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1.
J Appl Microbiol ; 126(1): 87-101, 2019 Jan.
Article in English | MEDLINE | ID: mdl-30329212

ABSTRACT

AIMS: The aim of this study was to develop a new class of gallium (Ga)-doped chitosan (CS) coatings fabricated by electrophoretic deposition (EPD) in staphylococcal infection therapy. METHODS AND RESULTS: Biofilm formation on EPD CS/Ga coatings by Staphylococcus epidermidis and Staphylococcus aureus, which are the main strains involved in postarthroplasty infections, was assessed. The codeposition of an antibacterial agent was effective; Ga loaded into CS matrix reduces biofilm viability by up to 86% and 80% for S. epidermidis and S. aureus strains respectively. Lastly, the influence of pulsed electromagnetic field (PEMF) on the bactericidal activity of CS/Ga coatings was investigated in vitro. To this end, the coatings were incubated with S. epidermidis and S. aureus and exposed to the PEMF using two different frequencies and times. Biofilm viability for S. epidermidis was decreased by 35-40% in the presence of low-frequency (LF) and high-frequency (HF) PEMF respectively. Biofilm viability by S. aureus was not further reduced in the presence of LF PEMF, but decreased by 38% at HF PEMF. CONCLUSIONS: This study has established that a combination of PEMFs with the antibacterial agent improves bactericidal activity of Ga against S. epidermidis strain 14990 and S. aureus strain 12600. SIGNIFICANCE AND IMPACT OF THE STUDY: This new integrated approach could reduce the incidence of infection in orthopaedic implant applications. It also clearly demonstrates that the combination of Ga treatment with PEMF could aid biofilm-associated infection therapy due to improved Ga efficiency.


Subject(s)
Anti-Bacterial Agents/pharmacology , Chitosan/chemistry , Gallium/pharmacology , Staphylococcal Infections/microbiology , Staphylococcus epidermidis/drug effects , Anti-Bacterial Agents/chemistry , Biofilms/drug effects , Gallium/chemistry , Humans , Staphylococcus epidermidis/growth & development , Staphylococcus epidermidis/physiology
2.
Mater Sci Eng C Mater Biol Appl ; 82: 130-140, 2018 Jan 01.
Article in English | MEDLINE | ID: mdl-29025641

ABSTRACT

In bone tissue regeneration, the use of biomineralized scaffolds to create the 3D porous structure needed for well-fitting with defect size and appropriate cell interactions, is a promising alternative to autologous and heterologous bone grafts. Biomineralized polyurethane (PU) foams are here investigated as scaffold for bone tissue regeneration. Biomineralization of the foams was carried out by activation of PU surface by a two steps procedure performed for different times (1 to 4 weeks). Scaffolds were investigated for morphological, chemico-physical and mechanical properties, as well as for in vitro interaction with rat Bone Marrow Mesenchymal Stem Cells (BMSCs). Untreated and biomineralized PU samples showed a homogenous morphology and regular pore size (average Ø=407µm). Phase and structure of formed calcium phosphates (CaPs) layer onto the PU foam were analyzed by Fourier Transform Infrared spectroscopy and X-ray diffraction, proving the formation of bone-like nano hydroxyapatite. Biomineralization caused a significant increase of mechanical properties of treated foams compared to untreated ones. Biomineralization also affected the PU scaffold cytocompatibility providing a more appropriate surface for cell attachment and proliferation. Considering the obtained results, the proposed scaffold can be considered suitable for bone tissue regeneration.


Subject(s)
Durapatite/chemistry , Polyurethanes/chemistry , Animals , Bone and Bones/cytology , Calcium Phosphates/chemistry , Cell Proliferation/drug effects , Polyurethanes/pharmacology , Rats , Spectroscopy, Fourier Transform Infrared , Tissue Engineering/methods , X-Ray Diffraction
3.
Sci Rep ; 7: 45018, 2017 03 23.
Article in English | MEDLINE | ID: mdl-28332587

