|Articles about Biomaterials|
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| Factors influencing bacterial adherence to biomaterials
Merritt, K. and C. C. Chang (1991), J Biomater Appl 5(3): 185-203.
Abstract: The adherence of bacteria to implanted medical devices is believed to be important in the development of implant associated infections. Measures which reduce bacterial adherence should reduce the incidence of these infections. However, in order to assess the importance of adherence, the effectiveness of methods to reduce adherence, and compare data from different laboratories, the conditions of the in vitro studies on adherence need to be specified. There are currently no correct and incorrect methods, however, methods used need to be carefully described. The studies reported here indicate that the definition of adherence needs to be established, with the use of polystyrene as the reference material recommended. Since the adherent organisms lose adherence traits with culture, cultures must be selected for adherence regularly. It is important to control the number of organisms/ml but the volume used is not important. The medium used to grow the organisms and the use of stationary, rocking or flow conditions will alter adherence and need to be specified and be consistent within a set of experiments. Culture conditions, methods of rinsing the material, methods of elution and counting, or direct counting of organisms on the material need to be specified. Finally, as much information as possible on the bulk and surface properties of the material should be provided. The handling of the material for the experiments should be careful and defined. Fingerprints, contact with protein, wet surfaces vs dry surfaces, etc., will all affect the subsequent adherence. The materials should not be re-used since the removal of the adherent proteins or the biofilm is very difficult. Progress can be made in this important area if the details of procedures are specified.
| Factors influencing stem cell differentiation into the hepatic lineage in vitro
Heng, B. C., H. Yu, et al. (2005), J Gastroenterol Hepatol 20(7): 975-87.
Abstract: A major area of research in transplantation medicine is the potential application of stem cells in liver regeneration. This would require well-defined and efficient protocols for directing the differentiation of stem cells into the hepatic lineage, followed by their selective purification and proliferation in vitro. The development of such protocols would reduce the likelihood of spontaneous differentiation of stem cells into divergent lineages upon transplantation, as well as reduce the risk of teratoma formation in the case of embryonic stem cells. Additionally, such protocols could provide useful in vitro models for studying hepatogenesis and liver metabolism. The development of pharmokinetic and cytotoxicity/genotoxicity screening tests for newly developed biomaterials and drugs, could also utilize protocols developed for the hepatic differentiation of stem cells. Hence, this review critically examines the various strategies that could be employed to direct the differentiation of stem cells into the hepatic lineage in vitro.
| Factors involved in the development of polymerization shrinkage stress in resin-composites: a systematic review
Braga, R. R., R. Y. Ballester, et al. (2005), Dent Mater 21(10): 962-70.
Abstract: OBJECTIVES: Polymerization shrinkage stress of resin-composite materials may have a negative impact on the clinical performance of bonded restorations. The purpose of this systematic review is to discuss the primary factors involved with polymerization shrinkage stress development. DATA: According to the current literature, polymerization stress of resin composites is determined by their volumetric shrinkage, viscoelastic behavior and by restrictions imposed to polymerization shrinkage. Therefore, the material's composition, its degree of conversion and reaction kinetics become aspects of interest, together with the confinement and compliance of the cavity preparation. SOURCES: Information provided in this review was based on original scientific research published in Dental, Chemistry and Biomaterials journals. Textbooks on Chemistry and Dental Materials were also referenced for basic concepts. CONCLUSIONS: Shrinkage stress development must be considered a multi-factorial phenomenon. Therefore, accessing the specific contribution of volumetric shrinkage, viscoelastic behavior, reaction kinetics and local conditions on stress magnitude seems impractical. Some of the restorative techniques aiming at stress reduction have limited applicability, because their efficiency varies depending upon the materials employed. Due to an intense research activity over the years, the understanding of this matter has increased remarkably, leading to the development of new restorative techniques and materials that may help minimize this problem.
| Factors that influence transgene expression and cell viability on DNA-PEI-seeded collagen films
Katz, J. M., C. M. Roth, et al. (2005), Tissue Eng 11(9-10): 1398-406.
