Cross-linking Reagent Research Articles

Repairing the ruptured annular fibrosus by using type I collagen combined with citric acid, EDC and NHS: an in vivo study

To explore the effect of citric acid (CA)-1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)/N-hydroxysuccinimide (NHS) collagen gel on repairing annular defects. Type I collagen was extracted from the rat-tail tendon and crosslinked with CA at different mass ratio using EDC and NHS as crosslinking reagents to prepare four kinds of collagen gels. Forty-eight adult SD rats were divided into first sham group (n=8), second group (n=10) which was punctured and injected with CA-EDC/NHS collagen gel, third group (n=10) which was punctured and injected with CA-EDC/NHS collagen gel, fourth group (n=10) which was punctured and injected with EDC/NHS collagen gel, and fifth group (n=10) which was punctured and untreated. X-ray images and magnetic resonance imaging images were obtained before puncture and at the 1st, 2nd, and 4thweek after puncture. At each time point, disc height index (%DHI), voxel count and modified MRI Pfirrmann grading were collected and analyzed. All animals were killed at the 4th week to study the morphology. The discs in the second group showed only slight degeneration compared with the healthy discs, and the results of %DHI (average 79%), voxel count (average 126.9), Pfirrmann grading (average grade 1.3) and morphology in the second group indicated less degeneration tendency compared with the other three puncture groups at the 4th week (P<0.05). The annular fibrosus was partially repaired by the collagen gels that bridged the defects. CA-EDC/NHS collagen gel is capable of repairing annular defects induced by needle puncture, which may be closely related to the dose of CA.

Improvement on Physical Properties of Pullulan Films by Novel Cross-Linking Strategy.

Pullulan based films possess several advantages, including high transparency, low toxicity, good biodegradability, good mechanical properties, and low oxygen permeability, are preferable for food packaging. The application of pullulan films on food packaging, however, has inherent disadvantage of high water solubility. In this study, glutaraldehyde and glycerol were used as the cross-linking reagent and the plasticizer respectively to improve water resistance and physical properties of the pullulan films. Effects of cross-linking degree on physical properties, including water absorptions, swelling behaviors, water vapor permeability and tensile strengths of films were evaluated. FTIR results demonstrated that the pullulan films were successfully cross-linked by glutaraldehyde. The tensile strength of pullulan films could be enhanced significantly (P < 0.05) when glutaraldehyde was between 1% and 5% (w/w); nevertheless, the amount of glutaraldehyde above 20% (w/w) led to films brittleness. With the addition of glycerol as a plasticizer enhanced the extensibility of films as well as the hydrophilicity, resulting in higher water vapor permeability.

An Integrated Mass Spectrometry Based Approach to Probe the Structure of the Full-Length Wild-Type Tetrameric p53 Tumor Suppressor.

We present an integrated approach for investigating the topology of proteins through native mass spectrometry (MS) and cross-linking/MS, which we applied to the full-length wild-type p53 tetramer. For the first time, the two techniques were combined in one workflow to obtain not only structural insight in the p53 tetramer, but also information on the cross-linking efficiency and the impact of cross-linker modification on the conformation of an intrinsically disordered protein (IDP). P53 cross-linking was monitored by native MS and as such, our strategy serves as a quality control for different cross-linking reagents. Our approach can be applied to the structural investigation of various protein systems, including IDPs and large protein assemblies, which are challenging to study by the conventional methods used for protein structure characterization.

The Development of a Portable SPR Bioanalyzer for Sensitive Detection of Escherichia coli O157:H7.

