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A method for the molecular imprinting of hemoglobin on silica surfaces using silanes
Shiomi, T., M. Matsui, et al. (2005), Biomaterials 26(27): 5564-71.
Abstract: A new molecular imprinting technique using covalently immobilized hemoglobin (Hb) is described for creating Hb-specific recognition cavities on silica. Two kinds of organic silane (3-aminopropyltrimethoxysilane: APTMS, and trimethoxypropylsilane: TMPS) were polymerized on a surface of porous silica after the Hb template was covalently immobilized by forming imine bonds, and their influence was analyzed. The results showed that not only the silane amount but also the relative proportions play an important role in protein imprinting. Pore size distribution on Hb imprinted silica was determined by nitrogen adsorption/desorption after removing the template Hb. The Hb-imprinted silica using covalently immobilized Hb (MIPi) as a template proved superior to silica using free Hb (MIPf) regarding displacement of template Hb, and selective re-adsorption as compared with other non-template proteins. The results suggested the capacity for selective adsorption of MIPi to be not only based on the isoelectric point (pI) and protein molecular weight, but also the characteristics of protein recognition cavities imprinted on base silica.

A method for toxicity screening of biomaterials using cells cultured on millipore filters
Wennberg, A., G. Hasselgren, et al. (1979), J Biomed Mater Res 13(1): 109-20.
Abstract: In the present paper a tissue culture technique is described whereby the toxicity of setting and solid materials may be evaluated. A cell monolayer was established on a millipore filter which was placed on an agar medium, cell side down. Test specimens were placed on top of the millipore filter and were allowed to influence the cells through the filter for two hours. The cell reaction was assessed by incubating the cells, still adherent to the filter, for the demonstration of succinate dehydrogenase activity. Materials with a cytotoxic effect caused a zone of inhibited enzyme activity in the cell-material contact area. The filters were examined macroscopically and scores from 0 to 3 were given to grade the severity of the cell response. Unset and set silicate cement, zinc phosphate cement and an acrylic resin were tested. The results obtained were consistent and in accordance with those of previous reports. The method was simple and rapid and appeared suitable for the assay of larger test series.

A method for toxicological evaluation of biomaterials based on colony formation of V79 cells
Kotoura, Y., T. Yamamuro, et al. (1985), Arch Orthop Trauma Surg 104(1): 15-9.
Abstract: This report describes a method for cytotoxicity screening of biomaterials based on colony formation of V79 cells. For this test, two metals (titanium and nickel), two ceramics (alumina ceramic and tricalcium phosphate), and two types of polymeric material [high density polyethylene (HDP) and polyvinylchloride (PVC)] were used. Each metal and ceramic was cast into a disk and semidisk 49 mm in diameter and 1 to 2 mm thick. The HDP was molded into a petri dish and PVC was used as a thin film. The materials were sterilized by heating or with ethylene oxide and placed in plastic petri dishes, after which 8 ml cell suspension containing 100 cells were added to each dish. After 1 week, the colonies formed on the materials were fixed, stained, and then the number of colonies was counted. Titanium, alumina ceramic, and HDP showed no differences from the controls in terms of colonies. On the disks and the semidisks of nickel and tricalcium phosphate and on the thin disks of PVC, however, no colonies were detected. The V79 cells used in this experiment showed a rapid and logarithmically stable growth curve and such a high rate of colony formation as to form visible noticeable colonies, and were therefore suitable cells for screening test the cytotoxicity of biomaterials. Unlike other previously reported methods of in vitro cytotoxicity testing, this method permits assay of colonies formed from a single cell after proliferation directly on the materials. Moreover, the test with semidisks permits simple screening to assess the cytotoxicity is caused by either the chemical substances or the physical properties of the materials.(ABSTRACT TRUNCATED AT 250 WORDS)

