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An electrochemical investigation of hemoglobin and catalase incorporated in collagen films
Li, M., P. He, et al. (2005), Biochim Biophys Acta 1749(1): 43-51.
Abstract: Collagen, an electrochemically inert protein, formed films on pyrolytic graphite (PG) electrodes, which provided a suitable microenvironment for heme proteins to transfer electron directly with the underlying electrodes. Hemoglobin (Hb) and catalase (Cat) incorporated in collagen films exhibited a pair of well-defined and quasi-reversible cyclic voltammetric peaks at around -0.35 V and -0.47 V (vs. SCE) in pH 7.0 buffers, respectively, characteristic of the protein heme Fe(III)/Fe(II) redox couples. UV-vis spectra showed that the heme proteins in collagen films retained their near-native conformations in the medium pH range. The results of scanning electron microscopy (SEM) demonstrated that the interaction between heme proteins and collagen made the morphology of dry protein-collagen films different from the collagen films alone. The electrochemical parameters such as apparent heterogeneous electron transfer rate constant (k(s)) and formal potential (E degrees ') of the films were estimated by using square wave voltammograms (SWV) and nonlinear regression analysis. The heme protein-collagen film electrodes were also used to catalyze the reduction of nitrite, oxygen and hydrogen peroxide, indicating potential applications of the films for the fabrication of a new type of biosensor that does not use mediators.

An evaluation of the surface changes in PMMA biomaterial formulations as a result of toothbrush/dentifrice abrasion
Richmond, R., T. V. Macfarlane, et al. (2004), Dent Mater 20(2): 124-32.
Abstract: OBJECTIVES: The objective of this study was to evaluate the abrasion resistance and surface roughness of two injection-molded poly(methylmethacrylate) denture base materials (SR-Ivocap 'Plus', Ipsyl 60 RV), and also one compression-molded material (Trevalon). A fourth group of specimens (prepared from Trevalon using the injection-molding procedure) was compared to the compression-molded specimens. METHODS: Ten specimens were prepared according to manufacturers' instructions. An experiment involving toothbrush and dentifrice abrasion was performed on all specimens from each of the materials and the cumulative percentage weight-loss was calculated after 100,000 brush strokes. A series of surface roughness profile measurements were also obtained from each specimen within the groups. The data were analysed by using the Kolmogorov-Smirnov test. A level of significance of 0.017 was set in order to adjust for multiple comparisons between the three sets of materials. RESULTS: There was found to be no statistically significant difference between the three groups of materials in terms of percentage weight-loss, and no statistically significant difference between the Trevalon specimens when injection-molded or compression-molded. In terms of surface roughness, however, SR-Ivocap 'Plus' recorded the lowest surface roughness profile of the three groups. It was found that there was a statistically significant difference (P<0.017) between this material and Ipsyl RV 60 (producing the highest roughness), and compression-molded Trevalon. Furthermore, there was found to be no statistically significant difference between injection-molded and compression-molded specimens of Trevalon with regard to surface texture. SIGNIFICANCE: From a clinical standpoint, the surface roughness results suggest SR-Ivocap 'Plus' to be the material most likely to produce the least suitable substrate for the accumulation of denture plaque.

An ex vivo original test using radiotracers for evaluating haemocompatibility of tubular biomaterials
Janvier, G., J. Caix, et al. (1994), Appl Radiat Isot 45(2): 207-18.
Abstract: The haemocompatibility of a vascular prosthesis can be estimated as the result of its interaction with blood components. The authors describe an ex vivo canine shunt for evaluating isotopic haemocompatibility in blood-wall interactions. Methods employing radioisotopic tracers can be used to dynamically monitor the adsorption of labelled blood cells and proteins on different biomaterial surfaces. This ex vivo test should enable materials to be assessed for quality according to two thrombogenic criteria: (i) number of adhered platelets mm-2 s-1; (ii) quantity of adsorbed fibrinogen expressed as microgram mm-2 s-1, which would provide the basis for a scale of haemocompatibility.

