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PubMed1998

Apatite-organic polymer composites prepared by a biomimetic process: improvement in adhesion of the apatite layer to the substrate by ultraviolet irradiation.

A dense and uniform layer of highly bioactive apatite can be formed in arbitrary thickness on any kind and shape of organic polymer substrates by the following biomimetic process. The substrate is first placed in contact with granular particles of CaO, SiO2-based glass soaked in a simulated body fluid with ion concentrations nearly equal to those of human blood plasma for forming apatite nuclei, and then soaked in another fluid highly supersaturated with respect to the apatite for making the apatite nuclei grow. In the present study, the polymer substrates were pretreated with ultraviolet (UV) light, and then subjected to the biomimetic process described above. By UV irradiation, the induction period for the apatite nucleation of poly(ethylene terephthalate) (PET), poly-ether sulphone (PESF), polyethylene (PE), poly(methyl methacrylate) (PMMA) and polyamide 6 (N6) substrates were reduced form 24 h to 10 h. The adhesive strengths of the apatite layer to the substrates increased from 2.5-3.2 MPa to 4.5-6.0 MPa for PET, PESF and PMMA, and from about 1.0 MPa to 4.0-6.5 MPa for PE and N6 substrates. These results have been explained by assuming that silicate ions, which induce apatite nucleation, are easily adsorbed on the substrates due to the formation of polar groups, with an improved hydrophilic nature, on the polymer surfaces by UV irradiation.

Journal Article↗
PubMed1999

Surface modification tailors the characteristics of biomimetic coatings nucleated on starch-based polymers.

This work describes the influence of surface pretreatments over the nucleation and growth of an apatite layer, formed by a biomimetic process, on which a bioactive glass is used as a precursor of the calcium-phosphate (Ca-P) formation on the materials surface. SEVA-C, a corn starch-based biodegradable blend, was used as substrate. The surfaces were pretreated during various periods by: (i) physical methods, namely ultraviolet radiation (u.v.), and over exposure to ethylene oxide sterilization (EtO); and (ii) chemical methods, namely potassium hydroxide (KOH) and acetic anhydride (CH3CO)2 etchings. The surface modifications, performed before the production of the biomimetic coatings, resulted in a faster formation of Ca-P nuclei during the first stages of SBF immersion, particularly in the case of the KOH etching. In this case, it was possible to observe a decrease in the average surface roughness, as measured by laser profilometry, and an increase of the hydrophilicity of the material, which was evident from a clear increment in the water-uptake ability and quantified by contact angle measurements. With this treatment it was possible not only to reduce the induction period for the formation of a well defined and dense apatite-like layer, as observed by scanning electron microscopy (SEM), but also to improve the adhesion of the Ca-P layer to the substrate, as confirmed by the adhesion strength tests. For all the studied pre-treatments, the composition of the films, analyzed by energy dispersive spectroscopy (EDS) and identified by thin-film X-ray diffraction (TF-XRD), seems to be very similar to that of human bone apatites.

Journal Article↗
PubMed2001

Biomimetic coatings on titanium: a crystal growth study of octacalcium phosphate.

The biomimetic approach allows the coating of metal implants with different calcium-phosphate (Ca-P) phases. Films elaborated at physiological conditions exhibited structures closely resembling those of bone mineral. For instance, octacalcium phosphate (OCP, Ca8(HPO4)2(PO4)4 . 5H2O) crystals have been deposited on titanium through a two-step procedure. After cleaning and etching, Ti6Al4V plates were immersed for 24 h into a simulated body fluid (SBF1). A thin amorphous carbonated Ca-P layer precipitated on the metal substrate. Secondly, these thinly Ca-P coated titanium substrates were immersed for 48 h into another simulated body fluid (SBF2). The thin amorphous carbonated Ca-P layer induced the fast precipitation of a second Ca-P layer of 55 microm in thickness composed of OCP crystals. The measurements of Ca and P concentrations versus soaking time in SBF2 showed that the carbonated Ca-P layer partially dissolved before the deposition of the OCP coating. X-ray diffraction (XRD) revealed that OCP crystals grew epitaxially on the substrate. OCP is known to be one of the precursors during the bone mineralization process, thereby, this new generation of biomimetic coatings are promising for orthopedic surgery.

