Vibrio cholerae -loaded poly(DL lactide co-glycolide) microparticles

Vibrio cholerae (VC)-loaded microparticles were prepared using poly(DL lactide-co-glycolide) with a water-in-oil-in-water emulsion/solvent extraction technique. Particle characteristics including size distribution, VC-loading efficiencies, and in-vitro release pattern were investigated. The dispersed phase was PLG dissolved in dichloromethane, and the continuous phase was water containing PVP as a stabilizer with varied sodium chloride concentrations. VC was successfully entrapped in the microparticles with trapping efficiencies up to 97.8%, a loading level of 55.4 g/mg, and particle size of 3.8 #181;m. Using 10% w/v PVP with 5% w/v NaCl in the continuous phase resulted in a higher loading level (55.4 #45 6.9 g/mg), loading efficiency (97.8%), core region content (25.7 #45 1.9 g/mg) and lower surface content (6.2 #45 0.9 g/mg) than without NaCl (loading content: 40.7 #45 5.1 g/mg; loading efficiency 52.1%; core region content: 8.3 #45 0.5 g/mg; surface content: 19.5 #45 1.1 g/mg). A linear release profile from VC-loaded microparticles was found. A preliminary animal oral administration study indicated that the VC-loaded microparticles, as an oral delivery system, have shown effective immunogencity in rats for 2 months. The VC incorporation and physicochemical characterization data obtained in this study may be relevant in optimising the vaccine incorporation and delivery properties of these potential vaccine targeting carriers.     

Inactive Vibrio cholerae whole-cell vaccine-loaded biodegradable microparticles: in vitro release and oral vaccination

An approach is proposed using Vibrio cholerae (VC)-loaded microparticles as oral vaccine delivery systems for improved vaccine bioavailability and increased therapeutic efficacy. The VC-loaded microparticles were prepared with 50:50 poly(DL-lactide-co-glycolide) (PLG), 75:25 poly(DL-lactide-co-glycolide) and poly(lactide acid) (PLA)/PEG blend copolymers by the solvent evaporation method. VC was successfully entrapped in three types of microparticles with loading efficiencies and loading levels as follows: 50:50 PLG systems: 97.8% and 55.4 +/- 6.9 micro g/mg; 75:25 PLG systems: 89.2% and 46.5 +/- 4.4 micro g/mg; PLA/PEG-blended systems: 82.6% and 53.7 +/- 5.8 micro g/mg. The different distributions of VC in the core region and on the surface were as follows: 50:50 PLG systems 25.7 +/- 1.9 and 6.2 +/- 0.9 micro g/mg; 75:25 PLG systems: 25.8 +/- 2.2 and 3.6 +/- 0.4 micro g/mg; PLA/PEG-blended systems: 32.4 +/- 2.1 and 5.2 +/- 1.0 micro g/mg, respectively. In vitro active release of VC was affected mainly by matrix type and VC-loaded location in microparticles. The therapeutic immunogenic potential of VC loaded with 50:50 PLG, 75:25 PLG and PLA/PEG-blended microparticles was evaluated in adult mice by oral immunization. Significantly higher antibody responses and serum immunoglobin Ig G, IgA and IgM responses were obtained when sera from both VC-loaded 75:25 PLG and PLA/PEG-blended microparticles immunized mice were titrated against VC. The most immunogenicity in evoking serum IgG, IgA and IgM responses was immunized by VC-loaded PLA/PEG-blended microparticles, and with VC challenge in mice, the survival rate (91.7%).     

