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Biomedical subjects

F Rustichelli

Publications and source records attributed to F Rustichelli.

At least 19 recordsLinked to original sources
PubMed2006

High-resolution X-ray microtomography for three-dimensional visualization of human stem cell muscle homing.

In the perspective of clinical translation of stem cell research, it would be advantageous to develop new techniques to detect donor cells after transplantation to track their fate and thus better understand their role in regeneration of damaged and diseased tissues. In this study we use X-ray computed microtomography for three-dimensional visualization of stem cells that were labeled with magnetic nanoparticles and transplanted via intra-arterial infusion. We show that X-ray computed microtomography offers the possibility to detect with high definition and resolution human cells after transplantation, and opens new possibilities for both experimental stem cell research.

AC133 Antigen↗
PubMed2006

Engineered bone from bone marrow stromal cells: a structural study by an advanced x-ray microdiffraction technique.

The mechanism of mineralized matrix deposition was studied in a tissue engineering approach in which bone tissue is formed when porous ceramic constructs are loaded with bone marrow stromal cells and implanted in vivo. We investigated the local interaction between the mineral crystals of the engineered bone and the biomaterial by means of microdiffraction, using a set-up based on an x-ray waveguide. We demonstrated that the newly formed bone is well organized inside the scaffold pore, following the growth model of natural bone. Combining wide angle (WAXS) and small angle (SAXS) x-ray scattering with high spatial resolution, we were able to determine the orientation of the crystallographic c-axis inside the bone crystals, and the orientation of the mineral crystals and collagen micro-fibrils with respect to the scaffold. In this work we analysed six samples and for each of them two pores were studied in detail. Similar results were obtained in all cases but we report here only the most significant sample.

Anisotropy↗
PubMed2005

Tissue engineering of bone: search for a better scaffold.

BACKGROUND: Large bone defects still represent a major problem in orthopedics. Traditional bone-repair treatments can be divided into two groups: the bone transport (Ilizarov technology) and the graft transplant (autologous or allogeneic bone grafts). Thus far, none of these strategies have proven to be always resolving. As an alternative, a tissue engineering approach has been proposed where osteogenic cells, bioceramic scaffolds, growth factors and physical forces concur to the bone defect repair. Different sources of osteoprogenitor cells have been suggested, bone marrow stromal cells (BMSC) being in most cases the first choice. METHODS AND RESULTS: In association with mineral tridimensional scaffolds, BMSC form a primary bone tissue which is highly vascularized and colonized by host hemopoietic marrow. The chemical composition of the scaffold is crucial for the osteoconductive properties and the resorbability of the material. In addition, scaffolds should have an internal structure permissive for vascular invasion. Porous bioceramics [hydroxyapatite (HA) and tricalcium phosphate] are osteoconductive and are particularly advantageous for bone tissue engineering application as they induce neither an immune nor an inflammatory response in the implanted host. Earlier, we first reported a cell-based tissue engineering procedure to treat three patients with long bone segmental defects. Cells were loaded on a 100% HA porous ceramic. These scaffolds proved to have good osteoconductive properties resulting in a good functional recovery, but they have not been resorbed after more than 5 years from the implant. In addition, due to the high density of the mineral and the relatively low porosity (50-60%), it was very difficult to monitor the patient recovery during the post-surgery time using X-rays. CONCLUSIONS: We report here some pre-clinical testing of new scaffolds. To compare these second generation ceramic scaffolds more suitable for a tissue engineering approach we had to first establish animal models and analysis procedures including the use of X-ray-computed microtomography associated with X-rays synchroton radiation.

Animals↗
PubMed2004

Osteointegration of bioactive glass-coated and uncoated zirconia in osteopenic bone: an in vivo experimental study.