The purpose of this study was to develop a portable surface plasmon resonance (SPR) bioanalyzer for the sensitive detection of Escherichia coli O157:H7 in comparison with an enzyme-linked immunosorbent assay (ELISA). The experimental setup mainly consisted of an integrated biosensor and a homemade microfluidic cell with a three-way solenoid valve. In order to detect Escherichia coli O157:H7 using the SPR immunoassay, 3-mercaptopropionic acid (3-MPA) was chemisorbed onto a gold surface via covalent bond for the immobilization of biological species. 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) were used as crosslinker reagents to enable the reaction between 3-MPA and Escherichia coli O157:H7 antibodies by covalent –CO–NH– amide bonding. The experimental results were obtained from the Escherichia coli O157:H7 positive samples prepared by 10-, 20-, 40-, 80-, and 160-fold dilution respectively, which show that a good linear relationship with the correlation coefficient R of 0.982 existed between the response units from the portable SPR bioanalyzer and the concentration of Escherichia coli O157:H7 positive samples. Moreover, the theoretical detection limit of 1.87 × 103 cfu/mL was calculated from the positive control samples. Compared with the Escherichia coli O157:H7 ELISA kit, the sensitivity of this portable SPR bioanalyzer is four orders of magnitude higher than the ELISA kit. The results demonstrate that the portable SPR bioanalyzer could provide an alternative method for the quantitative and sensitive determination of Escherichia coli O157:H7 in field.

A new covalent peroxidase conjugation method using bis(sulfosuccinimidyl) suberate as cross-linking reagent in a two-step procedure

ABSTRACTA new method has been developed to prepare horseradish peroxidase (HRP) conjugates using bis(sulfosuccinimidyl)suberate (BS3) as cross-linking reagent in a two-step procedure. The enzyme is reacted with BS3, introducing active ester molecules into the enzyme itself, and it is then used directly to label amino-containing compounds without further treatment. The proteins involved in the conjugation undergo minimal modifications. The reaction conditions are evaluated, as well as studies on the conservation of the biological activities of the conjugated proteins and the stability of the conjugates in time. These conjugates are found to be significantly improved compared with similar products prepared by conventional methods.

ChIP-seq Analysis of Condensin Complex in Cultured Mammalian Cells.

ChIP-seq, or chromatin immunoprecipitation combined with massively parallel DNA sequencing, is a powerful technique to investigate in vivo protein-DNA interactions on a genome-wide scale at high resolution. Here we describe a ChIP-seq protocol optimized for analysis of condensin I complex on human mitotic chromosomes. The protocol includes procedures of intensive cell fixation by two cross-linking reagents and thorough chromatin shearing by nuclease and sonication treatments, both of which contribute to improving the signal-to-noise ratio of condensin I ChIP-seq profiles. The optimized protocol may also be helpful to explore chromosomal binding sites of other "hard-to-see" proteins by ChIP-seq.

Cross-Linked Enzyme Aggregates for Applications in Aqueous and Nonaqueous Media.

Extensive cross-linking of a precipitate of a protein by a cross-linking reagent (glutaraldehyde has been most commonly used) creates an insoluble enzyme preparation called cross-linked enzyme aggregates (CLEAs). CLEAs show high stability and performance in conventional aqueous as well as nonaqueous media. These are also stable at fairly high temperatures. CLEAs with more than one kind of enzyme activity can be prepared, and such CLEAs are called combi-CLEAs or multipurpose CLEAs. Extent of cross-linking often influences their morphology, stability, activity, and enantioselectivity.

Effects of pectin structure and crosslinking method on the properties of crosslinked pectin nanofibers

We reported crosslinking of electrospun nanofibers of three representative pectins (high-methoxylated, low-methoxylated, low-methoxylated and amidated pectin) and characterization of the crosslinked nanofibers. One mono-crosslinking strategy and two dual-crosslinking strategies were developed. Mono-crosslinking is achieved using calcium ions (Ca2+) to crosslink carboxylate ions in galacturonic acid residues. Dual-crosslinking is achieved using covalent crosslinking reagents glutaraldehyde (GLU) or adipic acid dihydrazide (ADH) to further crosslink hydroxyl groups or carboxylate ions after Ca2+ crosslinking. Mechanical tests and degradation experiments indicated pectin structure affected mechanical and degradation properties of Ca2+-crosslinked nanofibers remarkably. Subsequent GLU crosslinking improved their mechanical strength moderately but did not inhibit their degradation, while subsequent ADH crosslinking improved their mechanical strength and slowed down their degradation dramatically. Cell studies demonstrated that most crosslinked pectin nanofibers were of no obvious cytotoxicity, and both ADH crosslinking and high degree of methoxylation facilitated cell adhesion and proliferation on pectin nanofiber mats.