Inactive Vibrio cholerae whole-cell vaccine-loaded biodegradable microparticles: in vitro release and oral vaccination

An approach is proposed using Vibrio cholerae (VC)-loaded microparticles as oral vaccine delivery systems for improved vaccine bioavailability and increased therapeutic efficacy. The VC-loaded microparticles were prepared with 50:50 poly(DL-lactide-co-glycolide) (PLG), 75:25 poly(DL-lactide-co-glycolide) and poly(lactide acid) (PLA)/PEG blend copolymers by the solvent evaporation method. VC was successfully entrapped in three types of microparticles with loading efficiencies and loading levels as follows: 50:50 PLG systems: 97.8% and 55.4 ± 6.9 µg/mg; 75:25 PLG systems: 89.2% and 46.5 ± 4.4?µg/mg; PLA/PEG-blended systems: 82.6% and 53.7 ± 5.8?µg/mg. The different distributions of VC in the core region and on the surface were as follows: 50:50 PLG systems 25.7 ± 1.9 and 6.2 ± 0.9?µg/mg; 75:25 PLG systems: 25.8 ± 2.2 and 3.6 ± 0.4?µg/mg; PLA/PEG-blended systems: 32.4 ± 2.1 and 5.2 ± 1.0?µg/mg, respectively. In vitro active release of VC was affected mainly by matrix type and VC-loaded location in microparticles. The therapeutic immunogenic potential of VC loaded with 50:50 PLG, 75:25 PLG and PLA/PEG-blended microparticles was evaluated in adult mice by oral immunization. Significantly higher antibody responses and serum immunoglobin Ig G, IgA and IgM responses were obtained when sera from both VC-loaded 75:25 PLG and PLA/PEG-blended microparticles immunized mice were titrated against VC. The most immunogenicity in evoking serum IgG, IgA and IgM responses was immunized by VC-loaded PLA/PEG-blended microparticles, and with VC challenge in mice, the survival rate (91.7%).     

The mechanism of PLA microparticle formation by water-in-oil-in-water solvent evaporation method

Microparticles containing ovalbumin as a model protein drug were prepared using poly(L-lactide; PLA) with a water-in-oil-in-water emulsion/solvent evaporation technique. The dispersed phase was PLA dissolved in dichloromethane (DCM), and the continuous phase was water-containing polyvinyl pyrolidone (PVP) as stabilizer with sodium chloride. Microparticle characteristics, loading efficiencies, protein distribution in microparticles, and in-vitro release properties were investigated. The OVA leaking into the continuous phase during the formation of microparticle by DCM evaporation was also evaluated. Results show that OVA was successfully entrapped in the microparticles with trapping efficiencies up to 72%, loading level 8.7% w/v, and particle size 14 microm. The semi-solid suspension changes to a solid particle happened during a 10-min period. Total protein-leaking amount was reduced after addition of NaCl in the continuous aqueous phase, which resulted from reducing the solidification time and protein-leaking rate. Using 5% w/v NaCl in the continuous phase resulted in higher loading content (87.2 +/- 1.0 microg/mg), and loading efficiency (72.2%), which resulted from more protein in the deeper layer (50.2 +/- 2.3 microg/mg) and higher microparticle yield (75.2%) than without NaCl (loading content: 74.0 +/- 1.0 microg/mg; loading efficiency 51.8%; deeper layer content: 18.3 +/- 3.5 microg/mg; yield: 63.6%). These results constitute a step forward in the improvement of existing technology in controlling protein encapsulation and delivery from microparticles prepared by the multiple emulsion solvent evaporation method.     

