Surface modification with polyallylamines for adhesion of biopolymers and cells

Cationic polymers with NH₂-groups were used for modification of charged and uncharged surfaces of planar slides, wells of plates, and spherical nanoparticles. Our study was aimed at development of a simple functionalization method of plain surfaces and colloids of different chemical compositions for adhesion of native biopolymers, including proteins, and viable bacterial and eukaryotic cells. Poly(allylamine)s (pAA) and polylysines (pLys) of different molecular weights spontaneously formed interfaces convenient for adhesion of biopolymers and cells. Thickness of the pAA 65 kDa layer ∼1.5–2 nm was measured by two methods: 1) atomic force microscopy (AFM) on mica slides and 2) registration of the long range surface optical waves excitation angle and the critical angle of total internal reflection from the liquid on a photonic crystal surface by using the biosensor. The sorption capacity of 0.1 mg/ml pAA 65 kDa exceeded the values of other polyamines at different concentrations. Physisorption of proteins on pAA layer was reversible and up to 70% of attached proteins could be removed by subsequent washes. Additional treatment with glutaraldehyde (GA) provided stable chemical cross-linking of the compounds containing primary NH₂-groups with aminated surfaces. The proteins immobilized on the pAA-covered surface retained their ability to bind with specific monoclonal and polyclonal antibodies. Bacterial cells after adhesion on pAA65-covered surfaces maintained their morphology, could reproduce and express the green fluorescent protein (gfp) gene under control of the inducible lac promoter. Eukaryotic cells of human and mammalian origin also remained viable on pAA-treated slides as proven by their staining with fluorescent dyes and cell divisions until confluent monolayers. Mammalian cells could not attach onto silicon wafers but grew on pAA interface of the silicon slides until confluent monolayers. Thus, surface modification with polyallylamines provides adhesion of native biopolymers and living cells.     

3D Printing of Mechanically Stable Calcium‐Free Alginate‐Based Scaffolds with Tunable Surface Charge to Enable Cell Adhesion and Facile Biofunctionalization

For the 3D printing of bioscaffolds, the importance of a suitable bioink cannot be overemphasized. With excellent printability and biocompatibility, alginate (Alg) is one of the most used bioinks. However, its bioinert nature and insufficient mechanical stability, due to only crosslinking via cation interactions, hinder the practical application of Alg‐based bioinks in the individualized therapy of tissue defects. To overcome these drawbacks, for the first time, an ε‐polylysine (ε‐PL)‐modified Alg‐based bioink (Alg/ε‐PL) is produced. The introduction of ε‐PL improves the printability of the Alg‐based bioink due to increasing electrostatic interactions, which enhances the self‐supporting stability of the as‐printed scaffolds. The presence of the functional crosslinking –COOH and –NH₂ groups in Alg and ε‐PL under mild conditions further enhances the mechanical stability of the scaffolds, far exceeding that of Alg/Ca²⁺ scaffolds. The surface charge of the prepared scaffolds is finely tuned by the feed ratio of Alg to ε‐PL and postimmobilization of different quantities of additional ε‐PL, with a view to enhancing cell adhesion and further biofunctionalization. The results indicate that chondroitin sulfate, an extracellular matrix component, and vascular endothelial growth factor can be successfully applied to biofunctionalize the scaffolds via electrostatic adsorption for enhanced biological activity.     

Preparation and properties of bio‐based epoxy montomorillonite nanocomposites derived from polyglycerol polyglycidyl ether and ε‐polylysine

Glycerol polyglycidyl ether (GPE) and polyglycerol polyglycidyl ether (PGPE) were cured with ε‐poly(L ‐lysine) (PL) using epoxy/amine ratios of 1 : 1 and 2 : 1 to create bio‐based epoxy cross‐linked resins. When PGPE was used as an epoxy resin and the epoxy/amine ratio was 1 : 1, the cured neat resin showed the greatest glass transition temperature (T_g), as measured by differential scanning calorimetry. Next, the mixture of PGPE, PL, and montomorillonite (MMT) at an epoxy/amine ratio of 1 : 1 in water was dried and cured finally at 110°C to create PGPE‐PL/MMT composites. The X‐ray diffraction and transmission electron microscopy measurements revealed that the composites with MMT content 7–15 wt % were exfoliated nanocomposites and the composite with MMT content 20 wt % was an intercalated nanocomposite. The T_g and storage modulus at 50–100°C for the PGPE‐PL/MMT composites measured by DMA increased with increasing MMT content until 15 wt % and decreased at 20 wt %. The tensile strength and modulus of the PGPE‐PL/MMT composites (MMT content 15 wt %: 42 and 5300 MPa) were much greater than those of the cured PGPE‐PL resin (4 and 6 MPa). Aerobic biodegradability of the PGPE‐PL in an aqueous medium was ～ 4% after 90 days, and the PGPE‐PL/MMT nanocomposites with MMT content 7–15 wt % showed lower biodegradability. © 2009 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