In elderly and osteoporotic patients an age-related loss of osteoinductivity could be the biological cause of implant failure regardless of the high quality of the implanted device. yttria stabilized tetragonal zirconia (YSTZ), either coated with the bioactive glass named RKKP bioglaze (RKKP) or uncoated, was implanted in the distal femurs of sham-operated and ovariectomized female rats. Animals were sacrificed at 30 and 60 days. Histomorphometry and microhardness tests were performed to assess osteointegration rate as well as bone quality around the implants. Significant decreases (p < 0.0005) in trabecular bone volume, BV/TV (41%), trabecular bone surface BS/TV (33%), trabecular thickness Tb.Th (20%), and trabecular number Tb.N (32%), together with a significant increase in trabecular separation Tb.Sp (184%), were found for the osteopenic rats compared with the sham-operated rats. At both experimental times the RKKP coating ensured a better osteointegration rate with higher AI values than the uncoated YSTZ, even when osteopenic rats were used (48% at 30 days and 12% at 60 days). No differences were observed at the bone-biomaterial interfaces for either material when comparing sham-operated with osteopenic rats. The present results demonstrate that the RKKP bioactive glass used as a coating ensures a high osteointegration rate even in osteoporotic bone, which is already visible from postoperative day 30 and is still apparent on day 60.

Animals↗
PubMed2004

Influence of fluorapatite minor additions on behavior of hydroxyapatite ceramics.

Fluorinated hydroxyapatite is known to be less soluble by body fluids, resulting in enhanced resistance to biodegradation in vivo conditions, as compared to the pure hydroxyapatite ceramics. The present work was aimed at the investigation of the effect of minor additions of ultrafine fluorapatite (up to 10 wt%) on the sintering behavior and mechanical properties of hydroxyapatite ceramics. In vitro testing for the osteoblast-like cells viability and proliferation was performed with the samples of varying fluorapatite content. It was found that the fluorapatite addition hinders the sintering shrinkage and lowers the strength, but does not generally affect negatively the viability of the cells.

Apatites↗
PubMed2004

Phase-contrast microtomography of thin biomaterials.

Phase-contrast microtomography, performed at the beamline ID 22 of the European Synchrotron Radiation Facility (ESRF, Grenoble, France), is demonstrated for high-resolution 3-D imaging of a hydroxyapatite sample. The technique, which relies on phase contrast imaging, gives the possibility to observe features inside samples with negligible absorption contrast. The positive results obtained suggest a possible future investigation of the influence of the distribution of pores and defects inside biomaterial coatings, on the growth of osteoblast cells.

Biocompatible Materials↗
PubMed2003

X-ray micro-diffraction analysis of reconstructed bone at Zr prosthetic surface with sub-micrometre spatial resolution.

The purpose of the present investigation is to demonstrate the power of the x-ray micro-diffraction technique in biological studies. In particular the reported experiment concerns the study of the interface between a Zr prosthetic device implanted in a rat femur and the newly-formed bone, with a spatial resolution of 0.5 microm. The obtained results give interesting information on the Zr deformation and on the crystallographic phase, the grain size and the orientation of the new bone. Moreover the study reveals a marked difference in the structure of the reconstructed bone with respect to the native bone, which cannot be appreciated with other techniques.

Animals↗
PubMed2002

Improvement in zirconia osseointegration by means of a biological glass coating: An in vitro and in vivo investigation.

The biocompatibility and osseointegration of zirconia (ZrO(2)), either coated with RKKP bioglazeor uncoated, were evaluated in vitro and in vivo. The in vitro test was performed in human osteoblasts, whereas maximal sensitization was performed in 23 Dunkin Hurtley guinea pigs. RKKP bioglaze-coated and uncoated (controls) ZrO(2) cylinders were implanted in the distal femoral epiphyses of 14 Sprague-Dawley rats under general anesthesia, and animals were sacrificed at 30 and 60 days. Lactate dehydrogenase, alkaline phosphatase, and Thiazolyl Blue (MTT) were tested in vitro. A graded score was used for evaluating the results of the sensitization test. Histomorphometry and microhardness testing were performed to quantify the osseointegration rate, as well as bone quality around the implants. Neither in vitro cytotoxicity nor sensitization were observed. Histomorphometry demonstrated that at 30 days, the affinity index was significantly higher in coated implants than in uncoated ones (p < 0.05); at 60 days, the behavior of coated implants was better than that of uncoated ones, but differences were not significant. Significant increases in bone microhardness were found at 1000 microm from the interface area for both uncoated (p < 0.0005) and RKKP bioglaze-coated (p < 0.0005) ZrO(2), and also within 200 microm from the interface (p = 0.014) but only for coated ZrO(2.) These results suggest that RKKP bioglaze-coated ZrO(2) permits biocompatible devices with improved osseointegration properties to be manufactured.

Animals↗
PubMed2002

Osteointegration of bioactive glass-coated zirconia in healthy bone: an in vivo evaluation.