Divide and conquer: cleavable cross-linkers to study protein conformation and protein-protein interactions.

Chemical cross-linking combined with mass spectrometry (MS) and computational modeling has evolved as an alternative method to address fundamental questions in structural biology. The constraints revealed by the cross-links yield valuable distance information and allow one to deduce three-dimensional structural information on very large and transient protein complexes. During the past few years, technical advances in the cross-linking/MS approach have been enormous, mainly owing to the fantastic advances in MS technology, and it is easily overlooked that significant progress has been made in the design of novel cross-linking reagents. In this review, the advent of cleavable cross-linking reagents will be highlighted. In particular, gas-phase (MS-) cleavable cross-linkers offer unique properties for an automated, data-dependent assignment of cross-linked products based on the generation of characteristic fragment ion signatures in MS/MS and MS3 spectra. Therefore, MS-cleavable cross-linkers are envisioned to hold the key for proteome-wide applications of the chemical cross-linking/MS approach, not only to delineate the conformation of single proteins but also to decipher protein interaction networks. Graphical Abstract Outline of the analytical strategy using cleavable cross-linkers in combination with MS to conduct protein conformational and protein interaction studies.

TM4 of the glutamate transporter GLT-1 experiences substrate-induced motion during the transport cycle.

Excitatory amino acid transporter 2 (EAAT2), also known as glial glutamate transporter type 1 (GLT-1), plays an important role in maintaining the extracellular glutamate concentrations below neurotoxic levels. The highly conserved TM2 transmembrane domain of GLT-1 maintains a stable position during the transport cycle; however, the effect of the transport cycle on the topology of TM4 in not well established. To further reveal the function of TM4, two cysteine pairs between TM2 and TM4 were introduced using site-directed mutagenesis. A significant decrease of transport activity was observed in the I93C/V241C and I97C/V241C mutants upon application of the oxidative cross-linking reagent, copper (II) (1,10-phenanthroline)3 (CuPh), which suggests that a conformational shift is essential for transporter activity. Furthermore, the decrease in activity by CuPh crosslinking was enhanced in external media with glutamate or potassium, which suggests that TM2 and TM4 assume closer proximity in the inward-facing conformation of the transporter. Our results suggest that the TM4 domain of GLT-1, and potentially other glutamate transporters, undergoes a complex conformational shift during substrate translocation, which involves an increase in the proximity of the TM2 and TM4 domains in the inward-facing conformation.

Horseradish peroxidase-assisted approach to decolorize and detoxify dye pollutants in a packed bed bioreactor

In this study, horseradish peroxidase (HRP) was covalently immobilized on the calcium-alginate support using glutaraldehyde (GA) as a cross-linking reagent for detoxification and degradation of synthetic dyes. Immobilization procedure furnished significant immobilization efficiency (86.27 ± 3.43%) along with apparent and relative activity of 24.39 ± 1.03 U/g and 84.97 ± 3.54%, respectively, for immobilized-HRP. In comparison to free-state, immobilized-HRP catalyzed the substrate oxidation reaction in a slightly acidic and wider temperature range, with an optimum at 60 °C. After 10 and 60 min of incubation at 60 °C, the immobilized-HRP displayed 99.0% and 89.0% of residual activities, whereas the free counterpart retained only 34.0% and 18.0% of residual activities, respectively. Moreover, the immobilized-HRP showed potential efficiency for the decolorization of dyes in sequential dye-decolorizing batch reactions. Cytotoxicity analysis using a plant bioassay and acute test demonstrated that the Ca-alginate immobilized-HRP may effectively be used for detoxification of dyes and has a great potential for large-scale environmental remediation.

