Self-assembly of pH-responsive fluorinated dendrimer-based particulates for drug delivery and noninvasive imaging

Dendrimers are nanoscale macromolecules with well-defined branching chemical structures. Control over the architecture and function of these structures has enabled many advances in materials science and biomedical applications. Though dendrimers are directly synthesized by iteration of simple repetitive steps, generation of the larger, more complex structures required for many biomedical applications by covalent synthetic methods has been challenging. Here we demonstrate a spontaneous self-assembly of poly(amidoamine) dendrimers into complex nanoscopic and microscopic particulates following partial fluorination of the constituent dendrimer subunits. These dense particulates exhibit a stimulus-induced response to low external pH that causes their disassembly over time, enabling controlled release of encapsulated agents. In addition, we show that these assemblies offer a sufficiently high density of fluorine spins to enable detection of their site-specific accumulation in vivo by ¹⁹F magnetic resonance imaging (¹⁹F MRI). Fluorinated dendrimer-based particulates present new features and capabilities important for a wide variety of emerging biomedical applications.     

质谱技术研究进展

质谱技术具有高灵敏度、高精确度等特点,已广泛应用于生物学、生物医学、生物化学等学科的研究,特别是在蛋白质等生物大分子的研究中作用越来越重要。对质谱技术的研究现状及新技术的研究进展作一综述,并对未来予以展望。     

Collagen-based new bioartificial polymeric materials

Bioartificial polymeric materials, based on blends of biological and synthetic polymers, have been proposed as new materials for applications in the biomedical field. They should usefully combine the biocompatibility of the biological component with the physical and mechanical properties of the synthetic component. Blends of collagen with either poly(vinyl alcohol) or poly(acrylic acid) have been prepared by mixing aqueous solutions of the two polymers. Differential scanning calorimetry and dynamic mechanical thermal analysis has been carried out to investigate the miscibility properties of the polymers and the mechanical behaviour of the blends. Biomaterials (1994) 15, 1229–1233     

合成高分子材料诱导的细胞凋亡

在综合讨论了合成高分子材料的细胞凋亡之后认为，由于生物医用材料涉及人体疾病治疗，包括组织缺陷的替代与工程化修复、基因治疗、药物缓释等许多方面，对其诱导细胞凋亡现象应作系统深入的研究，包括凋亡机制的研究。     

Selective Surface Modification in Silicon Microfluidic Channels for Micromanipulation of Biological Macromolecules

Interactions between biological macromolecules and micrometer- and sub-micrometer-scale surface structures are directly influenced by the surface wettability, chemical reactivity and surface charge. Understanding these interactions is crucial for developing integrated microsystems for biological and biomedical processing and analysis. We report development of selective surface modification techniques based on microcontact printing and polyelectrolyte adsorption. These techniques were applied to lithographically patterned silicon microfluidic channels and flat silicon substrates to create surface microstructures with contrasting wetting properties and surface charges. These controls enabled us to devise various techniques for controlled loading and processing of biomaterials in the channels. Solutions containing long chain biological macromolecules DNA and microtubules were directly loaded into the microchannels by using a micromanipulator/microinjector system. Structural arrangements of these linear macromolecules, which were probed by using fluorescence and laser scanning confocal microscopy, were found to be quite different from bulk solutions. As expected, the filamentous molecules were observed to align linearly along the channels, with the degree of alignment dependent on channel width as well as the length of the molecule. This molecular alignment, which is induced by both the surface confinement effect and capillary flow during sample loading, may be used to enhance processing of biological materials in silicon biomedical microdevices. It also opens up the possibility of carrying out direct combinatorial structural characterization of proteins in the microchannels utilizing X-ray diffraction, which so far has not been possible.     

Biostability of medical elastomers: a review

Biostability of synthetic elastomers used for manufacturing artificial replacements of human organs or their parts is a critical property of such materials as it determines the long-term function of a specific biomedical device. This paper presents a critical review of the present knowledge of biostability of elastomeric biomedical materials used as artifacts functioning under the simultaneous effects of dynamic flexing and contact with body fluids. The main topics discussed are silicone rubber, elastomers for artificial blood pumps, and the methodology of model fatigue-life testing.     

