Lignin-Based Nonviral Gene Carriers Functionalized by Poly[2-(Dimethylamino)ethyl Methacrylate]: Effect of Grafting Degree and Cationic Chain Length on Transfection Efficiency

Lignin is a natural renewable biomass resource with great potential for applications, while its development into high value-added molecules or materials is rare. The development of biomass lignin as potential nonviral gene delivery carriers was initiated by our group through the “grafting-from” approach. Firstly, the lignin was modified into macroinitiator using 2-bromoisobutyryl bromide. Then cationic polymer chains of poly[2-(dimethylamino)ethyl methacrylate] (PDMAEMA) were grown from the lignin backbone using atom transfer radical polymerization (ATRP) to yield lignin-PDMAEMA graft copolymers (LPs) with branched structure. To gain a deep understanding of the relationship between the nonviral gene transfection efficiency of such copolymers and their structural and compositional factors, herein eight lignin-based macroinitiators with different modification degrees (MDs, from 3.0 to 100%) were synthesized. Initiated by them, a series of 20 LPs were synthesized with varied structural factors such as grafting degree (GD, which is equal to MD, determining the cationic chain number per lignin macromolecule), cationic chain length (represented by number of repeating DMAEMA units per grafted arm or degree of polymerization, DP) as well as the content of N element (N%) which is due to the grafted PDMAEMA chains and proportional to molecular weight of the LPs. The in vitro gene transfection capability of these graft copolymers was evaluated by luciferase assay in HeLa, COS7 and MDA-MB-231cell lines. Generally, the copolymers LP-12 (N% = 7.28, MD = 36.7%, DP = 13.6) and LP-14 (N% = 6.05, MD = 44.4%, DP = 5.5) showed good gene transfection capabilities in the cell lines tested. Overall, the performance of LP-12 was the best among all the LPs in the three cell lines at the N/P ratios from 10 to 30, which was usually several times higher than PEI standard. However, in MDA-MB-231 at N/P ratio of 30, LP-14 showed the best gene transfection performance among all the LPs. Its gene transfection efficiency was ca. 11 times higher than PEI standard at this N/P ratio. This work demonstrated that, although the content of N element (N%) which is due to the grafted PDMAEMA chains primarily determines the gene transfection efficiency of the LPs, it is not the only factor in explaining the performance of such copolymers with the branched structure. Structural factors of these copolymers such as grafting degree and cationic chain length could have a profound effect on the copolymer performance on gene transfection efficiency. Through carefully adjusting these factors, the gene transfection efficiency of the LPs could be modulated and optimized for different cell lines, which could make this new type of biomass-based biomaterial an attractive choice for various gene delivery applications.

Microwave-Assisted Synthesis of Modified Glycidyl Methacrylate–Ethyl Methacrylate Oligomers, Their Physico-Chemical and Biological Characteristics

In this study, well-known oligomers containing ethyl methacrylate (EMA) and glycidyl methacrylate (GMA) components for the synthesis of the oligomeric network [P(EMA)-co-(GMA)] were used. In order to change the hydrophobic character of the [P(EMA)-co-(GMA)] to a more hydrophilic one, the oligomeric chain was functionalized with ethanolamine, xylitol (Xyl), and L-ornithine. The oligomeric materials were characterized by nuclear magnetic resonance and Fourier transform infrared spectroscopy, scanning electron microscopy, and differential thermogravimetric analysis. In the final stage, thanks to the large amount of -OH groups, it was possible to obtain a three-dimensional hydrogel (HG) network. The HGs were used as a matrix for the immobilization of methylene blue, which was chosen as a model compound of active substances, the release of which from the matrix was examined using spectrophotometric detection. The cytotoxic test was performed using fluid extracts of the HGs and human skin fibroblasts. The cell culture experiment showed that only [P(EMA)-co-(GMA)] and [P(EMA)-co-(GMA)]-Xyl have the potential to be used in biomedical applications. The studies revealed that the obtained HGs were porous and non-cytotoxic, which gives them the opportunity to possess great potential for use as an oligomeric network for drug reservoirs in in vitro application.

Improvement of in vitro antioxidant activity of kaempferol by encapsulation in copolymer micelles

Antioxidant capacity of poorly soluble natural antioxidant kaempferol, in particular free or loaded in two types of cationic micelles, was studied on non-enzyme induced lipid peroxidation (LPO) in vitro. The micelles were based on triblock copolymers - poly(2-(dimethylamino)ethyl methacrylate-b-poly(propylene oxide)-b-poly(2-(dimethylamino)ethyl methacrylate (PDMAEMA-PPO-PDMAEMA) and poly(2-(dimethylamino)ethyl methacrylate-b-poly(ε-caprolactone)-b-poly(2-(dimethylamino)ethyl methacrylate (PDMAEMA-PCL-PDMAEMA). The lipid peroxidation was induced by incubating of rat liver microsomes with iron sulphate and ascorbic acid (Fe2+/AA). The effect of free and micellar kaempferol (at concentrations 25, 50 and 75 μg/ml) was assessed after 20 min incubation time. In the non-enzyme lipid peroxidation model, the kaempferol-loaded micelles significantly decreased the formation of malondialdehyde (MDA). The effect of kaempferol loaded in PDMAEMA-PCL-PDMAEMA micelles was more pronounced, showing an improved antioxidant activity in the conditions of oxidative stress and lipid peroxidation in vitro.

Preparation and application of poly (hydroxyl ethyl methacrylate) nanocomposite hydrogels containing iron oxide nanoparticles as wound dressing

In recent years, polymeric hydrogels are widespread in the field of biological materials such as wound dressing and wound care. In this work, we report for the first time the preparation and application of pHEMA nanocomposite hydrogels containing iron oxide nanoparticles as wound dressings. For this purpose, nanocomposite hydrogels based on poly (hydroxyl ethyl methacrylate) (pHEMA) and various amounts of 5, 10, and 15 wt% iron oxide nanoparticles were successfully prepared via radical polymerization. The structure and morphology of nanocomposite hydrogels were determined by Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscope (FE-SEM), respectively. The results of gel fraction and the degree of swelling of hydrogels demonstrated that the gel percentage of pHEMA increased, and the degree of swelling decreased with increasing the percentage of nanoparticles. The WVRT and the porosity of hydrogels decreased by increasing the quantity of nanoparticles and were suitable for wound dressing applications. The effect of iron oxide nanoparticles on the mechanical properties of nanocomposite hydrogels was also studied using compression test and hardness shore A durometer. The results indicated that the compression strength, modulus, strain, and hardness are steadily increasing compared to pure hydrogel by adding nanoparticles. The maximum increase was obtained for a hydrogel sample with 15 wt% iron oxide nanoparticles. Antibacterial properties and biocompatibility were determined by the disk-diffusion and MTT assay methods, respectively. Based on the results, nanocomposite hydrogels exhibited higher percentages of cell survival and better antibacterial properties compared to pure pHEMA.

Determination of dopamine based on a temperature-sensitive PMEO2MA and Au@rGO-MWCNT nanocomposite-modified electrode

First, the nanocomposite Au@rGO-MWCNT was synthesized by hydrothermal method. Next, a temperature-controlled composite sensing film was prepared by composite modification of poly(2-(2-methoxyethoxy)ethyl methacrylate) (PMEO2MA) and Au@rGO-MWCNT on a glassy carbon...