Inhibition of α-glucosidase activity and intestinal glucose transport to assess the in vivo anti-hyperglycemic potential of dodecyl-acylated phlorizin and polydatin derivatives.

A diet containing natural active compounds that can inhibit the hydrolytic activity of α-glucosidase on carbohydrates and intestinal glucose absorption is an effective means of controlling postprandial hyperglycemia. Phlorizin and polydatin as phenolic glycosides have a high affinity for the catalytic site of α-glucosidase, but exhibited unsatisfactory competitive inhibitory capacity, with an IC50 of 0.97 and >2 mM, respectively. However, dodecyl-acylated derivatives of phlorizin and polydatin exerted α-glucosidase inhibitory capacity, with an IC50 of 55.10 and 70.95 µM, respectively, which were greatly enhanced and much stronger than that of acarbose with an IC50 of 2.46 mM. The SPR assay suggested the high affinity of dodecyl phlorizin and dodecyl polydatin to α-glucosidase with equilibrium dissociation constant (KD) values of 12.0 and 7.9 µM, respectively. Both dodecyl phlorizin and dodecyl polydatin reduced the catalytic ability of α-glucosidase by reversible noncompetitive and uncompetitive mixed inhibition, which bind noncovalently to the allosteric site 2 through hydrogen bonds and hydrophobic interactions, thereby inducing the secondary structure unfolding and intrinsic fluorescence quenching of α-glucosidase. Confocal microscopy detection visually showed significant inhibitory effects on FITC-labeled glucose uptake in intestinal Caco-2 cells by phlorizin, polydatin, dodecyl phlorizin and dodecyl polydatin. In addition, based on the differentiated Caco-2 cell monolayer model, dodecyl phlorizin and dodecyl polydatin suppressed intestinal glucose transport more effectively than phlorizin and polydatin, suggesting that they were promising in vivo hypoglycemic active compounds.

Antibacterial Nanomaterials: Mechanisms, Impacts on Antimicrobial Resistance and Design Principles.

Antimicrobial resistance (AMR) is one of the biggest threats to the environment and health. AMR rapidly invalidates conventional antibiotics, and antimicrobial nanomaterials have been increasingly explored as alternatives. Interestingly, several antimicrobial nanomaterials show AMR-independent antimicrobial effects without detectable new resistance and have therefore been suggested to prevent AMR evolution. In contrast, some are found to trigger the evolution of AMR. Given these seemingly conflicting findings, a timely discussion of the two faces of antimicrobial nanomaterials is urgently needed. This review systematically compares the killing mechanisms and structure-activity relationships of antibiotics and antimicrobial nanomaterials. We then focus on nano-microbe interactions to elucidate the impacts of molecular initiating events on AMR evolution. Finally, we provide an outlook on future antimicrobial nanomaterials and propose design principles for the prevention of AMR evolution.

Physicochemical aspects of design of ultrathin films based on chitosan, pectin, and their silver nanocomposites with antiadhesive and bactericidal potential.

Implant-related infection is one of the serious problems in regenerative medicine. Promising approach to overcome the problems caused by bacterial growth on the medical implants is their modification by bioactive coatings. A versatile technique for designing multilayer films with tailored characteristics at the nanometer scale is layer-by-layer assembly. In this study, multilayer films based on biopolymers (pectin and chitosan) and their nanocomposites with silver nanoparticles have been prepared and evaluated. The buildup of multilayers was monitored using the quartz crystal microbalance with dissipation technique. The morphology of the obtained films was investigated by atomic force microscopy. We have demonstrated that pectin-Ag-containing films were characterized by the linear growth and smooth defect-free surface. When pectin-Ag was substituted for the pectin in the multilayer systems, the properties of the formed coatings were significantly changed: the film rigidity and surface roughness increased, as well as the film growth acquired the parabolic character. All prepared multilayer films have shown antibacterial activity against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria. The significant decrease in the number of the adhered E. coli on the multilayer surface has been determined; moreover, many of the cells were misshapen with cytoplasm leaking. The prepared multilayer films showed a mild activity against S. aureus predominantly due to the antiadhesive effect. Our results indicate that antibacterial activity of biopolymer multilayers is determined by the film composition and physicochemical characteristics and can be associated with their antiadhesive and bactericidal behaviors.

