Dental restorative materials containing quaternary ammonium compounds have sustained antibacterial action

BackgroundThis narrative review addresses dental restorative materials with sustained antibacterial action, especially those containing quaternary ammonium compounds. Secondary caries occurs around restorations, causing further loss of mineral and breakdown of the restoration. Lesions adjacent to restorations account for more than 40% of needed restorations. Restorative materials with antibacterial properties will potentially solve this problem.Types of Studies ReviewedSeveral groups are researching composite restorative materials that incorporate antibacterial agents. These agents are mostly exhausted over time. Newer studies involve materials that incorporate antibacterial microparticles that remain active and do not leach out.ResultsOne such antibacterial agent, quaternary ammonium coupled with inorganic silica into minute particles (QASi), has been studied in the laboratory and in humans. QASi particles incorporated into dental materials retain their antibacterial action over time without leaching or loss of activity. A clinical in situ study in humans using dental composite containing QASi resulted in highly significantly less demineralization in the adjacent enamel than the control composite material.Conclusions and Practical ImplicationsDental restorative materials that contain QASi have sustained antibacterial properties, have mechanical properties comparable to those of presently marketed materials, and have been cleared by the US Food and Drug Administration. Clinical studies have shown that composites incorporating QASi have the potential to markedly reduce the occurrence of caries around restorations. Because caries around restorations is a major problem, restorative materials with sustained antibacterial properties will have an important effect in reducing secondary caries around restorations.     

Lipid coated chitosan-DNA nanoparticles for enhanced gene delivery

Chitosan as a polycationic non-viral vector for gene delivery has the advantage of being a biocompatible and biodegradable polymer. However, without laborious chemical modifications to its structure, it is of limited use as a gene delivery vehicle due to its low ability to efficiently transfect under physiological conditions. To address this problem, we developed novel liposome encapsulated chitosan nanoparticles; lipochitoplexes (LCPs). Chitosan nanoparticles (CsNPs) were obtained using the ionic gelation technique. For this purpose, an ultrapure low molecular weight chitosan with a high degree of deacetylation was cross-linked using polyanionic tripolyphosphate resulting in efficient entrapment of plasmid DNA (pDNA) inside the nanoparticles. LCPs were prepared by incubating chitosan nanoparticles together with anionic liposomes (DPPC/Cholesterol). The LCPs offered better pDNA protection, reduced cytotoxicity and at least twofold increase in the transfection efficiency under physiological conditions. The efficiency of our delivery vehicle was also proved in vivo in the chorioallantoic membrane model (CAM). LCPs were able to transfect the CAM without traumatising the surrounding blood vessels. This new biocompatible composite system devoid of chemical modifications, organic solvents and harsh production conditions makes it an optimal gene delivery vehicle for in vivo applications offering new insights into the field of non-viral gene therapy.     

Effective PEI-mediated delivery of CRISPR-Cas9 complex for targeted gene therapy

The-state-of-art CRISPR/Cas9 is one of the most powerful among the approaches being developed to rescue fundamental causes of gene-based inheritable diseases. Several strategies for delivering such genome editing materials have been developed, but the safety, efficacy over time, cost of production, and gene size limitations are still under debate and must be addressed to further improve applications. In this study, we evaluated branched forms of the polyethylenimine (PEI) – branched PEI 25 kDa (BPEI-25K) – and found that it could efficiently deliver CRISPR/Cas9 plasmids. Plasmid DNA expressing both guide RNA and Cas9 to target the Slc26a4 locus was successfully delivered into Neuro2a cells and meditated genome editing within the targeted locus. Our results demonstrated that BPEI-25K is a promising non-viral vector to deliver the CRISPR/Cas9 system in vitro to mediate targeted gene therapy, and these findings contribute to an understanding of CRISPR/Cas9 delivery that may enable development of successful in vivo techniques.     

Characterization and compatibility of hydroxyethyl starch–polyethylenimine copolymers for DNA delivery

Hydroxyethyl starch (HES) has been proposed as a biodegradable polymer for shielding of DNA polyplexes, where the feasibility of this approach was shown both in vitro and in vivo. In this study, we report on the physicochemical characterization, the in vitro cytocompatibility and hemotoxicity of HES-decorated polyplexes. For this purpose, various HES molecules were coupled to a 22?kDa linear polyethylenimine (LPEI22) to produce a library of nine different HES–PEI conjugates. Particle analysis using dynamic light scattering showed that, neither the molar mass of HES nor the amount of HES in the polyplexes affected the particle diameter, as it was consistently around 70–80?nm. Imaging using atomic force microscopy and transmission electron microscopy showed that, both naked and HESylated polyplexes were in the same size range and had a spherical morphology. Meanwhile, the HES-mediated particle-shielding effect, manifested as reduction in the surface charge, strongly correlated with the molar mass of HES, where the charge decreased linearly with the increase in molar mass. Ethidium bromide binding assay showed that HES–PEI did not negatively affect DNA condensation at N/P ratios higher than 4. HES conjugation also showed a stabilizing effect against salt-induced particle disassembly, and particle aggregation in protein-containing media. Compatibility tests included cellular viability, as well as erythrocyte aggregation and hemolysis assays. HES–PEI conjugates showed lower cytotoxicity, no aggregation, and much lower hemolysis compared to unmodified PEI. In conclusion, these results show that the HES–PEI conjugates are promising gene delivery polymers with favorable physicochemical properties and compatibility profile.     