An experimental study of apico-aortic valved conduit (AAVC) for surgical treatment of aortic stenosis in dogs
Hirao, H., T. Inoue, et al. (2005), J Vet Med Sci 67(4): 357-62.
Abstract: A new valved conduit was developed using a canine aortic valve. The bioprosthetic valve was fixed with glutaraldehyde and epoxy compound (Denacol-EX313/810). A vascular graft composed of ultra-fine polyester fiber (10 mm in diameter, 200 mm in length) was used. Four dogs underwent apico-aortic valved conduit (AAVC) implantation and aortic banding (bypass group, BG), while another 4 dogs underwent aortic banding without AAVC implantation (control group, CG). Cardiac catheterization and angiocardiography were performed for assessment of hemodynamics 2 weeks and 6 months after surgery. Left ventricular systolic pressure, left ventricular end-diastolic pressure and the left ventricular-aortic pressure gradient differed significantly (P<0.01) between the BG and CG dogs. Left ventricular angiocardiography showed patency of the valved conduit in all the BG dogs. Echocardiography was performed before and 2, 4 and 6 months after surgery, and showed that while pressure overload caused concentric myocardial hypertrophy in the CG dogs, the left ventricle dilated eccentrically in the BG dogs. Furthermore, relief of left ventricular pressure overload by AAVC was maintained.

An image analysis method for the study of cell adhesion to biomaterials
Dubois, J. C., C. Souchier, et al. (1999), Biomaterials 20(19): 1841-9.
Abstract: This fluorescence image analysis method for the quantitative determination of cell adhesion on biomaterials allows bone cells labelled with propidium iodide to be counted automatically, directly on their support. The reliability of the estimation by fluorescence image analysis was validated by comparison with visual counting and with results obtained by an electronic particle counter. In this way it was possible to demonstrate that the adhesive properties of bone cells are dependent on the type of substrate--enstatite (MgO, SiO2, CaO-P2O5-Al2O3), Thermanox (modified polyethyleneterephthalate), or glass. In contrast, the spread of the cell cytoplasm, labelled with fluorescein isothiocyanate and measured by image analysis, does not vary significantly according to the substrate. The characterisation by SKIZ tessellation of the spatial cell arrangement shows that the bone cells have a random organisation on Thermanox and glass, whereas they form aggregates on enstatite.

An immunoproteomic approach for identification of clinical biomarkers for monitoring disease: application to cystic fibrosis
Pedersen, S. K., A. J. Sloane, et al. (2005), Mol Cell Proteomics 4(8): 1052-60.
Abstract: Circulating antibodies can be used to probe protein arrays of body fluids, prepared by two-dimensional gel electrophoresis, for antigenic biomarker detection. However, detected proteins, particularly low abundance antigens, often remain unidentifiable due to proteome complexity and limiting sample amounts. Using a novel enrichment approach exploiting patient antibodies for isolation of antigenic biomarkers, we demonstrate how immunoproteomic strategies can accelerate biomarker discovery. Application of this approach as a means of identifying biomarkers was demonstrated for cystic fibrosis (CF) lung disease by isolation and identification of inflammatory-associated autoantigens, including myeloperoxidase and calgranulin B from sputum of subjects with CF. The approach was also exploited for isolation of proteins expressed by the Pseudomonas aeruginosa strain PA01. Capture of PA01 antigens using circulating antibodies from CF subjects implicated in vivo expression of Pseudomonas proteins. All CF subjects screened, but not controls, were immunoreactive against immunocaptured Pseudomonas proteins, representing stress (GroES and ferric iron-binding protein HitA), immunosuppressive (thioredoxin), and alginate synthetase pathway (nucleoside-diphosphate kinase) proteins, implicating their clinical relevance as biomarkers of infection.

An impending crisis involving biomaterials
Hill, J. D. (1994), Ann Thorac Surg 58(6): 1571.

An important lesson about biomaterials in the TMJ
Kent, J. N. (1991), J Oral Maxillofac Surg 49(4): 442-3.