Patent Briefing

Japanese encephalitis virus (JEV)-loaded poly(lactide) (PLA) lamellar and poly(DL-lactide-co-glycolide) (PLG) microparticles were successfully prepared with low molecular weight PLA by the precipitate method and with 6% w/v PLG in the organic phase, 10% w/v PVP and 5% w/v NaCl in the continuous phase, by using a water-in-oil-in-water emulsion/solvent extraction technique, respectively. JEV was entrapped in the PLG microparticles by a solvent extraction technique with trapping efficiencies up to 98%, loading level 5.5% w/w, and mean particle size 3.8 #181;m. The distribution (%) of JEV on the PLG microparticles surface, outer layer, and core were 11.2, 41.7 and 46.4%, respectively. The cumulative release of JEV had an upper limit of ~ 58% of the JEV load at 24 days. The steady release rate was 1.33 #181;g JEV/mg microparticles/day of JEV release maintained for 24 days. The corresponding virus loading of the PLA lamellae is ~ 0.78% w/w and the loading efficiency (77.8%), JEV content (7.84 #181;g/mg), and yield (96.3%), respectively. The distribution (%) of JEV on the microparticles surface, outer layer, and core were 82.1, 13.3 and 2.2%, respectively. The live JEV challenge in mice test, in which mice received one dose of 20 mg JEV-loaded PLG microparticles, 20 mg JEV-loaded PLA lamellar in comparison with JEV or PBS solution, was evaluated after IP immunization of BALB/c mice. The study results show that the greatest survival was observed in the group of mice immunized with 20 mg JEV-loaded PLG microparticles and 20 mg JEV-loaded PLA microparticles group (80%). The JEV incorporation, physicochemical characterization data, and the animal results obtained in this study may be relevant in optimizing the vaccine incorporation and delivery properties of these potential vaccine targeting carriers.     

The mechanism of PLA microparticle formation by waterin-oil-in-water solvent evaporation method

Microparticles containing ovalbumin as a model protein drug were prepared using poly(L-lactide; PLA) with a water-in-oil-in-water emulsion/solvent evaporation technique. The dispersed phase was PLA dissolved in dichloromethane (DCM), and the continuous phase was water-containing polyvinyl pyrolidone (PVP) as stabilizer with sodium chloride. Microparticle characteristics, loading efficiencies, protein distribution in microparticles, and in-vitro release properties were investigated. The OVA leaking into the continuous phase during the formation of microparticle by DCM evaporation was also evaluated. Results show that OVA was successfully entrapped in the microparticles with trapping efficiencies up to 72%, loading level 8.7% w/v, and particle size 14 #181;m. The semi-solid suspension changes to a solid particle happened during a 10-min period. Total protein-leaking amount was reduced after addition of NaCl in the continuous aqueous phase, which resulted from reducing the solidification time and protein-leaking rate. Using 5% w/v NaCl in the continuous phase resulted in higher loading content (87.2 1.0 #181;g/mg), and loading efficiency (72.2%), which resulted from more protein in the deeper layer (50.2 2.3 #181;g/mg) and higher microparticle yield (75.2%) than without NaCl (loading content: 74.0 1.0 #181;g/mg; loading efficiency 51.8%; deeper layer content: 18.3 3.5 #181;g/mg; yield: 63.6%). These results constitute a step forward in the improvement of existing technology in controlling protein encapsulation and delivery from microparticles prepared by the multiple emulsion solvent evaporation method.     

Japanese encephalitis virus vaccine formulations using PLA lamellar and PLG microparticles

Japanese encephalitis virus (JEV)-loaded poly(lactide) (PLA) lamellar and poly(DL-lactide-co-glycolide) (PLG) microparticles were successfully prepared with low molecular weight PLA by the precipitate method and with 6% w/v PLG in the organic phase, 10% w/v PVP and 5% w/v NaCl in the continuous phase, by using a water-in-oil-in-water emulsion/solvent extraction technique, respectively. JEV was entrapped in the PLG microparticles by a solvent extraction technique with trapping efficiencies up to 98%, loading level 5.5% w/w, and mean particle size 3.8 microm. The distribution (%) of JEV on the PLG microparticles surface, outer layer, and core were 11.2, 41.7 and 46.4%, respectively. The cumulative release of JEV had an upper limit of approximately 58% of the JEV load at 24 days. The steady release rate was 1.33 microg JEV/mg microparticles/day of JEV release maintained for 24 days. The corresponding virus loading of the PLA lamellae is approximately 0.78% w/w and the loading efficiency (77.8%), JEV content (7.84 microg/mg), and yield (96.3%), respectively. The distribution (%) of JEV on the microparticles surface, outer layer, and core were 82.1, 13.3 and 2.2%, respectively. The live JEV challenge in mice test, in which mice received one dose of 20 mg JEV-loaded PLG microparticles, 20 mg JEV-loaded PLA lamellar in comparison with JEV or PBS solution, was evaluated after IP immunization of BALB/c mice. The study results show that the greatest survival was observed in the group of mice immunized with 20 mg JEV-loaded PLG microparticles and 20 mg JEV-loaded PLA microparticles group (80%). The JEV incorporation, physicochemical characterization data, and the animal results obtained in this study may be relevant in optimizing the vaccine incorporation and delivery properties of these potential vaccine targeting carriers.     