ABSTRACT

Autologous chondrocyte implantation for cartilage repair represents a challenge because strongly limited by chondrocytes' poor expansion capacity in vitro. Mesenchymal stem cells (MSCs) can differentiate into chondrocytes, while mechanical loading has been proposed as alternative strategy to induce chondrogenesis excluding the use of exogenous factors. Moreover, MSC supporting material selection is fundamental to allow for an active interaction with cells. Here, we tested a novel thermo-reversible hydrogel composed of 8% w/v methylcellulose (MC) in a 0.05 M Na2SO4 solution. MC hydrogel was obtained by dispersion technique and its thermo-reversibility, mechanical properties, degradation and swelling were investigated, demonstrating a solution-gelation transition between 34 and 37 °C and a low bulk degradation (<20%) after 1 month. The lack of any hydrogel-derived immunoreaction was demonstrated in vivo by mice subcutaneous implantation. To induce in vitro chondrogenesis, MSCs were seeded into MC solution retained within a porous polyurethane (PU) matrix. PU-MC composites were subjected to a combination of compression and shear forces for 21 days in a custom made bioreactor. Mechanical stimulation led to a significant increase in chondrogenic gene expression, while histological analysis detected sulphated glycosaminoglycans and collagen II only in loaded specimens, confirming MC hydrogel suitability to support load induced MSCs chondrogenesis.


Subject(s)
Biocompatible Materials , Cell Culture Techniques , Cell Differentiation , Chondrogenesis , Hydrogels , Mesenchymal Stem Cells/cytology , Methylcellulose , Animals , Biocompatible Materials/chemistry , Biomarkers , Bioreactors , Cell Differentiation/genetics , Chondrogenesis/genetics , Gene Expression Profiling , Humans , Materials Testing , Mesenchymal Stem Cells/metabolism , Mice
4.
J Biomater Sci Polym Ed ; 26(15): 1002-12, 2015.
Article in English | MEDLINE | ID: mdl-26230911

ABSTRACT

Injectable and resorbable hydrogels are an extremely attractive class of biomaterials. They make it possible to fill tissue defects accurately with an undoubtedly minimally invasive approach and to locally deliver cells that support repair or regeneration processes. However, their use as a cell carrier is often hindered by inadequate diffusion in bulk. A possible strategy for overcoming this transport limitation might be represented by injection of rapidly degradable cell-loaded microcapsules, so that maximum material thickness is limited by sphere radius. Here, the possibility of achieving programmable release of viable cells from alginate-based microcapsules was explored in vitro, by evaluating variations in material stability resulting from changes in hydrogel composition and assessing cell viability after encapsulation and in vitro release from microcapsules. Degradation of pure alginate microspheres was varied from a few days to several weeks by varying sodium alginate and calcium chloride concentrations. The addition of poloxamer was also found to accelerate degradation significantly, with capsule breakdown almost complete by two weeks, while chitosan was confirmed to strengthen alginate cross-linking. The presence of viable cells inside microspheres was revealed after encapsulation, and released cells were observed for all the formulations tested after a time interval dependent on bead degradation speed. These findings suggest that it may be possible to fine tune capsule breakdown by means of simple changes in material formulation and regulate, and eventually optimize, cell release for tissue repair.


Subject(s)
Cell- and Tissue-Based Therapy/methods , Hydrogels , Microspheres , Alginates/chemistry , Animals , Biocompatible Materials/chemistry , Calcium Chloride/chemistry , Cell Count , Cell Line , Cell Survival , Cell- and Tissue-Based Therapy/instrumentation , Chitosan/chemistry , Equipment Design , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Hydrogels/chemistry , Materials Testing , Mice , Myoblasts/cytology , Myoblasts/physiology , Poloxamer/chemistry , Pressure , Time Factors
5.
J Mater Sci Mater Med ; 24(10): 2439-47, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23793565

ABSTRACT

Microenvironmental cues, such as surface topography and substrate stiffness, may affect stem cells adhesion, morphology, alignment, proliferation and differentiation. Adipose derived stem cells (ASCs) have attracted considerable interest in regenerative medicine due to their easy isolation, extensive in vitro expandability and ability to differentiate along a number of different tissue-specific lineages. The aim of this work was to investigate ASCs adhesion, alignment and differentiation into myogenic lineage on nanofibrous polymeric scaffolds with anisotropic topography. Nanostructured scaffolds with randomized or parallel fibers were fabricated by electrospinning using polycaprolactone (PCL) and the polycarbonate-urethane ChronoFlex AL 80A (CFAL). Cells expressed myosin (fast skeletal) and tropomyosin in all surface topographies 7 days after seeding but myotube formation was only observed on CFAL scaffolds and only few myotubes were formed on PCL scaffolds. The different cell behavior could be ascribed to two main parameters: fibers dimensions and fibers orientation of the substrates that could result in a better myotube formation on CFAL scaffolds.


