A novel cyclen-based linear polyamine (POGEC) was designed and synthesized from the reaction between 1,3-propanediol diglycidyl ether and 1,7-bis(diethoxyphosphory)-1,4,7,10-tetraazacyclod-odecane. High-affinity bindi...A novel cyclen-based linear polyamine (POGEC) was designed and synthesized from the reaction between 1,3-propanediol diglycidyl ether and 1,7-bis(diethoxyphosphory)-1,4,7,10-tetraazacyclod-odecane. High-affinity binding between POGEC and DNA was demonstrated by agarose gel electrophoresis and scanning electron microscopy (SEM). Moreover,the formed POGEC/DNA complex (termed polyplex) could be disassociated to release the free DNA through addition of the physiological concentration of NaCl solution. Fluorescence spectrum was used to measure the high-affinity binding and DNA condensation capability of POGEC. Circular dichroism (CD) spectrum indicates that the DNA conformation did not change after binding to POEGC.展开更多
Many experimental cancer vaccines are exploring toll-like receptor agonists （TLRas） such as CpG, a DNA motif that agonizes toll-like receptor 9 （TLR9）, to trigger immune responses that are potent and molecularly-s...Many experimental cancer vaccines are exploring toll-like receptor agonists （TLRas） such as CpG, a DNA motif that agonizes toll-like receptor 9 （TLR9）, to trigger immune responses that are potent and molecularly-specific. The ability to tune the immune response is especially important in the immunosuppressive microenvironments of tumors. Because TLR9 is located intracellularly, CpG must be internalized by immune cells for functionality. Polyplexes can be self- assembled through electrostatics using DNA （anionic） condensed by a positively charged carrier. These structures improve cell delivery and have been widely explored for gene therapy. In contrast, here we use cationic poly （^-amino esters） （PBAEs） to assemble polyplexes from CpG as an adjuvant to target and improve immune stimulation in cells and mouse models. Polyplexes were formed over a range of PBAE：CpG ratios, resulting in a library of complexes with increasingly positive charge and stronger binding as PBAE：CpG ratio increased. Although higher PBAE：CpG ratios exhibited improved CpG uptake, lower ratios of PBAE：CpG--which condensed CpG more weakly, activated DCs and tumorspecific T cells more effectively. In a mouse melanoma model, polyplexes with lower binding affinities improved survival more effectively compared with higher binding affinities. These data demonstrate that altering the polyplex interaction strength impacts accessibility of CpG to TLRs in immune cells. Thus, physiochemical properties, particularly the interplay between charge, uptake, and affinity, play a key role in determining the nature and efficacy of the immune response generated. This insight identifies new design considerations that must be balanced for engineering effective immunotherapies and vaccines.展开更多
Over the past decade, several natural and synthetic cationic polymers have been utilized for gene delivery into cells. Among them, polyethylenimine(PEI) was used for gene therapy successfully. The present study invest...Over the past decade, several natural and synthetic cationic polymers have been utilized for gene delivery into cells. Among them, polyethylenimine(PEI) was used for gene therapy successfully. The present study investigated the effect of PEI and ultrasound waves on ssD NA delivery into saffron cells. Gel retardation, dynamic light scattering(DLS) and scanning electron microscopy(SEM) assays were employed to determine the physicochemical properties of PEI/f-DNA polyplex(complex of PEI and fluorescently labeled DNA). Moreover, the cytotoxicity of PEI, PEI/f-DNA polyplex and ultrasound were investigated on saffron cells at different concentrations. The gel retardation results indicated that the formation and neutralization of the PEI/f-DNA polyplex were completed at N/P=5. The particle size distribution of the polyplexes was from 50 to 122 nm. The experimental results revealed that the cytotoxicity of the PEI/f-DNA polyplex was lower than that of PEI alone, hence the cells showed both dose-and exposure duration-dependent responses. Furthermore, the viability of saffron cells declined extremely after 5 and 10 min sonication but this reduction was not significant at 2 min exposure duration. The results also indicated that the combined utilization of ultrasound and PEI nanoparticles increased the transfection efficiency of saffron cells up to two times higher than those obtained by PEI or ultrasound separately.展开更多