A microfabrication method of a biodegradable polymer chip for a controlled release system
Ito, Y., H. Hasuda, et al. (2005), J Biomater Sci Polym Ed 16(8): 949-55.
Abstract: A simple microfabrication method for a controlled-release drug-delivery system has been designed using biodegradable polymeric microchips. Microholes were made in a poly(L-lactic acid) plate and dyes were cast in each well. After drying, the wells were sealed with polymers having different biodegradation rates using a mold that had hollows corresponding to the wells. The polymers were prepared by mixing polylactides with the co-polymers. The sealing was confirmed by ultrasonication. The plate was incubated in phosphate-buffered saline and the dye released from the plate as the degradation proceeded was detected spectrophotometrically. The higher the degradation rate of the polymer sealing, the faster the sealed dye was released. This biodegradable biochip is useful for the design of controlled-release drug-delivery systems.

A microfluidic biomaterial
Cabodi, M., N. W. Choi, et al. (2005), J Am Chem Soc 127(40): 13788-9.

A miniaturized nebulization catheter for improved gene delivery to the mouse lung
Koping-Hoggard, M., M. M. Issa, et al. (2005), J Gene Med 7(9): 1215-22.
Abstract: BACKGROUND: The available methods for administration of gene delivery systems to the lungs of small animals via nebulization have several drawbacks. These include lack of control over the delivered dose and a negative impact on the stability of the formulation. This paper describes a new nebulization catheter device for the administration of plasmid-based gene delivery systems (polyplexes) as aerosols to the mouse lung in vivo. METHODS: The physical stability of naked pDNA and polyplexes formulated with chitosan oligomers and PEI was examined following nebulization with the catheter device. We also examined the in vitro transfection efficiency of the polyplexes recovered after nebulization. Lung distribution and gene expression after administration of the selected gene delivery systems to the mouse lung were also investigated. RESULTS: In contrast to previously described nebulization methods, the structural integrity of the unprotected naked pDNA was maintained following nebulization by the catheter device, which indicates relatively mild nebulization conditions. In addition, the nebulization procedure did not affect the physical stability of the formulated polyplexes. Small volumes of the pDNA aerosol (10-20 microl) were delivered in a highly controlled and reproducible manner. The aerosol droplet size varied with the molecular weight of the polycations. Aerosol delivery via this method resulted in improved lung distribution of pDNA polyplexes and a six-fold increase in the efficiency of gene delivery in vivo over that seen with the commonly used intratracheal instillation method. CONCLUSION: The use of the nebulization catheter device provides a promising alternative for aerosol gene delivery to the mouse lung.

A mixed mode fracture toughness test of bone-biomaterial interfaces
Wang, X. and C. M. Agrawal (2000), J Biomed Mater Res 53(6): 664-72.
Abstract: Tissue-biomaterial interfacial bonding plays a significant role in the success of biomaterials used for load-bearing orthopedic and dental prostheses. The objective of this study was to develop a physically sound and practically effective technique for assessment of the strength of bone-biomaterial interfaces under mixed mode loading. A single-edge notched sandwich specimen was developed for this purpose, wherein a bilayer specimen comprising the interface between tissue and biomaterial was sandwiched between two holders and loaded under mixed modes. First, a closed form solution was derived for the sandwich specimen under the assumption of linear elasticity, based on a general solution for sandwich structures reported in the literature. Then, a correction factor was determined for the solution using finite element models to compensate for errors induced by finite interlayer thickness. Moreover, using the same FEA models, it was found that crack closure may occur when the shear component is dominant at the crack. However, its effects were estimated to be limited and negligible. Furthermore, as an example, the strength of a bone/dental cement interface under different loading modes was tested using this sandwich technique. It is expected that the mixed mode technique can provide an effective means for investigators to study the mechanical integrity of bone-biomaterial interfaces under complex loading conditions.