Abstract: Gene delivery from tissue-engineering devices has the potential to improve healing, but better regulation of the level and duration of gene expression is needed. We hypothesized that transgene expression could be controlled by varying the fabrication and soaking parameters used in making collagen- based gene delivery scaffolds. Collagen films were made from acid-insoluble type I bovine dermal collagen and seeded with plasmid DNA encoding firefly luciferase, complexed with polyethylenimine. By varying the thickness of the films, the volume of the DNA soak solution, and the pH of the DNA soak solution, and by cross-linking the films, we identified variable combinations that produce significantly different levels of cell number and transgene expression in L-929 cells in vitro. Increasing film thickness or soak volume increased overall reporter gene expression. Decreasing film thickness or soak volume decreased cell number but did not significantly change reporter gene expression per cell. Cross-linking by ultraviolet irradiation (before adding the DNA) significantly decreased transgene expression, probably because of decreased swelling of the collagen film. These results suggest that collagen-based biomaterials may be designed and fabricated to induce, in a controlled fashion, various levels of cellularity and transgene expression.
| Fatal encephalopathy after otoneurosurgery procedure with an aluminum-containing biomaterial
Hantson, P., P. Mahieu, et al. (1995), J Toxicol Clin Toxicol 33(6): 645-8.
Abstract: Refractory status epilepticus was observed in two patients who underwent vestibular neurectomy. We investigated the relationship with the use of an aluminum containing bone cement during the procedure. Two patients developed focal and thereafter generalized seizures in the late postoperative period of vestibular neurectomy (respectively after 42 and 35 days). A cement (1 g aluminum-calcium fluorosilicate) was used during the procedure to bridge bone defects. Both patients presented cerebrospinal fluid fistula. Investigations excluded common etiologies, in particular infections, and a toxic origin was suspected. Aluminum concentration was determined repeatedly in serum urine, cerebrospinal fluid and retroauricular fistula. The highest aluminum values were respectively in case 1 and 2, 112 and 63 micrograms/L for the cerebrospinal fluid, 495 and 1440 micrograms/L for the fistula, 4.4 and 4.4 micrograms/L in serum. Desferrioxamine was used as chelating agent and aluminum elimination was analyzed in the urine. Status epilepticus became refractory to intensive care therapy. The patients never recovered normal consciousness. Case 1 died 143 days after the procedure and case 2 at 80 days from brain failure. Brain post-mortem examination was obtained in Case 2. Brain aluminum concentration was 2.5 micrograms/g (wet weight) (0.85 micrograms/g in a control non exposed cadaver). The cement (0.2 g) was incubated in vitro (16 h-37 degrees C) with the cerebrospinal fluid of a control patient (cerebrospinal fluid aluminum 8 micrograms/L): aluminum concentration reached 2750 micrograms/L. A close contact between an aluminum containing cement and the cerebrospinal fluid may have resulted in encephalopathy and fatal status epilepticus in these two patients.
| Fate of thrombin and thrombin-antithrombin-III complex adsorbed to a heparinized biomaterial: analysis of the enzyme-inhibitor complexes displaced by plasma
Hatton, M. W., G. Rollason, et al. (1983), Thromb Haemost 50(4): 873-7.
Abstract: Heparin covalently-linked to polyvinyl alcohol (PVA) is a biomaterial which is of potential value as a non-thrombogenic coating. 125I-labelled thrombin adsorbed to heparin-PVA beads was not dislodged by phosphate-buffered saline, pH 7.4, although radioactivity was progressively displaced from the adsorbent by fibrinogen-free human plasma. Analysis by gel filtration and affinity chromatography showed that the released radioactivity was distributed between (thrombin-antithrombin-III) complex (approx. 70%) and, probably, (thrombin-alpha-2-macroglobulin) complex (approx. 30%). Less efficient thrombin displacement was obtained by either bovine serum albumin (5% w/v) or antithrombin-III-free human plasma: in the latter case, the dislodged enzyme was presumably associated with alpha-2-macroglobulin. Purified alpha-2-macroglobulin did not displace thrombin from heparin-PVA. The quantity of thrombin displaced by an alpha-2-macroglobulin-free plasma fraction compared well with fibrinogen-free plasma: The eluted enzyme was largely associated with antithrombin-III. Purified antithrombin-III did not displace thrombin from heparin-PVA despite causing greater than 70% inactivation of the bound enzyme. Subsequent treatment with fibrinogen-free plasma dislodged (thrombin-antithrombin-III) at a similar rate to that of bound thrombin. We conclude that plasma contains a component(s) which displaces (thrombin-antithrombin-III) complex from immobilised heparin: presumably this leaves the heparin sites free for further use in enzyme inactivation.
| Favorable chitosan/cellulose film combinations for copper removal from aqueous solutions
Lima, I. S., A. M. Lazarin, et al. (2005), Int J Biol Macromol 36(1-2): 79-83.