目的:为降低聚阳离子基因载体polyplex的正电荷和毒性,在其表面构建中性磷脂膜制备lipopolyplex,并测定lipopolyplex对小鼠结肠癌细胞CT26和人乳腺癌细胞MCF-7的细胞毒性。方法:采用PEI25KDa与DNA复合制备polyplex,在polyplex体系中加...目的:为降低聚阳离子基因载体polyplex的正电荷和毒性,在其表面构建中性磷脂膜制备lipopolyplex,并测定lipopolyplex对小鼠结肠癌细胞CT26和人乳腺癌细胞MCF-7的细胞毒性。方法:采用PEI25KDa与DNA复合制备polyplex,在polyplex体系中加入中性脂质体和SADGE制备lipopolyplex。采用琼脂糖凝胶电泳考察lipopolyplex对质粒DNA的包裹能力;采用激光粒度仪和zeta电位分析仪测定lipopolyplex的粒径与zeta电位;采用透射电镜观察lipopolyplex的形态;采用CCK-8试剂盒考察lipopolyplex对CT26和MCF-7的细胞毒性。结果:琼脂糖凝胶电泳显示lipopolyplex可以完全包裹质粒DNA;lipopolyplex的粒径在200 nm左右,电位在-20 m V左右;透射电镜下为较为规则的球状颗粒;lipopolyplex在CT26和MCF-7细胞中的毒性明显低于聚阳离子基因载体polyplex。结论:在polyplex表面成功构建中性磷脂膜制备的lipopolyplex,可以完全的包裹DNA并且细胞毒性明显低于polyplex,在基因输送载体领域具有潜在应用价值。展开更多
Oligonucleotide therapeutics have great potential to target the currently undruggable genes and to generate entirely new therapeutic paradigms in multiple types of disease,thus having attracted much attention in recen...Oligonucleotide therapeutics have great potential to target the currently undruggable genes and to generate entirely new therapeutic paradigms in multiple types of disease,thus having attracted much attention in recent years.However,their applications are greatly hindered by a lack of safe and efficient oligonucleotide-delivery vectors.Polyplex nanovesicles formed from oligonucleotides and the cationic block have shown exceptional features for the delivery of therapeutic oligonucleotides and other biopharmaceuticals.Nevertheless,these polyplex nanovesicles are deeply fraught with difficulty in tolerating physiological ionic strength.Inspired by the high binding ability between the dipicolylamine(DPA)/zinc(Ⅱ)complex and the phosphodiester moieties of oligonucleotides,herein,we designed a coordinative cationic block to solve the intrinsic stability dilemma.Moreover,we found the stability of the resulted polyplex nanovesicles could be easily tuned by the content of coordinated zinc ions.In vitro cellular studies implied that the prepared zinc(Ⅱ)-coordinative polyplex nanovesicles preferred to retain in the lysosomes upon internalization,making them ideal delivery candidates for the lysosome-targeting oligonucleotide therapeutics.展开更多
Superwetting membranes have emerged as promising materials for the efficient treatment of oily wastewater.Typically,superwetting membranes can be developed by ingeniously chemical modification and topographical struct...Superwetting membranes have emerged as promising materials for the efficient treatment of oily wastewater.Typically,superwetting membranes can be developed by ingeniously chemical modification and topographical structuration of microporous membranes.Herein,we report the hierarchical assembly of metal-phenolic-polyplex coating to manipulate membrane surface superwettability by integrating metal-phenolic(Fe^(Ⅲ)-tannic acid(TA))assembly with polyplex(tannic acid-polyethylenimine(PEI))assembly.The proposed Fe-TA-PEI coating can be deposited on microporous membrane via simply dipping into Fe^(Ⅲ)-TA-PEI co-assembly solution.Based on the catechol chemistry,the coordination complexation of Fe^(Ⅲ)and TA develops metal-phenolic networks to provide hydrophilic chemistries,and the electrostatic complexation of TA and PEI generates nanoconjugates to impart hierarchical architectures.Benefiting from the synergy of hydrophilic chemistries and hierarchical architectures,the resulting PVDF/Fe-TA-PEI membrane exhibits excellent superhydrophilicity(~0°)underwater superoleophobicity(~150°)and superior anti-oil-adhesion capability.The superhydrophilicity of PVDF/Fe-TA-PEI membrane greatly promotes membrane permeability,featuring water fluxes up to 5860 L m^(-2)h^(-1).The underwater superoleophobicity of PVDF/Fe-TA-PEI membrane promises potential flux(3393 L m^(-2)h^(-1)),high separation efficiency(99.3%)and desirable antifouling capability for oil-in-water emulsion separation.Thus,we highlight the reported hierarchical metal-phenolic-polyplex assembly as a straightforward and effective strategy that enables the synchronous modulation of surface chemistry and topography toward superwetting membranes for promising high-flux and antifouling oil-water separation.展开更多