In vivo and in vitro characteristics for insulin-loaded PLA microparticles prepared by w/o/w solvent evaporation method with electrolytes in the continuous phase

Insulin-loaded poly(lactide) (PLA) microparticles were successfully prepared by 6% w/v PLA in the organic phase, 10% w/v PVP and varied types of 5%w/v electrolytes in the continuous phase, by using a water-in-oil-in-water emulsion/solvent extraction technique. Addition of electrolytes such as NaCl, CaCl₂ into the external phase significantly improved insulin entrapment efficiency compared to the case of no additives. NaCl was the most effective for obtaining high entrapment efficiency, with microparticle yield 81.2%, trapping efficiencies 49%, insulin-loading level 5.5% w/w and mean particle size 14.8?µm. The distribution (%) of insulin on the PLA microparticles surface, outer layer and core were 8, 37 and 43%, respectively. The cumulative release of insulin had an upper limit of ～24% of the insulin load at 24 days. A steady release rate was 0.5?µg insulin/mg microparticles/day of insulin release maintained for 24 days. Total protein-leaking amount was reduced after addition of electrolytes in the continuous aqueous phase. Rabbit glucose levels were evaluated after subcutaneous 20?mg insulin-loaded PLA microparticles or PLA blank microparticles. Study results show that the insulin-loaded PLA microparticles significantly reduced the glucose level than PLA blank microparticles. The insulin-loaded PLA microparticles, physicochemical characterization data and the animal result obtained in this study may be relevant in optimizing the PLA microparticle formulation incorporation and delivery insulin carriers.     

In vivo and in vitro characteristics for insulin-loaded PLA microparticles prepared by w/o/w solvent evaporation method with electrolytes in the continuous phase

Insulin-loaded poly(lactide) (PLA) microparticles were successfully prepared by 6% w/v PLA in the organic phase, 10% w/v PVP and varied types of 5%w/v electrolytes in the continuous phase, by using a water-in-oil-in-water emulsion/ solvent extraction technique. Addition of electrolytes such as NaCl, CaCl2 into the external phase significantly improved insulin entrapment efficiency compared to the case of no additives. NaCl was the most effective for obtaining high entrapment efficiency, with microparticle yield 81.2%, trapping efficiencies 49%, insulin-loading level 5.5% w/w and mean particle size 14.8 microm. The distribution (%) of insulin on the PLA microparticles surface, outer layer and core were 8, 37 and 43%, respectively. The cumulative release of insulin had an upper limit of approximately 24% of the insulin load at 24 days. A steady release rate was 0.5 microg insulin/mg microparticles/day of insulin release maintained for 24 days. Total protein-leaking amount was reduced after addition of electrolytes in the continuous aqueous phase. Rabbit glucose levels were evaluated after subcutaneous 20 mg insulin-loaded PLA microparticles or PLA blank microparticles. Study results show that the insulin-loaded PLA microparticles significantly reduced the glucose level than PLA blank microparticles. The insulin-loaded PLA microparticles, physicochemical characterization data and the animal result obtained in this study may be relevant in optimizing the PLA microparticle formulation incorporation and delivery insulin carriers.     

