Chondrocyte-alginate bioconstructs: a nuclear magnetic resonance relaxation study.

Proton nuclear magnetic resonance (NMR) relaxometry can give informations about hydrogel scaffold properties. As these properties can be modified with culture time and conditions according to scaffold biodegradability and new tissue biosynthesis, the aim of this research was to test the efficiency of this noninvasive NMR technique in the follow-up of 3D cultures for tissue engineering. The distributions of proton relaxation times T1 and T2 have been measured on cylindrical gel samples of different types of alginate, in the presence or absence of hyaluronate, in gels or bioconstructs with encapsulated chondrocytes cultured for 30 days in normal or reduced weight conditions. It was found that T2 increases with the mannuronate/guluronate ratio in alginate samples and with the presence of hyaluronate. The distributions of both T1 and T2 result wider for bioconstructs cultured in normal gravity than for those cultured in reduced weight conditions. Neither cell growing nor collagen production but only GAG neosynthesis have been demonstrated in our experimental conditions. In conclusion, T2 is sensitive to the gel properties (possibly to the rigidity of macromolecular components). The homogeneity of bioconstructs can be monitored by the distribution of T1 and T2. We propose that nonspatially resolved NMR relaxometry can efficiently be used in monitoring tissue development in a biodegradable scaffold for tissue engineering.

NMR relaxometry and imaging of water absorbed in biodegradable polymer scaffolds.

Porous substrates made of poly(3-hydroxybutyrate-3-hydroxyvalerate) (PHBHV) were prepared by a particulate leaching method. After removing the salt by extraction in water, proton nuclear magnetic resonance (NMR) relaxometry and imaging were performed on sets of PHBHV substrates immersed in phosphate-buffered solution during 3 months at different time points. Polarized optical microscopy studies were performed on thin sections, 25 and 5 mum, of the PHBHV samples. The results of NMR relaxometry showed two (1)H nuclei populations, well distinguishable on the free induction decay (FID), due to the different decay time constants, a factor of 10(2) apart. Thus, it was possible to separate the two populations, giving separate distributions of T(1) relaxation times. One population could be associated with water protons in the pores and the other to macromolecular protons. The distributions of T(1) and T(2) of the water proton shifted to lower values with increasing immersion time to a constant value after 30 days. The results obtained by NMR imaging showed an initial increase in the apparent porosity, reaching a plateau after 25 days of immersion. This increase is attributed mainly to the absorption of water in the microporosity as supported by the results of the relaxometry measurements and shown by scanning electron microscopy. The average porosity measured by NMR imaging at the plateau, 78+/-3%, is slightly higher than that determined by optical microscopy, 73+/-9%, which may be due to the fact that the latter method did not resolve the microporosity. Overall, the results suggest that at early stages after immersing the scaffolds in the aqueous medium, first 30 days approximately, NMR imaging could underestimate the porosity of the substrate.

Magnetic resonance imaging and relaxation analysis to predict noninvasively and nondestructively salt-to-moisture ratios in dry-cured meat.

The current systems are unable to control and predict the cured meat composition nondestructively and in a reasonable time for production needs. In this work, T1 and T2 maps were obtained, with a monoexponential model, on internal sections of Longissimus dorsi muscle at increasing salting times. The maps allow one to visualize the salting process nondestructively and noninvasively. The method goes beyond the simple qualitative visualization, because, for each section of the sample and in any region of the section, it is possible to obtain quantitative information on the progress of salting and to predict salt-to-moisture ratios. In addition, detailed relaxation measurements were performed on samples cored after imaging in order to define better the relaxation properties of the dry-cured meat.

Preliminary 1H NMR study on archaeological waterlogged wood.

Magnetic Resonance Relaxation (MRR) and Magnetic Resonance Imaging (MRI) are powerful tools to obtain detailed information on the pore space structure that one is unlikely to obtain in other ways. These techniques are particularly suitable for Cultural Heritage materials, because they use water 1H nuclei as a probe. Interaction with water is one of the main causes of deterioration of materials. Porous structure in wood, for example, favours the penetration of water, which can carry polluting substances and promote mould growth. A particular case is waterlogged wood from underwater discoveries and moist sites; in fact, these finds are very fragile because of chemical, physical and biological decay from the long contact with the water. When wood artefacts are brought to the surface and directly dried in air, there is the collapse of the cellular structures, and wood loses its original form and dimensions and cannot be used for study and museum exhibits. In this work we have undertaken the study of some wood finds coming from Ercolano's harbour by MRR and MRI under different conditions, and we have obtained a characterization of pore space in wood and images of the spatial distribution of the confined water in the wood.