Abstract: Three well-formed film combinations of chitosan, in the beta form and cellulose acetate biopolymer, having different proportions, have been synthesized and characterized by X-ray diffraction and infrared spectroscopy. The film having a 1.0/0.50 proportion presented 6.87 mmol of nitrogen atoms per gram of synthesized hybrid, with the highest affinity for adsorbing copper from aqueous solutions at 298+/-1K. The isotherm obtained in this adsorption showed a saturation plateau that corresponds to 1.92 mmol of copper per gram of hybrid. The energetic effects caused by copper ion interaction were determined through calorimetric titration at the solid-liquid interface in aqueous solution and gave a net thermal effect that enabled the calculation of the exothermic enthalpic values and the equilibrium constant. The complete thermodynamic data showed that the system is favored by exothermic enthalpies, negative Gibbs free energies and positive entropies.
| Feasibility and development of a high-power real accommodating intraocular lens
Ben-Nun, J. and J. L. Alio (2005), J Cataract Refract Surg 31(9): 1802-8.
Abstract: PURPOSE: To develop an accommodating intraocular lens (IOL) that changes power with ciliary body action and generates a minimum of 8.0 diopters (D) by manipulation of a flexible material between a sulcus-fixated rigid plate and a ciliary muscle-operated capsular diaphragm. SETTING: Vissum-Instituto Oftalmologico de Alicante and Miguel Hernandez University, Alicante, Spain. METHODS: The feasibility of the concept was tested by a laboratory lens model. An implantable measuring device was constructed to simulate the lens action. The device was implanted in monkey eyes to measure the various parameters involved with such IOLs. Based on these measurements, an accommodating IOL prototype was built and implanted in monkeys' eyes. Pharmacologic agents were used to achieve ciliary relaxation and spasm. Ultrasound biomicroscopy (UBM) imaging was used to document the active changes of the IOL flexible lens curvature as related to the ciliary muscles status. RESULTS: The laboratory model produced more than 50.0 D of accommodation. The UBM demonstrated changes in lens curvature between cyclospasm to cycloplegia of calculated 9.0 to 53.0 D for flexible material with a refractive index of 1.41. CONCLUSIONS: Flexible material is capable of being manipulated when placed between a scleral-fixated rigid plane and the ciliary muscles-operated capsular diaphragm. Such manipulation as used by the IOL concept presented here has generated an active change of more than 40.0 D in the monkey eye.
| Feasibility of chitosan-based hyaluronic acid hybrid biomaterial for a novel scaffold in cartilage tissue engineering
Yamane, S., N. Iwasaki, et al. (2005), Biomaterials 26(6): 611-9.
Abstract: In this study, we hypothesized that hyaluronic acid could provide superior biological effects on the chondrocytes in a three-dimensional culture system. To test this hypothesis, we investigated the in vitro behavior of rabbit chondrocytes on a novel chitosan-based hyaluronic acid hybrid polymer fiber. The goal of the current study was to show the superiority of this novel fiber as a scaffold biomaterial for cartilage tissue engineering. Chitosan polymer fibers (chitosan group) and chitosan-based hyaluronic acid hybrid polymer fibers (HA 0.04% and HA 0.07% groups, chitosan coated with hyaluronic acid 0.04% and 0.07%, respectively) were originally developed by the wetspinning method. Articular chondrocytes were isolated from Japanese white rabbits and cultured in the sheets consisting of each polymer fiber. The effects of each polymer fiber on cell adhesivity, proliferation, morphological changes, and synthesis of the extracellular matrix were analyzed by quantitative a cell attachment test, DNA quantification, light and scanning electron microscopy, semi-quantitative RT-PCR, and immunohistochemical analysis. Cell adhesivity, proliferation and the synthesis of aggrecan were significantly higher in the hybrid fiber (HA 0.04% and 0.07%) groups than in the chitosan group. On the cultured hybrid polymer materials, scanning electron microscopic observation showed that chondrocytes proliferated while maintaining their morphological phenotype and with a rich extracellular matrix synthesis around the cells. Immunohistochemical staining with an anti-type II collagen antibody demonstrated rich production of the type II collagen in the pericellular matrix from the chondrocytes. The chitosan-based hyaluronic acid hybrid polymer fibers show great potential as a desirable biomaterial for cartilaginous tissue scaffolds.
| Femoropopliteal subintimal angioplasty and nitinol stenting: a marriage of technique and technology.but will it last?