Journal Article↗
PubMedDate not supplied

Biomimetic coatings functionalized with adhesion peptides for dental implants.

A complete biological integration into the surrounding tissues (bone, gingiva) is a critical step for clinical success of a dental implant. In this work biomimetic coatings consisting either of collagen type I (for the gingiva region) and hydroxyapatite (HAP) or mineralized collagen (for the bone interface) have been developed as suitable surfaces regarding the interfaces. Additionally, using these biomimetic coatings as a matrix, adhesion peptides were bound to further increase the specificity of titanium implant surfaces. To enhance cell attachment in the gingiva region, a linear adhesion peptide developed from a laminin sequence (TWYKIAFQRNRK) was bound to collagen, whereas for the bone interface, a cyclic RGD peptide was bound to HAP and mineralized collagen using adequate anchor systems. The biological potential of these coatings deduced from cell attachment experiments with HaCaT human keratinocytes and MC3T3-E1 mouse osteoblasts showed the best results for collagen and laminin sequence coating for the gingiva region and mineralized collagen and RGD peptide coatings for regions with bone contact. Our concept opens promising approaches to improve the biological integration of dental implants.

Journal Article↗
PubMed2002

Hydroxyapatite coating of titanium by biomimetic method.

The biomimetic method was used in order to deposit, on titanium substrates, an hydroxyapatite (HA) coating. The bioactive HA layer was obtained by using, in the first stage of the process, a glass having the composition 2.5CaO.2SiO(2) different from the one proposed for the application of the biomimetic method. This glass can be obtained via sol-gel, a method that allows one to obtain, easily, very pure products. The growth of HA crystals was confirmed by Fourier transform infrared, SEM, EDS and X-ray photoelectron spectroscopy (XPS) results. The experimental results suggest that, as reported in the literature for other supports, the silicate ions released from the glass in the first stage bind themselves to the titanium support. In particular, from XPS analysis it is evident that the titanium substrate is well covered by a calcium phosphate layer of the type of HA.

Journal Article↗
PubMed2003

Apatite deposition on polyamide films containing carboxyl group in a biomimetic solution.

The development of organic-inorganic hybrids composed of hydroxyapatite and organic polymers is attractive because of their novelty in being materials that show a bone-bonding ability, i.e. bioactivity, and because they have mechanical properties similar to those of natural bone. The biomimetic process has received much attention for fabricating such a hybrid, where bone-like apatite is deposited under ambient conditions on polymer substrates in a simulated body fluid (SBF) having ion concentrations nearly equal to those of human extracellular fluid or related solutions. It has been shown that the carboxyl group is effective for inducing heterogeneous nucleation of apatite in the body. In the present study, apatite deposition on polyamide films containing various numbers of carboxyl groups was investigated in 1.5 SBF, which had ion concentrations 1.5 times those of a normal SBF. The effect of incorporation of calcium chloride on the formation of apatite was examined. Polyamide films containing or=40 mass % CaCl(2) formed apatite on their surfaces in 1.5 SBF. The ability of the modified film to form an apatite layer increased, and the adhesion of the apatite layer bonded to the film improved, with increasing carboxyl group content. It is concluded that novel apatite-polyamide hybrids can be prepared by a biomimetic process.

Journal Article↗
PubMed1998

Biomimetic engineering of non-adhesive glycocalyx-like surfaces using oligosaccharide surfactant polymers.

The external region of a cell membrane, known as the glycocalyx, is dominated by glycosylated molecules, which direct specific interactions such as cell-cell recognition and contribute to the steric repulsion that prevents undesirable non-specific adhesion of other molecules and cells. Mimicking the non-adhesive properties of a glycocalyx provides a potential solution to the clinical problems, such as thrombosis, that are associated with implantable devices owing to non-specific adsorption of plasma proteins. Here we describe a biomimetic surface modification of graphite using oligosaccharide surfactant polymers, which, like a glycocalyx, provides a dense and confluent layer of oligosaccharides. The surfactant polymers consist of a flexible poly(vinyl amine) with dextran and alkanoyl side chains. We show that alkanoyl side chains assemble on graphite through hydrophobic interaction and epitaxial adsorption. This constrains the polymer backbone to lie parallel to the substrate, with solvated dextran side chains protruding into the aqueous phase, creating a glycocalyx-like coating. The resulting biomimetic surface is effective in suppressing protein adsorption from human plasma protein solution.