An improved peel test method for measurement of adhesion to biomaterials
Bundy, K., U. Schlegel, et al. (2000), J Mater Sci Mater Med 11(8): 517-21.
Abstract: Adhesion of tissues to biomaterials is desirable to prevent bacterial proliferation and for epithelial/transmucosal sealing of transcutaneous appliances, but can be counter-productive elsewhere, e.g. implants contacting tendons or maxillofacial subcutaneous tissue. It is therefore important to gauge adhesion strength of tissues to biomaterials before clinical use. Peel-testing is widely used for industrial product adhesion monitoring, but has rarely been applied biomedically. Here we describe peel-testing instrumentation designed for testing adherence of soft tissues to biomaterials. It offers the advantage that a 90 degrees angle between peel and substrate is maintained, simplifying determination of applied normal forces separating tissue layers from material surfaces. The device is portable and can be brought directly to the specimen removal site. This minimizes time delays between explantation and testing, maintaining the tissue/biomaterial interface in the freshest possible state closely approximating in vivo conditions, and so avoids measurement artifacts. So far, the instrument has been used to test adhesion of tape to a biomaterial surface (for determining the device's technical performance), assess strength of tissue adhesives, and measure adhesion of subcutaneous tissue to orthopaedic biomaterials. However, its versatility suggests additional applications for the peel-tester where adhesion of soft tissue to biomaterials is of interest.

An improved video-based computer tracking system for soft biomaterials testing
Downs, J., H. R. Halperin, et al. (1990), IEEE Trans Biomed Eng 37(9): 903-7.
Abstract: We present an improved video-based computer system for on-line tracking of small markers moving in a plane. The system consists of a CCD camera, a video monitor, a dedicated 80386/20 microcomputer, a video frame grabber, and custom software. Up to four markers can be tracked at the 30-Hz video frame rate using a two-step, correlation-based search procedure. We discuss the requisite hardware and software requirements and illustrate how this tracking system can be used to collect strain data during biaxial stretching tests on planar soft tissues. Operating at 30 Hz, this system is an improvement over those previously reported, which are either slower or yield less information.

An improvement of double emulsion technique for preparing bovine serum albumin-loaded PLGA microspheres
Zhang, J. X. and K. J. Zhu (2004), J Microencapsul 21(7): 775-85.
Abstract: A modified double emulsion technique was adopted to prepare bovine serum albumin (BSA) loaded poly (D,L-lactic-co-glycolic acid) (PLGA) microspheres. In the formulations, polysorbates (Tween) such as Tween20, Tween40 or Tween80, instead of frequently used poly (vinyl alcohol) (PVA), was used as the emulsifier. Microspheres with porous surface, large particle size, low microsphere yield (approximately 65.4%) and BSA entrapment efficiency (approximately 25.2%) were obtained when Tween80 aqueous solution alone was used as the outer aqueous phase. However, microspheres with smooth surface, high yield and BSA entrapment efficiency could be produced successfully by introducing sodium chloride or glucose into the outer aqueous phase. Adding 5.0%(w/v) sodium chloride into the continuous phase led to increase in microsphere yield and BSA entrapment efficiency from 65.4% and 25.2% to approximately 100% and 76.6%, respectively. Microsphere yield and BSA entrapment efficiency increased from 64.5% and 25.2% to 97.2% and 89.3%, respectively, when 15.0%,(w/v) glucose was added into the continuous phase. In constrast to the microspheres prepared in the presence of additive, a more marked burst release was observed for microspheres prepared without additive in the continuous phase, which may be attributed to the porous morphology of the latter.