Rocha-Singh, K. (2005), Catheter Cardiovasc Interv 65(4): 544-6.
| Fibrillar assembly and stability of collagen coating on titanium for improved osteoblast responses
Kim, H. W., L. H. Li, et al. (2005), J Biomed Mater Res A 75(3): 629-38.
Abstract: Collagen, as a major constituent of human connective tissues, has been regarded as one of the most important biomaterials. As a coating moiety on Ti hard-tissue implants, the collagen has recently attracted a great deal of attention. This article reports the effects of fibrillar assembly and crosslinking of collagen on its chemical stability and the subsequent osteoblastic responses. The fibrillar self-assembly of collagen was carried out by incubating acid-dissolved collagen in an ionic-buffered medium at 37 degrees C. The degree of assembly was varied with the incubation time and monitored by the turbidity change. The differently assembled collagen was coated on the Ti and crosslinked with a carbodiimide derivative. The partially assembled collagen contained fibrils with varying diameters as well as nonfibrillar aggregates. On the other hand, the fully assembled collagen showed the complete formation of fibrils with uniform diameters of approximately 100-200 nm with periodic stain patterns within the fibrils, which are typical of native collagen fibers. Through this fibrillar assembly, the collagen coating had significantly improved chemical stability in both the saline and collagenase media. The subsequent crosslinking step also improved the stability of the collagen coating, particularly in the unassembled collagen. The fibrillar assembly and the crosslinking of collagen significantly influenced the osteoblastic cell responses. Without the assembly, the collagen layer on Ti adversely affected the cell attachment and proliferation. However, those cellular responses were improved significantly when the collagen was assembled to fibrils and the assembly degree was increased. After crosslinking the collagen coating, these cellular responses were significantly enhanced in the case of the unassembled collagen but were not altered much in the assembled collagen. Based on these observations, it is suggested that the fibrillar assembly and the crosslinking of collagen require careful considerations in the collagen administration as a coating moiety. (c) 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005.
| Fibrillar collagen assembled in the presence of glycosaminoglycans to constitute bioartificial stem cell niches in vitro
Salchert, K., J. Oswald, et al. (2005), J Mater Sci Mater Med 16(6): 581-5.
Abstract: Fibrillar collagen was reconstituted from mixtures of monomeric tropocollagen and heparin or hyaluronic acid, respectively. Turbidity measurements were utilized to follow the fibrillar assembly and demonstrated the influence of the concentration of the glycosaminoglycan on the maximum optical densities. Thin film coatings of maleic anhydride copolymers were utilized for the covalent immobilization of the fibrillar assemblies to solid supports. Quantification of surface-bound collagen was accomplished by ellipsometry and HPLC-based amino acid analysis indicating that less collagen was immobilized in the presence of the glycosaminoglycans. SEM and AFM revealed various sizes and shapes of the immobilized fibrillar assemblies if collagen fibrils were prepared in the presence of heparin or hyaluronic acid. Human hematopoietic stem cells (HSCs) were cultivated on the surface-bound collagen fibrils and the migration of adherent cells was studied by time-lapse microscopy. Migration rates on fibrillar structures were significantly lower then on tropocollagen indicating a more intimate contact of HSCs to the fibrillar substrates.
| Fibrin gel-immobilized primary osteoblasts in calcium phosphate bone cement: in vivo evaluation with regard to application as injectable biological bone substitute
Kneser, U., A. Voogd, et al. (2005), Cells Tissues Organs 179(4): 158-69.