ε-聚赖氨酸高产菌株的选育

依据传统诱变理论和白色链霉菌生物合成ε 聚赖氨酸的特点 ,确立了以枯草芽孢杆菌为敏感菌株 ,抗性突变株筛选和抑菌圈法相结合的初筛方法。在优化诱变条件的基础上 ,以白色链霉菌为出发菌株 ,经紫外线与硫酸二乙酯诱变 ,选育出一株遗传性状稳定 ,遗传标记为AECr+Glyr的ε 聚赖氨酸生产菌株 ,其产量可达 0 79g/L。     

Activity of ε–Polylysine Against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes

ABSTRACT: ε–polylysine is a homopolymer of L-lysine, an essential amino acid, with a reportedly wide antimicrobial spectrum. This study evaluated the antimicrobial activity of ε–polylysine, as compared with known preservatives and organic acids, against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes , in culture broth. The compounds tested included ε–polylysine (0.0025% to 0.05%), sodium diacetate (0.25%), sodium lactate (3.0%), lactic acid (0.1%), and acetic acid (0.1%), alone, as well as in combination with ε– polylysine (0.0025% to 0.03%); all treatments were evaluated in tryptic soy broth supplemented with 0.6% yeast extract. Treatments were inoculated (approximately 2 log colony-forming units [CFU]/mL) with 5-strain ( E. coli O157:H7, S. Typhimurium) or 10-strain ( L. monocytogenes ) mixtures of the pathogens. Survival/growth of the inoculated bacteria was periodically monitored during incubation at 4 °C (30 d) and 24 °C (48 h). Bactericidal effects of ε–polylysine were obtained against E. coli O157:H7 and S. Typhimurium at 4 °C. At the same temperature (4 °C), ε–polylysine alone or in combination with other compounds tested inhibited growth or was bactericidal against L. monocytogenes. All 3 pathogens were inhibited by ε–polylysine at 24 °C; however, L. monocytogenes was the most sensitive and S. Typhimurium the most resistant. The antimicrobial activity of ε–polylysine against E. coli O157:H7 and S. Typhimurium was enhanced ( P < 0.05) when tested in combination with sodium diacetate or acetic acid. Combination treatments with sodium lactate resulted in loss of ε–polylysine activity by the end of the incubation period. Overall, under the conditions of this study, ε–polylysine exhibited antimicrobial effects against the 3 pathogens tested.     

Polylysine as an adhesive for the attachment of nanoplankton to substrates for electron microscopy

Nanoplankton can be readily attached to polylysine-coated substrates for examination by transmission and scanning electron microscopy. This simple technique gives high yields of organisms which are consistently better preserved and less obscured by detritus than samples dried directly on to substrates.     

Stimuli-triggered structural engineering of synthetic and biological polymeric assemblies

Response to external stimuli is a fundamental and intrinsic behavior of living systems. There has been increasing interest for designing and constructing responsive polymeric superstructures by self-assembly. Stimuli-induced self-assembly and post-assembly triggering strategies provide an alternative approach for the manipulation of self-assembled architectures of either biological or synthetic polymeric materials. Stimuli-induced structural transformations may produce ensembles with new topologies or materials with exceptionally complex features inaccessible via conventional self-assembly processes. This is in contrast to materials that simply undergo stimuli-induced degradation, or disassembly processes. Since a variety of cellular processes depend on responses to environmental stimuli that lead to more complexity and increased function, and are related to structural transitions over the nano- to microscale, insights into stimuli-triggered morphogenesis can further deepen our understanding of cellular behaviors. Indeed, an understanding of these processes will likely inspire scientists to develop materials with advanced and tailored architectures for biosensing, diagnosis and therapy as well as other biomedical applications. Herein, we highlight state-of-the-art achievements in the stimuli-triggered structural manipulation of polymer assemblies. Furthermore, future developments in this nascent and growing field are briefly discussed.     

Chitosan utilized for bacterial preparation for scanning electron microscopy

Bacterial sample preparation is crucial for its observation by scanning electron microscopy (SEM). However, the current polylysine (PLL) method leads to bacterial morphological changes. To overcome this problem, we employed chitosan (CS) to coat coverslips to prepare bacteria for SEM and compared it with the PLL method. Coverslips coated with 0.025% (w/v) CS showed satisfactory bacterial binding ability. Within 30 min of binding time, the number of bacteria on CS-coated and PLL-coated coverslips exhibited no differences. Four bacteria strains were employed to compare the differences in SEM images between the two methods. Most of the bacteria showed irregular surface or sticky substances after settling on PLL-coated coverslips, while bacteria with clear surface texture were observed on CS-coated coverslips. Transmission electron microscopy (TEM) images showed deformed bacterial envelope on PLL-coated coverslips; meanwhile, similar intact envelope was observed from the bacteria on CS-coated coverslips and the bacteria without any treatment. The TEM results verified the morphological differences of SEM between the two methods. Except for morphology, the length of the rod-shaped bacteria was longer on CS-coated coverslips than that on PLL-coated coverslips, less shrinkage of the sample was observed, and CS could preserve the length of the rod-shaped bacteria better than PLL in its preparation for SEM. It is demonstrated that the low-cost CS could be utilized in bacterial preparation for SEM to acquire preferable images. Bacterial suspension with optical density at 600 nm of about 0.5, deposited on 0.025% CS-coated coverslips for 30 min, and followed by routine fixation, dehydration, and drying are optimal parameters.