Coating With Chitooligosaccharides Enhances the Cytokine Induction of Listeria ivanovii–Based Vaccine Strain

Listeria ivanovi (LI) is an available live bacterial vaccine vector. This work attempted to coat LI-based vaccine candidates (LI-Rv0129c) with chitooligosaccharides (COSs) as an adjuvant to enhance the cellular immune responses induced. COS-bacteria composite was achieved by mixing the bacteria suspension with equal volume of COS solution, and this process accompanied with the increase of bacteria superficial zeta potential and formation of special superficial configurations. COS coating improved the ratio swallowed by the macrophage-like RAW264.7 cells from 0.54% to 2.88% (p < 0.001). In vivo, the COS-coated LI-Rv0129c strain did elicit significantly higher specific CD4⁺ IFN-γ, CD4⁺ TNF-α or CD8⁺ IFN-γ secretion (0.91%, 1.00%, 0.30%, respectively) than naked LI-Rv0129c (0.32%, 0.38%, 0.07%, respectively) (p < 0.01). These results demonstrated that COS is a promising adjuvant to enhance the protective cellular immune responses induced by LI-based vaccine strains. Our work provided a notion for developing adjuvant for Listeria and other bacterial vector-based vaccines.     

Cardiac RNAi therapy using RAGE siRNA/deoxycholic acid-modified polyethylenimine complexes for myocardial infarction

Inflammatory response in myocardial ischemia-reperfusion injury plays a critical role in ventricular remodeling. To avoid deleterious effects of overwhelming inflammation, we blocked the expression of receptor for advanced glycation end-products (RAGE), a key mediator of the local and systemic inflammatory responses, via RNAi mechanism. Herein, a facial amphipathic deoxycholic acid-modified low molecular weight polyethylenimine (DA-PEI) was used as a siRNA delivery carrier to myocardium. The DA-PEI conjugate formed a stable complex with siRNA via electrostatic and hydrophobic interactions. The siRAGE/DA-PEI formulation having negligible toxicity could enhance intracellular delivery efficiency and successfully suppress RAGE expression both in vitro and in vivo. Furthermore, the cardiac administration of siRAGE/DA-PEI reduced apoptosis and inflammatory cytokine release, subsequently led to attenuation of left ventricular remodeling in rat myocardial infarction model. The potential therapeutic effects of RAGE gene silencing on myocardial ischemia-reperfusion injury may suggest that the siRAGE/DA-PEI delivery system can be considered as a promising strategy for treating myocardial infarction.     

非病毒型载体介导基因转染

基因载体是制约基因转移技术发展的关键。近年来,非病毒载体由于其安全、低毒、低免疫原性等特点而备受青睐。文章以脂质体和聚乙烯亚胺为代表,介绍了非病毒载体的性质、介导转染的机制。随着人们对细胞转染机制了解的深入以及生物材料科学的迅速发展,非病毒型载体将有望实现高效、低毒、靶向特异等特点,从而成为基因治疗中的理想载体。     

A Simple Scheme for Grafting an Ion‐Imprinted Layer onto the Surface of Poly(propylene) Fibers

A simple method for surface ion‐imprinting is described in which poly(glycidyl methacrylate) is grafted onto PP fibers to act as a binder to a Cu²⁺ ion‐imprinted polymer layer. Mechanical and thermal analysis shows excellent tensile and thermal properties of the ion‐imprinted fibers. Competitive adsorption of Cu²⁺ and other ions such as Zn²⁺, Ni²⁺, Cd²⁺, Mn²⁺, and Mg²⁺ on the fibes is examined and a highly selective adsorption of Cu²⁺ is observed. The selectivity coefficient for Cu²⁺ with respect to Cd²⁺ can be as high as 20.2. Dynamic adsorption results indicate that the fibers are able to thoroughly remove Cu²⁺ from a solution. Regeneration performance tests show that the fibers maintain almost the same adsorption capacity for copper ions after 10 adsorption/desorption cycles.

     

新型非病毒载体聚乙烯亚胺介导基因转染参数的研究

聚乙烯亚胺是一种新型阳离子多聚物基因释放载体,可结合浓缩DNA,通过粘附内吞,进入细胞,使携带的质粒表达。本研究目的通过对聚乙烯亚胺各种转染参数的测定,为合成以聚乙烯亚胺为骨架的人工载体积累数据。方法:本研究利用聚乙烯亚胺分别结合含β半乳糖甙酶报告基因的pSVβ表达质粒、含绿色荧光蛋白报告基因的pEGFP质粒转染Cos-7细胞,通过组织化学法测定细胞抽提产物中β半乳糖甙酶的表达量、流式细胞仪法测定绿色荧光蛋白阳性细胞的表达比例,来测定影响转基因效率的各种参数。结果:在培养液中,6μg/ml聚乙烯亚胺作用24h,NIH 3T3细胞生存率为64.2%,7μg/ml聚乙烯亚胺细胞生存率为54.4%。电泳阻滞试验,聚乙烯亚胺在N/P比在3.0以上方可完全结合DNA。溶酶体抑制剂氯喹可增加聚乙烯亚胺的转染效率。培养液中的白蛋白、血清可降低转染效率。作为配制聚乙烯亚胺/DNA复合物的溶媒,HEPES缓冲液优于生理盐水,生理盐水优于5%葡萄糖。配制聚乙烯亚胺/DNA复合物的溶媒中加入Mg2+降低转染效率。聚乙烯亚胺转染效率优于SuperFectTM(断裂型树突状多聚物),而毒性低于SuperFectTM。结论:本研究首次报道了聚乙烯亚胺与DNA结合配伍的N/P比计算公式,N/P=7.75×b/c,这里b是PEI的质量(μg),c是质粒的质量(μg);PEI的工作终浓度应≤6μg/ml。通过体外细胞试验证明,聚乙烯亚胺是一种有效的真核细胞转染剂和人工合成基因载体的骨架。