Abstract: Osteogenic injectable bone substitutes may be useful for many applications. We developed a novel injectable bone substitute based on osteoblast-fibrin glue suspension and calcium phosphate bone cement (BC). Human osteoblasts were isolated from trabecular bone samples and cultured under standard conditions. Osteoblasts were suspended in fibrinogen solution (FS). BC was cured with thrombin solution. 8 x 4 mm injectable bone discs were prepared using silicon molds and a custom-made applicator device. Discs containing BC, BC/FS, or BC/FS/osteoblasts were implanted subcutaneously into athymic nude mice. After 3, 9 and 24 weeks, specimens were explanted and subjected to morphologic and biomechanical evaluation. In vitro fibrin gel-embedded osteoblasts displayed a differentiated phenotype as evidenced by alkaline phosphatase, collagen type 1 and von Kossa stains. A proportion of osteoblasts appeared morphologically intact over a 3-day in vitro period following application into the BC. BC/FS and BC/FS/osteoblast discs were sparsely infiltrated with vascularized connective tissue. There was no bone formation in implants from all groups. However, positive von Kossa staining only in BC/FS/osteoblast groups suggests engraftment of at least some of the transplanted cells. Biomechanical evaluation demonstrated initial stability of the composites. Young's modulus and maximal load did not differ significantly in the BC/FS and BC/FS/osteoblast groups. The practicability of osteoblast-containing injectable bone could be demonstrated. The dense microstructure and the suboptimal initial vascularization of the composites may explain the lack of bone formation. Modifications with regard to enhanced osteoblast survival are mandatory for a possible application as injectable osteogenic bone replacement system.
| Fibrin(ogen) mediates acute inflammatory responses to biomaterials
Tang, L. and J. W. Eaton (1993), J Exp Med 178(6): 2147-56.
Abstract: Although "biocompatible" polymeric elastomers are generally nontoxic, nonimmunogenic, and chemically inert, implants made of these materials may trigger acute and chronic inflammatory responses. Early interactions between implants and inflammatory cells are probably mediated by a layer of host proteins on the material surface. To evaluate the importance of this protein layer, we studied acute inflammatory responses of mice to samples of polyester terephthalate film (PET) that were implanted intraperitoneally for short periods. Material preincubated with albumin is "passivated," accumulating very few adherent neutrophils or macrophages, whereas uncoated or plasma-coated PET attracts large numbers of phagocytes. Neither IgG adsorption nor surface complement activation is necessary for this acute inflammation; phagocyte accumulation on uncoated implants is normal in hypogammaglobulinemic mice and in severely hypocomplementemic mice. Rather, spontaneous adsorption of fibrinogen appears to be critical: (a) PET coated with serum or hypofibrinogenemic plasma attracts as few phagocytes as does albumin-coated material; (b) in contrast, PET preincubated with serum or hypofibrinogenemic plasma containing physiologic amounts of fibrinogen elicits "normal" phagocyte recruitment; (c) most importantly, hypofibrinogenemic mice do not mount an inflammatory response to implanted PET unless the material is coated with fibrinogen or the animals are injected with fibrinogen before implantation. Thus, spontaneous adsorption of fibrinogen appears to initiate the acute inflammatory response to an implanted polymer, suggesting an interesting nexus between two major iatrogenic effects of biomaterials: clotting and inflammation.
| Fibrin-based biomaterials to deliver human growth factors
Wong, C., E. Inman, et al. (2003), Thromb Haemost 89(3): 573-82.
Abstract: Fibrin-based biomaterial preparations can be used as provisional growth matrices for cells important in tissue repair during wound healing in vivo. Their efficacy can be enhanced by including bioactive agents that promote specific cellular responses. This study examined the controlled delivery of the angiogenic growth factors bFGF, VEGF(165), and VEGF(121) using biomatrix preparations prepared from Fibrin Sealant product components. The growth factors were added prior to formation of the Fibrin Sealant clots, and the release kinetics of the proteins from the clots measured. The results indicated that the proteins were released from the clots more slowly in the order bFGF << VEGF(165) < VEGF(121). The biologic activity of the growth factors delivered from Fibrin Sealant clots was established by assaying growth stimulation of human microvascular endothelial cells (HMVEC) and angiogenesis in the chicken embryo chorioallantoic membrane (CAM) model of neovascularization. In the latter assay, clots containing bFGF, VEGF(165), or VEGF(121) all displayed angiogenic activity. However, delivery of either bFGF, VEGF(165), or VEGF(121) alone resulted in a significant percentage of clots becoming filled with blood, indicating that the newly developing vessels invading the clots were leaky and immature. In contrast, this hemorrhaging behavior did not occur with delivery of combinations, e.g., (VEGF(165) + VEGF(121)) or (VEGF(165) + bFGF), indicating that the vessels were more mature than those produced in response to single growth factors. Thus, delivering a combination of growth factors constituted an improvement over the delivery of individual growth factors for enhancing neovascularization.
