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Biocompatibility of NDGA-polymerized collagen fibers. I. Evaluation of cytotoxicity with tendon fibroblasts in vitro.

The material properties of tendon type I collagen fibers polymerized with nordihydroguaiaretic acid (NDGA) are equivalent to native tendon, suggesting that NDGA crosslinking may provide a viable approach to stabilizing collagenous materials for repairing ruptured, lacerated, or surgically transected fibrous tissues, such as tendon and ligament (Koob & Hernandez, Biomaterials, in press). Using standard cytotoxicity tests, the present study evaluated the biocompatibility of these fibers with cultured bovine tendon fibroblasts. Primary fibroblasts obtained from calf digital extensor tendons were exposed to NDGA, reaction products generated from the polymerization protocol, and the crosslinked fibers. NDGA was cytotoxic to these cells at concentrations above 100 microM. NDGA oxidation products were similarly cytotoxic. At concentrations below 100 microM, fibroblast viability was not affected by NDGA or its oxidation products. At these lower concentrations, fibroblast proliferation was unaffected compared to controls not exposed to NDGA. Fibers crosslinked with NDGA contained no unreacted NDGA, but they did contain soluble reaction products that were cytotoxic to tendon fibroblasts in both the elution and the direct contact tests. Washing the fibers in 70% ethanol and phosphate-buffered saline eliminated cytotoxicity of the fibers. Ethanol simultaneously sterilized the fibers. Tensile tests established that the ethanol/phosphate buffer wash did not adversely affect the material properties of the fibers. The results of these experiments indicate that NDGA-crosslinked fibers can be rendered nontoxic to tendon fibroblasts and may provide a novel approach for producing biologically based, biocompatible, tendon bioprostheses.

Animals↗

Proteomic analysis of human skin fibroblasts grown on titanium: novel approach to study molecular biocompatibility.

Despite the growing use of titanium as an implant material, there is a lack of consensus on what constitutes the molecular basis of its biocompatibility. In the present study we compared the response of skin fibroblasts to two different growth supporting surfaces: commercially pure titanium (cpTi) and tissue culture polystyrene (TCPS). Proteins from extracts of whole cells and adsorbed serum were separated and identified by 2-dimensional gel electrophoresis. In all, 40 proteins (46 spots) were identified by matrix-assisted laser desorption/ionization and mass spectrometry, database searching, immunoblotting, running a standard, or a combination of these techniques. Many of the proteins collected from the two surfaces were found to derive from the serum used in the culture medium. The surface properties of titanium appeared to promote the formation of a more concentrated carpet of serum proteins. Several proteins from bovine or human serum, such as albumin, alpha2-HS-glycoprotein, alpha-fetoprotein, plasminogen, thrombospondin 1, and serotransferrin, along with a few unidentified serum components, were found to adsorb onto cpTi in comparatively high concentrations. The adsorption of serum proteins did not appear to be selective on either substrate. We found that among the major cellular proteins, fibronectin and a cytoskeletal protein (non-muscle myosin heavy chain type A) were expressed at lower levels by fibroblasts grown on cpTi compared to TCPS. By analyzing the changes in the entire proteome of cells in response to different growth substrates, we may gain a better understanding of the molecular basis of biocompatibility.

Animals↗

Application of self-assembly techniques in the design of biocompatible protein microarray surfaces.

This review focuses on the application of novel technologies for generating biocompatible surfaces for high-throughput screening (HTS) of proteins. Various methods of coupling and spotting proteins on self-assembled monolayer (SAM) surfaces will be described along with the protein chip challenges pertaining to spot homogeneity, morphology, biocompatibility and reproducibility.

Biocompatible Materials↗

Assembly-controlled biocompatible interface on a microchip: strategy to highly efficient proteolysis.