An in vitro evaluation of PCL-TCP composites as delivery systems for platelet-rich plasma
Rai, B., S. H. Teoh, et al. (2005), J Control Release 107(2): 330-42.
Abstract: In this study, we first investigated the in vitro degradation properties of biodegradable, bioresorbable polycaprolactone-20% tricalcium phosphate (PCL-TCP) composites immersed in simulated body fluid (SBF) and phosphate buffered saline (PBS). Then, the release profiles of the growth factors present in platelet-rich plasma (PRP) loaded onto the composites incubated in SBF and PBS were compared. Composites immersed in both buffers showed water uptake of 13.7%+/-0.75 at day 1, followed by a constant uptake of 12.1%+/-0.3 until day 12. Henceforth the water uptake declined for SBF- and increased for PBS-soaked composites. The weight loss data did not reveal any trend. SBF- and PBS-soaked samples displayed 1-2% weight loss for 2 and 5 of the ten time points measured respectively. The original protein retention (PR) of the composites was 49.1%+/-1.50. After immersion in SBF and PBS for 4 weeks, the PR was augmented to 88.5%+/-1.40 and 69.1%+/-1.40 correspondingly. PRP after activation contained 164.7+/-24.8, 194+/-43 and 18.3+/-4.75 ng/ml of TGF-beta1, PDGF-BB and IGF-1. Microscopic analysis verified the attachment of PRP to the rods and pores of the composites. Interestingly, the buffers played an important role in determining the release profiles of TGF and PDGF. Firstly, PBS-soaked composites manifested a tri-phasic burst-like profile that was absent in SBF. Secondly, SBF-soaked composites experienced delayed release of the growth factors and total release was not achieved (64.4% for TGF and 60.5% for PDGF), whereas total release was realized for PBS-soaked composites. Lastly, release profiles from SBF-soaked composites were growth factor mediated in terms of their amounts and sizes. This was not observed for PBS-soaked composites. IGF-1, on the other hand, exhibited a progressive reduction in levels over the entire experimental period for both buffers. The mechanisms of release were theorized to be a combination of diffusion, degradation and bioactivity. Since SBF is analogous to our body fluids in terms of its ionic constituents, we expect the elution profiles derived from SBF-soaked samples to more accurately emulate the in vivo situation. In conclusion, this study has deemed PCL-TCP composites as suitable delivery systems for platelet-rich plasma.

An in vitro investigation of the PEMA/THFMA polymer system as a biomaterial for cartilage repair
Wyre, R. M. and S. Downes (2000), Biomaterials 21(4): 335-43.
Abstract: A polymer system consisting of poly(ethyl methacrylate)/tetrahydrofurfuryl methacrylate (PEMA/THFMA) was investigated as a biomaterial for cartilage repair using chondrocyte culture. The PEMA/THFMA system and Thermanox control were shown to support chondrocytes seeded directly onto the surface for up to 28 days in culture. Differences were seen between the PEMA/THFMA system and Thermanox in DNA content, proliferation and glycosaminoglycon (GAG) synthesis. There was a significantly greater medium: cell GAG ratio for the PEMA/THFMA system compared to Thermanox. A greater number of chondrocytes isolated from the superficial zone of bovine cartilage attached to the PEMA/THFMA system compared to cells isolated from the deep zone, whereas the converse was seen for Thermanox. Matrix constituents including collagen type II were synthesised indicating that the differentiated phenotype was maintained for some of the chondrocytes, although the production of type I collagen indicated that dedifferentiation of some of the chondrocytes had occurred. In conclusion, this study has shown that the PEMA/THFMA system can support chondrocytes in vitro and together with further investigations could lead to the development of the polymer as an ideal candidate for articular cartilage repair.

An in vitro model of chlorhexidine-induced tooth staining
Carpenter, G. H., R. Pramanik, et al. (2005), J Periodontal Res 40(3): 225-30.
Abstract: BACKGROUND: Tooth staining is a common feature of chlorhexidine treatment for periodontal disease and there is a large variation between patients as to the degree of their tooth staining. Although the mechanism of tooth staining is uncertain, diet, smoking and oral hygiene appear probable factors. OBJECTIVES: This study investigated the role of saliva in chlorhexidine-induced tooth staining and used tea as the staining agent in an in vitro model with hydroxyapatite mimicking teeth. METHODS: Saliva has been used to create an acquired pellicle and in solution to mimic its effects in vivo. Using different combinations of tea, chlorhexidine and parotid saliva, substances binding to hydroxyapatite were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Using this system, tea, chlorhexidine and salivary proteins were clearly identifiable following staining by Coomassie Brilliant Blue. RESULTS: The results indicated that tea interacted with many salivary proteins, in particular proline-rich proteins and histatins. Chlorhexidine did not appear to complex with or precipitate salivary proteins nor prevent the formation of an acquired pellicle on the hydroxyapatite. In isolation, tea and chlorhexidine bound in small amounts to hydroxyapatite, but when added in combination, binding of both to hydroxyapatite was greatly increased. The acquired pellicle reduced chlorhexidine and tea binding, but conversely increased the binding of either tea or chlorhexidine alone to hydroxyapatite. CONCLUSION: In conclusion, salivary proteins play an important role in the staining process and the combination of tea and chlorhexidine appears to be a very potent staining factor.