Formulation factors for preparing ocular biodegradable delivery system of 5-fluorouracil microparticles

Microparticles containing 5-fluorouracil (5-FU) were prepared using poly(dllactide-co-glycolide) with an oil-in-oil emulsion/solvent extraction technique. Particle characteristics including size distribution, 5-FU loading efficiencies, in vitro release and degradation were investigated. The dispersed phase was composed of PLG dissolved in dichloromethane, and the continuous phase was paraffin oil containing lecithin. 5-FU was successfully entrapped in the microparticles with trapping efficiencies up to 76%, loading level 10% w/v, and particle size 3 µm. Release profiles of 5-FU loaded microparticles were determined to follow a first-order-time relationship. An optimized preparation of 5-FU microparticles was achieved and was capable of controlling the release of 5-FU over 21 days with an in vitro delivery rate of 0.4 µg 5-FU/mg particles/ day in the study. Preliminary animal studies indicated that the 5-FU loaded microparticles as an ocular delivery system showed no ocular toxicity and no significant inflammatory response in rabbits for 2 months. The 5-FU loaded microparticles approach, with PLG, might be a potential for the application of long-term delivery of hydrophilic drugs in the eye.     

Formulation factors for preparing ocular biodegradable delivery system of 5-fluorouracil microparticles.

Microparticles containing 5-fluorouracil (5-FU) were prepared using poly(DL-lactide-co-glycolide) with an oil-in-oil emulsion/solvent extraction technique. Particle characteristics including size distribution, 5-FU loading efficiencies, in vitro release and degradation were investigated. The dispersed phase was composed of PLG dissolved in dichloromethane, and the continuous phase was paraffin oil containing lecithin. 5-FU was successfully entrapped in the microparticles with trapping efficiencies up to 76%, loading level 10% w/v, and particle size 3 microm. Release profiles of 5-FU loaded microparticles were determined to follow a first-order-time relationship. An optimized preparation of 5-FU microparticles was achieved and was capable of controlling the release of 5-FU over 21 days with an in vitro delivery rate of 0.4 microg 5-FU/mg particles/day in the study. Preliminary animal studies indicated that the 5-FU loaded microparticles as an ocular delivery system showed no ocular toxicity and no significant inflammatory response in rabbits for 2 months. The 5-FU loaded microparticles approach, with PLG, might be a potential for the application of long-term delivery of hydrophilic drugs in the eye.     

The stability of insulin in biodegradable microparticles based on blends of lactide polymers and polyethylene glycol

Insulin-loaded microparticles were produced from blends of poly(ethylene glycol) (PEG) with poly (L-lactide) (PLA) homopolymer and poly (DL-lactide co-glycolide) copolymers (PLG) using a water-in-oil solvent extraction method. The dispersed phase was composed of PLG/PEG or PLA/PEG dissolved in dichloromethane, and the continuous phase was methanol containing 10% PVP. Characteristics, including particle size distribution, insulin loading capacity and efficiencies, in vitro release, degradation and stability, were investigated. The stability of insulin associated with microparticles prepared using PEG and 50:50 PLG and PLA was analysed by HPSEC and quantified by peak area following incubation in PBS at 37 degrees C for up to 1 month. Insulin was successfully entrapped in the PLG/PEG and PLA/PEG microparticles with trapping efficiencies up to 56 and 48%, loading levels 17.8 and 10.6% w/w, and particle sizes 8 and 3 microm, respectively. The insulin-loaded PLG/PEG and PLA/PEG microparticles were capable of controlling the release of insulin over 28 days with in vitro delivery rates of 0.94 and 0.65 microg insulin/mg particles/day in the first 4 days and a steady release with rate of 0.4 and 0.43 microg insulin/mg particles/day over the following 4 weeks, respectively. Extensive degradation of the PLG/PEG microparticles also occurred over 4 weeks, whereas the use of PLA/PEG blends resulted in a stable microparticle morphology and much reduced fragmentation and aggregation of the associated insulin.     

