Influence of the composition and preparation method on the morphology and swelling behavior of alginate–chitosan hydrogels

In this study, a 2⁴ factorial experimental design was employed in order to evaluate the influence of the reaction conditions and preparation method on alginate–chitosan hydrogel properties. Alginate content, pH, chitosan molecular weight and the hydrogel preparation method were the independent variables and the reaction yield, particle size, swelling degree and point of zero surface charge were the dependent variables. The results showed that hydrogels were spherical with an average diameter of 5.0 ± 2.0 μm. Reaction yield varied according to the parameters, and chitosan molecular weight showed the greatest influence. Furthermore, the swelling degree and point of zero surface charge showed a linear dependence on the alginate content. In this regard, the study showed that hydrogels with a specific charge and swelling degree can be obtained by controlling the alginate content using the equation here provided to give an enhanced and site-specific controlled drug release.     

Poly(ethylene glycol acrylate)‐functionalized hydrogels for heparan sulfate oligosaccharide recognition

Heparan sulfates are complex polysaccharides belonging to the family of glycosaminoglycans that participate to the regulation of cell behavior and tissue homeostasis. The biological activities conferred to heparan sulfates are largely dependent on the content and positioning of the sulfate groups along their saccharidic units. At present, identification of particular sulfation patterns in biologically relevant heparan sulfate sequences remains challenging. Although several approaches for structure analysis exist, the complexity of heparan sulfates makes new and original approaches still required. Here, we used molecular imprinting technologies to prepare a library of polyethylene glycol acrylate functionalized hydrogels with the aim to investigate their applicability as specific recognizing systems for fondaparinux, a synthetic pentasaccharide analog to the antithrombin binding site of heparin. Adequate choice of the hydrogel composition and controlling rebinding conditions were important determinants for improving the sulfated oligosaccharide recognition specificity and selectivity. Our results suggest that molecular imprinting approaches could be a possibility for the specific recognition of biologically active sequences in heparan sulfates.     

Density Gradient Multilayered Polymerization (DGMP): A Novel Technique for Creating Multi-compartment,Customizable Scaffolds for Tissue Engineering

Complex tissue culture matrices, in which types and concentrations of biological stimuli (e.g. growth factors, inhibitors, or small molecules) or matrix structure (e.g. composition, concentration, or stiffness of the matrix) vary over space, would enable a wide range of investigations concerning how these variables affect cell differentiation, migration, and other phenomena. The major challenge in creating layered matrices is maintaining the structural integrity of layer interfaces without diffusion of individual components from each layer¹. Current methodologies to achieve this include photopatterning^2-3, lithography⁴, sequential functionalization5, freeze drying⁶, microfluidics⁷, or centrifugation⁸, many of which require sophisticated instrumentation and technical skills. Others rely on sequential attachment of individual layers, which may lead to delamination of layers⁹. DGMP overcomes these issues by using an inert density modifier such as iodixanol to create layers of varying densities¹⁰. Since the density modifier can be mixed with any prepolymer or bioactive molecule, DGMP allows each scaffold layer to be customized. Simply varying the concentration of the density modifier prevents mixing of adjacent layers while they remain aqueous. Subsequent single step polymerization gives rise to a structurally continuous multilayered scaffold, in which each layer has distinct chemical and mechanical properties. The density modifier can be easily removed with sufficient rinsing without perturbation of the individual layers or their components. This technique is therefore well suited for creating hydrogels of various sizes, shapes, and materials.A protocol for fabricating a 2D-polyethylene glycol (PEG) gel, in which alternating layers incorporate RGDS-350, is outlined below. We use PEG because it is biocompatible and inert. RGDS, a cell adhesion peptide¹¹, is used to demonstrate spatial restriction of a biological cue, and the conjugation of a fluorophore (Alexa Fluor 350) enables us to visually distinguish various layers. This procedure can be adapted for other materials (e.g. collagen, hyaluronan, etc.) and can be extended to fabricate 3D gels with some modifications¹⁰.     