An in vitro test system for estimation of blood compatibility of biomaterials
Mason, R. G., R. W. Shermer, et al. (1974), J Biomed Mater Res 8(6): 341-56.

An in vivo model for evaluating the response of pulp to various biomaterials
McClugage, S. G., Jr., J. O. Holmstedt, et al. (1980), J Biomed Mater Res 14(5): 631-8.
Abstract: An in vivo model has been designed to study the acute response of exposed or unexposed dental pulp to the topical application of various biomaterials. This model permits sequential microscopic observations of the microvascular system of dental pulp before and after application of pulp capping agents, cementing agents, or cavity liners. The use of this experimental model provides useful information related to the tolerability of dental pulp to various biomaterials used in dentistry. Furthermore, this model serves as a useful supplement to more traditional long term methods for evaluating the biocompatability of dental materials.

An in vivo model to study the pathobiology of infectious biofilms on biomaterial surfaces
Buret, A., K. H. Ward, et al. (1991), J Biomed Mater Res 25(7): 865-74.
Abstract: This study examines the morphology, ultrastructure, and microbiology of the intact biofilm developing on an implant surface. Silastic subdermal implant material was colonized with P. aeruginosa and surgically inserted into the peritoneal cavity of adult rabbits. After 4, 8, 28, and 42 days implants were recovered and the intact biofilms examined. P. aeruginosa colonized the implant throughout the entire experimental time. Microcolonies of glycocalyx-coated bacteria were observed within the biofilm. However, the bulk of the biofilm was host-generated and typically contained phagocytes trapped within a thick mesh of fibrin. Polymorphonuclear neutrophils were the predominant cell type. Isolated erythrocytes, macrophages, and fibroblasts were also observed. By day 28, the biofilm was enclosed in a fibrous capsule of vascularized connective tissue. The low numbers of neutrophils seen in biofilms from sterile Silastic sheets implanted into control animals suggested that neutrophilia may represent a specific cellular response to the bacterial colonization. The results indicate that the cell-mediated immune response provides for most of the biofilm mass on colonized implant surfaces. Inactivated phagocytes trapped in fibrin may "wall-off" the embedded bacterial microcolonies and thus shield them from live phagocytic leucocytes. Such a mechanism may play an important role in the pathogenesis of prosthetic device infections.

An in vivo study of the host response to starch-based polymers and composites subcutaneously implanted in rats
Marques, A. P., R. L. Reis, et al. (2005), Macromol Biosci 5(8): 775-85.
Abstract: Implant failure is one of the major concerns in the biomaterials field. Several factors have been related to the fail but in general these biomaterials do not exhibit comparable physical, chemical or biological properties to natural tissues and ultimately, these devices can lead to chronic inflammation and foreign-body reactions. Starch-based biodegradable materials and composites have shown promising properties for a wide range of biomedical applications as well as a reduced capacity to elicit a strong reaction from immune system cells in vitro. In this work, blends of corn starch with ethylene vinyl alcohol (SEVA-C), cellulose acetate (SCA) and polycaprolactone (SPCL), as well as hydroxyapatite (HA) reinforced starch-based composites, were investigated in vivo. The aim of the work was to assess the host response evoked for starch-based biomaterials, identifying the presence of key cell types. The tissues surrounding the implant were harvested together with the material and processed histologically for evaluation using immunohistochemistry. At implant retrieval there was no cellular exudate around the implants and no macroscopic signs of an inflammatory reaction in any of the animals. The histological analysis of the sectioned interface tissue after immunohistochemical staining using ED1, ED2, CD54, MHC class II and alpha/beta antibodies showed positively stained cells for all antibodies, except for alpha/beta for all the implantation periods, where it was different for the various polymers and for the period of implantation. SPCL and SCA composites were the materials that stimulated the greatest cellular tissue responses, but generally biodegradable starch-based materials did not induce a severe reaction for the studied implantation times, which contrasts with other types of degradable polymeric biomaterials.

