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Biocompatibility of glass-crystalline materials obtained by the sol-gel method: effect on macrophage function.

The aim of this work was to confirm in vitro biocompatibility of a new gel-derived glass-crystalline material containing hydroxyapatite and wollastonite phases. For the purpose of comparison, studies were also carried out for a material of the same chemical composition obtained by the traditional melting method. We examined the behaviour and response of cells cultured in the presence of the studied materials. The level of activation of macrophages in culture was determined using three different methods: measurement of respiratory burst by chemiluminescence, nitrite assay and by bioassay of secreted cytokines after immunoelectrophoresis of acute phase proteins from hepatoma cells. All our results show a relatively low, close to control level, activation of macrophages exposed to the studied materials. This indicates a good biocompatibility of both the gel-derived material and the material obtained by the traditional melting method.

Animals↗

Raman spectroscopy in the study of biocompatibility.

This review deals with the application of Raman spectroscopy to the study of the biocompatibility of orthopaedic and ophthalmological materials and includes an introduction, a brief theory on the fundamental concepts of the technique, a description of the method and of the traditional and non-traditional instrumentation and a brief treatment of the surface properties of the different classes of materials. In the orthopaedic field, Raman studies on the structures of glasses, ceramic materials, carbon fibres and polymers before and after implantation are reviewed, particularly as far as structural modifications at the biomaterial-tissues interface are concerned. In the ophthalmological field, the chemical biocompatibility of hydrophobic and hydrophilic polymers for intraocular and intrastromal implants and for soft contact lenses is considered with respect to the presence of monomeric reactive centres in hydrophobic materials and the water amount in hydrophilic ones. The progress of the multichannel Raman technique for 'in-vivo' measurements is also described.

Biocompatible Materials↗

Biocompatibility issues in hemodialysis.

Hemodialysis, as a life-saving treatment modality for uremic patients, implies a repeated and compulsory contact of blood with foreign materials. As a consequence, biocompatibility problems are unavoidable. The same applies for the material used for the creation of vascular access, and for the alternative dialysis method, CAPD (continuous ambulatory peritoneal dialysis), although each system might cause its own and specific problems. Although in early dialysis the focus has been on maintenance of life and elimination of toxins, later on the important morbid implications of this lack of biocompatibility have been recognized. Eight major problems will be discussed, especially in the perspective of recent new findings in this field: (1) coagulation and clotting; (2) complement and leukocyte activation; (3) susceptibility to infection; (4) leaching or spallation; (5) surface alterations of solid materials; (6) allergic reactions; (7) shear; (8) transfer of compounds from contaminated dialysate. After description of the major biochemical and clinical implications of these problems, ways to prevent morbid events and future perspectives will be described.

Biocompatible Materials↗

Cell culture methods for testing biocompatibility.

Cell culture systems may be of value in testing the biocompatibility of prosthetic materials before they are introduced into clinical use. In recent years, in vitro methods for assaying biomaterials have gained in importance owing to the growing concern over the use of animals for biomaterials testing. Significant effort is therefore being focused toward developing predictive and quantitative, but also simple and reliable, methods of testing using cultured cells. At present, a number of methods for measuring both the cytotoxicity and the specific cytocompatibility of different materials are available. The usefulness of these systems is no longer confined to screening new materials; they can be used to study the mechanisms of action of various materials during tissue/material interaction. This paper reviews the published literature on the use of cell culture models in evaluating biocompatibility and reports on the personal experience of the authors, who have been using cell culture systems for many years and for different purposes.

Biocompatible Materials↗

Biocompatibility of hydroxyapatite ceramic: response of chondrocytes in a test system using low temperature scanning electron microscopy.

An investigation is described of R37W, one of a number of porous ceramics being developed for maxillofacial surgery and the restoration of periodontal bone defects. Sensitive and precise methods are needed to assess the biocompatibility of these new materials. Mammalian chondrocytes are known to form colonies on and within porous ceramics; therefore, the tissue formed has been evaluated to gauge the response of these proliferating mesenchymal cells to the hydroxyapatite. Cell colonies, grown on R37W, have been rapidly frozen at -210 degrees C (63K) in nitrogen slush and examined by low temperature scanning electron microscopy (SEM). This method enables unfixed, fully hydrated cells to be viewed in detail and demonstrates the three-dimensional surface structure of chondrocytes in a life-like state. Features such as complex pericellular fenestrations and papilliform surface processes are believed to indicate cell viability and normality: they are not detectable by the light microscopy and SEM of fixed, dehydrated preparations. This recently recognized fine structure, together with the determination of rates of increase of cell numbers and histochemical and immunological tests of cell synthetic and secretory behaviour, provides a new guide to biocompatibility. It is concluded that the low temperature SEM of chondrocytes grown on ceramics is a valuable addition to the procedures available for the testing of hard materials before their adoption in oral surgery.

