Resomer rg 503h

The survival of the donor cells of the recipient animals was supported by a single subcutaneous injection of tacrolimus-eluting polymer microspheres. We used tacrolimus in a dose of 0.25 mg/kg BW. Tacrolimus-eluting polymer microspheres were prepared from tacrolimus powder (Prograf^®, Astellas Pharma, Deerfield, IL, USA) and poly(D,L-lactide-co-glycolide) (Resomer RG 503H, Evonik, Germany) by adopting a procedure previously described by Wang et al. (Citation). The concentration of tacrolimus in polymer microspheres was 51.3 mg/g, as determined by HPLC. The proper application of tacrolimus-containing microspheres was performed six days before eye surgery and only once at a time. The tacrolimus levels in the blood of all minipigs are shown in Figure 2.

Ivacaftor was purchased from Matrix Scientific (Columbia, SC). Poly lactide-co-glycolide (PLGA, Resomer^® RG 503 H, MW= 24,000 – 38,000, viscosity= 0.32–0.44 dL/g in chloroform and Resomer^® RG 502 H, MW = 7,000 – 17,000, viscosity = 0.16–0.24 dL/g in chloroform) were purchased from Evonik (Parsippany, NJ). Polyvinyl alcohol (PVA, Mowiol 8–88, M.Wt 67,000), screw-capped dialysis tube (Spectra/PorTM Float-A-LyzerTM G2, MWCO 50 kDa), and phosphate-buffered saline (PBS) tablets were purchased from Sigma Aldrich (St. Louis, MO). Tween^® 80, methylene chloride (dichloromethane, DCM), and acetonitrile were obtained from Fisher Chemicals (Waltham, MA). Balb C Mouse plasma (Na Heparin) was obtained from Innovative research, Inc., (Novi, MI). All other chemicals and reagents used were of analytical or HPLC grade.

Medium molecular weight chitosan, was purchased from Sigma Aldrich (USA), pullulan was purchased from Alfa Aesar (USA), and Carbopol (Carbomer 941, NF) was purchased from Spectrum (USA). Poly(lactic‐co‐glycolic acid), with lactide and glycolide at a 50/50 molar ratio (Resomer® RG 503 H), was purchased from Evonik (Germany), acetic acid glacial was purchased from Carlo Erba (Italy), and SYLGARD™ 184 silicone elastomer kits were purchased from Dow Corning (USA). Acetonitrile was purchased from JT Baker (USA), and 2‐octyl‐CA (Dermabond®), Evicel®, and Surgicel® were purchased from Ethicon Inc. (USA). Duraseal® was purchased from Integra LifeSciences Corporation (France). Live/dead cell viability assay kits were purchased from Abcam (England), and CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS) kits were purchased from Promega (USA). Penicillin–streptomycin, fetal bovine serum, and l‐glutamine were purchased from IM Beit HaEmek (Israel). Pre‐hydrated collagen membranes were purchased from Vista International Packaging (USA), POLYDINE® was purchased from Dr. Fisher (Israel), and buprenorphine (1.3 mg/mL) was purchased from LogiPharm Ltd. (Israel).

PLGA-Nanoparticles were prepared by a double emulsion solvent evaporation method [14 ] using an ultra turrax^® (T25, IKA Labortechnik, Staufen, Germany) for emulsification steps. For the inner emulsion, Ovalbumin (albumin from chicken egg white, lyophilised powder ≥98%, Sigma-Aldrich, St. Louis, MO, USA) was dissolved in phosphate-buffered saline pH 7.4 (European Pharmacopoeia. 10.0/4.1.3; 4005000), obtaining the internal aqueous phase (W1). PLGA (Resomer^® RG 503 H, Evonik, Darmstadt, Germany) was dissolved in ethyl acetate (HPLC grade, Carl Roth, Karlsruhe, Germany) for the organic phase (O). W1 and O were combined and emulsified in a 15-mL-centrifuge tube before they were added to the external aqueous phase (W2), consisting of polyvinyl alcohol (PVA) (Mowiol 4-88, Sigma-Aldrich, St. Louis, MO, USA) in ultrapure water (Direct-Q^®, Merck, Darmstadt, Germany), in a 50-mL-centrifuge tube. Then, the inner emulsion and W2 were emulsified resulting in a W/O/W double emulsion. The double emulsion was added to the stabiliser solution and stirred at 400 rpm on a magnetic stirrer (IKA^® Ro 15, IKA Labortechnik, Staufen, Germany). Ethyl acetate was evaporated under continued stirring at 400 rpm overnight. The product was a nanoparticle suspension. Table 1 lists all compositions and quantities, all of which were held constant.