A biocompatible interface was constructed on a microchip by using the layer-by-layer (LBL) assembly of charged polysaccharides incorporating proteases for highly efficient proteolysis. The controlled assembly of natural polyelectrolytes and the enzyme-adsorption step were monitored by using a quartz-crystal microbalance and atomic force microscopy (AFM). Such a multilayer-assembled membrane provides a biocompatible interconnected network with high enzyme-loading capacity. The maximum digestion rate of the adsorbed trypsin in a microchannel was significantly accelerated to 1600 mM min(-1) microg(-1), compared with the tryptic digestion in solution. Based on the Langmuir isotherm model, the thermodynamic constant of adsorption K was calculated to be 1.6 x 10(5) M(-1) and the maximum adsorption loading Gammamax was 3.6 x 10(-6) mol m(-2), 30 times more than a monolayer of trypsin on the native surface. The tunable interface containing trypsin was employed to construct a microchip reactor for digestion of femtomoles of proteins and the produced peptides were analyzed by MALDI-TOF mass spectroscopy. The efficient on-chip proteolysis was obtained within a few seconds, and the identification of biological samples was feasible.

Biocompatible Materials↗

A PEG-based biocompatible block catiomer with high buffering capacity for the construction of polyplex micelles showing efficient gene transfer toward primary cells.

Nonviral gene vectors from synthetic catiomers (polyplexes) are a promising alternative to viral vectors. In particular, many recent efforts have been devoted to the construction of biocompatible polyplexes for in vivo nonviral gene therapy. A promising approach in this regard is the use of poly(ethylene glycol) (PEG)-based block catiomers, which form a nanoscaled core-shell polyplex with biocompatible PEG palisades. In this study, a series of PEG-based block catiomers with different amine functionalities were newly prepared by a simple and affordable synthetic procedure based on an aminolysis reaction, and their utility as gene carriers was investigated. This study revealed that the block catiomers carrying the ethylenediamine unit at the side chain are capable of efficient and less toxic transfection even toward primary cells, highlighting critical structural factors of the cationic units in the construction of polyplex-type gene vectors. Moreover, the availability of the polyplex micelle for transfection with primary osteoblasts will facilitate its use for bone regeneration in vivo mediated by nonviral gene transfection.

Animals↗

Poly(vinyl alcohol) hydrogel as an artificial articular cartilage: evaluation of biocompatibility.

We have developed a new poly(vinyl alcohol) hydrogel (PVA-H) of increased physical strength through a new manufacturing process. Its mechanical properties have been found to be preferable as a substitute for articular cartilage. To evaluate its biocompatibility as an artificial articular cartilage, a series of in vivo tests within the intraarticular, as well as the intramuscular, environment were conducted. Tissue reactions of cartilage, bone, synovium, and muscle to PVA-H were studied histologically. In the experimental group, in which PVA-H was implanted, inflammatory reactions of all of these tissues were very slight. In the control group, in which ultra-high molecular weight polyethylene (UHMWPE) was implanted, although tissue reactions of bone and muscle were as slight as in the experimental group, those of cartilage and synovium were somewhat more conspicuous. By way of these findings, the better biocompatibility of PVA-H was documented.

Animals↗

Biocompatibility studies on biodegradable polyester-based composites of human osteoblasts: a preliminary screening.

A series of biodegradable composites with natural hydroxyapatite, designed for possible use in orthopedics applications, were preliminarily screened for biocompatibility by employing primary cultures of human osteoblasts in a direct contact method. The cells were seeded at low density onto the materials under investigation and allowed to grow for 2 weeks. They then were analyzed for morphology, proliferation, viability, alkaline phosphatase activity (AP), osteocalcin (OC) production, and extracellular matrix mineralization. The results showed that all materials have good biocompatibility. Cell viability tests demonstrated that in all cases the values were comparable to the control, and the addition of hydroxyapatite always resulted in an enhancement of performance with respect to the plain polymer. AP and OC analysis confirmed that all composites allowed the expression of phenotypic markers. Scanning electron microscopy provided direct evidence of intense cell adhesion and proliferation on the tested materials.

Biocompatible Materials↗

Preparation of calcium aluminate cement for hard tissue repair: effects of lithium fluoride and maleic acid on setting behavior, compressive strength, and biocompatibility.