小鼠胎肝细胞透明质酸3D培养体系的建立

目的 利用透明质酸建立小鼠胎肝细胞3D培养体系。 方法 分离获得胚胎12-14天胎肝细胞,利用KM培养基进行初步2D肝干/祖细胞的筛选培养,并利用透明质酸及KM培养基配制水凝胶建立3D细胞培养体系。 结果 胎肝细胞在2D体系中呈现克隆状生长。分离培养获得的肝干/祖细胞克隆在透明质酸建立的3D培养体系保持增殖活性,并进一步获得肝细胞功能特性,表现为3D培养上清中白蛋白合成和尿素水平显著增加。Q-PCR结果显示随着3D培养时间的延长,其肝细胞干性标志如AFP、CK19、EpCAM、Prox1等表达水平都大幅度降低且接近成年小鼠肝脏表达水平。 结论 本研究成功建立基于透明质酸的小鼠胎肝细胞的3D无血清培养体系,并可促进小鼠胎肝细胞肝细胞功能进一步成熟。     

Characterization of thin gelatin hydrogel membranes with balloon properties for dynamic tissue engineering

Cell or tissue stretching and strain are present in any in vivo environment, but is difficult to reproduce in vitro. Here, we describe a simple method for casting a thin (about 500 μm) and soft (about 0.3 kPa) hydrogel of gelatin and a method for characterizing the mechanical properties of the hydrogel simply by changing pressure with a water column. The gelatin is crosslinked with mTransglutaminase and the area of the resulting hydrogel can be increased up 13-fold by increasing the radial water pressure. This is far beyond physiological stretches observed in vivo. Actuating the hydrogel with a radial force achieves both information about stiffness, stretchability, and contractability, which are relevant properties for tissue engineering purposes. Cells could be stretched and contracted using the gelatin membrane. Gelatin is a commonly used polymer for hydrogels in tissue engineering, and the discovered reversible stretching is particularly interesting for organ modeling applications.     

Heterochiral tetrapeptide self-assembly into hydrogel biomaterials for hydrolase mimicry

Self-assembling short peptides have attracted great interest as enzyme mimics, especially if the catalytic activity resides solely in the supramolecular structure so that it can be switched on/off as needed by controlling assembly/disassembly. Among the various enzyme classes, hydrolases find wide application in biomaterials, and their mimetics often contain His residues, in addition to either divalent cations or other amino acids to mimic the catalytic site. This work reports two self-assembling tetrapeptides based on the Ser-His motif for catalysis and the Phe-Phe motif to drive amyloid structure formation. Both peptides form thermoreversible hydrogels in phosphate buffer at neutral pH that display a mild esterase-like activity, as demonstrated on the hydrolysis of 4-nitrophenyl acetate as a model substrate, although presence of Ser did not enhance catalytic activity. The systems are characterised by circular dichroism, transmission electron microscopy, oscillatory rheology and Thioflavin T fluorescence as an amyloid stain, to provide further insights that may assist the future design of improved supramolecular catalysts.     

生物可降解嵌段共聚物在给药载体中的应用

生物可降解嵌段聚合物因具有双亲性 ,靶向药物到特定部位等优点大大推动了作为给药载体系统的发展。本文综述了生物可降解嵌段聚合物在表面修饰、水凝胶、胶束、生物大分子载体系统中的应用     

硝酸铵水凝胶分子印迹聚合物的制备

目的：目前安全问题成为世界各国的首要问题,尤其是对炸药分子的检测。硝酸铵是硝铵炸药的主要成分。研究水凝胶分子印迹法对硝铵炸药分子的检测。方法：水凝胶分子印迹方法制备硝酸铵水凝胶分子印迹聚合物,运用静态结合实验对其结合率进行了测定。结果：聚合物对硝酸铵具有良好的识别和吸附性能。印迹聚合物的解离常数为4.08g/L,最大吸附量为3.51mg/g。结论：水凝胶分子印迹法可合成水溶性炸药分子印迹聚合物,并且识别及吸附性能良好。