Animals↗

Animal tests for biocompatibility of dental materials--relevance, advantages and limitations.

Biocompatibility refers to the ability of a material to perform with an appropriate host response, in a specific application. Usage tests have been developed specifically for the evaluation of the biocompatibility of dental materials. The pulp and dentine test, the pulpotomy test and the endodontic usage test are widely recognized. The relevance, advantages and limitations of these tests are discussed with particular reference to the pulp and dentine usage test. Controversies remain in the understanding of the role of several variables in determining the outcome of this test. These are highlighted by reference to the significance of the residual dentine thickness, the smear layer and microbial microleakage at the material-cavity wall interface. Limitations in the relevance of these animal models to man due to species variations in the structure and composition of the dental tissues and their oral environment may be more perceived than real. The nature of an appropriate host response may be different in the consideration of different dental materials, for example lining materials and filling materials, and there is a need for these criteria to be reviewed.

Animal Population Groups↗

Biocompatibility of extracorporeal immunoadsorption systems.

Following the first publication of a technique aiming for clinical application in 1969, extracorporeal immunoadsorption is increasingly finding a place among haemapheresis techniques. The apparent advantages outweigh the technical difficulties and economical drawbacks still connected with this treatment modality. At present three different developments are under clinical investigation: Staphylococcus Protein-A based immunoadsorption (SpA), the antibody-mediated adsorption (therapeutic affinity chromotography) and hydrophobic interaction-based immunoglobulin adsorption. The exact binding capacity and mechanism has not been completely elucidated for SpA and amino acid-based techniques. The lack of knowledge about pathogenic substrates exclusively responsible for the pathogenicity of many diseases, appears to be the major drawback for the prospective development of affinity chromatography-based extracorporeal systems. The practical application of molecular-biologic diagnostic procedures with a high grade of detection specificity (monoclonal antibodies) appears to be promising for the development of extracorporeal immunoadsorption. The immunomodulatory effect that results from the interaction of blood and plasma with artificial surfaces is an interesting subject of investigation, derived mainly from biocompatibility studies. The careful clinical and laboratory investigation of the biocompatibility of extracorporeal immunoadsorption systems contribute considerably to the prevention of undesired side effects.

Anticoagulants↗

Effects of surface treatment of Ti-6Al-4V titanium alloy on biocompatibility in cultured human umbilical vein endothelial cells.

Among the titanium alloys employed as implant materials, the Ti-6Al-4V alloy is still widely used. Ti-6Al-4V titanium alloy samples, in untreated state and subjected to treatments in air by furnace or glow-discharge processes, were put in contact with human umbilical vein endothelial cells (HUVEC) in order to evaluate their effects on biocompatibility. In HUVEC kept for 48 h in the presence of the three sample types neither cell proliferation nor protein content nor lactate dehydrogenase release in the culture medium are affected, while apoptosis is induced after 48- and 96-h contact of the cells with the untreated sample type, and after 96-h contact with the plasma treated one, the furnace treated sample type being ineffective. The expression of two adhesion molecules, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was also studied. The incubation of HUVEC with the three sample types for 48 or 96 h induces a significant increase in ICAM-1 protein levels, in comparison with control cells, while VCAM-1 expression is not detectable. In the same way, TNF-alpha release in the culture medium, assayed after 48- and 96-h contact of the cells with the three sample types, is significantly higher, in comparison with control, even if the highest values are registered in the presence of the untreated samples. Taken together, these data indicate that, although Ti-6Al-4V alloy samples, and in particular the treated ones, show a good biocompatibility, attention must be given to the first signs of inflammation.

Alloys↗

Synthesis of biocompatible segmented polyurethanes from aliphatic diisocyanates and diurea diol chain extenders.

