusted pH7.1 (TM7). Five ml of suspension were delivered to each well of a 6 well tissue culture plate (#353046, Corning Inc., Corning, NY, USA) at 15 °C. One seedling (5 day old for tomato or 3 day old for Medicago) was introduced in the center of each well, and the plates were transferred to room temperature. After 4 h, the number of J2 touching the terminal 7 mm of the root was counted. Each treatment was replicated six times, and the experiments repeated at least four times.
3
Sterilization and Germination of Cucumber Seeds
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Cucumis sativus ’Jinyan No.4’ was used in this experiment. Before germination, seeds were treated with 10% (w/v) trisodium phosphate for 30 min to sterilize, followed by washing with sterile water several times, and soaking in sterile water for approximately 3 h. Finally, the sterilized seeds were placed on a Petri dish covered with wet Whatman paper and cultivated for 3–4 days at 28 °C in the dark for the next experiment when the lateral roots grew out.
Pluronic F-127 (PF-127) is a copolymer of propylene oxide and ethylene oxide, with negligible toxicity to nematodes or plant tissues. The nematodes suspended in PF-127 gel can move freely in three-dimensional space, which is more similar to their movement in the soil. Therefore, using this system to simulate the natural soil environment can more effectively assess host–pathogen interactions [54 (link)]. Based on these properties, the assay used PF-127 as the medium for nematode infection. PF-127 was purchased from Sigma-Aldrich (USA). The preparation was performed according to the protocol of Xing et al. [55 ]. In brief, 23 g of Pluronic F-127 powder was added to 80 mL of pre-cooled sterile water dissolved under stirring at 4 °C for 24 h, and then stored at 15 °C for later experimental use.
Pluronic F-127 (PF-127) is a copolymer of propylene oxide and ethylene oxide, with negligible toxicity to nematodes or plant tissues. The nematodes suspended in PF-127 gel can move freely in three-dimensional space, which is more similar to their movement in the soil. Therefore, using this system to simulate the natural soil environment can more effectively assess host–pathogen interactions [54 (link)]. Based on these properties, the assay used PF-127 as the medium for nematode infection. PF-127 was purchased from Sigma-Aldrich (USA). The preparation was performed according to the protocol of Xing et al. [55 ]. In brief, 23 g of Pluronic F-127 powder was added to 80 mL of pre-cooled sterile water dissolved under stirring at 4 °C for 24 h, and then stored at 15 °C for later experimental use.
4
AG490 Incorporation in P-F127 Micelles
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5
Cytotoxicity Evaluation of Drug Formulations
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PCL was from Polysciences, Inc. (Warrington, PA, USA) and PF127 was from Sigma-Aldrich (St. Louis, MO, USA); DOX was from MedChemExpress (Monmouth Junction, NJ, USA) and CYP was from Sigma-Aldrich; The alamarBlue solution was obtained from Invitrogen (AlamarBlue™ Cell Viability Reagent; Carlsbad, CA, USA); d -luciferin was from GoldBio, St. Louis, (MO, USA).
6
Synthesis of PF127-Diacrylate Conjugate
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The PF127-DA was prepared by acrylation of PF127 (poly(ethylene glycol)-b-poly(propylene glycol)-b-poly(ethylene glycol)) using acryloyl chloride according to reference13 (link). A volume of 2.54 g (0.2 mmol) of PF127 (Sigma-Aldrich) and 0.061 g of triethylamine (J&K Chemical) (0.6 mmol) were dissolved in 20 mL anhydrous dichloromethane in an ice bath and then degassed by pouring nitrogen for 20 min. After that, 0.05 mL of acryloyl chloride (0.6 mmol) was slowly injected into the above solution under nitrogen environment. The reaction was performed at room temperature for 24 h. Following the reaction, the solvent was removed by rotational evaporation, and the crude product was dissolved in DI water, and dialyzed exhaustively (MWCO 3500) against deionized water for three days. The pure product was obtained by lyophilization. The chemical structure of PF127-DA was confirmed by 1H NMR (Supplementary Fig. 16 ) and FT-IR (Supplementary Fig. 17 ) spectra.