Subject(s)
Adipose Tissue/metabolism , Stem Cells/cytology , Tissue Scaffolds/chemistry , Adipose Tissue/pathology , Biocompatible Materials/chemistry , Cell Adhesion , Cell Differentiation , Cell Proliferation , Humans , Immunohistochemistry , Microscopy, Electron, Scanning , Muscle Development , Muscle, Skeletal/metabolism , Myosins/metabolism , Nanofibers/chemistry , Nanostructures/chemistry , Nanotechnology , Polyesters/chemistry , Tissue Engineering/methods , Tropomyosin/chemistry
6.
J Mater Sci Mater Med ; 21(1): 365-75, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19756969

ABSTRACT

Autologous and eterologous cell encapsulation has been extensively studied for clinical application in functional organs substitution, recombinant cell transplantation in gene therapy or in muscle and cartilage regeneration to treat degenerative pathologies. In this work, calcium alginate, calcium alginate/chitosan, calcium alginate/gelatin and pectin/chitosan microcapsules were prepared to be used as innovative injectable scaffolds for soft issue regeneration by a simple extrusion method from aqueous solutions. Prepared microcapsules had spherical morphology, whereas their size was deeply influenced by the polymeric composition. When incubated in a physiological-like environment up to 30 days, they underwent an initial swelling, followed by weight loss at different rates, depending on the microcapsules formulation. The encapsulation of mouse myoblast cells (C2C12 cell line) was obtained in calcium alginate, calcium alginate/chitosan, calcium alginate/gelatin microcapsules. Cells were alive throughout the encapsulation procedure, and were recovered by a mechanical rupture of the microcapsules. After 7 days, fractured microcapsules led cells to migrate gradually out.


Subject(s)
Capsules/chemistry , Guided Tissue Regeneration/methods , Polysaccharides/chemistry , Alginates/chemistry , Alginates/pharmacology , Animals , Biocompatible Materials/chemical synthesis , Biocompatible Materials/chemistry , Capsules/chemical synthesis , Cells, Cultured , Dose-Response Relationship, Drug , Drug Compounding/methods , Drug Stability , Guided Tissue Regeneration/instrumentation , Mice , Models, Biological , Muscles/physiology , Particle Size , Pectins/chemistry , Pectins/pharmacology , Wettability
7.
Acta Biomater ; 6(6): 1948-57, 2010 Jun.
Article in English | MEDLINE | ID: mdl-20040385

ABSTRACT

During tissue formation, skeletal muscle precursor cells fuse together to form multinucleated myotubes. To understand this mechanism, in vitro systems promoting cell alignment need to be developed; for this purpose, micrometer-scale features obtained on substrate surfaces by photolithography can be used to control and affect cell behaviour. This work was aimed at investigating how differently microgrooved polymeric surfaces can affect myoblast alignment, fusion and myotube formation in vitro. Microgrooved polymeric films were obtained by solvent casting of a biodegradable poly-l-lactide/trimethylene carbonate copolymer (PLLA-TMC) onto microgrooved silicon wafers with different groove widths (5, 10, 25, 50, 100microm) and depths (0.5, 1, 2.5, 5microm), obtained by a standard photolithographic technique. The surface topography of wafers and films was evaluated by scanning electron microscopy. Cell assays were performed using C2C12 cells and myotube formation was analysed by immunofluorescence assays. Cell alignment and circularity were also evaluated using ImageJ software. The obtained results confirm the ability of microgrooved surfaces to influence myotube formation and alignment; in addition, they represent a novel further improvement to the comprehension of best features to be used. The most encouraging results were observed in the case of microstructured PLLA-TMC films with grooves of 2.5 and 1microm depth, presenting, in particular, a groove width of 50 and 25microm.