Surface modification by poly(ethylene glycol)(PEGylation)has been acknowledged as a powerful strategy in minimizing non-specific reactions for biomedical devices.Once applied into manufacture of drug/gene delivery sys...Surface modification by poly(ethylene glycol)(PEGylation)has been acknowledged as a powerful strategy in minimizing non-specific reactions for biomedical devices.Once applied into manufacture of drug/gene delivery systems,PEGylation has demonstrated to significantly improve their biocompatibility and stealthiness in physiological environment.Nonetheless,reluctant cell membrane affinities thus cellular uptake efficiencies owing to PEGylation brought up further issues that are imperative to be resolved.Pertain to this PEGylation dilemma,we attempted to introduce peptide(GPLGVRG)linkage between block copolymer of PEG-poly{N’-[N-(2-aminoethyl)-2-aminoethyl]aspartamide}PAsp(DET),wherein the cationic PAsp(DET)could self-assemble with pDNA into nanoscaled complex core.Noteworthy was the peptide linkage whose amino acids sequence could be specifically recognized and degraded by matrix metalloproteinases(MMPs)(overexpressed in extracellular milieu of tumors).Therefore,our subsequent studies validated facile detachment of PEGylation from the aforementioned polyplex micelles upon treatment of MMPs,which elicited improved cytomembrane affinities and cellular uptake efficiencies.In addition,promoted escape from endosome entrapment was also confirmed through direct endosome membrane destabilization by PAsp(DET),which was further elucidated to be attributable to dePEGylation as well as elevated charged density of PAsp(DET)in acidic endosomes.These benefits from dePEGylation eventually contributed to promoted gene expression at the affected cells and potent tumor growth suppression based on anti-angiogenic approach.Therefore,our developed strategy has provided a facile approach in overcoming the dilemma of PEGylation,which could be informative in design of drug/gene delivery systems.展开更多
This study demonstrates that our previously reported polywraplex, a synthetic siRNA carrier consisting of a uni-molecular polyplex core of customizable size and a self-assembled triblock copolymer envelop, may be cons...This study demonstrates that our previously reported polywraplex, a synthetic siRNA carrier consisting of a uni-molecular polyplex core of customizable size and a self-assembled triblock copolymer envelop, may be constructed using dendrimers as the crosslinking junctions. Replacing the branched low molecular weight PEI with polyamidoamine(PAMAM) dendrimer in the zeta potential regulated polymerization resulted in the similar network structured cationic polymer with electron microscopically visible crosslinking junctions. This visibility may offer a convenient way to characterize the molecular structure of the rationally designed networked siRNA-packing cationic polymer without altering its chemical properties and biologic functions. A series of physical-chemical characterizations and biological assays, comprising size, zeta potential, pre-phagocytic siRNA leaking and degradation, and silencing of functional genes, confirmed that the advanced properties of polywraplexes remained with the dendrimer junctions. Although sixth generation PAMAM dendrimer was used as the crosslinking junctions in the size-customizable polymerization for electron microscopic observation, lower generation dendrimer should also work in case more practical and structurally defined cationic polymer is needed.展开更多
基金Supported by the National Natural Science Foundation of China (Grant Nos. 20725206 and 20732004)Specialized Research Fund for the Doctoral Program of Higher EducationScientific Fund of Sichuan Province for Outstanding Young Scientist
文摘A novel cyclen-based linear polyamine (POGEC) was designed and synthesized from the reaction between 1,3-propanediol diglycidyl ether and 1,7-bis(diethoxyphosphory)-1,4,7,10-tetraazacyclod-odecane. High-affinity binding between POGEC and DNA was demonstrated by agarose gel electrophoresis and scanning electron microscopy (SEM). Moreover,the formed POGEC/DNA complex (termed polyplex) could be disassociated to release the free DNA through addition of the physiological concentration of NaCl solution. Fluorescence spectrum was used to measure the high-affinity binding and DNA condensation capability of POGEC. Circular dichroism (CD) spectrum indicates that the DNA conformation did not change after binding to POEGC.