A model for CD2/CD58-mediated adhesion strengthening
Shao, J. Y., Y. Yu, et al. (2005), Ann Biomed Eng 33(4): 483-93.
Abstract: Stable cell adhesion is vital for structural integrity and functional efficacy. Yet how low affinity adhesion molecules such as CD2 and CD58 can produce stable cell adhesion is still not completely understood. In this paper, we present a theoretical model that simulates the accumulation of CD2 and CD58 in the contact area of a Jurkat T lymphoblast and a CD58-containing substrate. The cell is assumed to have a spherical shape initially and it is allowed to spread gradually on a circular substrate. Mobile CD2 and CD58 can diffuse freely on both the cell and substrate. Their binding in the contact area is controlled by first-order kinetics. The contact area grows linearly with the total number of CD2/CD58 bonds. Cellular deformation and cytoskeleton involvement were not considered. This time-dependent moving-boundary problem was solved with the Crank-Nicolson finite difference scheme and the variable space grid method. Our simulated results are in reasonable agreement with the experimental observations. The role of diffusion becomes more and more prominent during the contact area increase, which is not sensitive to the kinetic rate constants tested in this study. However, it is very sensitive to the dissociation equilibrium constant and the concentrations of CD2 and CD58.

A model for studying epithelial attachment and morphology at the interface between skin and percutaneous devices
Knowles, N. G., Y. Miyashita, et al. (2005), J Biomed Mater Res A 74(3): 482-8.
Abstract: Percutaneous devices are indispensable in modern medicine, yet complications from their use result in significant morbidity, mortality, and cost. Bacterial biofilm at the device exit site accounts for most infections in short-term devices. We hypothesize that advanced biomaterials can be developed that facilitate attachment of skin cells to percutaneous devices, forming a seal to preclude bacterial invasion. To study the skin/biomaterial interface systematically, we first identified biomaterials with physical properties compatible with histological processing of skin. Second, we developed an organ culture system to study skin response to implants. Organ cultures implanted with porous poly(2-hydroxyethyl methacrylate) [poly(HEMA)] or polytetrafluoroethylene (PTFE) could easily be evaluated histologically with preservation of the skin/biomaterial interface. Epithelial cells migrated down the cut edges of the biomaterial in a pattern seen in marsupialization of percutaneous devices in vivo. This in vitro model maintains skin viability and allows histologic evaluation of the skin/biomaterial interface, making this a useful, inexpensive test-bed for studies of epidermal attachment to modified biomaterials.

A model for the preliminary biological screening of potential keratoprosthetic biomaterials
Sandeman, S. R., A. W. Lloyd, et al. (2003), Biomaterials 24(26): 4729-39.
Abstract: A series of in vitro screening assays for the preliminary selection of biomaterials for use in the fabrication of artificial corneas (keratoprostheses) (KPros) have been investigated. These screening assays assessed the initial binding of inflammatory and cell adhesive proteins, activation of inflammatory proteins, adhesion of keratocytes, epithelial cells and macrophages and the production of inflammatory cytokines by keratocytes contacting biomaterials. Central optic biomaterials were selected on the basis of low-inflammatory and cell adhesion potential. Peripheral skirt materials were selected on the basis of low-inflammatory potential but good cell adhesion to anchor the implant within the host cornea. Green fluorescent protein (GFP) gene transfer was used in a novel context to investigate cell invasion in the absence of external staining techniques. Confocal laser scanning microscopy and scanning electron microscopy were used to investigate GFP positive keratocyte invasion of porous materials. The results of in vitro assays were compared to a corneal organ culture system in which the biomaterials were assessed within a stromal environment. A range of polyurethane-based interpenetrating polymers with a range of water contents were screened. All materials showed low-inflammatory potential. A reduction in biomaterial water content induced an increase in complement C3 and fibronectin binding and in cell adhesion to materials, whilst differences in co-monomer formulation had little impact. The screening methods used in the current study provide a suitable preliminary assessment regime for the in vitro evaluation of potential KPro materials.

