Search PubMed⌕ Search

Biomedical subjects

L Ambrosio

Publications and source records attributed to L Ambrosio.

At least 19 recordsLinked to original sources

Human bone marrow stromal cells: In vitro expansion and differentiation for bone engineering.

Stromal cells from marrow hold a great promise for bone regeneration. Even if they are already being exploited in many clinical settings, the biological basis for the source and maintenance of their proliferation/differentiation potential after in vitro isolation and expansion needs further investigation. Most studies on osteogenic differentiation of marrow stromal cells (MSC) have been performed using bone marrow from the iliac crest. In this study, MSC were derived from spare femoral bone marrow obtained during hip replacement surgery from 20 adult donors. After in vitro isolation the cells were grown in osteogenic medium, and their proliferation and differentiation analysed during in vitro expansion. We found that MSC isolated from the femur of adult patients consistently maintain an osteogenic potential. Using biochemical signals, these cells turn to fully differentiated osteoblasts with a predictable set of molecular and phenotypic events of in vitro bone deposition. When seeded on polycaprolactone-based scaffold or surfaces, the proliferation and mineralization of femur-derived MSC were modulated by the surface chemistry/topography. Despite remarkable differences between individual colony-forming ability, alkaline phosphatase production, and mineralization ability, these cells are a potential source for bone engineering, either by direct autologous reimplantation or by ex vivo expansion and reimplantation combined to a proper scaffold.

Adult↗

A tissue engineering approach to meniscus regeneration in a sheep model.

OBJECTIVE: Regeneration of the meniscal tissue occurs to a limited extent, and the loss of meniscal tissue leads to osteoarthritis. A new biomaterial consisting of hyaluronic acid and polycaprolactone was used as a meniscus substitute in sheep to evaluate the properties of the implant material with regard to size, biomechanical stability, tissue ingrowth, and integration. METHODS: Eight sheep (right stifle joints) were treated with three total and three partial meniscus replacements while two meniscectomies served as empty controls. The animals were euthanized after 6 weeks. The specimens were assessed by gross inspection and histology, and compared with the nonoperated left joints. RESULTS: The surgical technique was found to be feasible. The implants remained in position, did not tear, and showed excellent tissue ingrowth to the capsule. Tissue integration was also observed between the original meniscus and the implant. However, graft compression and extrusion occurred. The histological investigation revealed tissue formation, cellular infiltration and vascularization. Cartilage degeneration was more severe in the operated joints. CONCLUSION: The present study shows promising results concerning the qualities of this biomaterial with regard to implantation technique, stability and tissue ingrowth.

Adjuvants, Immunologic↗

Poly-epsilon-caprolactone/hydroxyapatite composites for bone regeneration: in vitro characterization and human osteoblast response.

Polycaprolactone (PCL), a semicrystalline linear resorbable aliphatic polyester, is a good candidate as a scaffold for bone tissue engineering, due to its biocompatibility and biodegradability. However, the poor mechanical properties of PCL impair its use as scaffold for hard tissue regeneration, unless mechanical reinforcement is provided. To enhance mechanical properties and promote osteoconductivity, hydroxyapatite (HA) particles were added to the PCL matrix: three PCL-based composites with different volume ratio of HA (13%, 20%, and 32%) were studied. Mechanical properties and structure were analysed, along with biocompatibility and osteoconductivity. The addition of HA particles (in particular in the range of 20% and 32%) led to a significant improvement in mechanical performance (e.g., elastic modulus) of scaffold. Saos-2 cells and osteoblasts from human trabecular bone (hOB) retrieved during total hip replacement surgery were seeded onto 3D PCL samples for 1-4 weeks. Following the assessment of cell viability, proliferation, morphology, and ALP release, HA-loaded PCL was found to improve osteoconduction compared to the PCL alone. The results indicated that PCL represents a potential candidate as an efficient substrate for bone substitution through an accurate balance between structural/ mechanical properties of polymer and biological activities.