文摘Many experimental cancer vaccines are exploring toll-like receptor agonists （TLRas） such as CpG, a DNA motif that agonizes toll-like receptor 9 （TLR9）, to trigger immune responses that are potent and molecularly-specific. The ability to tune the immune response is especially important in the immunosuppressive microenvironments of tumors. Because TLR9 is located intracellularly, CpG must be internalized by immune cells for functionality. Polyplexes can be self- assembled through electrostatics using DNA （anionic） condensed by a positively charged carrier. These structures improve cell delivery and have been widely explored for gene therapy. In contrast, here we use cationic poly （^-amino esters） （PBAEs） to assemble polyplexes from CpG as an adjuvant to target and improve immune stimulation in cells and mouse models. Polyplexes were formed over a range of PBAE：CpG ratios, resulting in a library of complexes with increasingly positive charge and stronger binding as PBAE：CpG ratio increased. Although higher PBAE：CpG ratios exhibited improved CpG uptake, lower ratios of PBAE：CpG--which condensed CpG more weakly, activated DCs and tumorspecific T cells more effectively. In a mouse melanoma model, polyplexes with lower binding affinities improved survival more effectively compared with higher binding affinities. These data demonstrate that altering the polyplex interaction strength impacts accessibility of CpG to TLRs in immune cells. Thus, physiochemical properties, particularly the interplay between charge, uptake, and affinity, play a key role in determining the nature and efficacy of the immune response generated. This insight identifies new design considerations that must be balanced for engineering effective immunotherapies and vaccines.
基金supported by the University of Mohaghegh Ardabili, India under Grant (51-487)
文摘Over the past decade, several natural and synthetic cationic polymers have been utilized for gene delivery into cells. Among them, polyethylenimine(PEI) was used for gene therapy successfully. The present study investigated the effect of PEI and ultrasound waves on ssD NA delivery into saffron cells. Gel retardation, dynamic light scattering(DLS) and scanning electron microscopy(SEM) assays were employed to determine the physicochemical properties of PEI/f-DNA polyplex(complex of PEI and fluorescently labeled DNA). Moreover, the cytotoxicity of PEI, PEI/f-DNA polyplex and ultrasound were investigated on saffron cells at different concentrations. The gel retardation results indicated that the formation and neutralization of the PEI/f-DNA polyplex were completed at N/P=5. The particle size distribution of the polyplexes was from 50 to 122 nm. The experimental results revealed that the cytotoxicity of the PEI/f-DNA polyplex was lower than that of PEI alone, hence the cells showed both dose-and exposure duration-dependent responses. Furthermore, the viability of saffron cells declined extremely after 5 and 10 min sonication but this reduction was not significant at 2 min exposure duration. The results also indicated that the combined utilization of ultrasound and PEI nanoparticles increased the transfection efficiency of saffron cells up to two times higher than those obtained by PEI or ultrasound separately.
