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Biomedical subjects

L Ambrosio

Publications and source records attributed to L Ambrosio.

At least 37 records · Page 2Linked to original sources

Osteoblast growth and function in porous poly epsilon -caprolactone matrices for bone repair: a preliminary study.

Current methods for the replacement of skeletal tissue involve the use of autografts, allografts and, recently, synthetic substitutes, which provide a proper amount of material to repair large bone defects. Engineered bone seems a promising approach, but a number of variables have to be set prior to any clinical application. In this study, four different poly caprolactone-based polymers (PCL) were prepared and tested in vitro using osteoblast-like Saos-2 cells. Differences among three-dimensional polymers include porosity, addition of hydroxyapatite (HA) particles, and treatment with simulated body fluid. Biochemical parameters to assess cell/material interactions include viability, growth, alkaline phosphatase release, and mineralization of osteoblastic cells seeded onto three-dimensional samples, while their morphology was observed using light microscopy and SEM. Preliminary results show that the polymers, though degrading in the medium, have a positive interaction with cells, as they support cell growth and functions. In the short-term culture (3-7 days) of Saos-2 on polymers, little differences were found among PCL samples, with the presence of HA moderately improving the number of cells onto the surfaces. In the long term (3-4 weeks), it was found that the HA-added polymers obtained the best colonization by cells, and more mineral formation was observed after coating with SBF. It can be concluded that PCL is a promising material for three-dimensional scaffold for bone formation, and the presence of bone-like components improves osteoblast activity.

Biocompatible Materials↗

Dynamic mechanical behavior of PMMA based bone cements in wet environment.

The mechanical properties of three wet commercial bone cements, namely Braxel (from Bioland), Simplex-P (from Howmedica) and CMW1-G (from DePuy) are investigated by means of stress relaxation and dynamic mechanical analysis (DMA). The geometry of loading that was used is the three point bending method (ASTM D790); all the tests were performed in a water chamber by means of temperature sweeps between 17 and 57 degrees C and spanning four frequency decades. The results show that viscoelastic properties are strongly dependent on specimen conditioning (i.e. water uptake and heat treatment). The results also show that all the cements that were analyzed show mechanical properties which are intermediate between the ones of the cancellous bone and of the metals of which prostheses are normally made. As a consequence, the cement is able to reduce the stress concentrations due to the interfacing of materials which have very different stiffnesses. Moreover, the results of the DMA, particularly the ones concerning the damping factor (tan delta), indicate that at body temperature the bone cements tested show an increased capacity of dissipation, the higher is the loading frequency, thus displaying shock absorbing properties.

Journal Article↗

In vitro biocompatibility of titanium oxide for prosthetic devices nanostructured by low pressure metal-organic chemical vapor deposition.

Metal-Organic Chemical Vapor Deposition (MOCVD) has recently been proposed to coat orthopedic and dental prostheses with metal nanostructured oxide films through the decomposition of oxygenated compounds (single-source precursors) or the reaction of oxygen-free metal compounds with oxygenating agents. The present study was carried out to assess the in vitro biocompatibility in terms of cell proliferation and activation, of commercially pure Ti (control material: TI/MA) coated with nanostructured TiO2 film by MOCVD (Ti/MOCVD) using osteoblast-like cell cultures (MG-63). Evaluations were performed at 3, 7 and 14 days. Cell proliferation showed a similar trend for Ti/MA and TilMOCVD compared to polystyrene; cell number increased with time from seeding to day 7 (p < 0.005), and then decreased progressively until day 14 (ranging from -14% to -47%). The ALP level and OC production showed no significant differences between Ti/MOCVD and Ti/MA at each experimental time. Significantly higher ALP levels were found in Ti/MA at 3 days and in Ti/MOCVD at 7 and 14 days when compared to the polystyrene group. OC production decreased over time and the highest values were observed at 3 days, when it was significantly higher in the Ti/MA than in the polystyrene group (50%, p < 0.05). CICP synthesis was positively affected by the presence of Ti/MOCVD and was higher in Ti/MOCVD than in the polystyrene group. No significant differences were found between Ti/MOCVD and Ti/MA in terms of IL-6 and TGF-beta1 synthesis at any experimental time. In conclusion, the current findings demonstrate that the nanostructured TiO2 coating positively affects the osteoblast-like cell behavior in terms of cell proliferation and activity, thus confirming its high level of in vitro biocompatibility in accordance with expectations.

Biocompatible Materials↗

Hyaluronic acid hydrogel in the treatment of osteoarthritis.

