Polyhydroxyalkanoate (PHBV) fibers obtained by a wet spinning method: Good in vitro cytocompatibility but absence of in vivo biocompatibility when used as a bone graft.

Polyhydroxyalkanoates (PHAs) are biomaterials widely investigated for tissue-engineering applications. In this regard, we describe a method to prepare fibers of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by a wet-spinning technique. Polymer fibers were used to test the cytocompatibility of the material in vitro. We have investigated their behavior in vitro in presence of the osteoblast-like (SaOs2) and macrophage (J774.2) cell lines. The PHBV fibers used were 100-200µm in diameter and offered a large surface for cell adhesion, similar to that they encounter when apposed onto a bone trabeculae. The fiber surface possessed a suitable roughness, a factor known to favor the adherence of cells, particularly osteoblasts. PHBV fibers were degraded in vitro by J774.2 cells as erosion pits were observable by transmission electron microscopy. The fibers were also colonisable by SaOs2 cells, which can spread and develop onto their surface. However, despite this good cytocompatibility observed in vitro, implantation in a bone defect drilled in rabbit femoral condyles showed that the material was only biotolerated without any sign of osteoconduction or degradation in vivo. We can conclude that PHBV is cytocompatible but is not suitable to be used as a bone graft as it does not favor osteoconduction and is not resorbed by bone marrow macrophages.

Effects of the length of crosslink chain on poly(2-hydroxyethyl methacrylate) (pHEMA) swelling and biomechanical properties.

Polymers are widely used in medicine for vascular prostheses, bone substitutes, and devices for controlled release. Among these polymers, poly(2-hydroxyethyl methacrylate) (pHEMA) is the most employed. To confer particular properties, pHEMA can be copolymerized with other monomers or in the presence of plasticizers or crosslinking agents. The influence of the length of crosslink chains on swelling, surface rugosity, hardness, and stiffness of crosslinked pHEMA were studied by several techniques, including fractal analysis and AFM. Four crosslinking agents (divinyl benzene, DVB; ethylene glycol dimethacrylate, EGDMA; tetraethylene glycol diacrylate, TEGDA; and polyethylene glycol diacrylate, PEGDA) were added to the bulk polymerization mixture. Only linear and PEGDA-pHEMA presented a significant decrease in surface roughness confirmed by fractal analysis. Differences in hardness and biomechanical properties were evidenced on dried polymers but the highest differences were exhibited for hydrated pHEMA. Correlations between the length of the crosslink chain and hardness or stiffness of hydrated crosslinked pHEMA were evidenced. TEGDA and PEGDA appeared to be the two most suitable crosslinking agents for controlled release of bioactive molecules in bone.

Synthesis of methacryloyloxyethyl phosphate copolymers and in vitro calcification capacity.

Methacryloyloxyethyl phosphate is a methacrylic monomer used to modify different substrates by copolymerisation, in order to enhance hydroxyapatite deposition onto their surfaces. We report the synthesis of two copolymers series using increasing concentrations of methacryloyloxyethyl phosphate with (diethylamino) ethyl methacrylate and 1-vinyl-2-pyrrolidinone. Reactivity ratios were evaluated for the two copolymer systems. The influence of phosphate content and distribution on the capacity to form a calcium-rich layer was evaluated after immersion for 15 days in a synthetic body fluid. Corresponding homopolymers were synthesised as controls. Calcium-phosphorus globules were developed only on samples containing (diethylamino) ethyl methacrylate, and presenting a low density of phosphate groups. The amounts of calcium increased when higher concentrations of methacryloyloxyethyl phosphate were used. The use of 1-vinyl-2-pyrrolidinone was associated with greater calcium amounts, (compared to (diethylamino) ethyl methacrylate). The amine groups may favour the attraction of phosphorus, thus creating another way for the nucleation of calcium/phosphate crystals.

The in vivo calcification capacity of a copolymer, based on methacryloyloxyethyl phosphate, does not favor osteoconduction.