Effect of silica coating on Fe3O4 magnetic nanoparticles for lipase immobilization and their application for biodiesel production

The scope of this study was to prepare different organosilane-modified Fe3O4@SiO2 core magnetic nanocomposites with immobilized lipase and explore their potential application in biodiesel field. Fe3O4 magnetic nanoparticles prepared by co-precipitation method were coated with various ratios of SiO2 as per Stöber method and further functionalized by different organosilane compounds APTES (3-aminopropyltriethoxysilane) and MPTMS (3-mercaptopropyltrimethoxysilane). Functionalized Fe3O4@SiO2 magnetic nanoparticles were immobilized with free lipase NS81006 by glutaraldehyde cross-linking reagent. The functional groups, structure, morphology and magnetic susceptibility of synthesized and modified Fe3O4@SiO2 magnetic nanoparticles were characterized by FTIR, XRD, SEM, TEM, and VSM techniques. The immobilization efficiency and activity recovery were reduced by increasing the ratio of silica coating on Fe3O4. Maximum activity recovery (84% by APTES, 83% by MPTMS) and biodiesel yield (>90%) were obtained by lipase immobilized on Fe3O4@SiO2 support when the Fe3O4 and SiO2 (TEOS) ratio was low (1:0.25).

Measurement of Ionic Conductivity and Mechanical Property of Slide-Ring Gel Swollen with Propylene Carbonate Including Lithium Ions

The slide-ring (SR) gel is made from polyrotaxane (PR), in which cyclic cyclodextrins (CDs) are threaded on an axial poly(ethylene glycol) (PEG) polymer chain capped by bulky end groups, through intermolecular crosslinking between the CDs. The CDs form mobile cross-linking junctions that slide along the PEG, so that these SR gels exhibit unique characteristics, ”pulley effect ”, such as unique swelling and mechanical properties. This research was directed to use SR gel to achieve polymer gel electrolytes with both high ionic conductivity and mechanical ductility, which was one of the indexes of mechanical strength, at the same time. By using the SR gels swollen with propylene carbonate containing lithium ion, this research explores the use of the SR gel to achieve polymer gel electrolytes with both high ionic conductivity and mechanical ductility. First, by modifying the hydroxyl groups of the CDs by methyl groups, the affinity with several solvents, particularly mixed electrolysis solution (ES) of propylene carbonate (PC) and lithium salt, was improved, and the SR gel was swollen with ES. In the present research, we report the ionic conductivity of PR or methylated polyrotaxanes (MePR)-SR gels containing lithium ES with varying the crosslinking density of SR gels to clarify the ion transport of ES in the SR gel network. Furthermore, we report compressive measurement of SR gel containing ES to show that SR gels have high ionic conductivity as well as high mechanical ductility. MePRs were prepared with a degree of substitution (DS) at 27.5 and 74.2 %. Subsequently, MePR-SR gels were prepared with various number ratios of crosslinking reagents, such as divinyl sulfone where the sum of hydroxyl and methyl group of MeCD was varied from 6 to 10 %. The SR gel was swollen with pure PC or ES, which was 1.0 M PC solution of LiTFSI, followed by evaluation of the swelling ratio of the SR gel. For investigating electrical properties, we measured ionic conductivity, molar conductivity and potential window of SR gels swollen with ES. Furthermore, the relationship of ionic conductivity and temperature of those swollen gel was examined to obtain activation energy. Moreover, by the compaction test of MePR-SR gel with DS of 74.2 %, the mechanical property was investigated. A pure PC could hardly make the SR gel swell both for PR-SR gel and MePR-SR gel with DS of 27.5 %, while MePR-SR gels with DS of 74.2 % could be slightly swollen with pure PC. Since PEG, which is component of PR, can be soluble in PC, the ability of swelling depends on the affinity of PC to CDs or MeCDs. Increasing DS of MeCDs make MePR soluble to pure PC, and consequently MePR-SR gels with DS of 74.2 % was slightly swollen with pure PC. Next, ES could not make PR-SR gel swell at all, while MePR-SR gels could be swollen with ES very well. From the results of conductivity measurements, MePR-SR gel with DS of 74.2 % and the number ratio of crosslinking reagent of 6 mol% had 95 % of molar conductivity for pure ES, and decreased with increasing number ratio of crosslinking reagent, which confirmed that the polymer matrix distrurbed the ionic transport of the ES. Whereas, the potential window of MePR-SR gels with DS of 74.2 % were identically 5.5 V, which agreed with the value of PC-LiTFSI. This result suggested that SR gels do not affect potential window. The mechanical property of MePR-SR gel with DS of 74.2 %, in all gels, Young’s modulus was quite small that meant that these gels were quite soft. With swelling ratio, that is, decreasing number ratios of crosslinking reagents, Young’s modulus decreased and gel became a bit soft. MePR-SR gels with high number ratios of crosslinking reagents were fractured, while MePR-SR gel with low number ratios of crosslinking reagents did not fractured under the compression of almost half thickness than its original thickness. This result suggested that gels with a small crosslinking density were very soft, hence the stress was dispersed and cracks were difficult to generate, so gels were not fractured under high compression. Figure 1