In order to overcome the problem of rapid clearance of the polysaccharide hyaluronic acid (Hyal) in the treatment of osteoarthritis (OA), a 50% cross-linked Hyal hydrogel (Hyal 50%) was synthesised. The 50% refers to the amount of COOH groups of the polysaccharide involved in the cross-linking reaction. i.e. 50% of the total amount. The rheological behaviour of the Hyal 50% hydrogel, and in particular the possibility to inject it through a needle, was studied. The results obtained demonstrated that the hydrogel injected through the needle still behaved like a gel, although it showed a reduction of the dynamic moduli. The most appropriate sterilisation technique for this kind of hydrogel was also evaluated. Liophilised Hyal 50% samples were sterilised by steam, Ethylene Oxide (EtO) and gamma-rays. EtO and gamma-rays did not modify the characteristics of the hydrogel in terms of swellability and morphology. Lastly, the in vivo effect of Hyal 50% hydrogel in the treatment of chondral defect in rabbit knee was also studied. The results obtained showed the Hyal 50% injections improved chondrocytes density and matrix appearance. Furthermore, the permanence in situ of the hydrogel was longer than that of the linear Hyal.

3T3 Cells↗

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↗

Spatial and structural dependence of mechanical properties of porcine intervertebral disc.

Structure-function relationship of natural tissues is crucial to design a device mimicking the structures present in human body. For this purpose, to provide guidelines to design an intervertebral disc (IVD) substitute, in this study the influence of the spatial location and structural components on the mechanical properties of porcine IVD was investigated. Local compressive stiffness (LCS) was measured on the overall disc, also constrained between the two adjacent vertebrae: the dependence on the lumbar position was evaluated. The compliance values in the anterior position (A) were higher than both in the central posterior (CP) and in the lateral-posterior (RP, LP) locations. The values of Young's Modulus (74.67+/-6.03 MPa) and compression break load (1.36x10(4)+/-0.09x10(4)N) of the disc were also evaluated by distributed compression test. The NP rheological behavior was typical of weak-gels, with elastic modulus G' always higher than viscous modulus G" all over the frequency range investigated (G' and G" respectively equal to 320 and 85 Pa at 1 Hz) and with the moduli trends were almost parallel to each other.

Journal Article↗

Mechanical strength of tooth fragment reattachment.

The aim of this study was static and fatigue test investigation of the strength of a tooth fragment reattached with adhesives to the tooth body. Central bovine incisor teeth were used, and standardized fragments were obtained by cutting the incisal edge of the selected teeth. All the fragments were reattached using a multistep dentine adhesive system, and the specimens were randomly divided into two groups (A and B). Group B specimens underwent a further dental treatment: a circumferential double chamfer prepared around the external cut interface was filled with light cured composite restorative resin. Static and fatigue bending tests were performed and linear elastic equations were used to analyze and compare the strength of the treated teeth. The results indicated that the static and fatigue bending properties were improved by using reinforcement with composite restorative resin.

Acrylic Resins↗

Improvement of swelling properties of poly(2-hydroxyethyl methacrylate) hydrogel by means of biomimetic method.

Poly(2-hydroxyethyl methacrylates) (PHEMAs) structurally modified by means of polymer blends and random copolymers are intensively studied in order to improve mechanical properties. It was recently shown that a hydroxyapatite coating, which should improve the bonding of this biomaterial to the bone, can be obtained by means of the biomimetic method. When PHEMA is submitted to the biomimetic method, its swelling ratio is improved. This can be ascribed to the deposition of a silicatic layer, which improves the hydrophilicity, on the surface of the internal pores during the first stage of the method. This appears to be a valuable result for producing modified PHEMAs with improved mechanical properties and good swelling. The experimental results indicate the following: stronger interactions with the water molecules are set up, an induction period is observed that is linked to the rate of the reactions occurring at the surface of the glass and the establishment of a convenient concentration of the silicate ions at the external surface of the polymer, and the diffusion of simulated body fluid into the pores is the limiting stage of the process.

Biocompatible Materials↗

Biocompatibility and biodegradation of intravitreal hyaluronan implants in rabbits.