A model for the stability and creep of organic materials
Jager, I. L. (2005), J Biomech 38(7): 1459-67.
Abstract: A model is presented for the thermally assisted breaking of a number of bonds arranged in parallel and stressed by an individual soft spring each. Using a simplified potential for the bond it is shown that in equilibrium there are two definite regions of elastic behavior: one with all bonds intact, the other with a variable fraction of bonds broken, therefore with a tangent modulus steadily decreasing with applied stress. Criteria are given for the existence of these regions. Beyond these regions time-dependent creep to rupture is found, limited, in turn, by the theoretical fracture strength, the stress necessary for fracture without any thermal assistance, beyond which a bound state is impossible. The time-to-fracture for creep rupture is calculated and an example of the time evolution of the accelerating creep given. The results of the calculations are applied to experimental data on Wallaby tendons by Wang and Ker (J. Exp. Biol. 198 (1995) 831) and data estimated for the bond potential depth, the theoretical fracture strength and the number density of bonds involved as well as the elastic modulus of the ensemble. Values are derived under the assumption of one deformation mechanism being dominant--e.g., (sub-)fibril sliding or sliding of collagen molecules along one another--but the model cannot definitely distinguish between mechanisms.

A model for thromboembolization on biomaterials
Reynolds, L. O., W. H. Newren, Jr., et al. (1993), J Biomater Sci Polym Ed 4(5): 451-65.
Abstract: A model was developed to describe the kinetics of protein and platelet deposition and embolization on biomaterials. The model assumes that proteins can be adequately represented by fibrinogen, albumin, and Factor XII, that protein adsorption is Langmuir-type, that surfaces are homogeneous, and that all adsorption and deposition steps are first order. Eleven model parameters were determined from literature experimental data from ex vivo experiments utilizing canine and baboon blood on Silastic, one parameter came from adsorption of Factor XII on glass, and three parameters were obtained by minimizing differences between experimental and predicted fibrinogen adsorption, and platelet deposition and embolization behavior. The model well predicted observed behavior for fibrinogen adsorption, platelet deposition, and platelet embolization on Silastic, and platelet embolization from both polyacrylamide and HEMA-MAAC.

A model of deposition and embolization of proteins and platelets on biomaterial surfaces
Wilson, R. S., A. Marmur, et al. (1986), Ann Biomed Eng 14(4): 383-400.
Abstract: A theoretical model for the deposition and detachment of protein and platelets on biomaterial surfaces is presented here. This work is an extension of the model previously reported. Two mechanisms of protein and platelet removal are assumed: A characteristic time elapses before adsorbed protein detaches from the surface, carrying away platelets and protein which have deposited on top of it; and thrombi that attain a critical size are subject to hydrodynamic forces which embolize them from the surface. A theoretical distribution of thrombus sizes is assumed. Analysis of the effects of varying model parameters on predicted protein and platelet deposition reveals that the addition of the embolization process does not change the overall structure of the deposition profiles, but does significantly affect the finer details.

A model of lysosomal metabolism of dextran coated superparamagnetic iron oxide (SPIO) nanoparticles: implications for cellular magnetic resonance imaging
Arbab, A. S., L. B. Wilson, et al. (2005), NMR Biomed 18(6): 383-9.
Abstract: Ferumoxides, dextran-coated superparamagnetic iron oxide (SPIO) particles, form ferumoxide-transfection agent (FE-TA) complexes that are internalized into endosomes/lysosomes and have been used to label cells for in vivo MRI tracking and localization studies. A better understanding of the physical state of the FE-TA complexes during endocytosis could improve their use. The purpose of this study was to measure the rate of the degradation of iron particles under varying physiological conditions. FE-TA complexes were incubated in seven different buffers containing different chelates with different pH. Reducible iron concentrations, T2 relaxation rates and gradient echo (GRE) magnetic resonance images (MRI) were obtained from each condition immediately after incubation and at 6, 24, 48, 72 and 96 h and days 7, 14 and 21. The dynamics of FE-TA in the endosome/lysosomes within the cells were visualized with electron microscopy. Sodium citrate buffer at pH 4.5 rapidly dissolved FE-TA complexes. However, FE-TA complexes were less soluble in the same buffer at pH 5.5. Similarly, FE-TA complexes were not readily soluble in any of the other buffers with or without chelates, regardless of pH. Electron microscopic images showed degraded FE-TA in some intracellular endosome/lysosomes between days 3 and 5. In the cellular environment, some of the FE-TA-containing endosomes were found to fuse with lysosomes, causing rapid dissociation at low pH and exposing the iron core to chelates that resulted in soluble Fe(III) within the lysosomes. The studies presented represent a first step in identifying the important cellular environmental parameters affecting the integrity of FE-TA complexes.