ChemInform Abstract: Polysaccharides/Silica Hybrid Materials: New Perspectives for Sustainable Raw Materials

AbstractReview: 79 refs.

Dissociation Behavior of a TEMPO-Active Ester Cross-Linker for Peptide Structure Analysis by Free Radical Initiated Peptide Sequencing (FRIPS) in Negative ESI-MS.

We have synthesized a homobifunctional amine-reactive cross-linking reagent, containing a TEMPO (2,2,6,6-tetramethylpiperidine-1-oxy) and a benzyl group (Bz), termed TEMPO-Bz-linker, to derive three-dimensional structural information of proteins. The aim for designing this novel cross-linker was to facilitate the mass spectrometric analysis of cross-linked products by free radical initiated peptide sequencing (FRIPS). In an initial study, we had investigated the fragmentation behavior of TEMPO-Bz-derivatized peptides upon collision activation in (+)-electrospray ionization collision-induced dissociation tandem mass spectrometry (ESI-CID-MS/MS) experiments. In addition to the homolytic NO-C bond cleavage FRIPS pathway delivering the desired odd-electron product ions, an alternative heterolytic NO-C bond cleavage, resulting in even-electron product ions mechanism was found to be relevant. The latter fragmentation route clearly depends on the protonation of the TEMPO-Bz-moiety itself, which motivated us to conduct (-)-ESI-MS, CID-MS/MS, and MS3 experiments of TEMPO-Bz-cross-linked peptides to further clarify the fragmentation behavior of TEMPO-Bz-peptide molecular ions. We show that the TEMPO-Bz-linker is highly beneficial for conducting FRIPS in negative ionization mode as the desired homolytic cleavage of the NO-C bond is the major fragmentation pathway. Based on characteristic fragments, the isomeric amino acids leucine and isoleucine could be discriminated. Interestingly, we observed pronounced amino acid side chain losses in cross-linked peptides if the cross-linked peptides contain a high number of acidic amino acids. Graphical Abstract ᅟ.