To study the biocompatibility and the biodegradation rate in vivo of new intravitreal implants made with three different hyaluronic acid esters: Hyaff7, Hyaff11 and Hyaff11p75 (100% ethyl ester, 100 and 75% benzyl esters, respectively), the plugs were implanted through a sclerotomy at 3.5 mm from the limbus of rabbit eyes. In order to evaluate the in vivo biodegradation the shaft diameter of the plugs was measured by ultrasound biomicroscopy. Slit lamp microscopy, ophthalmoscopy and ERG were performed periodically. The effects of the implants on ocular tissues were also evaluated histologically. All the plugs showed a good biocompatibilitv. Plugs of both the total esters, Hyaff7 and Hyaff11, were found to undergo a slow dissolution process for 60 and 150 days, respectively. The partial benzyl ester, Hyaff11p75, was completely reabsorbed after 15 days. Analysis of variance showed a high correlation between biodegradation rate and the time of resorption (F = 90.5; p < 0.001). The biodegradation rate of each implant is related to the chemical structure of the three types of Hyaff (F = 4.51; p = 0.005). The present data suggest that intravitreal implants based on hyaluronic acid esters represent useful biocompatible and biodegradable devices for a potential drug delivery system in the treatment of posterior segment ocular diseases.

Animals↗

Morphology and metabolism of hepatocytes cultured in Petri dishes on films and in non-woven fabrics of hyaluronic acid esters.

Polymers of hyaluronic acid (Hyal) esters exhibit good tissue compatibility and are available in various geometrical configurations. These properties can be exploited for the design of innovative bioartificial liver support devices (BALSDs) using primary hepatocytes. In this paper, we report a preliminary investigation of the polymer feasibility of the ethyl and the benzyl Hyal ester in the form of films and non-woven fabrics for the in vitro culture of primary rat hepatocytes. Cell function was evaluated daily in Petri dishes with respect to the rate of ammonia elimination (AER) and urea synthesis (USR). Cells cultured in non-woven fabrics of the ethyl ester of Hyal (HYAFF7nw) exhibited an initial AER about 32% lower and synthesised urea 33% faster than that of cells on collagen films. After a week in culture, cells on collagen films retained only a minor fraction of their initial rates. Cells cultured in non-woven fabrics of HYAFF7nw retained about 62 and 44% of their initial AER and USR, respectively, and exhibited an AER approximately equal to and a USR 3.6 times greater than those of cells adherent to collagen. These results suggest that non-woven fabrics of HYAFF7nw are promising substrata for hepatocyte culture in BALSDs.

Ammonia↗

Drosophila-raf acts to elaborate dorsoventral pattern in the ectoderm of developing embryos.

In the early Drosophila embryo the activity of the EGF-receptor (Egfr) is required to instruct cells to adopt a ventral neuroectodermal fate. Using a gain-of-function mutation we showed that D-raf acts to transmit this and other late-acting embryonic Egfr signals. A novel role for D-raf was also identified in lateral cell development using partial loss-of-function D-raf mutations. Thus, we provide evidence that zygotic D-raf acts to specify cell fates in two distinct pathways that generate dorsoventral pattern within the ectoderm. These functional requirements for D-raf activity occur subsequent to its maternal role in organizing the anterioposterior axis. The consequences of eliminating key D-raf regulatory domains and specific serine residues in the transmission of Egfr and lateral epidermal signals were also addressed here.

Animals↗

Network formation of low molecular weight hyaluronic acid derivatives.

The oscillatory and steady shear rheological properties of the benzyl esters of hyaluronic acid (HA), partially esterified (Hyaff 11p50), at low molecular weight (150 kDa) were evaluated and compared to the properties of HA at the same molecular weight. At concentrations up to 40 mg cm(-3) both Hyaff 11p50 solutions and HA solutions, behaved as viscous fluids. At higher concentrations, HA ester solutions exhibited an elastic response typical of weak gels, whereas HA exhibited a viscous behaviour. A solid-like response was also observed by lowering the temperature. These results indicate that hyaluronic acid ester solutions can form a weak gel network. The rheological properties of HA derivatives changed significantly compared to HA solutions. The improved elasticity and residence times of these solutions expand the possible applications of hyaluronic acid in the biomedical field.

Dose-Response Relationship, Drug↗

Polymer-based composite hip prostheses.