Osteointegration of yttria stabilised tetragonal zirconia (YSTZ), either coated with bioactive glass named RKKP bioglaze (RKKP) or uncoated, was evaluated in an animal model. RKKP-coated and uncoated (controls) YSTZ cylinders were implanted in the distal femoral epiphyses of 14 Sprague Dawley rats under general anaesthesia. At the experimental times of 30 and 60 days after sacrifice, histomorphometry and SEM microanalysis were performed on methylmethacrylate-embedded undecalcified sections to determine the osteointegration rate. At 30 days, a significantly higher affinity index was demonstrated in vivo by histomorphometric evaluation in RKKP-coated versus uncoated YSTZ implants p < 0.05); at 60 days, the coated implants behaved better than controls (affinity index of + 32%), but the difference observed lay within the statistical uncertainty. SEM analysis demonstrated better bone adhesion to the material in RKKP-coated YSTZ at both 30 and 60 days. These findings suggest that YSTZ coated with the bioactive glass named RKKP enhances osteointegration of ceramics.

Adsorption↗
PubMed2001

Sugar-induced stabilization of the monoolein Pn3m bicontinuous cubic phase during dehydration.

To explore the molecular mechanism of the protective function of sugars on cubic lipidic systems, the mesomorphic properties of the monoolein-water system, dehydrated in the presence of a series of sugars, have been studied by osmotic stress experiments. Two bicontinuous inverse cubic structures (Pn3m and Ia3d) and a lamellar L(alpha) phase form under dehydration in pure water. In sugar solutions, the Pn3m phase shows an extraordinary stability: as a function of sugar concentration, the lattice parameter decreases to very low values, but no phase transitions occur. Instead, the Pn3m to Ia3d phase transition is obtained by equilibrating the lipid phase with aqueous polymer solutions of increasing osmotic pressure. As a result, the pressure at which the phase transition occurs strongly depends on sugar concentration. The free-energy curves obtained from the osmotic-pressure unit-cell data show that the sugar exerts an additional stabilization on both the cubic phases. The analysis of the structural parameters indicates that sugars alter the interface geometry. We suggest that a consequent release of stretching contributions in the chain packing or a reduction of the inhomogeneity in molecular splay mainly stabilize the Pn3m phase and prevent the transition to the Ia3d phase on dehydration.

Biophysics↗
PubMed2001

Phase-contrast imaging of thin biomaterials.

The necessity of information about the inner microscopical features of low absorbing materials is one of the most important goals in the structural research field. So far, non destructive analysis have been performed using contact radiography giving the scope for great advances in the production and application of new materials. However, the nature of interaction, namely X-ray absorption, limited the observations only to materials having sufficient heavy elements content. The adoption of a different X-ray interaction with matter which involves refractive properties of materials is at the basis of phase-contrast imaging. The novel method allows the use of high X-ray energies, for a deeper penetration and a lower released dose, without losing any information on the nature of the sample. A demonstration study, performed at the third generation European Synchrotron Radiation Facility (ESRF)-Grenoble, to show the potential of the new technique applied to biomaterials characterization is presented here. The test samples are a commercial matrix barrier (GUIDOR) intended to aid the healing process after periodontal surgery and a hydroxyapatite thin slab originally deposited by plasma spray technique on a TA6V alloy substrate. Phase-contrast images showed significant advantages revealing features that have negligible absorption contrast. The technique can be successfully used for the characterization of biomaterials.

Absorptiometry, Photon↗
PubMed2001

Biological glass coating on ceramic materials: in vitro evaluation using primary osteoblast cultures from healthy and osteopenic rat bone.

ZrO2 and Al2O3 substrates were successfully coated by a double layer of a silica-based glass named RKKP, using a low-cost firing technique. RKKP is a glass well known for its bioactivity; therefore, a RKKP coating on Al2O3 or ZrO2, allows to combine the excellent mechanical properties of these strong ceramic substrates with its bioactivity. ZrO2 samples were easily coated using a double layer of RKKP by a simple enamelling technique. To accommodate the thermal expansion coefficient mismatch between Al2O3 and RK K P, this substrate was coated using a multilayered composite approach. All of the coatings were characterised from a morphological and compositional point of view, and an extensive biological evaluation was performed using fresh rat osteoblasts. Osteoblast primary cultures were derived from the trabecular bone of femoral condyles harvested from intact (NB) and osteopenic (OB) rats. After characterisation of their phenotype, osteoblasts were seeded on material samples of ZrO2 or Al2O3 coated with RKKP, and cultured for 7 days. Cell proliferation (MTT test) and cell differentiation (alkaline phosphatase activity) were evaluated at the end of the experiment, to assess osteoblast behaviour in the presence of biomaterials and determine if the results were related to the host bone quality. Results of both materials showed a good level of biocompatibility. In particular, MTT significant higher values were detected in NB cultures on ZrO2-RKKP samples; ALP activity significantly increased in NB cultures on Al2O3-RKKP and in OB cultures on both coated samples.

