PLGA MSs were prepared according to a solid-in-oil-in-water (S/O/W) emulsion-solvent evaporation technique previously described [30 (link)]. Briefly, 20 μg of GDNF (R&D, Systems, Minneapolis, MN) were first suspended in 20 μl of VitE (Sigma-Aldrich, Schnelldorf, Germany) via gentle sonication for 30 s at low temperature (0 °C; Sonicator XL, Heat System, Inc., Farmingdale, NY). Once formed, the suspension was added to 1 ml of PLGA (Resomer®503, Boehringer Ingelheim Pharma GmbH & Co, Ingelheim, Germany) solution in methylene chloride (20% w/v) to elaborate the GDNF-loaded PLGA/VitE MSs or a solution of 20 mg of melatonin (Sigma-Aldrich) and 200 mg of PLGA in 1 ml of methylene chloride to prepare the GDNF/melatonin-loaded PLGA/VitE MSs. The organic phase was then emulsified in both cases with 5 ml of polyvinyl alcohol (PVA) 72,000 g/mol (Merck KGaA, Darmstadt, Germany) MilliQ® water solution (2% w/v) in a homogenizer (Polytron® RECO, Kinematica GmbH PT 3000, Lucerna, Switzerland) at 5,000 rpm for 1 min and 6,000 rpm for 1 min. This emulsion was subsequently poured on 100 ml of an aqueous PVA solution (0.1%) and kept under constant stirring for 3 h, to allow evaporation of the organic solvent. Once formed, microspheres were washed to eliminate PVA, filtered, freeze-dried, and kept at −20 °C under dry conditions until use.
Resomer 503
Manufactured by Boehringer Ingelheim
Sourced in Germany
Resomer®503 is a biodegradable polymer used in the production of laboratory equipment. It is composed of polylactic acid (PLA) and has a molecular weight of approximately 50,000 g/mol.
Automatically generated - may contain errors
3 protocols using resomer 503
1
Preparation of GDNF-Loaded PLGA Microspheres
2
PLGA Polymer-Based Drug Delivery
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PLGA (50:50) polymer (Resomer®503) was purchased from Boehringer Ingelheim Pharma GmbH & Co. (Ingelheim, Germany). Tauroursodeoxycholic acid (TUDCA) and polyvinyl alcohol 72000 g/mol (PVA) were obtained from Merck KGaA (Darmstadt, Germany). All organic solvents were HPLC-grade and used as received.
3
Spironolactone-Loaded PLGA Microspheres
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PLGA ratio 50:50 (Resomer®503) was purchased from Boehringer Ingelheim Pharma GmbH & Co. (Ingelheim, Germany). Spironolactone was obtained from Sigma-Aldrich (Schnelldorf, Germany). Polyvinyl alcohol 72,000 g/mol (PVA) was obtained from Merck KGaA (Darmstadt, Germany). All organic solvents were high performance liquid chromatography (HPLC) grade and used as received.
Spironolactone-loaded and non-loaded PLGA MSs were elaborated using an oil-in-water (O/W) emulsion solvent evaporation technique35 (link). Briefly, The O-phase was prepared by suspension of 40 mg of Spironolactone in 1 ml of PLGA solution in methylene chloride (20% w/v) resulting in a Spiro:PLGA ratio of 2:10. This phase was emulsified with the W-phase composed by 5 ml of PVA MilliQ® water solution (2% w/v). The emulsification was performed at 5,000 rpm for 1 min (Polytron® RECO, Kinematica GmbH PT 10–35, Lucerna, Switzerland). This O/W emulsion was subsequently poured onto 100 ml of an aqueous PVA solution (0.1%) and maintained under constant stirring for 3 h to allow MSs hardening. After that, the MSs were washed, filtered, freeze-dried, and kept at −20 °C under dry conditions until used.
Spironolactone-loaded and non-loaded PLGA MSs were elaborated using an oil-in-water (O/W) emulsion solvent evaporation technique35 (link). Briefly, The O-phase was prepared by suspension of 40 mg of Spironolactone in 1 ml of PLGA solution in methylene chloride (20% w/v) resulting in a Spiro:PLGA ratio of 2:10. This phase was emulsified with the W-phase composed by 5 ml of PVA MilliQ® water solution (2% w/v). The emulsification was performed at 5,000 rpm for 1 min (Polytron® RECO, Kinematica GmbH PT 10–35, Lucerna, Switzerland). This O/W emulsion was subsequently poured onto 100 ml of an aqueous PVA solution (0.1%) and maintained under constant stirring for 3 h to allow MSs hardening. After that, the MSs were washed, filtered, freeze-dried, and kept at −20 °C under dry conditions until used.
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