Improved bioavailability of orally delivered insulin using Eudragit-L30D coated PLGA microparticles

Large porous microparticles of PLGA entrapping insulin were prepared by solvent evaporation method and evaluated in diabetes induced rat for its efficacy in maintaining blood sugar level from a single oral dose. Incorporation of Eudragit L30D (0.03% w/v) in the external aqueous phase resulted in formation of pH responsive enteric coated polymer particles which release most of the entrapped insulin in alkaline pH. At acidic pH, release of insulin from uncoated PLGA microparticles and Eudragit L30D coated PLGA microparticles was 31.62 +/- 1.8% and 17.5 +/- 1.29%, respectively, for initial 30 min. However, in 24 h, in vitro released insulin from uncoated PLGA and Eudragit coated particles was 96.29 +/- 1.01% and 88.30 +/- 1%, respectively. Released insulin from composite polymer particles were mostly in monomer form without aggregation and was stable for a month at 37 degrees C. Oral administration of insulin loaded PLGA (50 : 50) and Eudragit L30D coated PLGA (50 : 50) microparticles (equivalent to 25 IU insulin/kg of animal weight) in alloxan induced diabetic rats resulted in 37.3 +/- 11% and 62.7 +/- 3.8% reduction in blood glucose level, respectively, in 2 h. This effect continued up to 24 h in the case of Eudragit L30D coated PLGA microparticles. Results demonstrate that use of stabilizers during PLGA particle formulation, large porous particle for quick release of insulin and coating with Eudragit L30D resulted in a novel oral formulation for once a day delivery of insulin.     

Preparation and in vitro release of zein microparticles loaded with prednisolone for oral delivery

Zein has been proposed as a polymer for targeted-drug delivery via the oral route. Zein microparticles were loaded with prednisolone and evaluated as an oral delivery system. Microparticles were formulated using phase separation. Starting quantities of zein and prednisolone, along with the agitation method and temperature were found to significantly impact drug loading and loading efficiency. Vortex mixing produced the highest drug loading and loading efficiency. Drug release was measured in simulated conditions of the stomach and small intestine using the microparticles made with the method that best improved drug loading. In simulated stomach and small intestine conditions, prednisolone release reached almost 70% over 3 and 4?h, respectively. While a clinically relevant dose may be delivered using c. 100?mg of zein microparticles, prednisolone release from the microparticles indicates that they may not be suited as a controlled- or targeted-delivery system.     

Optimizing formulation factors in preparing chitosan microparticles by spray-drying method

The chitosan only, chitosan/Pluronic F68, chitosan/gelatin, chitosan/Pluronic F68/gelatin microparticles and betamethasone-loaded chitosan/Pluronic F68/gelatin microparticles were successfully prepared by a spray-drying method. Microparticle characteristics (yield rate, zeta potential, particle size and tap density), loading efficiencies, microparticle morphology and in-vitro release properties were investigated. By properly choosing excipient type, concentration and varying the spray-drying parameters, a high degree of control was achieved over the physical properties of the dry chitosan powders. SEM micrograph shows that the particle sizes of the varied chitosan composed microparticles ranged from 2.12-5.67 microm and the external surfaces appear smooth. Using betamethasone as model drug, the spray-drying is a promising way to produce good spherical and smooth surface microparticles with a narrow particle size range for controlled delivery of betamethasone. The positively charged betamethasone-loaded microparticles entrapped in the chitosan/Pluronic F68/gelatin microparticles with trapping efficiencies up to 94.5%, yield rate 42.5% and mean particle size 5.64 microm varied between 4.32-6.20 microm and tap densities 0.128 g/cm(3). The pH of particle was increased with increasing betamethasone-loaded amount, but both zeta potential and tap density of the particles decreased with increasing betamethasone-loaded amount. The betamethasone release rates from chitosan/Pluronic F68/gelatin microparticles were influenced by the drug/polymer ratio in the manner that an increase in the release% and burst release% was observed when the drug loading was decreased. The in vitro release of betamethasone showed a dose-dependent burst followed by a slower release phase that was proportional to the drug concentration in the concentration range between 14-44%w/w.     