Subject(s)
Absorbable Implants , Biocompatible Materials/chemistry , Muscle Fibers, Skeletal/cytology , Muscle Fibers, Skeletal/physiology , Polymers/chemistry , Tissue Engineering/methods , Animals , Cell Culture Techniques/methods , Cell Enlargement , Cell Line , Cell Polarity , Cell Proliferation , Crystallization/methods , Materials Testing , Mice , Photography/methods , Porosity , Surface Properties
8.
J Mater Sci Mater Med ; 21(3): 1005-11, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20012164

ABSTRACT

In bone tissue reconstruction, the use of engineered constructs created by mesenchymal stem cells (MSCs) that differentiate and proliferate into 3D porous scaffolds is an appealing alternative to clinical therapies. Human placenta represents a possible source of MSCs, as it is readily available without invasive procedures and because of the phenotypic plasticity of many of the cell types isolated from this tissue. The scaffold considered in this work is a slowly degradable polyurethane foam (EF PU foam), synthesized and characterized for morphology and in vitro interaction with chorion mesenchymal cells (CMCs). These cells were isolated from human term placenta and cultured onto the EF PU foam using two different culture media (EMEM and NH osteogenic differentiation medium). Synthesized EF PU foam showed homogeneous pore size and distribution, with 89% open porosity. In vitro tests showed CMCs scaffold colonization, as confirmed by Scanning Electron Microscopy (SEM) observations and hematoxylin-eosin staining. Alizarin Red staining revealed the presence of a small amount of calcium deposition for the samples treated with the osteogenic differentiation medium. Therefore, the proposed EF PU foam appears to stimulate cell adhesion in vitro, sustaining CMCs growth and differentiation into the osteogenic lineage.


Subject(s)
Osteogenesis , Placenta/metabolism , Polyurethanes/chemistry , Bone Transplantation/methods , Cell Adhesion , Cell Differentiation , Chorion/chemistry , Chorion/pathology , Culture Media/metabolism , Female , Humans , Imaging, Three-Dimensional , Mesoderm/cytology , Microscopy, Electron, Scanning/methods , Pregnancy , Tomography, X-Ray Computed/methods
9.
Acta Biomater ; 5(4): 1126-36, 2009 May.
Article in English | MEDLINE | ID: mdl-19147418

ABSTRACT

In bone tissue reconstruction, the use of engineered constructs created by mesenchymal stem cells (MSCs) that differentiate and proliferate into three-dimensional porous scaffolds is an appealing alternative to autologous and heterologous bone grafts. Scaffolds considered in this work are represented by polyurethane (PU) foams. Two PU foams (EC-1 and EC-2) were synthesized and characterized for morphology, mechanical properties and in vitro interaction with the osteoblast-like cell line MG63 and MSCs from human bone marrow. EC-1 and EC-2 showed similar densities (0.20 g cm(-3)) with different morphologies: EC-1 showed a more homogeneous pore size (average Phi = 691 microm) and distribution, with a 35% open porosity, whereas EC-2 evidenced a wide range of pore dimension, with an average pore size of 955 microm and a 74% open porosity. The compressive properties of the two foams were similar in the dry condition and both showed a strong decrease in the wet condition. In vitro tests showed good MG63 cell proliferation, as confirmed by the results of the MTT assay and scanning electron microscopy (SEM) observations, with a higher cell viability on EC-2 foam 7 days post-seeding. In the experiments with MSCs, SEM observations showed the presence of an inorganic phase deposition starting day 7 onto EC-1, day 14 onto EC-2. The inorganic particles (CaP) deposition was much more evident onto the pore surface of both foams at day 30, indicating good differentiation of MSCs into osteoblasts. Both PU foams therefore appeared to stimulate cell adhesion and proliferation in vitro, sustaining the MSCs' growth and differentiation into osteoblasts.


