Vibrational spectroscopic characterization of new calcium phosphate bioactive coatings.

In this work calcium phosphate (CaP) compounds with different PO(3-)(4)/HPO(2-)(4) R molar ratios in the 0.65-149 range were synthesized. In fact, all these CaPs contain different amounts of HPO(2-)(4) and PO(3-)(4) ions as well as the amorphous precursors (tricalcium phosphate and octacalcium phosphate) of hydroxyapatite deposition, which was shown by in vitro and in vivo measurements. Spectroscopical IR and Raman results showed the presence of bands whose intensity ratio can be related to the molar ratio R; in particular, the Raman I(962)/I(987) and the IR I(1035)/I(1125) intensity ratios were characterized as markers of the molar ratio. For these CaP compounds a nucleation model, which was based on the ability of HPO(2-)(4) ions to form strong H bonds with PO(3-)(4) ions, was proposed.

Raman and solid state 13C-NMR investigation of the structure of the 1 : 1 amorphous piroxicam : beta-cyclodextrin inclusion compound.

The results of a Raman and solid state 13C-NMR spectroscopic investigation aimed at studying the conformation of piroxicam (P) and its interaction with beta-cyclodextrin (betaCD) in 1 : 1 amorphous PbetaCD inclusion compound are reported. The 1700-1200 cm(-1) FT-Raman and the 13C CP/MAS NMR spectra of 1 : 1 PbetaCD inclusion compound are discussed and assigned in comparison with those of the three main modifications of piroxicam (alpha, beta, and monohydrate). The FT-Raman and 13C-NMR results show that in 1 : 1 PbetaCD inclusion compound piroxicam mainly assumes the zwitterionic structure typical of monohydrate, even if the presence of a different structure, that is, beta form, is not excluded. Piroxicam monohydrate, differently from alpha and beta forms, is characterized by a zwitterionic structure with an internal proton transfer and an increased charge delocalization, as shown by our spectroscopic results. The charge delocalization characteristic of this zwitterionic structure gives rise to the interaction with betaCD via electrostatic and hydrogen bonds. The possibility of a host-guest interaction between piroxicam and betaCD is not excluded; the guest molecule can be accommodated in betaCD cavity by interaction via hydrophobic bonds.

High-performance liquid chromatographic analysis of physiological amino acids in human brain tumors by pre-column derivatization with phenylisothiocyanate.

A reversed-phase high-performance liquid chromatographic technique for the determination of free amino acids in five biopsies of human brain tumors (two meningiomas, one glioblastoma and two oligodendrogliomas) is described. The frozen tissues were homogenized, deproteinized with perchloric acid and neutralized with potassium hydroxide. Aliquots of the supernatant containing the physiological amino acids are used for pre-column derivatization with phenylisothiocyanate. The derivatized PTC-amino acids (phenylthiocarbamyl derivatives) are stable for a five day period if stored as a powder at -20 degrees C in an inert atmosphere and they can be analyzed on a reversed-phase column (PicoTag) using a gradient of two eluents with absorption detection at a wavelength of 254 nm. Good resolution of several amino acids (>30) is achieved within ca. 60 min. For most amino acids this method is suitable for an accurate measurement over a wide range of physiological concentrations (50-400 pmol) starting from a very small amount of sample.

Applications of Raman spectroscopy to ophthalmology: spectroscopic characterization of disposable soft contact lenses.

Disposable soft contact lenses based on HEMA-MAA hydrogels are examined using Raman and ATR/FTIR vibrational spectroscopies and thermal analysis. The main factors dealing with physical, chemical, and biological biocompatibility are evaluated in relation to those of long-wear soft contact lenses with the aim of individuating the most biocompatible lens. The Raman spectra of HEMA-MAA lenses show that some biocompatibility factors are affected by environmental conditions and, in particular, by changes in pH and ionic strength values.

Magnetic resonance spectroscopy study of low grade extra and intracerebral human neoplasms.

In this study we report an in vitro 1H magnetic resonance spectroscopy (MRS) characterization of water soluble metabolites obtained from 17 low grade extra and intracerebral human tumors (8 meningiomas, 5 oligodendrogliomas, 2 subependimomas and 2 ependimomas). In addition, the in vivo localized 1H MRS results for 4 of the meningiomas are reported. The main metabolic features characterizing low grade tumors were investigated. Meningiomas are characterized by high Cho/Cr and Ala/Cr ratios; in many cases both in the in vitro and the in vivo spectra these ratios cannot be evaluated, due to the absence of the Cr metabolite. Low grade oligodendrogliomas are characterized by low Cho/Cr ratios. High amounts of myo-inositol are found in the spectra of ependimomas and subependimomas which are distinguished by different Cho/Cr ratio values and by a different Ala content.