We investigated lithium fluoride (LiF) and maleic acid (MA) containing calcium aluminate cement (CAC) for hard tissue repair. The objective of this study is to estimate the addition effects of LiF and MA on setting behavior, compressive strength, and biocompatibility of CAC and to find the most compatible composition of LiF and MA for using CAC as a new bone cement. The CAC was composed mainly of CaO. Al(2)O(3). Samples of LiF and MA containing CAC were formed along with recording of setting time and peak temperature and then set cement was analyzed by X-ray diffraction (XRD). Agar diffusion test, tetrazolium bromide (MTT) assay, and hemolysis test were used to detect initial in vitro biocompatibility of LiF and MA containing CAC. It was revealed from the results that LiF shortened setting time and decreased compressive strength, whereas MA delayed setting time and increased compressive strength. However, LiF and MA showed no or little influence on maximum temperature of CAC. CAC containing 0.5 g of LiF and 8.75 g of MA showed the highest compressive strength (111.64 +/- 7.74 MPa) across all the experimental compositions. The CACs containing 0.5 g of LiF/8.75 g of MA and 1.01 g LiF/8.75 g of MA had no cytotoxicity and hemolysis. In this study, CAC with 0.5 g of LiF and 8.75g of MA showed the most compatible properties for using bone cement, and thus it was assessed a candidate for a new bone cement along with CAC.

Aluminum Compounds↗

The anticorrosion ability of titanium nitride (TiN) plating on an orthodontic metal bracket and its biocompatibility.

Typically, an orthodontic metal bracket is made from stainless steel. It has been shown that such metal may corrode in an acid- and chloride-rich environment. The purpose of the current study was to investigate a titanium nitride (TiN) ion-plated stainless steel orthodontic bracket's anticorrosion properties and compare its biocompatibility with that of non-TiN-plated brackets. The stainless-steel brackets studied here were tested in acidic artificial saliva. The plated metal bracket was produced by the titanium nitride (TiN) ion-plating method. The TiN-plating on the bracket surface was demonstrated to be successful by EDX analysis. The quantity of metallic-ion release under test immersion solutions was analyzed by atomic absorption spectrophotometry. Both TiN- and non-TiN-plated brackets may release detectable ions into the test solution, including nickel, chromium, manganese, copper, and iron (ferric). The anticorrosion ability of the plated bracket was analyzed by means of inductively coupled plasma atomic emission spectroscopy. The results revealed that the TiN-plated metal bracket did not increase the anticorrosion ability of the standard bracket. The biocompatibility of the TiN plating versus the standard bracket material resulting from bracket immersion in the test solution revealed no toxicity on U2OS cells using a methylthiazole tetrazolium (MTT) colorimetric assay. Clearly, the search for an improved technique for enhancing the anticorrosion ability of the normal metal orthodontic bracket should be continued.

Cell Line↗

Biocompatibility of lipid-protein-sugar particles containing bupivacaine in the epineurium.

Novel lipid-protein-sugar particles (LPSPs) are potentially biocompatible because they are composed of naturally occurring ingredients and their expected tissue dwell times are relatively short. In this research, we used histological sections to study tissue reaction to LPSPs (4.4-microm median diameter) when used for sciatic nerve block in the rat. As a reference, we compared LPSPs to 60-microm median diameter poly(lactic-co-glycolic) acid (PLGA) microspheres (110,000 MW PLGA, glycolic/lactic ratio 65:35). Four days after injection, both particle types produced acute inflammation within the confines of the injectate, inflammation in adjacent tissues, and myotoxicity. Bupivacaine-free particles did not display myotoxicity, and inflammation in adjacent tissues was reduced. At 2 weeks, inflammation from LPSPs had almost disappeared, whereas PLGA microspheres had a foreign-body giant cell reaction until at least 8 weeks after injection. In contrast, 3.6-microm median diameter, 20,000-MW PLGA microspheres produced a primarily histiocytic reaction 2 weeks after injection. In summary, the LPSPs and PLGA microspheres studied herein have excellent biocompatibility, but tissue reaction to the former is of much shorter duration. Myotoxicity and inflammation of surrounding tissue is largely attributed to bupivacaine. Foreign-body giant cells may be attributed to particle size rather than a specific reaction to PLGA.

1,2-Dipalmitoylphosphatidylcholine↗

Evaluation of the biocompatibility of a chitosan scaffold in mice.