Subject(s)
Cell Differentiation , Mesenchymal Stem Cells/cytology , Osteoblasts/cytology , Polyurethanes , Cell Line , Cell Proliferation , Humans , Microscopy, Electron, Scanning , Stress, Mechanical
10.
J Biomed Mater Res A ; 87(3): 770-9, 2008 Dec 01.
Article in English | MEDLINE | ID: mdl-18200541

ABSTRACT

New porous scaffolds, with a suitable hydrolytic and enzymatic degradation, useful for tissue engineering applications have been obtained by a carbodiimide mediated reaction between hyaluronan (HA) and a synthetic polymer with a polyaminoacid structure such as alpha,beta-polyaspartylhydrazide (PAHy). Scaffolds with a different molar ratio between PAHy repeating units and HA repeating units have been prepared and characterized from a chemical and physicochemical point of view. Tests of indirect and direct cytotoxicity, cell adhesion, and spreading on these biomaterials have been performed by using murine L929 fibroblasts. The new biomaterials showed a good cell compatibility and ability to allow cell migration into the scaffolds as well as spreading on their surface.


Subject(s)
Hyaluronic Acid/chemistry , Peptides/chemistry , Tissue Engineering , Tissue Scaffolds/chemistry , Animals , Cell Adhesion , Cell Movement , Cells, Cultured , Compressive Strength , Materials Testing , Mice , Polymers
11.
J Appl Biomater Biomech ; 6(3): 132-43, 2008.
Article in English | MEDLINE | ID: mdl-20740457

ABSTRACT

It is well known that surface features, such as topographical or chemical cues, can affect cell behavior from initial attachment and migration to differentiation and production of new tissues; this phenomenon is called contact guidance. A great improvement in studies concerning this phenomenon comes from progress in microfabrication techniques such as photolithography or soft lithography. Due to these techniques, a wide variety of micro-patterned surfaces can be realized to control cell size, shape, spatial organization and proliferation. These studies promoted the development of cellular bioassays that provide entirely new insights into the factors that control cell adhesion, proliferation and differentiation onto material surfaces and molecular signaling pathways. The ability to control shape and spreading is also important to direct stem cells to different specific lineages, but it is also of great importance for the design of cell culture substrates for tissue engineering. In this work, the possibility of patterning surfaces is investigated, with particular focus on the micrometric scale. The response of different types of cells is also investigated.

12.
Biomed Pharmacother ; 60(8): 386-92, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16930938

ABSTRACT

A non-porous poly-DL-lactide tubular chamber filled by demineralised bone matrix (DBM) and bone marrow stromal cells (BMSC) in combination, was evaluated as a scaffold for guided bone regeneration (GBR) in an experimental model using the rabbit radius. The tubular chamber had an internal diameter of 4.7 mm, a wall thickness of 0.4 mm and a length of 18 mm. Autologous BMSC were obtained, under general anaesthesia from rabbit iliac crest and isolated by centrifugation technique. Allogenic DBM was obtained from cortico-cancellous bone of rabbits. In general anaesthesia, a 10-mm defect was bilaterally created in the radii of 10 rabbits. On the right side (experimental side) the defect was bridged with the chamber filled with both BMSC and DBM. On the left side (control side) the defect was treated by positioning DBM and BMSC between the two stumps. At an experimental time of 4 months histology and histomorphometry demonstrated that the presence of a tubular chamber significantly improved bone regrowth in the defect The mean thickness of newly-formed bone inside the chamber was about 56.7+/-3.74% of the normal radial cortex, in comparison with 46.7+/-10.7% when DBM and BMSC without the chamber were placed in the defect, P<0.05). These results confirmed the effectiveness of the chamber as a container for factors promoting bone regeneration.