原子力显微镜在细胞与生物大分子超微结构和力学研究中的应用

作为微纳米科学理论与技术迅猛发展的代表,原子力显微镜(atomic force microscopy,AFM)在其25年的发展过程中极大地推进了生物学在微纳米尺度上的拓展,为微纳米生物学的诞生与发展提供了重要技术手段。本文在介绍AFM基本原理和检测模式的基础上,结合作者在该领域的研究成果和工作经验,从生物结构与形态学研究、表面物化性质表征、生物大分子的力学操纵三方面综述了AFM在细胞与生物大分子超微结构与生物力学特性研究中的具体应用,并重点探讨了AFM在细胞与生物大分子科学研究中亟待改进和解决的科学与技术问题,提出了一些探讨性的见解和建议。     

Synthesis of new monomers by addition reactions of diethylamine to 1,4-divinylbenzene catalyzed by lithium diethylamide

Addition reactions of diethylamine onto 1,4-divinylbenzene (DVB) catalyzed by lithium diethylamide were examined to find a synthetic route to a group of new monomers possessing dialkylamino substituents, which is expected to be a starting material for biomedical polymers such as ionic polymer complexes, polycations, and others. It was found that 1-(2-diethylaminoethyl)-4-vinylbenzene ( 1 ) and 1,4-bis(2-diethylaminoethyl)-benzene ( 2 ) can be prepared selectively, because the rate of the first step of the addition reaction was twenty times as large as that of the second step. Results of kinetic studies on the addition reactions towards DVB are discussed with the aid of ¹³C NMR data and compared with other p-vinylbenzene derivatives.     

Porting a lexicalized-grammar parser to the biomedical domain

This paper introduces a state-of-the-art, linguistically motivated statistical parser to the biomedical text mining community, and proposes a method of adapting it to the biomedical domain requiring only limited resources for data annotation. The parser was originally developed using the Penn Treebank and is therefore tuned to newspaper text. Our approach takes advantage of a lexicalized grammar formalism, Combinatory Categorial Grammar (ccg), to train the parser at a lower level of representation than full syntactic derivations. The ccg parser uses three levels of representation: a first level consisting of part-of-speech (pos) tags; a second level consisting of more fine-grained ccg lexical categories; and a third, hierarchical level consisting of ccg derivations. We find that simply retraining the pos tagger on biomedical data leads to a large improvement in parsing performance, and that using annotated data at the intermediate lexical category level of representation improves parsing accuracy further. We describe the procedure involved in evaluating the parser, and obtain accuracies for biomedical data in the same range as those reported for newspaper text, and higher than those previously reported for the biomedical resource on which we evaluate. Our conclusion is that porting newspaper parsers to the biomedical domain, at least for parsers which use lexicalized grammars, may not be as difficult as first thought.     

PEG and Cholesterol‐Containing Pyromellitates: Synthesis and Self‐Assembly

A new synthetic route for two different amphiphilic macromolecular structures containing hydrophilic poly(ethylene glycol) (PEG) and lipophilic cholesterol fragments is described. The novelty of the synthesis is in combining hydrophilic and lipophilic fragments in a backbone using functional groups of the pyromellitic dianhydride (PMDA) molecule. The ability of the new oligomers to undergo self‐assembly and solubilization of lipophilic molecules in an aqueous medium is compared. The new oligomers form self‐assemblies capable of solubilizing lipophilic “guest” molecules in an aqueous medium. Their high solubilization capacity and biodegradability, as well as other properties (size distribution, ζ‐potential), make the synthesized macromolecules good candidates for biomedical applications. Using the developed approach, amphiphilic macromolecules with regulated surface activity can be synthesized by combining PEG of varying lengths with different hydrophobic fragments (for example, fatty alcohols) in the backbone.

     

Absorption performance of iodixanol-imprinted polymers in aqueous and blood plasma media

This paper presents the preparation and absorption performance of iodixanol-imprinted polymers in aqueous and blood plasma media in vitro for biomedical applications. The imprinted polymers were prepared by non-covalent imprinting of iodixanol in a matrix of poly(4-vinylpyridine) crosslinked by ethylene glycol dimethacrylate. The binding capacities (BCs) were investigated as a function of template-to-monomer, as well as monomer-to-crosslinker, ratios in the polymerization, and the solvent type. The highest BC of iodixanols achieved from the optimized imprinted polymer in the aqueous solution is 284 mg g^?1 dry polymer with an imprinting effect (IE) 8.8 times higher than that of the non-imprinted polymer. In blood plasma, the BC of this polymer is slightly reduced to 232 mg g^?1 with a smaller IE 4.3 times higher than that of the control polymer. The BCs of molecularly imprinted polymers as a function of the initial assay solution concentration as well as the examination time are also addressed. Surface analyses were additionally performed to characterize the surface morphologies and porosities of synthetic polymers. This work has demonstrated the feasibility of molecular imprinting of iodixanol, and the observed absorption performance of the imprinted polymers is encouraging for biomedical applications.     