Alkaline Phosphatase↗

Release kinetics of ampicillin, characterization and bioactivity of TiO2/PCL hybrid materials synthesized by sol-gel processing.

Poly(epsilon-caprolactone) (PCL 6, 12, and 24 wt %) and titanium (TiO2) organic-inorganic hybrid materials have been synthesized by the sol-gel method from a multicomponent solution containing titanium butoxide, poly(epsilon-caprolactone) (PCL), water, and chloroform (CHCl3). Sodium ampicillin was incorporated in the hybrid material to verify the effect as local controlled drug delivery system. The structure of a hybrid materials interpenetrating network is realized by hydrogen bonds between Ti-OH group (H-donator) in the sol-gel intermediate species and carboxylic group (H-acceptor) in the repeating units of the polymer. The presence of hydrogen bonds between organic/inorganic components of the hybrid materials was proved by FTIR analysis. The morphology of the hybrid materials was studied by scanning electron microscope (SEM). The structure of a molecular level dispersion has been disclosed by atomic force microscope (AFM), pore size distribution and surface measurements. The bioactivity of the synthesized hybrid materials has been showed by the formation of a layer of hydroxyapatite on the surface of TiO2/PCL samples soaked in a fluid simulating the composition of the human blood plasma. The amount of sodium ampicillin released has been detected by UV-vis spectroscopy and SEM. The release kinetics seems to occur in more than one stage. HPLC analysis has also been taken to ensure the integrity of ampicillin after the synthetic treatment.

Ampicillin↗

Hyaluronic acid based materials for intestine tissue engineering: a morphological and biochemical study of cell-material interaction.

A wide number of gastro-intestinal disorders are associated with structural alterations of this district leading to an impaired gastrointestinal function. The study of cell material interactions represents one of the major issues for the development of tissue engineering purposes. Benzyl esters of hyaluronic acid are promising materials because they exhibit good tissue compatibility and are available in various configurations. In this work they have been studied for the possible application of intestinal cell growth and functioning. The preliminary investigation on the morphologic and biochemistry data obtained by monitoring the growth and differentiation of intestinal epithelial cells on two hyaluronic acid benzyl esters is reported. Two types of materials structures were studied: a three dimensional matrix and a macroporous flat sheet membrane. Caco-2 cell line was used: these cells undergo spontaneous enterocytic differentiation after several days in culture. The differentiation status of these cells grown on different materials was used as a parameter of biocompatibility and cell functioning. The status of cell growth and differentiation was monitored by studying cell morphology using scanning electron microscopy. The results obtained were confirmed by biochemical determinations. Although both the configurations of the two polymers exhibited good compatibility with respect to intestinal cells, only the flat sheet membrane proved to induce cell differentiation, leading us to the conclusion that it is a promising substrate for the proposed application.

Biocompatible Materials↗

Biodegradable and semi-biodegradable composite hydrogels as bone substitutes: morphology and mechanical characterization.

Biodegradable and semi-biodegradable composite hydrogels are proposed as bone substitutes. They consist of an hydrophilic biodegradable polymer (HYAFF 11) as matrix and two ceramic powders (alpha-TCP and HA) as reinforcement. Both components of these composites have been of great interest in biomedical applications due to their excellent biocompatibility and tissue interactions, however they have never been investigated as bone substitute composites. Morphological and mechanical analysis have shown that the two fillers behave in a very different way. In the HYAFF 11/alpha-TCP composite, alpha-TCP is able to hydrolyze in contact with water while in the HYAFF 11 matrix. As a result, the composite sets and hardens, and entangled CDHA crystals are formed in the hydrogel phase and increases in the mechanical properties are obtained. In the HYAFF11/HA composite the ceramic reinforcement acts as inert phase leading to lower mechanical properties. Both mechanical properties and microstructure analysis have demonstrated the possibility to design hydrophilic biodegradable composite structures for bone tissue substitution applications.