Subject(s)
Absorbable Implants , Bone Marrow Cells/cytology , Bone Matrix/cytology , Bone Regeneration , Polyesters/chemistry , Animals , Bone Demineralization Technique , Male , Rabbits , Radius , Stromal Cells/cytology
13.
J Appl Biomater Biomech ; 4(3): 165-71, 2006.
Article in English | MEDLINE | ID: mdl-20799202

ABSTRACT

The long-term survival of total joint prostheses is influenced by many factors. Among these factors, the most critical one is the presence of wear debris, particularly from the joint articulating surfaces. While ultra high molecular weight polyethylene (UHMWPE) has shown to be extremely promising in vitro and short-term clinical results, shelf-aged, oxidized components have been extremely unsuccessful clinically. Although shelf-aged components have been frequently tested in the laboratory, few studies have compared the properties of clinical retrievals to shelf-aged UHMWPE components. In this study, a thorough analysis method was developed and applied to both UHMWPE components retrieved at the time of revision, and non-implanted, shelf-aged UHMWPE components with the aim to understand better the significance of the in vivo factors on the material properties of the retrievals. The UHMWPE components were analyzed and characterized by visual observation of the surfaces, mechanical properties were assessed by small punch tests, wear resistance was analyzed by a multidirectional pin-on-disk test, and the oxidation level was detected by the Fourier transformed infrared (FT-IR) technique. The results indicated a strong correlation between the UHMWPE oxidation level, mechanical properties and wear resistance.

14.
Int J Artif Organs ; 28(7): 701-10, 2005 Jul.
Article in English | MEDLINE | ID: mdl-16049904

ABSTRACT

In the present study, twelve explanted mechanical heart valves (MHVs)with pyrolitic carbon tilting disc and 14 bileaflet MHVs were analyzed to investigate the effects of material properties on valve performance and patients' general health conditions. Optical and scanning electron microscopy was used to investigate material imperfections, wear patterns or damages to housing and occluder components. All analyzed tilting disc valves exhibited wear effects, particularly due to abrasion and impact to both disc and housing. Wear of pyrolitic carbon disc and housing did not influence their in vivo performance. In the bileaflet MHVs, breakaway of the pyrolitic carbon coating sometimes caused malfunctioning and required surgical retrieval of the valve. In all cases, occurrence of clinical symptoms was more likely when wear effects were located in critical areas. The study supports a correlation between the properties of the MHVs material and patients' symptoms.


Subject(s)
Aortic Valve , Heart Valve Prosthesis , Mitral Valve , Prosthesis Failure , Adult , Aged , Blood Pressure , Carbon , Child , Coated Materials, Biocompatible , Device Removal , Equipment Failure Analysis , Female , Humans , Male , Middle Aged , Stress, Mechanical , Surface Properties
16.
J Appl Biomater Biomech ; 1(1): 58-66, 2003.
Article in English | MEDLINE | ID: mdl-20803473

