Zinc ions (Zn2+), known to be a novel intracellular second messenger related to various biological functions, have been delivered inside cells. For the intracellular Zn2+ delivery, Zn2+ has been chelated to carboxymet...Zinc ions (Zn2+), known to be a novel intracellular second messenger related to various biological functions, have been delivered inside cells. For the intracellular Zn2+ delivery, Zn2+ has been chelated to carboxymethyl poly(1-vinylimidazole) (CM-PVIm) by mixing zinc chloride (ZnCl2) or zinc acetate (Zn(OAc)2) with CM-PVIm. The resulting Zn2+-chelated CM-PVIm, that is, Zn2+/CM-PVIm complex by mixing ZnCl2 exhibited smaller particle size below 10 nm and possessed larger amount of Zn2+ ions, as compared to the Zn2+/CM-PVIm by mixing Zn(OAc)2. The both Zn2+/CM-PVIm complexes exhibited no significant cytotoxicity, leading to intracellular Zn2+ delivery. The Zn2+/CM-PVIm by mixing ZnCl2 delivered larger amount of intracellular Zn2+ ions than that by mixing Zn(OAc)2. These results suggest that the optimal Zn2+/CM-PVIm complex is a useful tool for intracellular Zn2+ delivery to control various biological functions.展开更多
Polymeric amines are being studied intensively as components of systems for gene delivery in genetic engineering and gene therapy of genetic disorders, including cancer. Despite remarkable achievements in the field, p...Polymeric amines are being studied intensively as components of systems for gene delivery in genetic engineering and gene therapy of genetic disorders, including cancer. Despite remarkable achievements in the field, polymeric amines, such as polyethyleneimine, show some disadvantages. Strong interaction between the amine-containing polymer and nucleic acid hampers the release of nucleic acid in the cell cytoplasm. Amine groups can interact with the cell membrane which results in cell death. These limitations of polymeric amines stimulated an investigation of new structures for gene delivery. Imidazole-containing polymers have attracted attention as lesser basic substances, while they are able to interact with polymeric acids. Further development of imidazole-based gene delivery agents requires knowledge about some fundamental aspects of interaction between nucleic acids, and polymeric imidazoles. In this work, we studied the complexation of poly(1-vinylimidazole) and oligomeric DNA. We found that the number of active sites capable of binding with negatively charged phosphate groups is comparable with the number of protonated imidazole units in the case of high molecular weight polymer. The increase in polymer charge by 1-bromopropane quaternizating 1%?5% imidazole units or by decreasing the pH to 6.5?7 considerably increased the ability of poly(1-vinylimidazole) to interact with oligonucleotides. The pH sensitivity of this interaction is interesting for cancer gene therapy because the tumours have a lowered intercellular pH (stable oligonucleotide complex) and a higher extracellular pH which can lead to complex dissociation. Minimal critical length for complexation of quaternized poly(1-vinylimidazole) and DNA is below eight units which corresponds to polymers with amine groups. Fluorescence-tagged poly(1-vinylimidazole) samples were obtained and their potential for monitoring the polymer and polymer-oligonucleotide complex internalization into living cells was demonstrated.展开更多
文摘Zinc ions (Zn2+), known to be a novel intracellular second messenger related to various biological functions, have been delivered inside cells. For the intracellular Zn2+ delivery, Zn2+ has been chelated to carboxymethyl poly(1-vinylimidazole) (CM-PVIm) by mixing zinc chloride (ZnCl2) or zinc acetate (Zn(OAc)2) with CM-PVIm. The resulting Zn2+-chelated CM-PVIm, that is, Zn2+/CM-PVIm complex by mixing ZnCl2 exhibited smaller particle size below 10 nm and possessed larger amount of Zn2+ ions, as compared to the Zn2+/CM-PVIm by mixing Zn(OAc)2. The both Zn2+/CM-PVIm complexes exhibited no significant cytotoxicity, leading to intracellular Zn2+ delivery. The Zn2+/CM-PVIm by mixing ZnCl2 delivered larger amount of intracellular Zn2+ ions than that by mixing Zn(OAc)2. These results suggest that the optimal Zn2+/CM-PVIm complex is a useful tool for intracellular Zn2+ delivery to control various biological functions.
基金financial support from a joint grant of the Russian Science Foundation (16-45-02001)the Department of Science Technology of the Ministry of Science and Technology of the Republic of India (INT/RUS/RSF/10)
文摘Polymeric amines are being studied intensively as components of systems for gene delivery in genetic engineering and gene therapy of genetic disorders, including cancer. Despite remarkable achievements in the field, polymeric amines, such as polyethyleneimine, show some disadvantages. Strong interaction between the amine-containing polymer and nucleic acid hampers the release of nucleic acid in the cell cytoplasm. Amine groups can interact with the cell membrane which results in cell death. These limitations of polymeric amines stimulated an investigation of new structures for gene delivery. Imidazole-containing polymers have attracted attention as lesser basic substances, while they are able to interact with polymeric acids. Further development of imidazole-based gene delivery agents requires knowledge about some fundamental aspects of interaction between nucleic acids, and polymeric imidazoles. In this work, we studied the complexation of poly(1-vinylimidazole) and oligomeric DNA. We found that the number of active sites capable of binding with negatively charged phosphate groups is comparable with the number of protonated imidazole units in the case of high molecular weight polymer. The increase in polymer charge by 1-bromopropane quaternizating 1%?5% imidazole units or by decreasing the pH to 6.5?7 considerably increased the ability of poly(1-vinylimidazole) to interact with oligonucleotides. The pH sensitivity of this interaction is interesting for cancer gene therapy because the tumours have a lowered intercellular pH (stable oligonucleotide complex) and a higher extracellular pH which can lead to complex dissociation. Minimal critical length for complexation of quaternized poly(1-vinylimidazole) and DNA is below eight units which corresponds to polymers with amine groups. Fluorescence-tagged poly(1-vinylimidazole) samples were obtained and their potential for monitoring the polymer and polymer-oligonucleotide complex internalization into living cells was demonstrated.
