Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method

Biodegradable poly(D,L-lactide-co-glycolide) (PLGA) microspheres is a promising gene carrier system to deliver the gene in gene therapy. This study describes fabrication and characterization of plasmid DNA-loaded PLGA microspheres using a versatile double-emulsion (w/o/w) solvent-evaporation method....

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Main Authors:	Doolaanea, Abd. Almonem, Harun Ismail, Ahmad Fahmi, Mohamed, Farahidah
Format:	Conference or Workshop Item
Language:	English
Published:	2010
Subjects:	RB Pathology
Online Access:	http://irep.iium.edu.my/20634/ http://irep.iium.edu.my/20634/ http://irep.iium.edu.my/20634/1/Effect_of_different_parameters.pdf

id	iium-20634
recordtype	eprints
spelling	iium-206342013-01-14T05:22:36Z http://irep.iium.edu.my/20634/ Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method Doolaanea, Abd. Almonem Harun Ismail, Ahmad Fahmi Mohamed, Farahidah RB Pathology Biodegradable poly(D,L-lactide-co-glycolide) (PLGA) microspheres is a promising gene carrier system to deliver the gene in gene therapy. This study describes fabrication and characterization of plasmid DNA-loaded PLGA microspheres using a versatile double-emulsion (w/o/w) solvent-evaporation method. For microsphere fabrication, plasmid DNA in TE buffer was added to PLGA solution, previously dissolved in DCM, and was homogenized to form the primary water-in-oil (w/o) emulsion. This w/o emulsion was immediately injected into 1% aqueous PVA and homogenized to form the secondary water-in-oil-in-water (w/o/w) emulsion. The w/o/w emulsion was then transferred to a continuously stirred hardening tank of 1% aqueous PVA and the stirring was continued for 2 hours to allow complete evaporation of DCM. The hardened microspheres were collected by centrifugation, washed and freeze-dried. Several parameters have been investigated including PVA concentration, different types of surfactants and surfactant blends and PLGA molecular weight. Resultant microsoheres were characterized for size distribution and external morphology by laser and scanning electron microscopy, respectively. Encapsulation efficiency was also calculated and the DNA was quantified by UV absorbance (NanoQuant). it was found that, increasing the PVA concentration from 1% w/v to 5% w/v reduced that mean particle size from 11.39um to 3.74um. These size were also evident by the microimages that depicted smooth surfaces of microspheres yielded for the range of PVA concentration. This research is still ongoing the future aims include transfection on neuro cell line to see feasibility of using this carrier system to deliver relevant gene to treat neurodegenerative diseases at molecular level. 2010 Conference or Workshop Item NonPeerReviewed application/pdf en http://irep.iium.edu.my/20634/1/Effect_of_different_parameters.pdf Doolaanea, Abd. Almonem and Harun Ismail, Ahmad Fahmi and Mohamed, Farahidah (2010) Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method. In: 3rd International Conference on Advancement in Science and Technology (iCAST) 2010, 27-29 Nov 2010, Vistana Hotel, Kuantan, Pahang. http://www.iium.edu.my/icast/2010/index.php?option=com_content&view=article&id=14&Itemid=17
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	RB Pathology
spellingShingle	RB Pathology Doolaanea, Abd. Almonem Harun Ismail, Ahmad Fahmi Mohamed, Farahidah Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method
description	Biodegradable poly(D,L-lactide-co-glycolide) (PLGA) microspheres is a promising gene carrier system to deliver the gene in gene therapy. This study describes fabrication and characterization of plasmid DNA-loaded PLGA microspheres using a versatile double-emulsion (w/o/w) solvent-evaporation method. For microsphere fabrication, plasmid DNA in TE buffer was added to PLGA solution, previously dissolved in DCM, and was homogenized to form the primary water-in-oil (w/o) emulsion. This w/o emulsion was immediately injected into 1% aqueous PVA and homogenized to form the secondary water-in-oil-in-water (w/o/w) emulsion. The w/o/w emulsion was then transferred to a continuously stirred hardening tank of 1% aqueous PVA and the stirring was continued for 2 hours to allow complete evaporation of DCM. The hardened microspheres were collected by centrifugation, washed and freeze-dried. Several parameters have been investigated including PVA concentration, different types of surfactants and surfactant blends and PLGA molecular weight. Resultant microsoheres were characterized for size distribution and external morphology by laser and scanning electron microscopy, respectively. Encapsulation efficiency was also calculated and the DNA was quantified by UV absorbance (NanoQuant). it was found that, increasing the PVA concentration from 1% w/v to 5% w/v reduced that mean particle size from 11.39um to 3.74um. These size were also evident by the microimages that depicted smooth surfaces of microspheres yielded for the range of PVA concentration. This research is still ongoing the future aims include transfection on neuro cell line to see feasibility of using this carrier system to deliver relevant gene to treat neurodegenerative diseases at molecular level.
format	Conference or Workshop Item
author	Doolaanea, Abd. Almonem Harun Ismail, Ahmad Fahmi Mohamed, Farahidah
author_facet	Doolaanea, Abd. Almonem Harun Ismail, Ahmad Fahmi Mohamed, Farahidah
author_sort	Doolaanea, Abd. Almonem
title	Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method
title_short	Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method
title_full	Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method
title_fullStr	Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method
title_full_unstemmed	Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method
title_sort	effect of different parameters on the pdna-loaded microspheres prepared by double emulsion solvent evaporation method
publishDate	2010
url	http://irep.iium.edu.my/20634/ http://irep.iium.edu.my/20634/ http://irep.iium.edu.my/20634/1/Effect_of_different_parameters.pdf
first_indexed	2023-09-18T20:31:06Z
last_indexed	2023-09-18T20:31:06Z
_version_	1777408741170216960

Effect of different parameters on the pDNA-loaded microspheres prepared by double emulsion solvent evaporation method

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