ABSTRACT

This work reports preliminary results on the development of biointegrable scaffolds, composed of biostable 3D polymer matrices and bioabsorbable inorganic salts, to be used for cell anchorage in bone regeneration. Three crosslinked polyurethane foams (PUFs), prepared by one-step bulk polymerisation from a polyether-polyol mixture, polymeric MDI and water as expanding agent, were tested for their ability to promote adhesion and growth of bone-derived cells. The open porosity of these foams ranged from 16 to 31% with an average pore size of 470 /600 microm, compressive strength (at 10% ε ) of 0.28/0.38 MPa and elastic moduli of 4.88/6.61 MPa. The human osteosarcoma line Saos-2, and primary cultures of normal human articular chondrocytes and bone marrow-derived (HBM) stromal cells were used for in vitro cytocompatibility tests. For cell adhesion and proliferation analysis, DNA synthesis was evaluated by 3 H-thymidine uptake. Osteoblastic differentiation of Saos-2 adherent cells was determined by measuring the enzymatic activity of alkaline phosphatase (ALP). All cell types were able to adhere to all tested PUFs and to synthesize DNA. At 48 hr culture, HBM stromal cells showed the maximal rate of adhesion with the highest rate of proliferation onto PUFs with the largest pore size, whereas both chondrocytes and Saos-2 appeared to adhere preferentially onto foams exhibiting the highest percentage of open porosity. Up to 8 days in culture Saos-2 cells were able to proliferate into all PUFs, with a time-dependent increase of DNA synthesis and ALP activity. At SEM, the morphology of cells adherent to PUF pores was spread with cytoplasmatic extroflessions, indicating a good metabolic activation. These results demonstrate a good cytocompatibility of the proposed 3D matrices, suggesting that their use in the preparation of composite scaffolds is worth further investigation. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 58-66)ABSTRACT: This work reports preliminary results on the development of biointegrable scaffolds, composed of biostable 3D polymer matrices and bioabsorbable inorganic salts, to be used for cell anchorage in bone regeneration. Three crosslinked polyurethane foams (PUFs), prepared by one-step bulk polymerisation from a polyether-polyol mixture, polymeric MDI and water as expanding agent, were tested for their ability to promote adhesion and growth of bone-derived cells. The open porosity of these foams ranged from 16 to 31% with an average pore size of 470 /600 microm, compressive strength (at 10% ε ) of 0.28/0.38 MPa and elastic moduli of 4.88/6.61 MPa. The human osteosarcoma line Saos-2, and primary cultures of normal human articular chondrocytes and bone marrow-derived (HBM) stromal cells were used for in vitro cytocompatibility tests. For cell adhesion and proliferation analysis, DNA synthesis was evaluated by 3 H-thymidine uptake. Osteoblastic differentiation of Saos-2 adherent cells was determined by measuring the enzymatic activity of alkaline phosphatase (ALP). All cell types were able to adhere to all tested PUFs and to synthesize DNA. At 48 hr culture, HBM stromal cells showed the maximal rate of adhesion with the highest rate of proliferation onto PUFs with the largest pore size, whereas both chondrocytes and Saos-2 appeared to adhere preferentially onto foams exhibiting the highest percentage of open porosity. Up to 8 days in culture Saos-2 cells were able to proliferate into all PUFs, with a time-dependent increase of DNA synthesis and ALP activity. At SEM, the morphology of cells adherent to PUF pores was spread with cytoplasmatic extroflessions, indicating a good metabolic activation. These results demonstrate a good cytocompatibility of the proposed 3D matrices, suggesting that their use in the preparation of composite scaffolds is worth further investigation. (Journal of Applied Biomaterials & Biomechanics 2003; 1: 58-66).

17.
J Mater Sci Mater Med ; 14(8): 683-6, 2003 Aug.
Article in English | MEDLINE | ID: mdl-15348408

ABSTRACT

Due to their similarity to natural soft tissues, water-swellable polymeric materials (hydrogels) are, in principle, ideal candidates for scaffolds/matrices in tissue engineering. Polyurethanes (PU), hydrophilic but water-insoluble, can be obtained by the incorporation of hydrophilic soft segments, e.g. poly(ethylene oxide) (PEO). These materials possess the favorable characteristics of the family of PUs as well as the ability to mimic soft tissues. In this work, new crosslinked PU-hydrogels were prepared in a one-step bulk polymerization process using an aliphatic diisocyanate, PEO, a low molecular weight diol, and a tri-functional crosslinking agent. A porous structure was also obtained by air-incorporation under mechanical stirring at a controlled high speed during the polymerization. Structural characteristics of the compact (PU-HyC) and the porous (PU-HyP) material were investigated. Molecular weight between cross-links, M(c), and crosslinking density, rho(x), were typical of a low crosslinking degree. A homogeneous distribution of non-interconnecting pores (phi100 microm) was observed in PU-HyP. Both materials showed a high water adsorption. The swelling behavior and weight loss in water was affected by porosity. For their mechanical behavior in the swollen state, the novel PU hydrogels can be considered for biomedical applications where good mechanical properties are required (i.e. 3D scaffold for tissue engineering).

18.
J Biomater Appl ; 14(4): 325-48, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10794506

ABSTRACT

The in vitro structural stability of poly-ether-urethanes (PEUs) and poly-carbonate-urethanes (PCUs) was examined under strong acidic (HNO3) or alkaline (NaClO) oxidative conditions and in presence of a constant strain state. Polyurethane (PU) samples were represented by sheets solvent-cast from commercial pellets or by tubular specimens cut from commercial catheters. The specimens were strained at 100% uniaxial elongation over appropriate extension devices and completely immersed into the oxidative solutions at 50 degrees C for 7-14 days. The changes induced by the oxidative treatments were then evaluated by molecular weight analysis, tensile mechanical tests, and scanning electron microscopy. In the experiments with solvent-cast samples, the PEU Pellethane was degraded more in the alkaline oxidative conditions and mainly in the absence of an applied uniaxial stress. All the tested PCUs were, on the contrary, more affected by the acidic oxidative agent. All the PCUs proved to have overall better stability than the PEU. The susceptibility to oxidation was also dependent on the shape and bulk/surface organisation acquired by the same polymer during its processing. When the oxidative test was applied to catheters made of a PEU and a PCU, the results confirmed the better stability of poly-carbonate-urethanes.