A model system to assess key vascular responses to biomaterials
Sprague, E. A. and J. C. Palmaz (2005), J Endovasc Ther 12(5): 594-604.
Abstract: PURPOSE: To establish a reproducible laboratory test to evaluate prospective vascular biomaterials with respect to their thromboinflammatory properties by examining fibrinogen, platelet, and monocyte binding. Endothelial migration onto these surfaces was used as an index of vascular healing. METHODS: To evaluate biomaterials for potential thrombogenicity and inflammation, binding assays of radiolabeled human fibrinogen, platelets, and monocytes were performed on standard pieces of vascular biomaterials, including metals and polymeric and ceramic-coated materials. Using an established in vitro endothelial cell migration model, the relative migration rate of cultured human aortic endothelial cells onto these vascular biomaterials was measured and compared. The fibrinogen, platelet, and monocyte binding results were combined along with the migration results to create an overall score of biocompatibility. RESULTS: A significant direct relation of platelet and monocyte binding to the amount of adsorbed fibrinogen was observed. In contrast, migration rates of cultured human aortic endothelial cells onto the same biomaterial surfaces were found to be inversely related the amount of bound fibrinogen. Among the materials tested, stainless steel received the highest score of biocompatibility, while turbostratic carbon scored the lowest. CONCLUSIONS: Fibrinogen, platelet, and monocyte binding levels, as well as endothelial migration rates onto vascular material surfaces, provide a basis for evaluating thrombogenicity, inflammatory potential, and endothelialization in the laboratory prior to in vivo testing.

A modular and supramolecular approach to bioactive scaffolds for tissue engineering
Dankers, P. Y., M. C. Harmsen, et al. (2005), Nat Mater 4(7): 568-74.
Abstract: Bioactive polymeric scaffolds are a prerequisite for the ultimate formation of functional tissues. Here, we show that supramolecular polymers based on quadruple hydrogen bonding ureido-pyrimidinone (UPy) moieties are eminently suitable for producing such bioactive materials owing to their low-temperature processability, favourable degradation and biocompatible behaviour. Particularly, the reversible nature of the hydrogen bonds allows for a modular approach to gaining control over cellular behaviour and activity both in vitro and in vivo. Bioactive materials are obtained by simply mixing UPy-functionalized polymers with UPy-modified biomolecules. Low-molecular-weight bis-UPy-oligocaprolactones with cell adhesion promoting UPy-Gly-Arg-Gly-Asp-Ser (UPy-GRGDS) and the synergistic UPy-Pro-His-Ser-Arg-Asn (UPy-PHSRN) peptide sequences are synthesized and studied. The in vitro results indicate strong and specific cell binding of fibroblasts to the UPy-functionalized bioactive materials containing both UPy-peptides. An even more striking effect is seen in vivo where the formation of single giant cells at the interface between bioactive material and tissue is triggered.

A multi-sample denaturation temperature tester for collagenous biomaterials
Lee, J. M., C. A. Pereira, et al. (1995), Med Eng Phys 17(2): 115-21.
Abstract: The temperature at which collagen denatures from a triple helix to a random coil structure is a useful measure of the degree of crosslinking. A new multi-sample denaturation temperature tester (DTT) has been constructed for rapid determination of the collagen denaturation temperature of natural tissues and collagenous biomaterials. To validate the system, the denaturation temperatures measured for the DTT are compared with results from differential scanning calorimetry (DSC). Data are presented for bovine pericardium in three states with denaturation temperatures ranging from 68 to 85 degrees C: fresh, or crosslinked with glutaraldehyde or the epoxide reagent Denacol EX-512 poly (glycidyl ether). Denaturation temperatures measured by DTT were not significantly different from those measured by differential scanning calorimetry (DSC); however, DSC onset systematically occurred at a slightly lower temperature than that measured by DTT. This result, seen only for fresh tissue is in agreement with earlier experiments using hydrothermal isometric tension (HIT) testing. By contrast, DTT and DSC onset were identical for the exogenously crosslinked materials. Since the measured transition temperature was independent of initial load, this variable may be chosen to yield sharper force-temperature transitions with a given sample geometry. This instrument allows accurate assessment of collagen denaturation temperatures for multiple samples in a fraction of the time required by other methods.