| Fibrin-based tissue-engineered blood vessels: differential effects of biomaterial and culture parameters on mechanical strength and vascular reactivity
Yao, L., D. D. Swartz, et al. (2005), Tissue Eng 11(7-8): 991-1003.
Abstract: We have shown that fibrin-based small-diameter tissue-engineered blood vessels (TEVs) exhibited considerable mechanical strength and could withstand implantation in the jugular veins of lambs, where they remained patent for 15 weeks. The microtopology of fibrin matrix is influenced by the concentration of fibrinogen and calcium, whereas fibrinolysis and matrix remodeling are affected by the presence of the fibrinolytic inhibitor aprotinin. Here we report the effects of these components on two key properties of TEVs, namely mechanical strength and vasoreactivity. We found that high concentrations of fibrinogen or calcium decreased significantly both strength and reactivity. Surprisingly, aprotinin increased mechanical strength but decreased vascular reactivity in a dose-dependent manner. Transforming growth factor beta(1) (TGF-beta(1)) and insulin had a moderate effect on mechanical strength but significantly enhanced reactivity, through receptor- and non-receptor- mediated pathways. In addition, the combination of TGF-beta(1), insulin, and aprotinin resulted in significant improvement of both properties. Our data suggest that the microtopology of fibrin matrix and the rates of fibrinolysis and extracellular matrix synthesis may affect the properties of TEVs significantly. They also indicate that biomaterial and culture parameters may have differential effects on mechanical properties versus vascular reactivity and, therefore, engineering blood vessels under conditions that maximize tissue strength may not always result in optimal function. Instead, strength and reactivity must be used in concert for more accurate evaluation of tissue-engineered vascular constructs.
| Fibrin-mediated endothelial cell adhesion to vascular biomaterials resists shear stress due to flow
Kumar, T. R. and L. K. Krishnan (2002), J Mater Sci Mater Med 13(8): 751-5.
Abstract: In vitro endothelial cell (EC) seeding onto biomaterials for blood-contacting applications can improve the blood compatibility of materials. Adhesive proteins adsorbed from serum that is supplemented with the culture medium intercede the initial cell adhesion and subsequent spreading on material surface during culture. Nevertheless, physical and chemical properties of vascular biomaterial surface fluctuate widely between materials resulting in dissimilarity in protein adsorption characteristics. Thus, a variation is expected in cell adhesion, growth and the ability of cell to resist shear stress when tissue engineering on to vascular biomaterials is attempted. This study was carried out with an objective to determine the significance of a matrix coating on cell adhesion and shear stress resistance when cells are cultured on materials such as polytetrafluoroethylene (PTFE, Teflon) and polyethyleneterephthalate (Dacron), ultra high molecular weight polyethylene (UHMWPE) and titanium (Ti), that are used for prosthetic devices. The study illustrates the distinction of EC attachment and proliferation between uncoated and matrix-coated surfaces. The cell attachment and proliferation on uncoated UHMWPE and titanium surfaces were not significantly different from matrix-coated surfaces. However, shear stress resistance of the cells grown on composite coated surfaces appeared superior compared to the cells grown on uncoated surface. On uncoated vascular graft materials, the cell adhesion was not supported by serum alone and proliferation was scanty as compared to matrix-coated surface. Therefore, coating of implant devices with a composite of adhesive proteins and growth factors can improve EC attachment and resistance of the cells to the forces of flow.
| Fibrinogen adsorption to receptor-like biomaterials made by pre-adsorbing peptides to polystyrene substrates
Grunkemeier, J. M. and T. A. Horbett (1996), J Mol Recognit 9(3): 247-57.