Subject(s)
Biocompatible Materials/chemistry , Polycarboxylate Cement/chemistry , Polyurethanes/chemistry , Catheterization , Drug Stability , Molecular Weight , Oxidation-Reduction , Stress, Mechanical , Surface Properties , Tensile Strength
19.
J Biomed Mater Res ; 45(1): 62-74, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10397959

ABSTRACT

The in vitro structural stability of polyetherurethanes (PEUs) and polycarbonateurethanes (PCUs and PCUUs) was examined under strong oxidative conditions (0.5N HNO3, pH 0.3; and NaClO, 4% Cl2 available, pH approximately 13) and in the presence of a constant strain state. Solvent-cast dog-bone shaped specimens were strained at 100% uniaxial elongation over extension devices and completely immersed in the oxidative solutions at 50 degrees C for 15 days. Unstrained polyurethane (PU) samples were treated in the same way for comparison. The modification of the PU molecular structure was determined by DSC, GPC, ATR-FTIR, static contact angle, and surface roughness analyses. The incubation in nitric acid and sodium hypochlorite brought about a greater degradation of samples tested under the applied strain with the exception of PEU treated with nitric acid. PEU was the most affected material, showing bulk deterioration in NaClO and significant modifications in nitric acid, with the appearance of new IR bands, which were assigned to oxidation products. A higher phase separation between soft and hard domains occurred in PCUs upon incubation in nitric acid, the treatment with NaClO gave rise to new bands in the IR spectra, denoting the presence of oxidation products at the surface. The surface roughness greatly increased in strained PCUs with SEM evidence of deep cracks and holes or ragged and stretched fractures perpendicular to the direction of stress. PCUU underwent complex chemical modifications with a marked decrease of N-H and urea IR absorptions and showed a lower degradation than PEU and PCUs under mechanical constraint. From these results, sodium hypochlorite appears to be able to create an ESC-like degradation for PUs that are resistant to other aggressive chemical environments.


Subject(s)
Polyurethanes/chemistry , Microscopy, Electron, Scanning , Molecular Weight , Nitric Acid/chemistry , Oxidants/chemistry , Oxidation-Reduction , Sodium Hypochlorite/chemistry , Spectrophotometry, Infrared , Spectroscopy, Fourier Transform Infrared , Stress, Mechanical , Surface Properties , Tensile Strength , Time Factors
20.
J Mater Sci Mater Med ; 10(12): 711-4, 1999 Dec.
Article in English | MEDLINE | ID: mdl-15347938

ABSTRACT

Several polyurethane-maleamides (PUMAs) containing polyether or polycarbonate soft segments, and aromatic or aliphatic hard segments were synthesized by solution or bulk polymerization, using maleic acid (MA) or a mixture of MA and butanediol as chain extenders. Using this process, activated double bonds are introduced into the polymer chains and the base polyurethanes may undergo further modification via specific grafting, thus improving their tissue compatibility. PUMAs chemicophysical properties were evaluated by gel permeation chromatography (GPC), intrinsic viscosity analyses, differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FT-IR) and tensile mechanical tests. Polycarbonate diol (PCU)-based PUMAs showed higher molecular weights than polyether diol (PEU)-based ones. The use of butanediol in mixture with maleic acid led to an increase of molecular weights. FT-IR confirmed the presence of the bands related to the amide groups and to the conjugated double bond, yet more evident for the polymer obtained in solution. The higher crystallinity shown by this polymer was also indicative of a better phase separation. All the PCU-PUMAs exhibited similar tensile properties with a higher stiffness than PEU-PUMAs. Among the PEU-PUMAs, the highest tensile properties were shown by the polymer obtained in solution, and by the one derived from a mixture of maleic acid and butanediol.

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