Many polyurethane elastomers display excellent mechanical properties and adequate biocompatibility. However, many medical-grade polyurethanes are prepared from aromatic diisocyanates and can degrade in vivo to carcinogenic aromatic diamines, although the question of whether the concentrations of these harmful degradation products attain physiologically relevant levels is currently unresolved and strongly debated. It is therefore desirable to synthesize new medical-grade polyurethanes from less toxic aliphatic diisocyanates. In this paper, biocompatible segmented polyurethane elastomers were synthesized from aliphatic diisocyanates (1,4-diisocyanatobutane (BDI) and lysine methyl ester diisocyanate (LDI)), novel diurea diol chain extenders based on tyrosine and tyramine, and a model poly(ethylene glycol) (PEG) diol soft segment. The objectives were to design a hard segment similar in structure to that of MDI-based polyurethanes and also investigate the effects of systematic changes in structure on mechanical and biological properties. The non-branched, symmetric polyurethane prepared from BDI and a tyramine-based chain extender had the highest modulus at 37 degrees C. Introduction of symmetric short-chain branches (SCBs) incorporated in the tyrosine-based chain extender lowered the modulus by an order of magnitude. Polyurethanes prepared from LDI were soft polymers that had a still lower modulus due to the asymmetric SCBs that hindered hard segment packing. Polyurethanes prepared from tyramine and tyrosine chain extenders thermally degraded at temperatures ranging from 110 to 150 degrees C, which are lower than that reported previously for phenyl urethanes. All four polyurethanes supported the attachment, proliferation, and high viability of MG-63 human osteoblast-like cells in vitro. Therefore, the non-cytotoxic chemistry of these polyurethanes make them good candidates for further development as biomedical implants.

Biocompatible Materials↗

Influence of the concentrations of hyaluronic acid on the properties and biocompatibility of Cs-Gel-HA membranes.

The object of this study was to investigate the relationship between the concentrations of HA solutions and the physicochemical properties and the biocompatibility of Cs-Gel-HA membranes. The addition of different concentrations of HA not only improved the wettability significantly and extended the degradation time of Cs-Gel-HA membranes, but also changed their mechanical properties. The concentration of HA had a significant influence on the biocompatibility of Cs-Gel-HA membranes. Results demonstrated that it was only the concentrations of HA in a certain range (0.01-0.1%), that could promote the cell adhesion, migration and proliferation, while the concentration of HA was above 0.1% it would either reduce or even inhibit these behaviors.

Absorbable Implants↗

Cellular biocompatibility and stimulatory effects of calcium metaphosphate on osteoblastic differentiation of human bone marrow-derived stromal cells.

In the present study, the in vitro biocompatibility of calcium metaphosphate (CMP) with human bone marrow stromal cells (HBMSCs) and its effect on osteoblastic differentiation have been investigated. Powder and disk forms of CMP do not exert a cytotoxic effect on the HBMSCs undergoing osteoblastic differentiation. In addition, the HBMSCs adhere to the surface of the CMP disk as successfully as to the culture plate or hydroxyapatite (HA) disk. The HBMSCs adhered to either the HA or CMP disk display an undistinguishable actin arrangement and cellular phenotypes, indicating that the CMP does not disrupt normal cellular responses. An analysis of the differentiation of the HBMSCs cultured on culture plate, the HA and the CMP disk shows that three matrices are capable of supporting osteoblastic differentiation of the HBMSCs as accessed by alkaline phosphatase (ALP) staining. Further molecular analysis of osteoblastic differentiation of HBMSCs reveals that the CMP disk has a better ability than the HA disk to induce an expression of osteoblast-related genes, including ALP, osteoprotegerin (OPG), a decoy receptor for RANK ligand, and osteopontin (OPN), a non-collagenous bone matrix protein. The results demonstrate that, in addition to favorable biocompatibility, the CMP can stimulate osteoblastic differentiation of the HBMSCs in vitro.

Biocompatible Materials↗

Basic study of corn protein, zein, as a biomaterial in tissue engineering, surface morphology and biocompatibility.

The zein films, were prepared for culturing human liver cells (HL-7702) and mice fibroblast cells (NIH3T3), while the Corning microplate and polylactic acid (PLA) were chosen as controls. The surface morphology of zein films prepared by two different methods was studied by scanning electron microscope (SEM), which revealed that the zein films were composed of particles of diameter 100-500 and 500-2500 nm, respectively. The biocompatibility of zein films was assessed by attachment, extensibility and proliferation of cells on them. Our study indicated that over 60% of both HL-7702 cells and NIH3T3 cells could attach to the Corning microplate, zein films and PLA at 3h after seeding. The concentration and particle sizes for preparing zein films did not seem to affect the proliferation of the cells tested. There were no significant differences in the proliferation of both HL-7702 cells and NIH3T3 cells between the Corning microplate and two kinds of zein films, except that the zein film composed of smaller particles at the lowest concentration exhibited a very good ability for proliferation of both the cells, while PLA was a poor matrix in the latter period of the cell proliferation. This preliminary study demonstrates that zein is a promising biomaterial with good biocompatibility for the development of tissue engineering.

