• Journal of Pharmaceutical Investigation
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• v.26 no.2
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• pp.105-112
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• 1996

The surface of albumin microspheres was modified with methotrexate(MTX) by using 1,3-dicyclohexylcarbodiimide (DCC). Surface-modified albumin microspheres entrapping no MTX (SAMS), free MTX (SAMSF) and MTX-bovine serum albumin(BSA) conjugates(SAMSC) were prepared. The organ-targeting ability of free $[^3H]MTX,\;[^3H]MTX-BSA$ conjugate and the above microspheres was evaluated after i.v. administration of the preparations, equivalent to 150 nCi via the tail vein of mice. The total radioactivity in the lung increased immediately in a few minutes after i.v. injection of the microspheres, and then declined for the period of 3-4 weeks. However, the radioactivity in the liver, spleen and kidney increased slowly during the rapid decrease in radioactivity in the lung. This suggested that the microspheres could be entrapped rapidly in the lung through mechanical filtration because of their large size and slowly redistributed to the liver, spleen and kidney due to either the microspheres being degraded enough for the size to allow passage through the capillary beds of the lung and/or the release of $[^3H]MTX\;or\;[^3H]MTX-BSA$ conjugates from the microspheres. The amount of $60{sim}70%$ of the dose was targeted to the liver after the i.v. injection of SAMS, SAMSF and SAMSC, and the values of $(R_e\;^*\;_{e)liver}$ from the microspheres were $5{\sim}7$ compared to free MTX. This suggested that the liver-targeting ability from surface-modified albumin microspheres could be $5{\sim}7$ times as that of free MTX. The liver-targeted drug was accumulated in the Kupffer cells at the initial stage, thereafter the drug in the Kupffer cell was slowly transferred into the hepatocytes. The value of AUQ for liver from SAMS was higher than that from SAMSF, but much lower than that from SAMSC. This suggest that MTX bound to their surface could be eliminated slower than the entrapped free MTX, and faster than the entrapped MTX-BSA conjugates. This is consistent with the in vitro release rates order in the presence of a proteolytic enzyme. Also, surface-modified MTX was scarcely released in the absence of a proteolytic enzyme. Therefore, the surface-modified MTX nay be released (or eliminated) rapidly from SAMSC at the target site, and thereafter MTX may be released (or eliminated) slowly from the entrapped MTX-BSA conjugates in SAMSC for a long period.

• Journal of Pharmaceutical Investigation
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• v.31 no.4
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• pp.241-256
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• 2001

Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6363-6368
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• 2012

Objective: The study aim was to prepare poly-DL-lactide-poly (PELA) microspheres encapsulating recombinant tissue inhibitors of metalloproteinase-1 (TIMP-1) in an adenovirus to investigate its inhibition on the proliferation of hepatocellular carcinoma cells HepG2. Methods: Microspheres were prepared by encapsulating the recombinant TIMP-1 adenovirus into biodegradable PELA. The particle size, viral load, encapsulation efficiency and in-vitro release were measured. Microspheres were used to infect HepG2 cells, then infection efficiency was examined under a fluorescent microscope and ultrastructural changes assessed by TEM. Expression of TIMP-1 mRNA in HepG2 cells was examined by semi-quantitative RT-PCR and proliferation by MTT and cell growth curve assays. Results: We successfully prepared microspheres encapsulating recombinant TIMP-1 adenovirus with a diameter of $1.965{\mu}m$, an encapsulation efficiency of 60.0%, a viral load of $10.5{\times}10^8/mg$ and approximate 60% of virus release within 120 h, the total releasing time of which was longer than 240 h. The microspheres were confirmed to be non-toxic with blank microspheres. Infected HepG2 cells could stably maintain in-vitro expression of TIMP-1, with significantly effects on biological behaviour Conclusion: PELA microspheres encapsulating a recombinant TIMP-1 adenovirus can markedly inhibit the proliferation of HepG2 cells, which provides an experimental basis for polymer/chemistry-based gene therapy of hepatocellular carcinomas.

• Journal of Pharmaceutical Investigation
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• v.40 no.4
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• pp.245-250
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• 2010

Porous poly(lactic-co-glycolic acid) microspheres (PLGA MS) have been utilized as an inhalation delivery system and a matrix scaffold system for tissue engineering. Here, gelatin (type A) is introduced as an extractable pH-responsive porogen, which is capable of controlling the porosity and pore size of PLGA microspheres. Porous PLGA microspheres were prepared by a water-in-oil-in-water ($w_1/o/w_2$) double emulsification/solvent evaporation method. The surface morphology of these microspheres was examined by varying pH (2.0~11.0) of water phases, using scanning electron microscopy (SEM). Also, their porosity and pore size were monitored by altering acidification time (1~5 h) using a phosphoric acid solution. Results showed that the pore-forming capability of gelatin was optimized at pH 5.0, and that the surface pore-formation was not significantly observed at pHs of < 4.0 or > 8.0. This was attributable to the balance between gel-formation by electrostatic repulsion and dissolution of gelatin. The appropriate time-selection between PLGA hardening and gelatin-washing out was considered as a second significant factor to control the porosity. Delaying the acidification time to ~5 h after emulsification was clearly effective to make pores in the microspheres. This finding suggests that the porosity and pore size of porous microspheres using gelatin can be significantly controlled depending on water phase pH and gelatin-removal time. The results obtained in this study would provide valuable pharmaceutical information to prepare porous PLGA MS, which is required to control the porosity.