Adhesiveness↗
PubMed2006

A biomimetic approach to the detection and identification of estrogen receptor agonists in surface waters using semipermeable membrane devices (SPMDs) and bioassay-directed chemical analysis.

GOAL, SCOPE AND BACKGROUND: Some anthropogenic pollutants posses the capacity to disrupt endogenous control of developmental and reproductive processes in aquatic biota by activating estrogen receptors. Many anthropogenic estrogen receptor agonists (ERAs) are hydrophobic and will therefore readily partition into the abiotic organic carbon phases present in natural waters. This partitioning process effectively reduces the proportion of ERAs readily available for bioconcentration by aquatic biota. Results from some studies have suggested that for many aquatic species, bioconcentration of the freely-dissolved fraction may be the principal route of uptake for hydrophobic pollutants with logarithm n-octanol/water partition coefficient (log Kow) values less than approximately 6.0, which includes the majority of known anthropogenic ERAs. The detection and identification of freely-dissolved readily bioconcentratable ERAs is therefore an important aspect of exposure and risk assessment. However, most studies use conventional techniques to sample total ERA concentrations and in doing so frequently fail to account for bioconcentration of the freely-dissolved fraction. The aim of the current study was to couple the biomimetic sampling properties of semipermeable membrane devices (SPMDs) to a bioassay-directed chemical analysis (BDCA) scheme for the detection and identification of readily bioconcentratable ERAs in surface waters. METHODS: SPMDs were constructed and deployed at a number of sites in Germany and the UK. Following the dialytic recovery of target compounds and size exclusion chromatographic cleanup, SPMD samples were fractionated using a reverse-phase HPLC method calibrated to provide an estimation of target analyte log Kow. A portion of each HPLC fraction was then subjected to the yeast estrogen screen (YES) to determine estrogenic potential. Results were plotted in the form of 'estrograms' which displayed profiles of estrogenic potential as a function of HPLC retention time (i.e. hydrophobicity) for each of the samples. Where significant activity was elicited in the YES, the remaining portion of the respective active fraction was subjected to GC-MS analysis in an attempt to identify the ERAs present. RESULTS AND DISCUSSION: Estrograms from each of the field samples showed that readily bioconcentratable ERAs were present at each of the sampling sites. Estimated log Kow values for the various active fractions ranged from 1.92 to 8.63. For some samples, estrogenic potential was associated with a relatively narrow range of log Kow values whilst in others estrogenic potential was more widely distributed across the respective estrograms. ERAs identified in active fractions included some benzophenones, various nonylphenol isomers, benzyl butyl phthalate, dehydroabietic acid, sitosterol, 3-(4-methylbenzylidine)camphor (4-MBC) and 6-acetyl-1,1,2,4,4,7-hexamethyltetralin (AHTN). Other tentatively identified compounds which may have contributed to the observed YES activity included various polycyclic aromatic hydrocarbons (PAHs) and their alkylated derivatives, methylated benzylphenols, various alkyl-phenols and dialkylphenols. However, potential ERAs present in some active fractions remain unidentified. CONCLUSIONS AND OUTLOOK: Our results show that SPMD-YES-based BDCA can be used to detect and identify readily bioconcentratable ERAs in surface waters. As such, this biomimetic approach can be employed as an alternative to conventional methodologies to provide investigators with a more environmentally relevant insight into the distribution and identity of ERAs in surface waters. The use of alternative bioassays also has the potential to expand SPMD-based BDCA to include a wide range of toxicological endpoints. Improvements to the analytical methodology used to identify ERAs or other target compounds in active fractions in the current study could greatly enhance the applicability of the methodology to risk assessment and monitoring programmes.

Biological Assay↗
PubMed2002

Biomimetic self-assembly of a functional asymmetrical electronic device.

This paper introduces a biomimetic strategy for the fabrication of asymmetrical, three-dimensional electronic devices modeled on the folding of a chain of polypeptide structural motifs into a globular protein. Millimeter-size polyhedra-patterned with logic devices, wires, and solder dots-were connected in a linear string by using flexible wire. On self-assembly, the string folded spontaneously into two domains: one functioned as a ring oscillator, and the other one as a shift register. This example demonstrates that biomimetic principles of design and self-organization can be applied to generate multifunctional electronic systems of complex, three-dimensional architecture.