Animals↗

Soluble eggshell membrane protein: preparation, characterization and biocompatibility.

The preparation, characterization and biocompatibility of soluble eggshell membrane (SEP) are reported. The dissolution process, which is the key step of the preparation of SEP, has been followed by scanning electron microscopy (SEM) to observe the changes of the surfaces and thickness of the eggshell membrane (ESM). The composition of SEP has been investigated by amino acid analysis and elemental analysis. Based on the fact that SEP losses significantly cystine, and that SEP has a higher content of sulfur, an assumption involving combination with 3-mercaptopropionic acid (the reagent used for reductive cleavage of disulfide bonds) following the cleavage of the original disulfide bonds has been proposed, which explains the solubility of SEP. The thermal and surface properties have been studied by thermogravimetric analysis (TGA) and contact angle measurement. The biocompatibility of SEP, as demonstrated by cell culture of NIH3T3, is comparable to collagen type I and superior to raw ESM either inside or outside surface.

Animals↗

Biocompatibility of beta-stabilizing elements of titanium alloys.

In comparison to the presently used alpha + beta titanium alloys for biomedical applications, beta-titanium alloys have many advantageous mechanical properties, such as an improved wear resistance, a high elasticity and an excellent cold and hot formability. This will promote their future increased application as materials for orthopaedic joint replacements. Not all elements with beta-stabilizing properties in titanium alloys are suitable for biomaterial applications-corrosion and wear processes cause a release of these alloying elements to the surrounding tissue. In this investigation, the biocompability of alloying elements for beta- and near beta-titanium alloys was tested in order to estimate their suitability for biomaterial components. Titanium (grade 2) and the implant steel X2CrNiMo18153 (AISI 316 L) were tested as reference materials. The investigation included the corrosion properties of the elements, proliferation, mitochondrial activity, cell morphology and the size of MC3T3-E1 cells and GM7373 cells after 7 days incubation in direct contact with polished slices of the metals. The statistical significance was considered by Weir-test and Lord-test (alpha = 0.05). The biocompatibility range of the investigated metals is (decreasing biocompatibility): niobium-tantalum, titanium, zirconium-aluminium-316 L-molybdenum.

3T3 Cells↗

Tethering poly(ethylene glycol)s to improve the surface biocompatibility of poly(acrylonitrile-co-maleic acid) asymmetric membranes.

To improve the surface biocompatibility, asymmetric membranes fabricated from poly(acrylonitrile-co-maleic acid)s (PANCMAs) synthesized by water-phase precipitation copolymerization were tethered (or immobilized) with poly(ethylene glycol)s (PEGs) by esterification reaction. Chemical changes on the membrane surface were characterized by Fourier transform infrared spectroscopy and elemental analysis to confirm the immobilization of PEG onto the PANCMA membranes. The hydrophilicity and blood compatibility of the PEG-tethered PANCMA membrane were investigated by water contact angle, water absorption, protein adsorption, plasma platelets adhesion and cell adhesion measurements, and the results were compared with the corresponding PANCMA membranes. It was found that, after the tethering of PEG, the hydrophilicity of the membrane can be improved significantly, and the protein adsorption, platelets adhesion and macrophage attachment on the membrane surface are obviously suppressed. Furthermore, not only the content of maleic acid in PANCMA, which influences the tethering density of PEG, but also the molecular weight of PEG has great effect on the surface modification of PANCMA membranes for biocompatibility.

Acrylic Resins↗

Fast setting calcium phosphate-chitosan scaffold: mechanical properties and biocompatibility.

