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Biomedical subjects

Silvano Lora

Publications and source records attributed to Silvano Lora.

10 recordsLinked to original sources

Electrospun polyphosphazene nanofibers for in vitro rat endothelial cells proliferation.

A large variety of natural and synthetic polymers have been explored as scaffolds for the seeding and growth of different types of cells. To fabricate a scaffold that can be used as a synthetic extracellular matrix (ECM), it is important to replicate the nanoscale dimensions of natural ECM. The electrospinning process allows to produce ultrathin fibers so that this method represents a suitable approach to scaffold fabrication for tissue engineering applications. In this work, the feasibility of obtaining flat or tubular matrices from biocompatible poly[(ethyl phenylalanato)(1.4) (ethyl glycinato)(0.6) phosphazene] by electrospinning was evaluated and the effect of process parameters on the diameter of nanofibers was examined. The adhesion and growth of rat neuromicrovascular endothelial cells cultured on sheets and tubes composed by the polymer with an average fiber diameter of 850 +/- 150 nm were also reported. Microscopic examination of the seeded tubes demonstrated that, after 16 days of incubation, endothelial cells formed a monolayer on the whole surface. These results are the first step to demonstrate that tubes of biodegradable polyphosphazenes might be a feasible model to construct human tissues such as vessels or cardiac valves.

Animals↗

Electrochemical, pulsed-field-gradient spin-echo NMR spectroscopic, and ESR spectroscopic study of the diffusivity of molecular probes inside gel-type cross-linked polymers.

The permeability of five gel-type synthetic resins, obtained by polymerization of 1-vinylpyrrolidin-2-one cross-linked with N,N'-methylenebisacrylamide (1, 2, 3, 4, and 5 wt %) and swollen by N,N-dimethylformamide (DMF), has been analyzed. The diffusion of 2,2,6,6-tetramethyl-4-oxo-1-piperidinyloxyl (TEMPONE) was studied by ultramicroelectrode voltammetry. Similar measurements were performed for solutions of non-cross-linked poly(vinylpyrrolidone) in DMF. To provide information on the rotational mobility of TEMPONE and the translational mobility of DMF, electron spin resonance (ESR) spectroscopic and pulsed-field-gradient spin-echo nuclear magnetic resonance (PGSE-NMR) spectroscopic experiments, respectively, were carried out. Comparative analysis of the results obtained by electrochemical, ESR spectroscopic, and PGSE-NMR spectroscopic measurements showed that diffusivity inside the polymer framework is significantly affected by the extent of cross-linking, the size of the diffusing probe, and the presence of electrolytes.

Journal Article↗

Bone marrow stromal cells and resorbable collagen guidance tubes enhance sciatic nerve regeneration in mice.

We evaluated peripheral nerve regeneration using a tubular nerve guide of resorbable collagen filled with either bone marrow-derived cells (BMDCs) in Dulbecco's cell culture medium (DMEM) or with DMEM alone (control). The control group received just the culture medium (vehicle). The left sciatic nerves of ten isogenic mice were transected and the tubular nerve guides were sutured to the end of the proximal and distal nerve stumps. Motor function was tested at 2, 4 and 6 weeks after surgery using the walking track test. The pawprints were analyzed and the print lengths (PL) were measured to evaluate functional recovery. After 6 weeks, mice were anesthetized, perfused transcardially with fixative containing aldehydes, and the sciatic nerves and tubes were dissected and processed for scanning and transmission electron microscopy. Scanning electron microscopy of the collagen tube revealed that the tube wall became progressively thinner after surgery, proving that the tube can be resorbed in vivo. Quantitative analysis of the regenerating nerves showed that the number of myelinated fibers and the myelin area were significantly increased in the experimental group. Also, motor function recovery was faster in animals that received the cell grafts. These results indicate that the collagen tube filled with BMDCs provided an adequate and favorable environment for the growth and myelination of regenerating axons compared to the collagen tube alone.

Animals↗

In vitro evaluation of poly[bis(ethyl alanato)phosphazene] as a scaffold for bone tissue engineering.

Polyphosphazenes with amino acid ester as side groups are biocompatible polymers that could provide valid scaffolds for cell growth. In the present study we investigate the adhesion and growth of osteoblasts obtained from rat bone marrow on matrices composed of thin fibers of poly[bis(ethyl alanato)phosphazene] (PAlaP), poly(d,l-lactic acid) (PDLLA), or PAlaP/PDLLA blend. Our data show that scaffolds of PAlaP or PAlaP/PDLLA blend enhanced the cell adhesion and growth in comparison with that observed in cultures seeded on polystyrene tissue culture plates. Although collagenase-digestible protein synthesis remained unchanged, all scaffolds induced a decrease in alkaline phosphatase activity, suggesting that osteoblasts are in the proliferation phase. Both PAlaP and PAlaP blended with PDLLA may represent a new and interesting substrate for bone tissue engineering.