• Nuclear Engineering and Technology
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• v.51 no.1
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• pp.257-262
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• 2019

The control of microstructure is critical for the porous fuel particles used for infiltrating actinide nuclides. This study concerns the effect of heating processes on properties and microstructure of the fuel particles. The uniform gel precursor beads were synthesized by a microfluidic sol-gel process and then the porous $CeO_2$ microspheres, as a surrogate for the ceramic nuclear fuel particles, were obtained by heating treatment of the gel precursors. The fabricated $CeO_2$ microspheres have a narrow size distribution and good sphericity due to the feature of microfluidics. The effects of heating processes parameters, such as heating mode and peak temperatures on the properties of microspheres were studied in detail. An optimized heating mode and the peak temperature of $650^{\circ}C$ were selected to produce porous $CeO_2$ microspheres. The optimized heating mode can avoid the appearance of broken or crack microspheres in the heating process, and as-prepared porous microspheres were of suitable pore size distribution and pore volume for loading minor actinide (MA) solution by an infiltration method that is used for fabrication of MA-bearing nuclear fuel beads. After the infiltration process, $1000^{\circ}C$ was selected as the final temperature to improve the compressive strength of microspheres.

• Membrane Journal
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• v.31 no.1
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• pp.52-60
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• 2021

When preparing calcium alginate microspheres using rotating membrane emulsification that rotates SPG (Shirasu porous glass) tubular membrane in the continuous phase, the optimal conditions of rotating membrane emulsification process parameters for producing monodisperse microspheres were determined. We determined the effects of process parameters of rotating membrane emulsification (the rotating speed of membrane module, the transmembrane pressure, the ratio of dispersed phase to continuous phase, the alginate concentration, the emulsifier concentration, the stabilizer concentration, the crosslinking agent concentration, and the membrane pore size) on the mean size and size distribution of alginate microspheres. As a result, the size of the microspheres decreased as the rotating speed of membrane module, the emulsifier concentration, and the crosslinking agent concentration increased among the process parameters of rotating membrane emulsification. On the contrary, as the ratio of dispersed phase to continuous phase, the transmembrane pressure, and the alginate concentration increased, the size of the microspheres increased. In the rotating membrane emulsification using an SPG membrane with a pore size of 3.2 ㎛, it was possible to finally prepare monodisperse alginate microspheres with a particle size of 4.5 ㎛ through the control of process parameters.

• Journal of Pharmaceutical Investigation
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• v.25 no.3
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• pp.33-35
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• 1995

Microspheres containing tetracaine or bupivacaine with poly-lactic-glycolic acid were prepared with a range of compositions. Using the rat scicatic nerve model in vivo it was found that prolonged blockade for periods of 2-7 days. depending on composition variables. Polymer-local anesthetics microspheres are feasible delivery vehicle for prolonged regional nerve blockade.

• Journal of Biomedical Engineering Research
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• v.14 no.3
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• pp.209-220
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• 1993

Microspheres are expected to be applied to biomedical areas such as solid-phase immunoassays, drug delivery systems, Immunomagnetic cell separation. To synthesize micro-spheres for biomedical application, "two stage shot growth method" was developed. The uniformity ratio or synthesized microspheres was always smatter than 1.05. And the surface charge density (or the number of ionizable functional groups) of the microspheres synthesized by "two stage shot growth method" was 6-13 times higher than thats of the ml crospheres synthesized by conventional seeded batch copolymerization. As a previous step for biomedical application, adsorption experiments of bovine albumin on microspheres were carried out under various conditions. The maximum adsorbed amount was obtained in the neighborhood of pH 4.5. Isoelectric point of bovine albumin Is pH 5.0, so experimental result shows that it shifted to acid area. The adsorption Isotherm was obtained, the plateau region was always reached at 2.Og/L (bulk concentration of bovine albumin ) . The effect of the kind and the amount of surface functional group was also examined.p was also examined.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.409.3-410
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• 2002

Microencapsulations of piroxicam using the mixture of Eudragit RS with RL or Eudragit L or E or S according to Eudragit RS were carried out. The Eudragit microspheres of piroxicam were prepared by solvent method. Piroxicam and Eudragit polymer were dissoved in methylene chloride and dispersed in 0.5% pOlyvinyl alcohol solution and solvent evaporated. The average diameters of various Eudragit microspheres were from 40 to 43${\mu}{\textrm}{m}$. A good and smooth surface of microspheres observed by SEM were shown in all type of microspheres. (omitted)

• Journal of Pharmaceutical Investigation
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• v.26 no.1
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• pp.13-22
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• 1996

Poly(D,L-lactic acid) (PLA) microspheres containing alprazolam(APZ) were prepared by a solvent-emulsion evaporation method and their release patterns were investigated in vitro. Various batches of microspheres with different size and drug content were obtained by changing the ratio of APZ to PLA, PLA concentration in the dispersed phase and stirring rate. Rod-like APZ crystals on microsphere surface, which were released rapidly and could act as a loading dose, were observed with increasing drug content. The release rate was increased with increase in drug contents and decrease in the molecular weight of PLA. The release rate of APZ for long-acting injectable delivery system in vitro, which would aid in predicting in vitro release profile, could be controlled by properly optimizing various factors affecting characteristics of microspheres.