Biocompatible Materials↗

In vitro bioactivity of poly(epsilon-caprolactone)-apatite (PCL-AP) scaffolds for bone tissue engineering: the influence of the PCL/AP ratio.

Porous poly(epsilon-caprolactone) (PCL) is used as long-term bioresorbable scaffold for bone tissue engineering. The bone regeneration process can be enhanced by addition of carbonated apatites (AP). This study was aimed at evaluating the influence of the PCL/AP ratio on the in vitro degradation and bioactivity of PCL-AP composites. To this purpose, PCL-AP samples were synthesised with the following PCL/AP weight/weight ratios: 50/50, 60/40 and 75/25. Vibrational IR and Raman spectroscopies coupled to thermogravimetry (TG) and differential scanning calorimetry (DSC) were used to investigate the in vitro degradation mechanism in different media: 0.01 M NaOH solution (pH=12), saline phosphate buffer at pH 7.5 (SPB), esterase in SPB and simulated body fluid (SBF) at pH 7.5. The latter medium was used to evaluate the bioactivity of the composites. A control PCL sample was analysed before the addition of the AP component. As regards the untreated samples, the method of synthesis utilised for preparing the composite was found to enhance the crystallinity degree. The AP component revealed to be constituted of a B-type carbonated hydroxyapatite with a 3% carbonate content. After 28 days of treatment, the samples showed different degradation patterns and extents depending on the degradation medium, the starting PCL crystallinity and composite composition. Weight measurements, Raman and TG analyses revealed deposition of an apatitic phase on all the composites immersed in SBF. Therefore, all the samples displayed a good bioactivity; the sample which showed the most pronounced apatitic deposition was 50/50, i.e. that containing the highest amount of AP.

Apatites↗

Chemical-physical characterization and in vitro preliminary biological assessment of hyaluronic acid benzyl ester-hydroxyapatite composite.

HYAFF11 is a biocompatible, biodegradable benzyl ester of hyaluronic acid. However, in order to use it for orthopedic application, its mechanical performance needs to be improved. In this study, a novel composite based on HYAFF11 polymer matrix reinforced with hydroxylapatite (HA) has been developed. Its advantage is having a similar component of the mineral phase of bone resulting in favorable osteoconductive properties. The present study has examined the compressive mechanical and surface chemical-physical properties of the novel HYAFF11-HA composite. Preliminary biological investigations, including pH and cytotoxicity studies of the material extracts, have also been performed using an in vitro primary human osteoblast-like cell model. Moreover, protein, especially fibronectin adsorption has been investigated following incubation in culture medium and human plasma. The results show a grainy surface topography composed mainly of C, P, and Ca, with a Ca/P atomic ratio indicating HA on the composite surface. Mechanical analysis shows an improvement of the compressive properties of HYAFF11 matrix, both in the dry and swollen states, with values in the range of that of spongy bone. No cytotoxic effects and no inhibition of cell proliferation have been observed in the presence of the material extracts with pH values within acceptable ranges for cell vitality. Protein studies reveal a similar pattern, but a higher amount of fibronectin following incubation in human plasma when compared with culture medium. The results show that the novel HYAFF11-HA composite shows a great potential for application in orthopedic fields, especially as vertebral trabecular bone substitute.

Biocompatible Materials↗

Evaluation of injection augmentation treatment of hyaluronic acid based materials on rabbit vocal folds viscoelasticity.

The viscoelastic properties of vocal folds after injection of hyaluronic acid (hyaluronan, HA) based materials have been studied in an animal model (rabbit) six months after injection. The results indicate that the viscoelastic properties of the vocal folds injected with the HA based materials are similar to the healthy vocal folds (non-injected samples) used as control. Histological analysis has been also performed to investigate on the fate of the injected materials after six months from the implant. The HA based materials remain up to six months and they recruited fibroblasts that induce the ingrowth of new connective tissue resulting in an endogenous soft tissue augmentation. The HA based compounds are good candidate for further studies aimed at restoring/preserving the vibratory capacity of the vocal folds with injection treatment in glottal insufficiency.