A composite hip prosthesis (CHP) made from poly(ether-imide) reinforced with carbon and glass fibres was manufactured and characterized. The main objective of the study was to evaluate the effect of fibre organization on the mechanical properties of the composite femoral implant and compare with the bone. A stacking sequence of drop-off plies of carbon/glass fibres reinforcing poly(ether-imide) (PEI) constitutes a symmetrical and balanced CHP. The hip was manufactured according to the finite element modelling (FEM) design and using the compression moulding and water-jet technologies. The measured stress-strain data according to tensile, flexural and torsional tests showed agreement with the numerical calculation. Young's modulus and the strength in tension are uniform along the stem axis (40 GPa and 600 MPa, respectively) while the elastic modulus in bending varies from 10 to 60 GPa in the tip-head direction. The composite stem showed a linear load-displacement relation up to 4500 N without breaking. Mechanical behaviour of the CHP is compared to that of a canine femur. Comparison with metal prostheses has also been undertaken. CHPs control stress-strain distributions, and hence the mechanical signals to bone, through a material-structure design.

Animals↗

Bone fracture analysis on the short rod chevron-notch specimens using the X-ray computer micro-tomography.

Mechanical fatigue of bone leads to micro-cracking which is associated with remodeling, establishing a balance in the microcrack population of the living tissue, thus, in the steady-state, the microstructure of bone provides sites of discontinuity acting as stress raisers. Hence fracture toughness plays a decisive role in bone functionality by determining the level to which the material can be stressed in the presence of cracks, or, equivalently, the magnitude of cracking which can be tolerated at a given stress level. Cortical bone, which behaves as a quasi-brittle solid when fractured, was tested as short-rod chevron-notched tension specimens (CNT). The main features of the CNT specimen are its geometry and the V shaped notch. The notch leads to steady-state crack propagation whilst the requested geometry allows a diameter 40% smaller than the thickness of a standard compact tension specimens (CT). These features are essential to distinguish the inhomogeneties in the fracture properties of materials like bone. Bone structure and crack propagation of the CNT specimens were analyzed using X-ray computed micro-tomography (XMT), which is a non-invasive imaging technique. The unique feature of the micro-CT is the high resolution three-dimensional image which consists of multi-sliced tomographs taken in a fine pitch along the rotational axis. Fracture toughness (K(IC)) computed according to the peak load was 4.8 MNm(-3/2) while that derived from experimental calibration tests using XMT was 4.9 MNm(-3/2).

Journal Article↗

Carbon fiber post adhesion to resin luting cement in the restoration of endodontically treated teeth.

Carbon fiber posts (CFP) are widely used in the restoration of endodontically treated teeth to enhance the mechanical behavior in spite of metallic posts and to prevent vertical fractures of the tooth under chewing loads. The post is cemented inside the canal lumen using polymer resins with Young's modulus lower than dentine. In this conditions the stress concentration is located at the post-cement interface and in the cement bulk itself, preserving radicular dentine from dangerous stress accumulation. The mechanical resistance of CFP posts cemented in human dentine was evaluated by the means of mechanical pull-out tests assisted by the finite element analysis. The average bond strength and the critical stress values of the CHP-cement interface were 25 MPa and 50 MPa respectively.

Journal Article↗

Synthesis, chemical and rheological characterization of new hyaluronic acid-based hydrogels.

New hyaluronic acid-based hydrogels have been synthesized. The carboxylate groups of hyaluronan were activated in order to bind the amino terminal groups of the di-amine cross-linking reagent. Different hydrogels were obtained according to the different di-amine cross-linking agents (1,3-diaminepropane, 1.6-diaminohexane, PEG500 di-amine. and PEG800 di-amine). The cross-linked polymer (C.L.Hyal) was then sulphated (C.L.HyalS) by a heterogeneous reaction using sulphur trioxide pyridine complex (SO3-Py). The thermo-mechanical properties and swelling degree were evaluated and are discussed in relation to the chemical structure and the hydrophilic character of the gels. The different behaviours of C.L.Hyal and C.L.HyalS indicate the important role of sulphated groups.

Biocompatible Materials↗

Response to polyetherimide based composite materials implanted in muscle and in bone.

The in-vivo response to a composite material obtained with polyetherimide (PEI) reinforced with carbon/glass fibers was investigated by histological methods by implanting cylinders in muscle and in bone of the New Zealand White rabbit. A common metallic alloy, widely used in orthopaedic surgery, was used as control (Stellite). The aim of the study was to analyze the biological response towards the surface of the material. Composite implants and metallic implants did not induce adverse or inflammatory reactions. The morphological picture produced was similar, in muscle and in bone, for both materials. In muscle, cylinders were confined by an extremely thin fibrous layer and the overall appearance of the muscular tissue was normal. In bone, cylinders were confined by a nearly annular rim of newly formed bone. From these data it is possible to derive that the response to PEI-based composite material is comparable with the response to metallic substrate and, then, the material can be suitable for clinical application.

Journal Article↗