Alkaline Phosphatase↗

Gel-type polyacrylic resins cross-linked with trimethylolpropanetrimethacrylate: the issue of their nanostructure and molecular accessibility unveiled with a combination of inverse steric exclusion chromatography (ISEC), and ESR and CP-MAS 13C NMR spectroscopy.

Six gel-type functional resins, that is, three poly-DMAA-co-TMPTP (DMAA = N,N-dimethylacrylamide, TMPTP = trimethylolpropyltrimethacrylate) samples with different degrees of cross-linking (0.6, 1.2, 1.7 % mol) and three poly-DMAA-co-MA-co-TMPTP (MA = methacrylic acid, ca. 5.5 % mol) samples with 1.7, 3.5, and 7 % mol cross-linking were investigated with ISEC (inverse steric exclusion chromatography), and ESR and CP-MAS (cross polarization magic angle spinning) 13C NMR spectroscopy after swelling in water and other solvents. This unprecedented combination of conceptually independent physicochemical techniques provides a thorough overall consistent picture of the morphology of the resins on the nanometer scale and of the molecular accessibility of the swollen polymer framework to the paramagnetic probe TEMPONE (2,2,6,6-tetramethyl-4-oxo-1-oxypiperidine) and to selected solvents.

Journal Article↗

In vitro culture of rat neuromicrovascular endothelial cells on polymeric scaffolds.

Polyphosphazenes are polymers possessing a skeleton composed of alternating phosphorous and nitrogen atoms, and two side-moieties linked to each phosphorous atom. Polyphosphazenes with amino acid esters as side-moieties are biocompatible and biodegradable polymers. Two polyphosphazenes, poly[bis(ethyl alanate) phosphazene] and poly[(ethyl phenylalanate)0.8(ethyl alanate)0.8(ethyl glycinate)0.4 phosphazene] (PPAGP) were synthesized, and processed to form small fibers. Their ability to support rat neuromicrovascular endothelial cell (EC) adhesion and growth has been studied, using poly(D,L-lactic acid) as reference compound. Scanning electron microscopy revealed that both poly[bis(ethyl alanate) phosphazene] and PPAGP fibers were thinner than poly(D,L-lactic acid) fibers, and possessed a more irregular and porous surface. All polymers increased EC adhesion, compared with polystyrene, but only polyphosphazenes were able to improve EC growth. The highest increase in EC proliferation was induced by PPAGP, which, as revealed by environmental scanning electron microscopy, was also able to induce ECs to arrange into tubular structures. The conclusion is drawn that PPAGP may provide the best scaffold for engineered blood vessels, because it promotes adhesion, growth, and organization of ECs into capillary-like structures.

Animals↗

Nanomorphology of polymer frameworks and their role as templates for generating size-controlled metal nanoclusters.

The microporous (gel-type) functional resin co-poly-N,N-dimethylacrylamide (DMAA) (88 % mol)/methacrylic acid (MAA) (8 % mol)/N,N'-methylenebisacrylamide (MBAA) (4 % mol) (MPIF(H)) is employed as the hosting framework for the production of resin-supported Pd(0) nanoclusters. The obtained composite MPIF(-)Na(+)/Pd(0) is prepared upon reducing, in ethanol, MPIF(-)Pd(2+) (0.5), obtained upon previous homogeneous dispersion of "Pd(2+)" inside the resin particles (XRMA control) through ion-exchange. Metal nanoclusters appear to be size-controlled (2.0+/-0.2 nm) and are seen to reasonably fit the predominant resin "nanopores" diameter, determined in ethanol (3.2 nm) by means of inverse steric exclusion chromatography (ISEC).

Journal Article↗

Metal catalysis inside polymer frameworks: evaluation of catalyst stability and reusability.

The polymer framework of a resin-based catalyst built up with Pd nanoclusters (ca. 3 nm) dispersed inside the nanoporous domains of a thermally stable gel-type polyacrylic resin exhibits a good chemical stability under 5 bar H(2) at 40 degrees C for reasonable contact times. Chemical and physico-chemical integrity of the polymer framework are checked with a variety of instrumental analytical methods. Catalyst reusability turns out to be quite good.

Journal Article↗

A new absorbable pressure-equalizing tube.

OBJECTIVE: We investigated pressure-equalizing (PE) tubes made of biodegradable, absorbable material in an animal model. METHODS: PE tubes, made of poly-bis(ethylanate)phosphazene (PBE) were inserted in 55 ears of 28 Hartley guinea pigs, with survival times of 10, 30, and 60 days after tube insertion. In vivo reactions between the PBE-PE tube and the tympanic membrane (TM) were studied. Tubes, TMs, and middle ears were examined by scanning electron microscopy and light microscopy. RESULTS: There was neither infection nor an inflammatory reaction to the tube within the middle ear in any animal. At 30 days, 53% of the tubes had disintegrated. At 60 days, tubes were still functioning in the 25% of ears. CONCLUSION: More research must be performed before these new PBE PE tubes can be considered for clinical use. Nonetheless, these tubes are promising. The disintegration rate can be controlled by varying the formulation of the polymer, so treatment can be adjusted to the needs of each patient.

Absorbable Implants↗