Animals↗

An experimental and theoretical composite model of the human mandible.

The purpose is to design and manufacture a composite mandible replicate suitable for testing the influence of prosthetic materials on the stress distribution of bone. Composite mandibles made of a poly(methylmethacrylate) core and a glass reinforced outer shell are manufactured and characterised through mechanical tests assisted by the finite element analysis. The mandible replicate has been conveniently equipped with strain gauges, moreover a video extensometer has also been used in order to measure the arch width change during loading. A close agreement is found between the experimental data and the theoretical predictions. By laterally loading the mandibles the maximum values of stress and strain take place in the premolar-incisal region. By varying technological parameters such as the fiber volume fraction and orientation, it is easy to replicate the behaviour of mandibles having different stiffnesses. The results obtained by laterally loading the composite mandibles through the condyles or through the gonion regions are consistent with literature data relative to the arch width decrease of natural jaws during opening and closing. This novel synthetic system coupled with the Finite Element model constitutes an experimental-theoretical model suitable to investigate the biomechanical effects of oral rehabilitations on mandibular bone.

Humans↗

Sol-gel processing of drug delivery materials and release kinetics.

Silica, calcium (5 mol%) silicate and silica/polycaprolactone hybrid inorganic/organic amorphous materials, all mixed with sodium ampicillin, a broad-spectrum antibiotic, have been synthesized by sol-gel method. The amorphous nature of the gels was ascertained by X-ray diffraction analysis. Release kinetics in a simulate body fluid (SBF) have been subsequently investigated. The amount of sodium ampicillin released has been detected by UV-Vis spectroscopy and SEM. The release kinetics seems to occur in more than one stage. Finally FTIR measurements and SEM micrograph showed the formation of a hydroxyapatite layer on the surface of the samples soaked in SBF. All data showed that these materials could be used as drug delivery bioactive systems.

Ampicillin↗

Development and cell response of a new biodegradable composite scaffold for guided bone regeneration.

Composites of biodegradable polymers with different calcium phosphate ceramics and glasses, have been developed as scaffolds for applications in bone-tissue engineering. In this work, phosphate glass particles have been incorporated into the polymer, poly(95L/5DL) lactic acid (PLA) and porous structures were elaborated. Their porosity, compressive mechanical properties and biological response were evaluated. Interconnected structures with evenly distributed pores and a porosity as high as 97% were obtained. The incorporation of glass particles into the polymer showed to have a positive effect in the mechanical properties of the foams. Indeed, the compressive modulus increased from 74.5 to 120 KPa and the compressive strength from 17.5 to 20.1 KPa for the PLA and the PLA/glass foams, respectively. The biological response was evaluated by means of the MTT test, the materials resulted to be noncytotoxic.

Absorbable Implants↗

Synthesis and characterisation of poly(2-hydroxyethyl methacrylate) polyelectrolyte complexes.

Copolymerisation of charged and neutral monomers is a well-known methodology to introduce charged moieties in a polymeric chain to obtain polyelectrolytes. New polyelectrolyte complexes have been synthesised by radical copolymerisation of neutral methacrylic monomer 2-hydroxyethyl methacrylate (HEMA) with cationic 2-methacryloyloxyethyltrimethyl ammonium chloride and anionic 2-acrylamido-2-methylpropane-sulphonic acid monomers in 10:1:1 and 10:1:2 stechiometric ratio. Chemical structure of the synthesised terpolymers was confirmed by FT-IR spectroscopy, moreover, X-ray photoelectron spectroscopy showed the presence of a cationic charge excess on the 10:1:2 terpolymer surface with respect to 10:1:1 terpolymer. Swelling studies for 10:1:2 terpolymers showed a high water content in the swollen state and a "smart behaviour" upon changes in external stimuli such as pH, while, 10:1:1 terpolymer presented the behaviour of a neutral polymer. Mechanical and differential scanning calorimetry analysis confirmed that terpolymer networks were stabilised by ionic co-operative interactions. Infact, the inclusion of oppositely ionic charges in the polymeric network of p(HEMA) represent a way to achieve higher elastic modulus as they stabilise the terpolymer networks. Cytotoxicity and cytocompatibility studies demonstrated that all materials were not toxic, moreover, the presence of a cationic charge excess on 10:1:2 terpolymer surface was able to promote fibroblast adhesion.