Amino Acid Motifs↗
PubMedDate not supplied

Bone morphogenetic protein 2 incorporated into biomimetic coatings retains its biological activity.

We have previously shown that proteins can be incorporated into the latticework of calcium phosphate layers when biomimetically coprecipitated with the inorganic components, upon the surfaces of titanium-alloy implants. In the present study, we wished to ascertain whether recombinant human bone morphogenetic protein 2 (rhBMP-2) thus incorporated retained its bioactivity as an osteoinductive agent. Titanium alloy implants were coated biomimetically with a layer of calcium phosphate in the presence of different concentrations of rhBMP-2 (0.1-10 microg/mL). rhBMP-2 was successfully incorporated into the crystal latticework, as revealed by protein blot staining. rhBMP-2 was taken up by the calcium phosphate coatings in a dose-dependent manner, as determined by ELISA. Rat bone marrow stromal cells were grown directly on these coatings for 8 days. Their osteogenicity was then assessed quantitatively by monitoring alkaline phosphatase activity. This parameter increased as a function of rhBMP-2 concentrations within the coating medium. rhBMP-2 incorporated into calcium phosphate coatings was more potent in stimulating the alkaline phosphatase activity of the adhering cell layer than was the freely suspended drug in stimulating that of cell layers grown on a plastic substratum. This system may be of osteoinductive value in orthopedic and dental implant surgery.

Alkaline Phosphatase↗
PubMed1999

Design of biomimetic habitats for tissue engineering with P-15, a synthetic peptide analogue of collagen.

In tissues, collagen forms the scaffold for cell attachment and migration, and it modulates cell differentiation and morphogenesis by mediating the flux of chemical and mechanical stimuli. We are constructing biomimetic environments by immobilizing a collagen-derived high-affinity cell-binding peptide P-15 in three-dimensional (3-D) templates. The cell-binding peptide can be expected to transduce mechanical forces. In their physiological environment, periodontal ligament fibroblasts (PDLF) are subject to significant mechanical forces. We have examined the behavior of human PDLF in culture on particulate bovine anorganic bone mineral (ABM) coated with P-15 (ABM-P-15). Greater numbers of cells associated with ABM-P-15 compared to ABM alone. Higher levels of incorporation of radiolabeled precursors in DNA and protein were consistent with the presence of larger numbers of cells on ABM-P-15 compared to ABM cultures. Scanning electron microscopic examination showed that cultures on ABM-P-15 generated highly oriented 3-D colonies of elongated cells and formed copious amounts of fibrous as well as membranous matrix reminiscent of ligamentous structures. PDLF cultured on ABM formed sparse monolayers with little order and a meager matrix. Alizarin Red stained the matrix of particle associated cells and inter-particle cellular bridges in P-15-associated cultures, indicating mineralization. 3-D colony formation and ordering of cells along with increased mineralization suggests that the coupling of cells to the ABM matrix through P-15 may provide a biomimetic environment permissive for cell differentiation and morphogenesis. Our studies suggest that ABM-P-15 templates may be effective as endosseous grafts, and, when seeded with PDLF, these matrices may serve as tissue engineered substitutes for autologous bone grafts.

Animals↗
PubMed2004

A biomimetic gelatin-calcium phosphate bone cement.

The interest in new bone substitutes is rapidly increasing in the field of orthopedic surgery. A variety of calcium phosphate bone cement has been developed and different additives have been used to improve their biocompatibility and bioactivity. Following a biomimetic strategy aimed at reproducing bone characteristics, this study investigates the biological properties of a new gelatin enriched calcium phosphate cement (GEL-CP) that exhibits improved mechanical properties with respect to cement prepared without gelatin (C-CP). Human osteoblast MG63 were cultured on the surfaces of GEL-CP and were compared to cells cultured on C-CP samples, and on polystyrene of plate culture as control (C). Cell attachment, proliferation and differentiation were evaluated up to 21 days. SEM revealed that osteoblasts grown on GEL-CP showed a normal morphology and biological tests demonstrated very good rate of proliferation and viability in every experimental time. The presence of gelatin stimulated alkaline phosphatase activity, collagen and transforming growth factor 31 production. The data indicate that the new cement GEL-CP favors osteoblast proliferation, activation of their metabolism and differentiation. The remarkable improvement of the setting properties of the calcium phosphate cement due to the presence of gelatin suggest that the biomimetic composite material could be successfully applied as bone substitute.