Controlled delivery of testosterone from smart polymer solution based systems: in vitro evaluation

The objective of this research is to develop injectable polymers solution based controlled release delivery systems for testosterone (TSN), using phase sensitive and thermosensitive polymers. A combination of poly(lactide) (PLA) and solvents mixture of benzyl benzoate (BB) and benzyl alcohol (BA) was used in the phase sensitive polymer delivery system. The effects of solvents system and drug loading on the in vitro TSN release were evaluated. In the case of thermosensitive polymer delivery systems, a series of low-molecular-weight poly(lactide-co-glycolide)-poly(ethylene glycol)-poly(lactide-co-glycolide) (PLGA-PEG-PLGA) triblock copolymers with varying ratio of lactide/glycolide (LA/GA, 2.0-3.5) were studied to control the release of TSN. The effects of varying block length of copolymers 1-4 on the in vitro TSN release were evaluated. Phosphate buffer saline (pH 7.4) containing 0.5% (w/v) Tween-80 was used as in vitro release medium. The amount of the released TSN was determined by an HPLC method. A controlled (zero-order) in vitro release of TSN was observed from both the phase sensitive and thermosensitive polymer delivery systems. Addition of BA (15%, v/v) in solvents system significantly (p<0.05) increased the release rate of TSN (0.33+/-0.01 mg/ml) from phase sensitive delivery system in comparison to solvent without BA (0.27+/-0.00 mg/day). Increasing drug loading also increased release rate. In the case of thermosensitive polymer delivery system, increasing the hydrophobic PLGA block length of copolymers significantly (p<0.05) decreased the release rate of TSN. It is evident from this study that the phase sensitive and thermosensitive polymers are suitable for developing prolong-release injectable implant delivery systems for TSN.     

Preparation of 3,4-diaminopyridine microparticles by solvent-evaporation methods

The present study compares two methods of preparation of microparticles of 3,4-diaminopyridine (3,4-DAP) for the treatment for multiple sclerosis and Lambert-Eaton myasthenia syndrome. Poly( epsilon -caprolactone) microparticles were prepared with a solvent-evaporation W/O method. The 3,4-DAP was dispersed in dichloromethane, leading to a suspension. The dispersion and the solidification of the dichloromethane droplets in an aqueous phase have led to microparticles of 55.3+/-34.7 microm. The incorporation of the drug by milligram of powder was very low (1.91 micrograms/mg) and the scanning electron microscopy (SEM) did not show any crystal but marks of dissolved crystals were observed on the polymeric surface. EudragitRS microspheres containing 3,4-DAP were prepared by a solvent-evaporation technique using light mineral oil as continuous phase. The drug and the polymer were completely dissolved in an acetone solution, used as discontinuous phase. This formulation have led to a higher incorporation of the drug (88.25 micrograms/mg). The particle size was 91.8+/-44.3 microm. The observation, by SEM, shows many crystals on the surface and inside the microparticles. A slow-release of the drug in a phosphate buffer pH 7.4 was observed (50% in 60 min and about 70% in 4 h).     

Improving Protein Delivery from Microparticles Using Blends of Poly(DL lactide co-glycolide) and Poly(ethylene oxide)-poly(propylene oxide) Copolymers