文摘目的:为降低聚阳离子基因载体polyplex的正电荷和毒性,在其表面构建中性磷脂膜制备lipopolyplex,并测定lipopolyplex对小鼠结肠癌细胞CT26和人乳腺癌细胞MCF-7的细胞毒性。方法:采用PEI25KDa与DNA复合制备polyplex,在polyplex体系中加入中性脂质体和SADGE制备lipopolyplex。采用琼脂糖凝胶电泳考察lipopolyplex对质粒DNA的包裹能力;采用激光粒度仪和zeta电位分析仪测定lipopolyplex的粒径与zeta电位;采用透射电镜观察lipopolyplex的形态;采用CCK-8试剂盒考察lipopolyplex对CT26和MCF-7的细胞毒性。结果:琼脂糖凝胶电泳显示lipopolyplex可以完全包裹质粒DNA;lipopolyplex的粒径在200 nm左右,电位在-20 m V左右;透射电镜下为较为规则的球状颗粒;lipopolyplex在CT26和MCF-7细胞中的毒性明显低于聚阳离子基因载体polyplex。结论:在polyplex表面成功构建中性磷脂膜制备的lipopolyplex,可以完全的包裹DNA并且细胞毒性明显低于polyplex,在基因输送载体领域具有潜在应用价值。
基金financially supported by the National Key Research and Development Program of China (No.2021YFA1201200)the National Natural Science Foundation of China (Nos. 51833008, 52173141 and 82102192)+1 种基金Zhejiang Provincial Key Research and Development Program (No.2020C01123)China Postdoctoral Science Foundation (No.2019M662059)
文摘Oligonucleotide therapeutics have great potential to target the currently undruggable genes and to generate entirely new therapeutic paradigms in multiple types of disease,thus having attracted much attention in recent years.However,their applications are greatly hindered by a lack of safe and efficient oligonucleotide-delivery vectors.Polyplex nanovesicles formed from oligonucleotides and the cationic block have shown exceptional features for the delivery of therapeutic oligonucleotides and other biopharmaceuticals.Nevertheless,these polyplex nanovesicles are deeply fraught with difficulty in tolerating physiological ionic strength.Inspired by the high binding ability between the dipicolylamine(DPA)/zinc(Ⅱ)complex and the phosphodiester moieties of oligonucleotides,herein,we designed a coordinative cationic block to solve the intrinsic stability dilemma.Moreover,we found the stability of the resulted polyplex nanovesicles could be easily tuned by the content of coordinated zinc ions.In vitro cellular studies implied that the prepared zinc(Ⅱ)-coordinative polyplex nanovesicles preferred to retain in the lysosomes upon internalization,making them ideal delivery candidates for the lysosome-targeting oligonucleotide therapeutics.
基金financially supported by National Natural Science Foundation of China(No.21706230)China Postdoctoral Science Foundation(No.2019M652141)+2 种基金National Natural Science Foundation of China(Nos.22075246 and 21776253)Public Welfare Project of the Science and Technology Committee of Zhejiang Province(No.LGF20B060002)Provincial Key R&D Program of Zhejiang Province(No.2019C03094)。
文摘Superwetting membranes have emerged as promising materials for the efficient treatment of oily wastewater.Typically,superwetting membranes can be developed by ingeniously chemical modification and topographical structuration of microporous membranes.Herein,we report the hierarchical assembly of metal-phenolic-polyplex coating to manipulate membrane surface superwettability by integrating metal-phenolic(Fe^(Ⅲ)-tannic acid(TA))assembly with polyplex(tannic acid-polyethylenimine(PEI))assembly.The proposed Fe-TA-PEI coating can be deposited on microporous membrane via simply dipping into Fe^(Ⅲ)-TA-PEI co-assembly solution.Based on the catechol chemistry,the coordination complexation of Fe^(Ⅲ)and TA develops metal-phenolic networks to provide hydrophilic chemistries,and the electrostatic complexation of TA and PEI generates nanoconjugates to impart hierarchical architectures.Benefiting from the synergy of hydrophilic chemistries and hierarchical architectures,the resulting PVDF/Fe-TA-PEI membrane exhibits excellent superhydrophilicity(~0°)underwater superoleophobicity(~150°)and superior anti-oil-adhesion capability.The superhydrophilicity of PVDF/Fe-TA-PEI membrane greatly promotes membrane permeability,featuring water fluxes up to 5860 L m^(-2)h^(-1).The underwater superoleophobicity of PVDF/Fe-TA-PEI membrane promises potential flux(3393 L m^(-2)h^(-1)),high separation efficiency(99.3%)and desirable antifouling capability for oil-in-water emulsion separation.Thus,we highlight the reported hierarchical metal-phenolic-polyplex assembly as a straightforward and effective strategy that enables the synchronous modulation of surface chemistry and topography toward superwetting membranes for promising high-flux and antifouling oil-water separation.