Calcium phosphate cement (CPC) sets in situ to form hydroxyapatite and is highly promising for a wide range of clinical applications. However, its low strength limits its use to only non-stress applications, and its lack of macroporosity hinders cell infiltration, bone ingrowth and implant fixation. The aim of this study was to develop strong and macroporous CPC scaffolds by incorporating chitosan and water-soluble mannitol, and to examine the biocompatibility of the new graft with an osteoblast cell line and an enzymatic assay. Two-way ANOVA identified significant effects on mechanical properties from chitosan reinforcement and powder:liquid ratio (p<0.001). The flexural strength of CPC-chitosan composite at a powder:liquid ratio of 2 was (13.6+/-1.2) MPa, which was significantly higher than (3.2+/-0.6) MPa for CPC control without chitosan (Tukey's at 0.95). At a powder:liquid ratio of 3.5, CPC-chitosan had a strength of (25.3+/-2.9) MPa, which was significantly higher than (10.4+/-1.7) MPa for CPC control. The scaffolds possessed total pore volume fractions ranging from 42.0% to 80.0%, and macroporosity up to 65.5%. At total porosities of 52.2-75.2%, the scaffold had strength and elastic modulus values similar to those of sintered porous hydroxyapatite and cancellous bone. Osteoblast mouse cells (MC3T3-E1) were able to adhere, spread and proliferate on CPC-chitosan specimens. The cells, which ranged from about 20 to 50 microm including the cytoplasmic extensions, infiltrated into the 165-271 microm macropores of the scaffold. In summary, substantial reinforcement and macroporosity were imparted to a moldable, fast-setting, biocompatible, and resorbable hydroxyapatite graft. The highly porous scaffold may facilitate bone ingrowth and implant fixation in vivo. In addition, the two to three times increase in strength may help extend the use of CPC to larger repairs in moderately stress-bearing locations.

Absorbable Implants↗

Short term in vivo biocompatibility testing of biodegradable poly(D,L-lactide)--growth factor coating for orthopaedic implants.

Fracture healing can be stimulated by exogenous application of growth factors. Using porcine and rat models the efficacy of locally delivered IGF-I and TGF-beta1 from an implant coating has been demonstrated. A thin and biomechanical stable biodegradable poly(D,L-lactide) was used to coat implants and serve as a drug carrier. Due to reports of possible foreign body reactions caused by polymer materials in orthopedic surgery, this study investigated the biocompatibility of the polylactide implant coating and the locally released growth factors during the time course of rat tibial fracture healing (days 5, 10, 15, and 28 after fracture). Monocytes/macrophages and osteoclast were detected using an monoclonal antibody against ED1 (comparable to CD68 in mice and human). The antibody ED1 stains monocytes, macrophages and osteoclast in the bone marrow and in the newly formed fracture callus. A moderate density of the monocytes/macrophages was seen in the proximal part of the medullary canal, but almost no cells were detectable in the region distal to the fracture. The amount of stained cells increased during the observation time with a maximum at days 10 and 15 followed by a decrease at day 28. No differences were detectable between the investigated groups from day 5 to 15 post fracture indicating, that the used poly(D,L-lactide) or the incorporated growth factors do not evoke an elevated immunological response compared to the uncoated titanium implant at the investigated time points. A significantly higher amount of ED1 positive cells was measured 28 days after fracture in the control group compared to the groups with the coated implants. In conclusion, no indication of a foreign body reaction due to the use of the polylactide or the growth factors was found indicating a good short-term biocompatibility of this bioactive coating.

Absorbable Implants↗

Morphological characterization and in vitro biocompatibility of a porous nickel-titanium alloy.

Disks consisting of macroporous nickel-titanium alloy (NiTi, Nitinol, Actipore) are used as implants in clinical surgery, e.g. for fixation of spinal dysfunctions. The morphological properties were studied by scanning electron microscopy (SEM) and by synchrotron radiation-based microtomography (SRmuCT). The composition was studied by X-ray diffractometry (XRD), differential scanning calorimetry (DSC), and energy-dispersive X-ray spectroscopy (EDX). The mechanical properties were studied with temperature-dependent dynamical mechanical analysis (DMA). Studies on the biocompatibility were performed by co-incubation of porous NiTi samples with isolated peripheral blood leukocyte fractions (polymorphonuclear neutrophil granulocytes, PMN; peripheral blood mononuclear leukocytes, PBMC) in comparison with control cultures without NiTi samples. The cell adherence to the NiTi surface was analyzed by fluorescence microscopy and scanning electron microscopy. The activation of adherent leukocytes was analyzed by measurement of the released cytokines using enzyme-linked immunosorbent assay (ELISA). The cytokine response of PMN (analyzed by the release of IL-1ra and IL-8) was not significantly different between cell cultures with or without NiTi. There was a significant increase in the release of IL-1ra (p<0.001), IL-6 (p<0.05), and IL-8 (p<0.05) from PBMC in the presence of NiTi samples. In contrast, the release of TNF-alpha by PBMC was not significantly elevated in the presence of NiTi. IL-2 was released from PBMC only in the range of the lower detection limit in all cell cultures. The material, clearly macroporous with an interconnecting porosity, consists of NiTi (martensite; monoclinic, and austenite; cubic) with small impurities of NiTi2 and possibly NiC(x). The material is not superelastic upon manual compression and shows a good biocompatibility.

Biocompatible Materials↗