Alkaline Phosphatase↗
PubMed2004

Poly(2-hydroxyethyl methacrylate) biomimetic coating to improve osseointegration of a PMMA/HA/glass composite implant: in vivo mechanical and histomorphometric assessments.

Bone implants must simultaneously satisfy many requirements, even though the surface properties remain a crucial aspect in osseointegration success. Since a single material with a uniform structure cannot satisfy all of these requirements, composite materials specifically designed for orthopedic or dental implant application should be envisaged. Two poly(methylmethacrylate)/hydroxyapatite composites reinforced by E-glass fibres, uncoated (PMMA/HA/Glass) and poly(2-hydroxyethyl methacrylate) (PMMA/HA/Glass+pHEMA) coated by the biomimetic method, were mechanically (push-out test) and histomorphometrically (Affinity Index, AI) investigated in an in vivo rabbit model. Cylindrical implants (diameter 2 mm x 5 mm length) were inserted into rabbit femoral cortical (mid-diaphysis) and cancellous (distal epiphysis) bone, under general anesthesia. The highest values of push-out force and ultimate shear strength were observed for the PMMA/HA/Glass at 12 weeks, which significantly (p < 0.001) differed from those of PMMA/HA/Glass+pHEMA at the same experimental time and from those of PMMA/HA/Glass at 4 weeks. At both experimental times, significantly (p < 0.0005) lower values of AI were observed in the PMMA/HA/Glass+pHEMA versus PMMA/HA/Glass (distal femoral epiphysis: 4 weeks = 33%; 12 weeks = 19%; femoral diaphysis: 4 weeks = 15%; 12 weeks = 11%). The good mechanical and histomorphometric results obtained with PMMA/HA/Glass should be followed by further evaluation of bone remodeling processes and mechanical strength around loaded PMMA/HA/Glass implants at longer experimental times. Finally, the biomimetic method applied to pHEMA needs to be further investigated in order to improve the positive effect of SBF on pHEMA and to enhance the coating adhesion.

Animals↗
PubMed2006

Biomimetic evolutionary analysis: testing the adaptive value of vertebrate tail stiffness in autonomous swimming robots.

For early vertebrates, a long-standing hypothesis is that vertebrae evolved as a locomotor adaptation, stiffening the body axis and enhancing swimming performance. While supported by biomechanical data, this hypothesis has not been tested using an evolutionary approach. We did so by extending biomimetic evolutionary analysis (BEA), which builds physical simulations of extinct systems, to include use of autonomous robots as proxies of early vertebrates competing in a forage navigation task. Modeled after free-swimming larvae of sea squirts (Chordata, Urochordata), three robotic tadpoles (;Tadros'), each with a propulsive tail bearing a biomimetic notochord of variable spring stiffness, k (N m(-1)), searched for, oriented to, and orbited in two dimensions around a light source. Within each of ten generations, we selected for increased swimming speed, U (m s(-1)) and decreased time to the light source, t (s), average distance from the source, R (m) and wobble maneuvering, W (rad s(-2)). In software simulation, we coded two quantitative trait loci (QTL) that determine k: bending modulus, E (Nm(-2)) and length, L (m). Both QTL were mutated during replication, independently assorted during meiosis and, as haploid gametes, entered into the gene pool in proportion to parental fitness. After random mating created three new diploid genotypes, we fabricated three new offspring tails. In the presence of both selection and chance events (mutation, genetic drift), the phenotypic means of this small population evolved. The classic hypothesis was supported in that k was positively correlated (r(2)=0.40) with navigational prowess, NP, the dimensionless ratio of U to the product of R, t and W. However, the plausible adaptive scenario, even in this simplified system, is more complex, since the remaining variance in NP was correlated with the residuals of R and U taken with respect to k, suggesting that changes in k alone are insufficient to explain the evolution of NP.