Abstract: Two peptides from the ligand-binding site of the platelet receptor GPIIb/IIIa, residues 296-306 of GPIIb, designated B12 by D'Souza et al. (1991), and 300-311 of GPIIb, designated G13 by Taylor et al., (1992), as well as two control peptides, designated C14 and C20, were adsorbed to treated polystyrene substrates. Fibrinogen adsorption to the peptide-coated substrates was characterized. The specificity of I-125 labeled fibrinogen binding to the peptide-coated substrates was investigated by measuring the amount of fibrinogen adsorbed to each substrate and the inhibition of fibrinogen binding by RGDS peptide, bovine serum albumin, a divalent ion chelator (ethylene diamine tetra-acetic acid disodium salt), unlabeled fibrinogen and the B12 peptide. The results show that non-specific binding of fibrinogen to the B12-coated substrate is predominant under most conditions. Binding of monoclonal antibodies to fibrinogen adsorbed to the peptide coated substrates was characterized. The failure of several antibodies to bind fibrinogen adsorbed to the B12 substrate suggested that adsorption of fibrinogen to the B12-coated substrate alters its conformation relative to fibrinogen adsorbed to the bare substrate.
| Fibrinogen and von Willebrand's factor adsorption are both required for platelet adhesion from sheared suspensions to polyethylene preadsorbed with blood plasma
Kwak, D., Y. Wu, et al. (2005), J Biomed Mater Res A 74(1): 69-83.
Abstract: Previous studies showed that platelet adhesion to biomaterials from static suspensions was greatly increased by the adsorption of even very small amounts (<5 ng/cm2) of fibrinogen (Fg). In this study, the sensitivity of platelet adhesion to Fg was reexamined by measuring platelet adhesion under flow conditions. The role of adsorbed von Willebrand's factor (vWf) was also studied. Polyethylene (PE) tubing was preadsorbed with Fg, vWf, vWf-deficient plasma, and Fg-deficient plasma or serum with added Fg, and Fg adsorption measured with 125I Fg. Platelets in a red blood cell suspension were passed through the tubes at either low (50 s(-1)) or high (500 or 1000 s(-1)) shear rates and adhesion measured with an improved LDH assay. Adhesion from flowing suspensions measured after preadsorption with afibrinogenemic plasma or serum was very low, but increased greatly with addition of Fg. Less than 10 ng/cm2 of adsorbed Fg was enough to greatly enhance platelet adhesion. Adhesion at high shear was also strongly affected by vWf, as platelet adhesion at 500 s(-1) to PE preadsorbed with vWf-deficient plasma decreased by more than tenfold compared to adhesion at 50 s(-1), but platelet adhesion to PE preadsorbed with normal plasma increased about eightfold when shear rate was increased. The results show that very low amounts of adsorbed Fg are able to support platelet adhesion under shear flow. However, adsorbed vWf also appears to play an important cofactor role in platelet adhesion to biomaterials, as its presence greatly augments platelet adhesion under high shear.
| Fibrinogen surface distribution correlates to platelet adhesion pattern on fluorinated surface-modified polyetherurethane
Massa, T. M., M. L. Yang, et al. (2005), Biomaterials 26(35): 7367-76.
Abstract: In previous work, it had been shown that platelet adhesion could be reduced by fluorinating surfaces with oligomeric fluoropolymers, referred to as surface-modifying macromolecules (SMMs). In the current study, two in vitro blood-contacting experiments were carried out on a polyetherurethane modified with three different SMMs in order to determine if altered platelet adhesion levels could be related to the pattern of adsorbed protein and more specifically to the manner in which fibrinogen (Fg) distribution occurs at the surface. In the first experiment, the materials were placed in whole human blood and the adherent platelets were viewed with high-resolution scanning electron microscopy (SEM). In a second experiment, the materials were incubated with human plasma with the absence of platelets. The plasma contained 5% fluorescent-Fg. The materials were then viewed with a fluorescence microscope and images were collected to define the distribution of high-density fluorescent-Fg areas. The SEM and fluorescent-Fg images were imported to Image Pro Plus imaging software to measure the area, length and circularity and a bivariate correlation test was conducted between the two sets of data. For area and length morphology parameters, there were high and significant correlations (r > 0.9, p < 0.05) between the platelets and Fg aggregates. The data suggest that the Fg distribution may serve as a predictor of platelet morphology/activation and provides insight into the non-thrombogenic character of biomaterials containing the fluorinated SMMs.
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