Chemical and physiological properties of polysucrose, a new marker of intestinal permeability to macromolecules.

To measure intestinal absorption of macromolecules we have developed a new technique employing synthetic polysucrose polymers as probe molecules. Polysucrose (PS) is water-soluble, nontoxic, resistant to intestinal enzymes, spherical and can be produced with a molecular weight distribution that relates to the size of many normal food proteins. Normally, a very small fraction of large molecules passes the exclusion barrier of the healthy intestine. Thus, quantification of resorbed macromolecules requires assays of high sensitivity. For detection of PS in various biological fluids, micro-ELISAs have been established. PS with a mean molecular weight of 14,700 daltons (PS 15,000) is rapidly excreted into the urine. Twenty-one healthy volunteers who orally ingested 1 g of this preparation showed a 12-hour urine excretion of 0.018% (interquartile range 0.014-0.022).     

Immobilization of RGD to < 1 1 1 > silicon surfaces for enhanced cell adhesion and proliferation

The ability of biomaterial surfaces to regulate cell behavior requires control over surface chemistry and microstructure. One of the greatest challenges with silicon-based biomedical microdevices such as those recently developed for neural stimulation, implantable encapsulation, biosensors, and drug delivery, is to improve biocompatibility and tissue integration. This may be achieved by modifying the exposed silicon surface with bioactive peptides. In this study, Arg-Gly-Asp (RGD) peptide conjugated surfaces were prepared and characterized. The effect of these surfaces on fibroblast adhesion and proliferation was examined over 4 days. Silicon surfaces coupled with a synthetic RGD peptide, as characterized with X-ray photoelectron spectroscopy and atomic force microscopy, display enhanced cell proliferation and bioactivity. Results demonstrate an almost three-fold greater cell attachment! proliferation on RGD immobilized surfaces compared to unmodified (control) silicon surfaces. Modulating the biological response of inorganic materials such as silicon will allow us to design more appropriate interfaces for implantable diagnostic and therapeutic silicon-based microdevices.     

Natural and synthetic sialic acid-containing inhibitors of influenza virus receptor binding

Influenza viruses attach to susceptible cells via multivalent interactions of their haemagglutinins with sialyloligosaccharide moieties of cellular glycoconjugates. Soluble macromolecules containing sialic acid from animal sera and mucosal fluids can act as decoy receptors and competitively inhibit virus-mediated haemagglutination and infection. Although a role for these natural inhibitors in the innate anti-influenza immunity is still not clear, studies are in progress on the design of synthetic sialic acid-containing inhibitors of receptor binding which could be used as anti-influenza drugs.     

Accelerated Micellization and Aggregation of Pluronic Micelles by Interaction with Heparin

The effect of attractive interaction between heparin and Pluronic co-polymer, two important macromolecules in biomedical applications, on the micellar state of Pluronic co-polymer was characterized. By the addition of heparin, the critical micellization temperature (CMT) of Pluronic solution decreased, indicating that heparin promoted micellization. Also, the Pluronic micelles were associated by the action of heparin, and the degree of aggregation became more pronounced by increasing the heparin amount. The association among Pluronic micelles by heparin enhanced the efficiency of chemical cross-linking of di-acrylated Pluronic solution by photo-polymerization, as well as the physical gelation by micelle packing. Thus, heparin can act as a sensitive modulator of Pluronic micelles in addition to its biological functions.     

Mesh-supported submicron parylene-C membranes for culturing retinal pigment epithelial cells

In this work, a mesh-supported submicron parylene-C membrane (MSPM) is proposed as an artificial Bruch's membrane for the therapy of age-related macular degeneration (AMD). Any artificial Bruch's membrane must first satisfy two important requirements. First, it should be as permeable as healthy human Bruch's membrane to support nutrients transportation. Secondly, it should be able to support the adherence and proliferation of retinal pigment epithelial (RPE) cells with in vivo-like morphologies and functions. Although parylene-C is widely used as a barrier layer in many biomedical applications, it is found that parylene-C membranes with submicron thickness are semipermeable to macromolecules. We first measure the permeability of submicron parylene-C and find that 0.15-0.30 μm parylene-C has similar permeability to healthy human Bruch's membranes. Blind-well perfusion cell viability experiments further demonstrate that nutrients and macromolecules can diffuse across 0.30 μm parylene-C to nourish the cells. A mesh-supported submicron parylene-C membrane (MSPM) structure is design to enhance the mechanical strength of the substrate. In vitro cells culture on the MSPM (with 0.30 μm ultrathin parylene-C) shows that H9-RPE cells are able to adhere, proliferate, form epithelial monolayer with tight intracellular junctions, and become well-polarized with microvilli, which exhibit similar characteristics to RPE cells in vivo. These studies have demonstrated the potential of the MSPM as an artificial Bruch's membrane for RPE cell transplantation.     