Animals↗
PubMed2002

Biomimetic solid-phase microextraction to predict body residues and toxicity of chemicals that act by narcosis.

A biomimetic extraction technique using solid-phase microextraction (SPME) fibers has been developed for the risk assessment of contaminants with a narcotic mode of action. Our goal is to apply this technique in the future for the prediction of total baseline toxicity of environmental water and effluent samples. Validation of this method requires establishing the relationship between contaminant accumulation and toxicity in biota and accumulation in the surrogate solid phase (the SPME fiber coating). For this purpose, we determined the median lethal concentration (LC50) values for Chironomus riparius midge larvae exposed to two halogenated aromatic compounds separately and measured body residues in the exposed larvae. Solid-phase microextraction fibers with an 85-microm polyacrylate (PA) coating served as the surrogate hydrophobic phase, mimicking the uptake of the compounds by midge larvae. The toxicant concentrations in SPME fibers measured directly by gas chromatography/mass spectrometry (GC-MS) or calculated from the SPME fiber-water partition coefficient, K(SPME) were related to the toxicant concentrations found in midge larvae. Our results demonstrated that the biomimetic SPME method enables the estimation of body residues in biota and prediction of the degree of baseline toxicity of a water medium.

Animals↗
PubMedDate not supplied

Porcelain veneers: dentin bonding optimization and biomimetic recovery of the crown.

PURPOSE: The purpose of this study was to investigate the biomimetic principle in porcelain veneer reconstruction, or in other words, to assess the extent to which the restoration can mimic the biomechanics and structural integrity of the original tooth. Using an optimized luting procedure, porcelain veneers are expected to present such features even when bonded to an extensive dentin surface. METHODS AND MATERIALS: Dentin-bonded porcelain veneers were assessed using functional and cyclic thermal loads with respect to two parameters: coronal stiffness (investigated using experimental strain gauges and finite element analysis) and morphology of the tooth-restoration interface (scanning electron microscope evaluation). Two different application modes of the same dentin-bonding agent, Optibond FL, were evaluated: a traditional method (dentin adhesive applied when proceeding to luting the veneer) and an alternative method (dentin adhesive applied to dentin and cured before taking the impression for the veneer). RESULTS: In the finite element model, the crown compliance increased by a factor of 2.16 after facial enamel removal and returned to 96% of its original value after the placement of the veneer. The finite element values showed a good correlation with strain gauge experimental results (one-sample t test, P > 0.35 after facial enamel removal and P > 0.19 after veneer placement). The dentin adhesive application mode was not critical to the recovery of tooth stiffness (analysis of variance, P = 0.10). However, qualitative scanning electron microscope observations demonstrated that the traditional dentin adhesive application was associated with bonding failures between the hybrid layer and the overlying resin, whereas unbroken and continuous interfaces were obtained with the new method using the same dentin adhesive. CONCLUSION: The results of this study definitely favor the biomimetic behavior of porcelain veneers bonded to teeth using an optimized application mode of dentin adhesives, because this treatment modality proved to restore both the mechanical behavior and microstructure of the intact tooth.

Analysis of Variance↗
PubMed1999

Biomimetic myoelectric hand with voluntary control of finger angle and compliance.

The purpose of this study was to develop a new type of myoelectrically controlled biomimetic prosthetic hand which has almost the same dynamics as that of the neuromuscular control system of the finger muscles, and in mechanical properties of the muscles and of the stretch reflex. One of the characteristic features of the neuromuscular control system in man is the increase in the compliance around the joint with decreasing activity of the muscle. Our prosthetic hand consisted of two surface EMG signal processing units, a digital servo system for a DC motor and 1 d.o.f. mechanical hand with three fingers. The dynamics of the neuromuscular control system including variation of the compliance around the joint was realized by using a position control system of the finger movement, force feedback and a variable gain which was modulated by the amplitude of IEMGs (rectified and smoothed EMG signals). EMG signals recorded from a pair of antagonistic muscles used to flex or to extend the wrist were used as control signals. It was shown that the finger angle and the compliance of the prosthetic hand could be controlled voluntarily with EMG signals, and that an amputee could grasp a soft object easily with the prosthetic hand. Utility of the biomimetic prosthetic hand was shown by executing myoelectric control experiments in one healthy subject and one amputee.

Artificial Limbs↗