Research Paper
Journal of Pharmacy And Pharmacology
Effects of dendrimer/cyclodextrin conjugates as gene transfer carriers on nitric oxide production from macrophages Keiichi Motoyamaa, Asumi Sakoa, Irhan Ibrahim Abu Hashima,b, Taishi Higashia and Hidetoshi Arimaa,c a
Department of Physical Pharmaceutics, Graduate School of Pharmaceutical Sciences, cProgram for Leading Graduate Schools ‘HIGO (Health life science: Interdisciplinary and Glocal Oriented) Program’, Kumamoto University, Kumamoto, Japan and bFaculty of Pharmacy, Mansoura University, Mansoura, Egypt
Keywords cyclodextrin; dendrimer; gene transfer carrier; nitric oxide; toll-like receptor ligands Correspondence Hidetoshi Arima, Department of Physical Pharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan. E-mail: [email protected] Received February 22, 2015 Accepted April 12, 2015 doi: 10.1111/jphp.12439
Abstract Objectives The development of safe gene transfer carriers with high transfection efficiency, which does not affect the cell function, is a challenging issue. In this study, we examined the effects of α-cyclodextrin (α-CyD)/dendrimer conjugate (α-CDE (G3)) on nitric oxide (NO) production in murine macrophages J774.1 cells stimulated with toll-like receptors (TLR) ligands. Methods NO production from macrophages stimulated with TLR ligands was determined by the Griess method. Transfection efficiency of α-CDE (G3)/plasmid DNA (pDNA) complex was quantified by a luminometer. Key findings α-CDE (G3) significantly inhibited NO production from J774.1 cells stimulated with TLR ligands. α-CyD molecules in α-CDE (G3) had no effect on NO production. The inhibitory effect of α-CDE (G3) on NO production might be attributed to the dendrimer (G3). Increasing the degree of substitution (DS) of α-CyD in the α-CDE (G3) molecule was accompanied by a significant decrease in the inhibition of NO production. Furthermore, higher gene transfection efficiency of α-CDE (G3)/pDNA complex was observed upon increasing the DS of α-CyD. Conclusions α-CDE (G3) with high DS value of α-CyD may be considered as a safe gene transfer carrier that does not adversely affect NO production from macrophages stimulated with TLR ligands.
Introduction In the concept of gene therapy, the selective delivery of therapeutic genes to the target cells with efficiency and safety is a crucial challenge. At the present time, viral and non-viral vectors are considered the two main categories of vectors used for gene delivery. Although viral vectors have demonstrated high transfection efficiencies, the concerns over their safety and immunogenicity have directed research towards the development of more efficient non-viral vectors. Starburst polyamidoamine (PAMAM) dendrimer (dendrimer) is a spherical, highly ordered, dendritic polymer with positively charged primary amino groups on the surface at physiological pH.[1,2] Dendrimer is one of the widely used non-viral vectors owing to its solubility in water, biocompatibility and ability to get cleared through 598
the kidneys.[3] These features have allowed their application in drug delivery, tissue engineering, gene transfer and in imaging.[4] Generally, it is evident that the nature of dendrimers as non-viral vectors depends significantly on their generations (G). Gene transfer activity of dendrimers with high generations is likely to be superior to that with low generations,[5,6] although their cytotoxicity concomitantly increases as the generation increases due to higher positive charge.[6] Therefore, there has been a growing interest in developing dendrimers with low generations (
Journal of Pharmacy And Pharmacology
Effects of dendrimer/cyclodextrin conjugates as gene transfer carriers on nitric oxide production from macrophages Keiichi Motoyamaa, Asumi Sakoa, Irhan Ibrahim Abu Hashima,b, Taishi Higashia and Hidetoshi Arimaa,c a
Department of Physical Pharmaceutics, Graduate School of Pharmaceutical Sciences, cProgram for Leading Graduate Schools ‘HIGO (Health life science: Interdisciplinary and Glocal Oriented) Program’, Kumamoto University, Kumamoto, Japan and bFaculty of Pharmacy, Mansoura University, Mansoura, Egypt
Keywords cyclodextrin; dendrimer; gene transfer carrier; nitric oxide; toll-like receptor ligands Correspondence Hidetoshi Arima, Department of Physical Pharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan. E-mail: [email protected] Received February 22, 2015 Accepted April 12, 2015 doi: 10.1111/jphp.12439
Abstract Objectives The development of safe gene transfer carriers with high transfection efficiency, which does not affect the cell function, is a challenging issue. In this study, we examined the effects of α-cyclodextrin (α-CyD)/dendrimer conjugate (α-CDE (G3)) on nitric oxide (NO) production in murine macrophages J774.1 cells stimulated with toll-like receptors (TLR) ligands. Methods NO production from macrophages stimulated with TLR ligands was determined by the Griess method. Transfection efficiency of α-CDE (G3)/plasmid DNA (pDNA) complex was quantified by a luminometer. Key findings α-CDE (G3) significantly inhibited NO production from J774.1 cells stimulated with TLR ligands. α-CyD molecules in α-CDE (G3) had no effect on NO production. The inhibitory effect of α-CDE (G3) on NO production might be attributed to the dendrimer (G3). Increasing the degree of substitution (DS) of α-CyD in the α-CDE (G3) molecule was accompanied by a significant decrease in the inhibition of NO production. Furthermore, higher gene transfection efficiency of α-CDE (G3)/pDNA complex was observed upon increasing the DS of α-CyD. Conclusions α-CDE (G3) with high DS value of α-CyD may be considered as a safe gene transfer carrier that does not adversely affect NO production from macrophages stimulated with TLR ligands.
Introduction In the concept of gene therapy, the selective delivery of therapeutic genes to the target cells with efficiency and safety is a crucial challenge. At the present time, viral and non-viral vectors are considered the two main categories of vectors used for gene delivery. Although viral vectors have demonstrated high transfection efficiencies, the concerns over their safety and immunogenicity have directed research towards the development of more efficient non-viral vectors. Starburst polyamidoamine (PAMAM) dendrimer (dendrimer) is a spherical, highly ordered, dendritic polymer with positively charged primary amino groups on the surface at physiological pH.[1,2] Dendrimer is one of the widely used non-viral vectors owing to its solubility in water, biocompatibility and ability to get cleared through 598
the kidneys.[3] These features have allowed their application in drug delivery, tissue engineering, gene transfer and in imaging.[4] Generally, it is evident that the nature of dendrimers as non-viral vectors depends significantly on their generations (G). Gene transfer activity of dendrimers with high generations is likely to be superior to that with low generations,[5,6] although their cytotoxicity concomitantly increases as the generation increases due to higher positive charge.[6] Therefore, there has been a growing interest in developing dendrimers with low generations (