基金the National Natural Science Foundation of China(No.21878041)the funding support from Talent Project of Revitalizing Liaoning(No.XLYC1807184)Dalian Science&Technology Innovation Fund(Nos.2020JJ26SN050,2020JJ26GX025)。
文摘Surface modification by poly(ethylene glycol)(PEGylation)has been acknowledged as a powerful strategy in minimizing non-specific reactions for biomedical devices.Once applied into manufacture of drug/gene delivery systems,PEGylation has demonstrated to significantly improve their biocompatibility and stealthiness in physiological environment.Nonetheless,reluctant cell membrane affinities thus cellular uptake efficiencies owing to PEGylation brought up further issues that are imperative to be resolved.Pertain to this PEGylation dilemma,we attempted to introduce peptide(GPLGVRG)linkage between block copolymer of PEG-poly{N’-[N-(2-aminoethyl)-2-aminoethyl]aspartamide}PAsp(DET),wherein the cationic PAsp(DET)could self-assemble with pDNA into nanoscaled complex core.Noteworthy was the peptide linkage whose amino acids sequence could be specifically recognized and degraded by matrix metalloproteinases(MMPs)(overexpressed in extracellular milieu of tumors).Therefore,our subsequent studies validated facile detachment of PEGylation from the aforementioned polyplex micelles upon treatment of MMPs,which elicited improved cytomembrane affinities and cellular uptake efficiencies.In addition,promoted escape from endosome entrapment was also confirmed through direct endosome membrane destabilization by PAsp(DET),which was further elucidated to be attributable to dePEGylation as well as elevated charged density of PAsp(DET)in acidic endosomes.These benefits from dePEGylation eventually contributed to promoted gene expression at the affected cells and potent tumor growth suppression based on anti-angiogenic approach.Therefore,our developed strategy has provided a facile approach in overcoming the dilemma of PEGylation,which could be informative in design of drug/gene delivery systems.
基金the grant of the Natural Science Foundation of China(Grant nos.81373352 and 81690262)。
文摘This study demonstrates that our previously reported polywraplex, a synthetic siRNA carrier consisting of a uni-molecular polyplex core of customizable size and a self-assembled triblock copolymer envelop, may be constructed using dendrimers as the crosslinking junctions. Replacing the branched low molecular weight PEI with polyamidoamine(PAMAM) dendrimer in the zeta potential regulated polymerization resulted in the similar network structured cationic polymer with electron microscopically visible crosslinking junctions. This visibility may offer a convenient way to characterize the molecular structure of the rationally designed networked siRNA-packing cationic polymer without altering its chemical properties and biologic functions. A series of physical-chemical characterizations and biological assays, comprising size, zeta potential, pre-phagocytic siRNA leaking and degradation, and silencing of functional genes, confirmed that the advanced properties of polywraplexes remained with the dendrimer junctions. Although sixth generation PAMAM dendrimer was used as the crosslinking junctions in the size-customizable polymerization for electron microscopic observation, lower generation dendrimer should also work in case more practical and structurally defined cationic polymer is needed.