Absorption↗

Sol-gel synthesis, structure and bioactivity of polycaprolactone/CaO . SiO2 hybrid material.

A method has been developed to cast novel organic/inorganic polymer hybrids from multicomponent solutions containing tetramethyl orthosilicate, calcium nitrate tetrahydrate, polycaprolactone, water, and methylethyketone via sol-gel process. The existence of the hydrogen bonds between organic and inorganic components of the hybrid and hydroxyapatite formation on the surface was proved by Fourier transform infrared analysis. The morphology of the hybrid material was studied by scanning electron microscopy. The structure of a molecular level dispersion was disclosed by atomic force microscopy, pore size distribution, and surface measurements. The infrared spectra of the hybrid relative to sample soaked in a fluid simulating the composition of human blood plasma suggests that polycaprolactone/CaO * SiO(2) hybrid material synthesised via sol-gel process is bioactive as well as the CaO * SiO(2) gel glass.

Biocompatible Materials↗

Hyaluronic-acid-based semi-interpenetrating materials.

In order to enhance the mechanical performances of hyaluronic acid (HA) without compromising its biological activity, HA has been interpenetrating with a fibrillar collagen scaffold. The semi-interpenetrating materials were obtained by mixing HA with different molecular weight and a pepsin-solubilized collagen (atelocollagen) solution, and then by inducing collagen fibrillogenesis. Results indicate that molecular weight of HA significantly influences the mechanical properties of the semi-interpenetrating materials and more specifically stronger material results from the use of low-molecular-weight (LMW) HA. According to the dynamic mechanical data the composite collagen-LMW HA has a higher elastic modulus than collagen, whereas the opposite is true for the high-molecular-weight (HMW) HA. This result highlights the role of specific interactions that occur between collagen and HA during the gel formation in controlling the network mechanical stability. LMW HA may, probably, interact more strongly with collagen during the fibrillogenesis process than HMW HA due to the higher mobility of the chains and the weaker homologous interactions. Moreover, morphological observations showed that LMW HA is intimately interdispersed within the collagen network and completely coated the fibrils, which act as mechanical support.

Biocompatible Materials↗

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↗

Biomedical application of a superabsorbent hydrogel for body water elimination in the treatment of edemas.

A novel system of body water elimination to be used for the treatment of edemas, based on superabsorbent cellulose derivatives able to absorb large amounts of water and water solutions, has been investigated. Hydrogels have been synthesized starting from water solutions of carboxymethylcellulose sodium salt and hydroxyethylcellulose, chemically crosslinked with divinylsulphone. Polyelectrolyte hydrogels displayed high sensitivity in sorption capacity to variations of the ionic strength and pH of the external solution, which is a key parameter for the application under investigation. Further, swelling properties have been modulated acting on the degree of crosslinking of the macromolecular network, and a direct method for the measurement of this parameter based on NMR solid-state analysis has been provided. The hydrogel biocompatibility has been studied in terms of its capacity either to induce nitric oxide and lactate dehydrogenase release by macrophages or influence their viability. The eventual release of toxic substances from the hydrogel was also investigated using Swiss 3T3 fibroblasts. The results obtained from the biocompatibility studies carried out in this work are consistent with the hypothesis that this gel may represent an alternative to diuretic therapies in those pathologic conditions in which edemas occur.

Absorption↗