Electrochemical detection of uric acid via uricase-immobilized graphene oxide

Measurement of the uric acid level in the body can be improved by biosensing with respect to the accuracy, sensitivity and time consumption. This study has reported the immobilization of uricase onto graphene oxide (GO) and its function for electrochemical detection of uric acid. Through chemical modification of GO using 1-ethyl-3-(dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysulfosuccinimide (NHS) as cross-linking reagents, the enzyme activity of the immobilized uricase was much comparable to the free enzyme with 88% of the activity retained. The modified GO-uricase (GOU) was then subjected to electrocatalytic detection of uric acid (UA) via cyclic voltammetry (CV). For that reason, a glassy carbon electrode (GCE) was modified by adhering the GO along with the immobilized uricase to facilitate the redox reaction between the enzyme and the substrate. The modified GOU/GCE outperformed a bare electrode through the electrocatalytic activity with an amplified electrical signal for the detection of UA. The electrocatalytic response showed a linear dependence on the UA concentration ranging from 0.02 to 0.49 mM with a detection limit of 3.45 μM at 3σ/m. The resulting biosensor also exhibited a high selectivity towards UA in the presence of other interference as well as good reproducibility.

Cross-linked enzyme aggregates (CLEAs) of Pencilluim notatum lipase enzyme with improved activity, stability and reusability characteristics

Cross-linked enzyme aggregates (CLEAs) are considered as an effective tool for the immobilization of enzyme. In this study, Pencillium notatum lipase (PNL) was immobilized as carrier free cross-linked enzyme aggregates using glutaraldehyde (GLA) and Ethylene glycol-bis [succinic acid N-hydroxysuccinimide] (EG-NHS) as cross-linking agents. The optimal conditions for the synthesis of an efficient lipase CLEAs such as precipitant type, the nature and amount of cross-linking reagent, and cross-linking time were optimized. The recovered activities of CLEAs were considerably dependent on the concentration of GLA; however, the activity recovery was not severely affected by EG-NHS as a mild cross-linker. The EG-NHS aggregates displayed superior hydrolytic (52.08±2.52%) and esterification (64.42%) activities as compared to GLA aggregates which showed 23.8±1.86 and 34.54% of hydrolytic and esterification activity, respectively. Morphological analysis by fluorescence and scanning electron microscope revealed that EG-NHS aggregates were smaller in size with larger surface area compared to GLA aggregates. The pH optima of both types of CLEAs were displaced to slightly alkaline region and higher temperature as compared to native enzyme. Highest enzyme activity of CLEAs was achieved at the pH of 9.0 and 42°C temperature. Moreover, a significant improvement in the thermal resistance was also recorded after immobilization. After ten reusability cycles in aqueous medium, GLA and EG-NHS cross-linked lipase CLEAs preserved 63.62% and 70.9% of their original activities, respectively. The results suggest that this novel CLEA-lipase is potentially usable in many industrial applications.

The Effect of Covalently-Attached ATRP-Synthesized Polymers on Membrane Stability and Cytoprotection in Human Erythrocytes.

Erythrocytes have been described as advantageous drug delivery vehicles. In order to ensure an adequate circulation half-life, erythrocytes may benefit from protective enhancements that maintain membrane integrity and neutralize oxidative damage of membrane proteins that otherwise facilitate their premature clearance from circulation. Surface modification of erythrocytes using rationally designed polymers, synthesized via atom-transfer radical polymerization (ATRP), may further expand the field of membrane-engineered red blood cells. This study describes the fate of ATRP-synthesized polymers that were covalently attached to human erythrocytes as well as the effect of membrane engineering on cell stability under physiological and oxidative conditions in vitro. The biocompatible, membrane-reactive polymers were homogenously retained on the periphery of modified erythrocytes for at least 24 hours. Membrane engineering stabilized the erythrocyte membrane and effectively neutralized oxidative species, even in the absence of free-radical scavenger-containing polymers. The targeted functionalization of Band 3 protein by NHS-pDMAA-Cy3 polymers stabilized its monomeric form preventing aggregation in the presence of the crosslinking reagent, bis(sulfosuccinimidyl)suberate (BS3). A free radical scavenging polymer, NHS-pDMAA-TEMPO˙, provided additional protection of surface modified erythrocytes in an in vitro model of oxidative stress. Preserving or augmenting cytoprotective mechanisms that extend circulation half-life is an important consideration for the use of red blood cells for drug delivery in various pathologies, as they are likely to encounter areas of imbalanced oxidative stress as they circuit the vascular system.