Chitosan scaffolds appear to be suitable for a variety of tissue engineering applications. This study addressed the biocompatibility of chitosan in a mouse implantation model. Porous chitosan scaffolds were implanted in mice, and animals were sacrificed after 1, 2, 4, 8, or 12 weeks. Macroscopic inspection of the implantation site revealed no pathological inflammatory responses. Histological assessment indicated marked neutrophil accumulation within the implant, which resolved with increasing implantation time. Gram staining and limulus assays revealed no evidence of infection or endotoxin. Collagen was observed within the chitosan pore spaces, indicating that connective tissue matrix was deposited within the implant. Angiogenic activity associated with the external implant surface was also observed. Cellular immune responses were determined by lymphocyte proliferation assays, and antibody responses were measured using ELISA techniques. These assays indicated a very low incidence of chitosan-specific reactions. Although there was a large migration of neutrophils into the implantation area, there were minimal signs of any inflammatory reaction to the material itself. This preliminary study demonstrates that chitosan has a high degree of biocompatibility in this animal model. Overall, the findings suggest that chitosan may be suitable for the development of implantable materials.

Animals↗

Biocompatibility of fluorinated polyimide.

Contact between blood and biomaterial triggers a complex series of events including protein adsorption, leukocyte adhesion and activation, and complement activation. In this article, a series of fluorinated polyimides cured at a different temperatures was prepared, and the biocompatibility of the membranes was evaluated using in vitro protein adsorption, neutrophil adhesion, and complement activation experiments under static conditions. We found that protein adsorption, neutrophil adhesion, and complement activation for the polyimides significantly depends on the curing temperature and decreases with an increase in the temperature and that the polyimide has a good biocompatibility compared with poly(styrene) and polydimethylsiloxane. We concluded that the rearrangement of molecules such as CF(3), sulfone, and ketone at the outermost surface occurs because of curing, which induces an increase in the hydrophobicity and that the cured polyimide suppresses protein adsorption, neutrophil adhesion, and complement activation because of its high hydrophobicity and low surface free energy.

Adsorption↗

Preparation and characterization of wear debris of orthopedic materials for biocompatibility studies.

In order to test the biocompatibility of wear debris of orthopedic materials, a method has been developed to produce artificial debris of stainless steel, cobalt-chromium alloy, and high-density polyethylene. In this process, called the accelerated rubbing process, two cylindrical blocks of the same alloy were held under normal pressure in triply distilled water. One block remained stationary while the other rotated at approximately 4000 rpm. The water with the metal debris was recirculated between the rubbing surfaces. To prepare debris from polyethylene, a stainless steel block was rubbed over a polyethylene block submersed in liquid nitrogen. The shape of the metal debris was granular, whereas the polyethylene debris had a shred-like shape. The size distributions were determined from scanning electron micrographs, and it was found that the particles of the metal debris (stainless steel or cobalt-chrome alloy) ranged in sizes from 0.1-10 mu with 75% of all particles less than 2 mu in diameter. The size of polyethylene debris, expressed as the volume of an ellipsoid, ranged from 10-20,000 mu3 with 45% of all particles in the range of 400-2500 mu3. The x-ray diffraction pattern revealed that the debris of stainless steel consisted primarily of the face-centered cubic (fcc) alloy with small amounts of body-centered cubic (bcc) alloy and Fe3O4, whereas the cobalt-chrome debris consisted of the fcc alloy with a small amount of Cr2O3. The structure of the polyethylene debris was quite similar to that of the solid sample used in its preparation. The percent crystallinity Pc and the size of the crystalline regions D were slightly smaller in the debris than in the solid sample, i.e., Pc = 52 and 55%, D = 90 and 130 A, in the debris and the solid, respectively. To test the suitability of the artificial debris for biocompatibility studies, stainless steel debris was implanted into knees of adult female New Zealand rabbits. Bacterial challenge studies indicated prolongation, but not increased susceptibility to infection.

Animals↗

Biocompatibility testing of polymers: in vivo implantation studies.

An in vivo method is described for screening polymeric materials for biocompatibility. The test is based on grading acute and subacute tissue reactions at 7 and 28 days, respectively, following implantation in rats. The methods is reproducible and reliable. It is designed to provide uniform test criteria for biocompatibility assessment in the early phases of the development of surgical implant materials.