Polyvinyl alcohol and pectin blended films: Preparation, characterization, and mesenchymal stem cells attachment.

The design of novel wound dressings for chronic wound treatment is still of great importance. One of the promising approaches is application of mesenchymal stem cells (MSCs), immobilized on a flexible polymer film, for healing. In this study, blended films based on polyvinyl alcohol (PVA) and pectin with different component ratio have been prepared by solution casting method and evaluated. Physicochemical properties of the formed PVA/pectin films, including their morphology, wettability, swelling, stability, mechanical characteristics, have been studied. We demonstrated that the surface of PVA/pectin films could be modified by ultraviolet or dielectric barrier discharge plasma exposure. After both ultraviolet and plasma treatment, the hydrophilicity of PVA/pectin films increased. It has been shown that additional crosslinking of PVA/pectin films with glutaraldehyde resulted in reinforcement of their structure. MSCs were cultured on neat and modified PVA/pectin samples to evaluate the effects of film characteristics and composition on cell behavior. It has been determined that MSCs effectively adhered to glutaraldehyde-crosslinked PVA/pectin films and formed on them the monolayer culture of fibroblast-like cells. The additional modification of PVA/pectin films with collagen resulted in enhancement of MSCs adhesion. Our results show that the obtained PVA/pectin films with adhered MSCs can be suggested for potential application as a part of novel complex wound dressings.

Fabrication and characterization of pectin-based three-dimensional porous scaffolds suitable for treatment of peritoneal adhesions.

Formation of peritoneal adhesions is common complication after abdominal and pelvic surgery. They bear a significant health problem with an influence to quality of life and health care expenses. Promising approach for their prevention is using of biodegradable barrier films for physical separation of peritoneal surfaces. In the present study, highly porous pectin-based three-dimensional (3D) scaffolds were obtained by freeze-drying technique. Physico-chemical properties of the formed materials, including their morphology, porosity, density, and stability, have been studied. The evaluation of their biocompatibility, biodegradation, and potential antiadhesion effect was studied by in vivo experiment. To reinforce the scaffolds structure and improve their stability in physiological solutions, pectin chains were cross-linked with divalent cations. We determined optimal cross-linking conditions, which allow obtaining scaffolds with desired biodegradation rate. These cross-linked scaffolds fully dissolved within 8 days in the peritoneal cavity with low presence of complications and some antiadhesive effect. It has also been determined that mesenchymal stem cells from adipose tissue could effectively adhere to the scaffolds with preservation of their viability. Our results show that obtained materials can be suggested as mechanical scaffold for delivery of the stem cells culture to peritoneal surfaces as a part of complex antiadhesive barrier system.

Layer-by-layer buildup of polysaccharide-containing films: Physico-chemical properties and mesenchymal stem cells adhesion.

Layer-by-Layer assembled polyelectrolyte films offer the opportunity to control cell attachment and behavior on solid surfaces. In the present study, multilayer films based on negatively charged biopolymers (pectin, dextran sulfate, carboxymethylcellulose) and positively charged polysaccharide chitosan or synthetic polyelectrolyte polyethyleneimine has been prepared and evaluated. Physico-chemical properties of the formed multilayer films, including their growth, morphology, wettability, stability, and mechanical properties, have been studied. We demonstrated that chitosan-containing films are characterized by the linear growth, the defect-free surface, and predominantly viscoelastic properties. When chitosan is substituted for the polyethyleneimine in the multilayer system, the properties of the formed films are significantly altered: the rigidity and surface roughness increases, the film growth acquires the exponential character. The multilayer films were subsequently used for culturing mesenchymal stem cells. It has been determined that stem cells effectively adhered to chitosan-containing films and formed on them the monolayer culture of fibroblast-like cells with high viability. Our results show that cell attachment is a complex process which is not only governed by the surface functionality because one of the key parameter effects on cell adhesion is the stiffness of polyelectrolyte multilayer films. We therefore propose our Layer-by-Layer films for applications in tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2093-2104, 2018.