Influence of natural and synthetic crosslinking reagents on the structural and mechanical properties of chitosan-based hybrid hydrogels

The objective of this work was to correlate the physical and chemical properties of chitosan/poly(vinyl alcohol)/genipin (CS/PVA/GEN) and chitosan/poly(vinyl alcohol)/glutaraldehyde (CS/PVA/GA) hydrogels with their structural and mechanical responses. In addition, their molecular structures were determined and confirmed using FTIR spectroscopy. The results indicated that the hybrid hydrogels crosslinked with genipin showed similar crystallinity, thermal properties, elongation ratio and structural parameters as those crosslinked with glutaraldehyde. However, it was found that the elastic moduli of the two hybrid hydrogels were slightly different: 2.82±0.33MPa and 2.08±0.11MPa for GA and GEN, respectively. Although the hybrid hydrogels crosslinked with GEN presented a lower elastic modulus, the main advantage is that GEN is five to ten thousand times less cytotoxic than GA. This means that the structural and mechanical properties of hybrid hydrogels crosslinked with GEN can easily be tuned and could have potential applications in the tissue engineering, regenerative medicine, food, agriculture and environmental industries.

Well-defined reducible cationic nanogels based on functionalized low-molecular-weight PGMA for effective pDNA and siRNA delivery

Nucleic acid-based gene therapy is a promising treatment option to cure numerous intractable diseases. For non-viral gene carriers, low-molecular-weight polymeric vectors generally demonstrate poor transfection performance, but benefit their final removals from the body. Recently, it was reported that aminated poly(glycidyl methacrylate) (PGMA) is one potential gene vector. Based on ethylenediamine (ED)-functionalized low-molecular-weight PGMA (denoted by PGED), a flexible strategy was herein proposed to design new well-defined reducible cationic nanogels (denoted by PGED-NGs) with friendly crosslinking reagents for highly efficient nucleic acid delivery. α-Lipoic acid (LA), one natural antioxidant in human body, was readily introduced into ED-functionalized PGMA and crosslinked to produce cationic PGED-NGs with plentiful reducible lipoyl groups. PGED-NGs could effectively complex plasmid DNA (pDNA) and short interfering RNA (siRNA). Compared with pristine PGED, PGED-NGs exhibited much better performance of pDNA transfection. PGED-NGs also could efficiently transport MALAT1 siRNA (siR-M) into hepatoma cells and significantly suppressed the cancer cell proliferation and migration. The present work indicated that reducible cationic nanogels involving LA crosslinking reagents are one kind of competitive candidates for high-performance nucleic acid delivery systems. Statement of SignificanceRecently, the design of new types of high-performance nanoparticles is of great significance in delivering therapeutics. Nucleic acid-based therapy is a promising treatment option to cure numerous intractable diseases. A facile and straightforward strategy to fabricate safe nucleic acid delivery nanovectors is highly desirable. In this work, based on ethylenediamine-functionalized low-molecular-weight poly(glycidyl methacrylate), a flexible strategy was proposed to design new well-defined reducible cationic nanogels (denoted by PGED-NGs) with α-Lipoic acid, one friendly crosslinking reagent, for highly efficient nucleic acid delivery. Such PGED-NGs possess plentiful reducible lipoyl groups, effectively encapsulated pDNA and siRNA and exhibited excellent abilities of nucleic acid delivery. The present work indicated that reducible cationic nanogels involving α-lipoic acid crosslinking reagents are one kind of competitive candidates for high-performance nucleic acid delivery systems.