Polymers can be interesting alternatives to bone grafts; they must present suitable mechanical and osteoconductive properties. Biomimetic properties may be a key factor for the recognition by bone cells. Methacryloyloxyethyl phosphate (MOEP) was found to enhance hydroxyapatite deposition. The copolymer containing MOEP and 1-vinyl-2-pyrrolidinone (50-50%) binds large amounts of calcium. Particles of the copolymer were used to fill large cranial bone defects in the rat. After a 12-week healing period, the animals were euthanized and the skulls examined by X-ray, histology, and electron microscopy (EM). The high phosphate content of the polymer conferred a marked calcium-binding capacity, and the particles were heavily calcified. They were embedded in a light fibrous stroma containing numerous capillaries and multinucleated giant cells. The osteoconductive properties were poor: only few trabeculae developed centripetally from the margins of the defects. There was no bone bonding and no osteoblast on the surface of the calcified material. Backscattered EM revealed that the degree of calcification was homogeneous in all particles. Calcium-phosphorus calcospherites were never observed. The material appeared to trap calcium but to impair nucleation because only small hydroxyapatite tablets were occasionally observed. Polyphosphated materials do not represent a suitable source of potentially usable bone substitutes.

Cushing's Syndrome : an overview.

Cushing's syndrome is the collection of symptoms and signs caused by chronic elevation of circulating free cortisol. Since the first description of the syndrome, results from long-term exposure to glucocorticoids has not been improved upon, but our understanding of its pathophysiologic features and our ability to diagnose and treat the disorder have increased dramatically. In this article we review the current literature about Cushing's syndrome.

Non-connected versus interconnected macroporosity in poly(2-hydroxyethyl methacrylate) polymers. An X-ray microtomographic and histomorphometric study.

Poly(2-hydroxyethyl methacrylate) (pHEMA) has potentially broad biomedical applications: it is biocompatible and has a hardness comparable to bone when bulk polymerized. Porous biomaterials allow bone integration to be increased, especially when the pores are interconnected. In this study, three types of porogens (sugar fibers, sucrose crystals, and urea beads) have been used to prepare macroporous pHEMA. The pore volume and interconnectivity parameters of the porosity were measured by X-ray microtomography and image analysis. Sucrose crystals, having a high volumetric mass, gave large pores that were located on the block sides. Urea beads and sugar fibers provided pores with the same star volume (2.65 +/- 0.46 mm3 and 2.48 +/- 0.52 mm3, respectively) but which differed in interconnectivity index, fractal dimension, and Euler-Poincarés number. Urea beads caused non-connected porosity, while sugar fibers created a dense labyrinth within the polymer. Interconnectivity was proved by carrying out surface treatment of the pHEMA (carboxymethylation in water), followed by von Kossà staining, which detected the carboxylic groups. Carboxymethylated surfaces were observed on the sides of the blocks and on the opened or interconnected pores. The disconnected pores were unstained. Macroporous polymers can be prepared with water-soluble porogens. X-ray microtomography appears a useful tool to measure porosity and interconnectedness.

Modification of some properties of polyamide-6 by electron beam induced grafting

Electron irradiation, applied on its own or in combination with other physical and chemical treatments, can result in radiation damage to materials. One category of such pursuits is electron beam radiation processing technologies, the intent being to produce favourable modification of materials. Study has been made herein of the use of accelerator-generated electron beams for the induced grafting of acrylic and methacrylic acids and acrylamide onto polyamide-6, the latter being popularly used in the textiles industry. Samples of polyamide-6, in the form of rectangular sheets of dimension, 10 cm x 10 cm or granules, have been irradiated using a 3 GHz travelling wave electron accelerator of energy 6.5 MeV. Results show that the degree of grafting is dependent upon radiation dose and extent of monomers dilution. Modifications have been observed in the melting temperature of the crystalline phase, glass transition temperature, mechanical parameters and the reticulation of polyamide.