An indium:calcium phosphate colloid that specifically targets fibrin
Makowski, G. S., M. L. Ramsby, et al. (2005), J Biomed Sci 12(2): 421-9.
Abstract: The ability of indium to target fibrin in vitro was evaluated. The radionuclide (114m)Indium (114mIn) was prepared as a soluble and colloidal (In:In) form, as well as, a mixed indium:calcium phosphate (In:CaP) colloid. Soluble 114mIn was prepared by maintaining acid pH (50 mM HCl). Colloidal 114mIn (In:In) was prepared under slightly basic conditions (50 mM Tris-Cl, pH 7.6). The mixed In:CaP colloid was prepared by incubation of 114mIn with calcium (10 mM) and phosphate (250 microM) under slightly basic conditions (50 mM Tris-Cl, pH 7.6). To assess fibrin binding, the three 114mIn preparations were mixed with diluted human plasma (source of fibrinogen). Fibrin polymerization was initiated by addition of calcium (5 mM) and thrombin (0.5 U/ml). Following incubation (15 min, 37 degrees C), the fibrin matrix was condensed, removed from the reaction mixture, and washed briefly. Fibrin uptake of 114mIn (soluble, colloidal, or In:CaP) was determined by gamma counting. Results demonstrated that soluble 114mIn exclusively bound a plasma protein electrophoretically and immunologically identified as transferrin. Although both colloidal 114mIn and 114mIn:CaP bound fibrin, the mixed 114mIn:CaP colloid demonstrated substantially higher fibrin binding activity (about 2-fold). The target of indium binding was confirmed as fibrin due to the presence of characteristic cross-linked gamma-gamma dimers (100 kDa) and beta-monomers (58 kDa) by SDS-PAGE. 114mIn colloid and the mixed 114mIn:CaP colloid demonstrated no ability to bind fibrin's precursor, fibrinogen. 114mIn:CaP fibrin binding was associated with formation of CaP, as evidenced by its dependence on phosphate concentration. The biocompatibility of CaP including its ability to bind 114mIn and specifically target fibrin may be of potential value for diagnostic imaging studies to identify regions of occult vascular stenosis (i.e., atherosclerotic plaques, deep vein thrombosis, pulmonary embolus).

An injectable porous poly(propylene glycol-co-fumaric acid) bone repair material as an adjunct for intramedullary fixation
Hile, D. D., M. P. Kowaleski, et al. (2005), Biomed Mater Eng 15(3): 219-27.
Abstract: A bioresorbable bone repair material made from the unsaturated polyester poly(propylene glycol-co-fumaric acid), PPF, was investigated for its potential to act as an adjunct to alleviate the disadvantages associated with wire fixation. The PPF bone repair material is an injectable system that can be delivered to the intramedullary site and crosslinked in the presence of a hydroxylapatite filler and effervescent agents. To test the feasibility of using a bioabsorbable material as an adjunct in fracture fixation, femoral osteotomies were created in two groups of 10 Sprague-Dawley rats. Osteotomies were fixed with a threaded Kirschner wire or stabilized with a Kirshner wire augmented with the PPF bone repair material. The quantity of new bone across the osteotomy site was assessed at 4 weeks postoperatively. Histologic analysis of the healing process revealed enhanced osteoconduction across the osteotomy with the PPF bone repair material. These findings were corroborated by histomorphometric analysis of new bone formation. These findings imply suitability of the PPF bone repair material to act as an adjunct to wire fixation, such as techniques used in hand surgery.

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Last Modified: 8 February 2006