Sr-substituted hydroxyapatites for osteoporotic bone replacement

Porous apatites, which during resorption can release in situ Sr ions, were prepared to associate an anti-osteoporotic action with the peculiar features of the inorganic phase constituting the bone. Sr-substituted hydroxyapatite (SrHA) powder was directly synthesized using the classical neutralization route, but including Sr ions, and characterized. The higher solubility of SrHA granules of 400–600 μm size, potentially usable as a bone filler, was assessed compared with that of analogous stoichiometric HA granules. The Sr released in synthetic body fluid became constant after 1 week. The Ca release is improved for SrHA compared with stoichiometric HA, due to the higher solubility of the first material. Porous scaffolds with micro–macro interconnected porosity, which mimic the morphology of the spongy bone, were prepared by the impregnation of cellulose sponges with suspensions of the powder and a specific sintering process. A compressive strength of 4.52 ± 1.40 MPa was obtained for SrHA scaffolds characterized with 45 vol.% of porosity. Promising biomedical applications, such as resorbable bone filler or bone substitute releasing in situ Sr ions for a prolonged time, can be hypothesized for the SrHA materials when pathologies related with Sr deficiency are present.     

Organoapatites: materials for artificial bone. I. Synthesis and microstructure.

We have synthesized a new family of materials we termed organoapatites which may be useful in the formulation of artificial bone. These materials are synthesized by nucleation and growth of apatite crystals in media containing poly(amino acids) or synthetic organic polyelectrolytes using strict atmospheric, temperature, and pH control. The macromolecules used to synthesize the organoapatites include poly(L-lysine), poly(L-glutamic acid), and poly(sodium acrylate). The products were characterized by x-ray diffraction, scanning electron microscopy, surface area measurements, elemental analysis, and spectroscopic techniques. Organoapatites were found to contain large surface area morphologies with small crystallites which mature slowly based on analysis of Ca/P ratios. The organic macromolecules are thought to induce nucleation of crystals but also to quench their growth, thus becoming intimately dispersed in a mineral network. The organomineral particles harvested from the reaction medium contain polymer-netted microcrystals, and for this reason the synthetic approach can be used to modulate crystal maturation and biological response. It is likely that the preparative approach mimics some aspects of natural bone matrix synthesis and could be specially useful in the preparation of mineral implants containing intimate dispersions of small amounts of biomolecules such as growth factors, special drugs, or bioadhesives.     

The relevance of the transfer of molecular information between natural and synthetic materials in the realisation of biomedical devices with enhanced properties

Past and recent attempts to introduce in synthetic polymers molecular information from natural substances through simple blending, template polymerization and molecular imprinting are reviewed. The most promising approaches that can open the way to the realisation of new materials with improved biocompatibility, antibody- or enzyme-like performances are analysed more deeply. The realisation of bioartificial blends from natural and synthetic polymers, molecularly imprinted nanospheres or membranes that can act as recognition element in (bio)sensing devices, as synthetic enzymes or as key constituents of body fluids purification tools is presented in order to make the reader aware of the fascinating possibilities that these techniques make available to the biomedical science and engineering in the close future. The last part of the paper describes recent attempts to insert recognition elements for large molecules as proteins, DNA segments, viruses or whole cells in synthetic polymer systems, in order to develop new systems in the treatments of diseases and for tissue-engineering applications.     

Capillary zone electrophoresis as a tool to monitor the last stages of the degradation of water-sensitive polymers

In order to monitor the formation of the water-soluble by-products from chain-scission of degradable polymers used in the biomedical field, four capillary electrophoresis methods are discussed with the aim of giving the limits and performance for each. Three of them (electroosmotic flow reversal by dynamic adsorption of a polycation, multilayer polyelectrolyte coatings and physical binding of polyethylene oxide) are based on the use of dynamic coatings onto the inner surface of a fused silica capillary, a simple means to adapt performance to specific separations via modification and control of the electroosmotic flow of fused capillary. Using oligomers of lactic acid considered as standards the methods are compared. Other examples of ester-containing macromolecules (poly(hydroxybutyrate)), as well as degradable polyanions are described, namely N-acetylneuraminate polymer and poly(beta-malic acid).