Aluminum Oxide↗
PubMed2001

Curved optics for x-ray phase contrast imaging by synchrotron radiation.

Conventional radiographic techniques have strong limitations when low-absorption contrast samples are imaged. Phase contrast radiography has been shown to produce high-quality images of soft tissues. In this technique the recorded intensity patterns are related to gradients in the refractive index of the sample. A critical point of this new technique is the need to employ crystal analysers, which results in an appreciable reduction in the beam intensity and consequently in rather long exposure times. In this paper the use of focused beams is suggested to overcome this aspect. Biological samples with small structures and low absorption variations were imaged using both flat and curved monochromator crystals, demonstrating that the use of curved optics leads to a decrease in the exposure time with only a limited degradation of the spatial resolution. This opens up the possibility of using the phase contrast technique with laboratory sources.

Animals↗
PubMed1992

Lipid-drug interaction: a structural analysis of pindolol effects on model membranes.

The ternary system constituted by distearoylphosphatidylcholine, pindolol (a vasodilator drug) and water has been investigated by using X-ray diffraction and calorimetric techniques. The structural modifications induced by the drug have been determined and a possible interaction model has been derived. In particular, the pindolol content-temperature dependent phase diagram shows the occurrence of two new phases: the first is an interdigitated gel, and the second is a lamellar structure presenting an unusual mixed disordered-ordered conformation of the hydrocarbon chains (L alpha beta). The comparative analysis of electron density profiles relative to the L alpha beta phase, reveals significant modifications in the paraffinic region of the lipid layer. In agreement with thermodynamic results, the structural data suggest that the drug induces a stiffening and a tightening of the hydrocarbon chains. Moreover, the hydrophilic properties of the membrane (particularly in P beta, and L alpha beta phases) present an evident dependence with the drug concentration.

Calorimetry↗
PubMed1990

Thermodynamic and structural effects of propranolol on DPPC liposomes.

Some thermodynamic and structural aspects of propranolol-DPPC liposomes interaction were investigated by DSC and X-ray diffraction: the lamellar arrangement of the lipid matrix remains intact even at high concentrations of the drug (until 1:1 drug/lipid molar ratio). However, the bilayer thickness increases significantly and the chains become perpendicular to the lamellar planes, for increasing drug content. At still higher propranolol concentrations a hexagonal phase occurs followed by a lamellar phase, in which the liposomes are destroyed. Moreover, the presence of propranolol has been found to impart fluidity to the lipid matrix.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1987

Studies of structural modifications induced by gamma-irradiation in distearoylphosphatidylcholine liposomes.

An investigation of the structural and thermodynamical modifications induced by gamma-irradiation in model membranes is reported. Differential scanning calorimetry and X-ray diffraction were used to study the different phases and associated transitions of distearoylphosphatidylcholine multilamellar liposomes after 60Co gamma-irradiation. Changes were observed in the shape of the calorimetric peaks and in the corresponding phase transition temperatures. In particular a shoulder was observed at about 20 kGy. The three phases characteristic of lecithins with identical acyl chains were detected also for the highest radiation dose. The formation of lysolecithin and stearic acid upon phospholipid degradation was observed. The lysolecithin concentration increases as a function of irradiation dose, until a saturation value is reached at 40 kGy. These results correlate quite well with those obtained for interlayer and interchain distances and for the width of the main phase transition calorimetric peak. At the highest dose (approximately equal to 80 kGy) cross-linked adjacent radicals and other molecular species are also formed. Appreciable differences, and some similarities, in the behaviour of DSPC and DPPC liposomes under gamma-irradiation were observed.

1,2-Dipalmitoylphosphatidylcholine↗