7
Formulation and Evaluation of Hybrid Nanogels
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PTX, DOX, hyaluronic acid (HA, molecular weight (MW) = 100-150 kDa), PF127, polyoxyethylene-polyoxypropylene-polyoxyethylene tri-block copolymer, MW = 12.5 kDa), and hydroxypropyl methylcellulose (HPMC) K4M were purchased from Sigma Chemical Co. (St. Louis, MO). TPGS was procured from Eastman Co. (USA). Potassium dihydrogen phosphate and sodium hydroxide were bought from Merck (Darmstads, Germany). HPLC grade methanol and acetonitrile were supplied by Caledon (Ontario, Canada).
8
Characterization of Nanoparticle Formulations
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EU-RL was provided by Akbarie Co. (from RÖhm Pharma GMBh, Weiterstadt, Germany). RHT was kindly provided by Tofigh-Daru (Tehran, Iran). PF-127 (molecular weight of 9840-14 600) was purchased from Sigma-Aldrich (St. Louis, USA). For cell culture tests, RPMI-1640 Medium, 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) and Fluorescein isothiocyanate (FITC) from Sigma-Aldrich (Poole, UK), Human lung adenocarcinoma cell line (A549) from the national cell bank (Tehran, Iran) and fetal bovine serum (FBS) from GIBCO/Invitrogen (Paisley, UK) were obtained. Dialysis membrane (mol wt cut off10 000-12 000 Da) was supplied by Biogen (Mashhad, Iran). Phosphate buffered saline (PBS) and all other chemicals and solvents were of analytical grade. Deionized water was used throughout the study.
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EU-RL was provided by Akbarie Co. (from RÖhm Pharma GMBh, Weiterstadt, Germany). RHT was kindly provided by Tofigh-Daru (Tehran, Iran). PF-127 (molecular weight of 9840-14 600) was purchased from Sigma-Aldrich (St. Louis, USA). For cell culture tests, RPMI-1640 Medium, 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazolium bromide (MTT) and Fluorescein isothiocyanate (FITC) from Sigma-Aldrich (Poole, UK), Human lung adenocarcinoma cell line (A549) from the national cell bank (Tehran, Iran) and fetal bovine serum (FBS) from GIBCO/Invitrogen (Paisley, UK) were obtained. Dialysis membrane (mol wt cut off10 000-12 000 Da) was supplied by Biogen (Mashhad, Iran). Phosphate buffered saline (PBS) and all other chemicals and solvents were of analytical grade. Deionized water was used throughout the study.
9
Lentiviral Vector Delivery in Brachial Plexus Avulsion
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PF-127 (Sigma-Aldrich, St. Louis, MO, USA) was prepared as a 25% (w/v) suspension in 0.1 M phosphate-buffered saline (PBS, pH 7.6). The mixture was shaken gently overnight at 4°C until complete dissolution and then filtered (0.22 μm aperture) and stored at 4°C until further use.
Before injecting into the brachial plexus avulsion model, lentiviral vectors (2 × 108 TU/mL) were mixed with 25% PF-127 on ice to obtain a final lentiviral vector concentration of 2 × 107 TU/mL. All procedures were conducted under sterile conditions.
Before injecting into the brachial plexus avulsion model, lentiviral vectors (2 × 108 TU/mL) were mixed with 25% PF-127 on ice to obtain a final lentiviral vector concentration of 2 × 107 TU/mL. All procedures were conducted under sterile conditions.
10
Osteoblast Differentiation Assay
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RSV calcium, CTS (deacetylation degree: 85%, viscosity: 20-300 cP, molecular weight (Mw): 50-190 kDa), CS, HA, (Mw = 100-150 kDa), PF127 (polyoxyethylene-poly oxypropylene-polyoxy ethylene tri-block copolymer, Mw = 12.5 kDa), G, and β-GP disodium salt were provided from Sigma Chemical Co. (St. Louis, MO, USA). Dulbecco’s modified eagle’s minimal essential medium (DMEM), fetal bovine serum (FBS) and penicillin-streptomycin (100 IU/mL and 100 μg/mL, respectively) were obtained from Biosera (France). Human osteoblast-like MG- 63 cells line was purchased from Pasteur institute (Tehran, I.R. Iran). 3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Alizarin red, and Triton™ 100X were supplied by Sigma-Aldrich (St. Louis, MO, USA).
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