Purpose. Microparticles containing ovalbumin as a model for protein drugs were formulated from blends of poly(DL lactide-co-glycolide) and poly(ethylene oxide)-poly(propylene oxide) copolymers (Pluronic). The objectives were to achieve uniform release characteristics and improved protein delivery capacity. Methods. The water- in oil -in oil emulsion/solvent extraction technique was used for microparticle production. Results. A protein loading level of over 40% (w/w) was attained in microparticles having a mean diameter of approximately 5 µm. Linear protein release profiles over 25 days in vitro were exhibited by certain blend formulations incorporating hydrophilic Pluronic F127. The release profile tended to plateau after 10 days when the more hydrophobic Pluronic L121 copolymer was used to prepare microparticles. A delivery capacity of 3 µg OVA/mg particles/ day was achieved by formulation of microparticles using a 1:2 blend of PLG:Pluronic F127. Conclusions. The w/o/o formulation approach in combination with PLG:Pluronic blends shows potential for improving the delivery of therapeutic proteins and peptides from microparticulate systems. Novel vaccine formulations are also feasible by incorporation of Pluronic L121 in the microparticles as a co-adjuvant.     

Formulation and evaluation of verapamil hydrochloride loaded solid lipid microparticles

The present study aimed to produce verapamil hydrochloride-loaded solid lipid microparticles (SLM) by the w/o/w emulsion solvent evaporation technique, using diethyl ether as solvent phase, glyceryl monostearate as biodegradable polymer and Span 60 as surfactant. SLM of spherical shape were prepared by simple dilution of the emulsion with water. To increase the lipid load the process was conducted at 50 degrees C, and in order to reach sub-micron size, a high-shear homogenizer was used. The encapsulation efficiency of prepared SLM reached 74.29 +/- 0.76%. Particle size (98.55 +/- 1.42 microm), surface morphology (spherical) and drug loading efficiency (18.57 +/- 1.25% w/w) were investigated. And optimization of drug polymer ratio (3:1), nature and concentration of emulsion stabilizer in the external aqueous (0.1%), phase viscosity of external aqueous phase (0.5%), volume of external aqueous phase and stirring rate (1000 rpm) were detected. Analysis of microsphere content after processing showed that verapamil did not undergo any chemical modification within the micro-particles. The in-vitro release of verapamil from the microparticles was very low and an initial burst effect of 17% of the dose was observed. The slow release may help to avoid a high frequency of administration. The prepared solid lipid microparticles appear to have interesting perspectives as delivery systems for the oral administration of verapamil hydrochloride with improved half-life, improved bioavailability, and minimized local and systemic gastrointestinal disturbances of the drug.     

The characteristics of betamethasone-loaded chitosan microparticles by spray-drying method

Betamethasone (BTM)-loaded microparticles prepared by a spray drying method using chitosan (CTS) as raw material, type-A gelatin and ethylene oxide-propylene oxide block copolymer (Pluronic F68) as modifiers. The BTM-loaded in varied chitosan/Pluronic F68/gelatin microparticle formulations was investigated. By properly choosing excipient type and concentration a high degree of control was achieved over the physical properties of the BTM-loaded microparticles. Microparticle characteristics (zeta potential, tap density, particle size and yield), loading efficiencies, microparticle morphology and in-vitro release properties were examined. Surface morphological characteristics and surface charges of prepared microparticles were observed by using scanning electron microscopy (SEM) and microelectrophoresis. A SEM micrograph shows that the particle sizes of the varied chitosan composed microparticles ranged from 1.1-4.7 microm and the external surfaces appear smooth. The BTM-loaded microparticles entrapped in the chitosan/Pluronic F68/gelatin microparticles with trapping efficiencies up to 93%, collected yield rate 44%, and mean particle size varied between 1-3 microm, positive surface charge (20-40 mv), and tap densities (0.04-0.40 g/cm3) were obtained. The collected BTM yield and size of particle was increased with increasing BTM-loaded amount but both zeta potential and tap density of the particles decreased with increasing BTM-loaded amount. The in vitro release of BTM showed a dose-dependent burst followed by a slower release phase that was proportional to the drug concentration in the concentration range between 5-30%w/w. The in vitro drug release from the chitosan/Pluronic F68/gelatin 1/0.1/0.4 microspheres had a prolong release pattern. These formulation factors were correlated to particulate characteristics for optimizing BTM microspheres in pulmonary delivery.