A new analysis of the depolymerized fragments of lignin polymer using ToF-SIMS
Saito, K., T. Kato, et al. (2005), Biomacromolecules 6(5): 2688-96.
Abstract: Lignin in plant cell walls is a complex, irregular polymer built from phenylpropanoid C6-C3 units that are connected via various C-C and C-O linkages. A recent study using time-of-flight secondary ion mass spectrometry (ToF-SIMS) with Ga primary ion bombardment showed that lignin polymers can be characterized by specific positive ions possessing a substituted aromatic ring (so-called guaiacyl or syringyl rings), which are the basic building units of lignin. To study the relationship between the characteristic ions of lignin and the common interunit linkages, various lignin dimer model compounds were investigated using ToF-SIMS. The resulting dimer spectra showed that the characteristic ions with a guaiacyl ring at m/z 137 and 151 result from rupture of most common interunit linkages, not only 8-O-4' linkages, which are the most abundant in lignin, but also 8-1', 8-5', and 8-8'. There was no evidence of rupture of 5-5' linkages. These results show that ToF-SIMS offers a new tool for the direct analysis of the depolymerized fragments of lignin polymers. The mechanisms for the fragmentation of lignin dimer models in ToF-SIMS were proposed that allow ToF-SIMS fragmentation rules to be deduced. Adduct ions such as [M + 13]+ ([M + CH]+) were also produced in fragmentation of the dimers and are thought to arise from the combination of the molecules with their stable fragments.

A new biomaterial for the control of infection in the burn wound
Nathan, P., E. J. Law, et al. (1976), Trans Am Soc Artif Intern Organs 22: 30-41.
Abstract: A synthetic dressing has been developed that isolates the burn wound to protect patients from microbial contamination. This dressing is unique as it is formed from a 2-component system directly on the wound, leaving no voids for microbial proliferation. The synthetic cover, HYDRON Burn Dressing, adheres to the entire wound surface so that additional dressings are not required. It is sufficiently flexible to permit patient mobility. The components used to form the dressing are an ultra-pure, high molecular weight form of HYDRON, a hydrophilic polymer, poly (2-hydroxyethyl methacrylate) and Polethylene Glycol-400. The dressing is intended to be applied directly to the wound immediately post-burn, prior to sloughing or removal of the eschar, a period in the burn therapy regimen for which satisfactory alternative dressings are not readily available. Decreased frequency of dressing changes compared to conventional procedure provides an additional benefit with a corresponding reduction in pain to the patient. Gross examination of the wounds under the dressing has shown that the healing process proceeds similarly to that of wounds under conventional treatment. There is no evidence of fluid accumulation or maceration or desiccation of the eschar. In our series of 32 patients the barrier dressing formed on the wound has offered a new, effective procedure for treatment of the burn wound.

A new biomaterial, hen egg shell membrane, to eliminate heavy metal ion from their dilute waste solution
Suyama, K., Y. Fukazawa, et al. (1994), Appl Biochem Biotechnol 45-46: 871-9.
Abstract: The egg shell membrane (ESM) is an intricate lattice network of stable and water-insoluble fibers with high surface area. ESM accumulates and eliminates various heavy metal ions from dilute aqueous solution with high affinity and in short contact time, depending on pH and characteristics of the individual ion. Under certain conditions, the level of precious ions, Au, Pt, and Pd accumulation approaches 55, 25, and 22% of dry wt of ESM, respectively. Also uranium uptake 30% of that of ESM. Experiments suggested that ESM is promising to use for the purpose of removal/recovery of metals and water pollution control.

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