Animals↗

In vitro calcification and in vivo biocompatibility of the cross-linked polypentapeptide of elastin.

The in vitro calcifiability and molecular weight dependence of calcification of the polypentapeptide, (L X Val1-L X Pro2-Gly3-L X Val4-Gly5)n, which had been gamma-irradiation cross-linked have been determined when exposed to dialyzates of normal, nonaugmented fetal bovine serum. The material was found to calcify: calcifiability was found to be highly molecular weight dependent and to be most favored when the highest molecular weight polymers (n approximately equal to 240) had been used for cross-linking. The in vivo biocompatibility, biodegradability, and calcifiability of the gamma-irradiation cross-linked polypentapeptide were examined in rabbits in both soft and hard tissue sites. The material was found to be biocompatible irrespective of its physical form and to be biodegradable but with n of 200 or less it was not shown to calcify or ossify in the rabbit tibial nonunion model.

Animals↗

The electrochemical behavior of metallic implant materials as an indicator of their biocompatibility.

This study introduces a simple in vitro arrangement to measure current densities of implant metals. The in vivo condition of a metallic implant lying in tissues exhibiting different redox potentials is simulated in so-called straddle tests by applying a constant potential difference of 250 mV in saline containing the stable, fast-reacting redox system K4Fe(CN)6/K3Fe(CN)6. From a variety of corrosion-resistant implant metals and alloys, gold showed the highest current densities, followed by the stainless steel, the cobalt-based alloy, and the TiAIV-alloy. The pure metals titanium, niobium, and tantalum showed the lowest values. This can be explained by the stable oxide layer on these base metals, preventing an exchange of electrons and thus any redox reaction. This rating of metallic implant materials based on in vitro measurements of current densities is in good accordance with their biocompatibility rating reported from in vivo experiences. It seems that simple and cheap electrochemical tests allow an even more precise differentiation of the suitability of metallic materials for implant purposes than most of the conventional implantation tests, considering that biocompatibility is not only determined by corrosion products, but also by exchange currents and reaction products of redox processes involving tissue compounds.

Biocompatible Materials↗

Tissue biocompatibility of cellulose and its derivatives.

Tissue biocompatibility of cellulose and its derivatives was examined in two in vivo tests, one for absorbance by living tissue and one for foreign body reaction. The samples examined were regenerated celluloses and cellulose derivatives: methyl cellulose, ethyl cellulose, aminoethyl cellulose, hydroxyethyl cellulose, and cellulosic polyion complexes. The in vivo absorbance by living tissue was found to depend on the degree of crystallinity and the chemical structure of the sample. The foreign body reaction was relatively mild for all the samples examined, showing that cellulose can be converted to biocompatible materials by physical and/or chemical transformation.

Animals↗

Brain biocompatibility of a biodegradable, controlled-release polymer in rats.

We report the biocompatibility in the rat brain of a controlled-release, biodegradable polymer, the polyanhydride poly-[bis(p-carboxyphenoxy)propane-sebacic acid] copolymer (PCPP-SA) in a 20:80 formulation. The biodegradable polyanhydride can be used for drug delivery directly into the brain, circumventing the difficulties posed by the blood-brain barrier and avoiding the consequences of having to administer toxic doses systematically to reach therapeutic doses in the central nervous system. The tissue reaction in the presence of PCPP-SA was compared to that seen with other standard neurosurgical implants. Fifty-six adult Sprague-Dawley rats were assigned to one of seven groups and underwent bilateral frontal lobe implantation of PCPP-SA (42 hemispheres), Surgicel (oxidized regenerated cellulose) (35 hemispheres), or Gelfoam (absorbable gelatin sponge) (35 hemispheres). None of the animals showed any behavioral changes or neurological deficits suggestive of either systemic or localized toxicity from the biodegradable polyanhydride, all surviving to the scheduled data of sacrifice. PCPP-SA evoked a well localized inflammatory reaction, comparable to that of Surgicel, which resolved as the PCPP-SA polymer degraded over five weeks. The biodegradable polyanhydride has been shown in this study to be nontoxic and biocompatible in the rat brain, when compared to standard neurosurgical implants.

Animals↗