In vivo single voxel 1H MRS of glial brain tumors: correlation with tissue histology and in vitro MRS.

Eighteen patients affected by glial brain tumors were examined by localized single voxel magnetic resonance spectroscopy (1H MRS). For all patients histological diagnosis was obtained after stereotactic biopsy or surgical removal of the tumor. Histological nuclear atypia, tumor cellularity, necrosis, vascular proliferation and percentage of Ki-67 positive cells in the total cell populations were evaluated. It was found that some of the in vivo 1H MRS parameters were related to the histological features of the tumor. In particular, the Choi/Cri metabolite ratio obtained from the neoplastic tissues paralleled the Ki-67 cell proliferation index. For some of the patients 1H and 13C in vitro high resolution magnetic spectroscopy of surgical specimens was also carried out. 1H MRS of aqueous extracts from different biopsy specimens of the same tumor furnished fine biochemical information on intratumor heterogeneity. The degree of histological vascular proliferation of the tumor was correlated to the presence of cholesteryl esters in the 13C spectrum of the lipid fraction of surgical specimens.

In vitro and in vivo MRS study of human glioma metabolites.

In this study an in vitro multinuclear magnetic resonance spectroscopy (MRS) characterization of water soluble metabolites and of the lipid fraction obtained from 19 human gliomas (12 glioblastomas, 3 anaplastic astrocytomas, 2 anaplastic oligodendrogliomas and 2 oligodendrogliomas) with a total of 27 surgical specimens, is reported. Moreover, some in vivo H-1 MRS results are shown. The regional metabolic heterogeneity of glioblastomas, according to their morphological heterogeneity, is documented. For glioblastomas a specific in vitro H-1 MRS metabolite profile cannot be defined. Low and high grade oligodendrogliomas showed characteristic choline/creatine and alanine/creatine metabolite ratios. The spectroscopical characterization of histopathological factors concerning malignant gliomas is shown.

In vivo 1H MRS and in vitro multinuclear MR study of human brain tumors.

Localized in vivo 1H MR spectra of four patients with brain tumors and in vitro multinuclear (1H, 13C and 31P) MR characterization of water soluble and lipid components of the same tumors after surgical removal are reported herein. The in vivo spectra of the neoplastic regions of all patients showed N-Acetyl-Aspartate/Choline (NAA/Cho), Choline/Creatine (Cho/Cr) and N-Acetyl-Aspartate/Creatine (NAA/Cr) metabolite ratios markedly different from those of the controlateral healthy tissue. In vitro 1H MR measurements allowed for the identification of a much larger number of metabolites than those measured in vivo. Indeed, well resolved 1H, 13C and 31P MR spectra were obtained for water soluble and lipid components in all cases. Our data confirm that some metabolites observed in vitro could be specific markers of brain tumors.

Synovial fluid parameters in normal and osteochondritic hocks of horses with open physis.

An investigation was carried out on most common synovial fluid parameters of normal and osteochondritic hocks of horses less than 12 months old in order to confirm the presence of an inflammatory process. Furthermore, a spectroscopic study was performed on synovial fluid from both normal and diseased hocks. A depolymerization of hyaluronic acid was demonstrated in synovial fluid from diseases joints, similar to that reported in human rheumatoid arthritis. A one month rest seem to normalize all parameters considered and in one joint, a return to normal infrared spectrum was demonstrated.

Raman spectroscopy in the study of biocompatibility.

This review deals with the application of Raman spectroscopy to the study of the biocompatibility of orthopaedic and ophthalmological materials and includes an introduction, a brief theory on the fundamental concepts of the technique, a description of the method and of the traditional and non-traditional instrumentation and a brief treatment of the surface properties of the different classes of materials. In the orthopaedic field, Raman studies on the structures of glasses, ceramic materials, carbon fibres and polymers before and after implantation are reviewed, particularly as far as structural modifications at the biomaterial-tissues interface are concerned. In the ophthalmological field, the chemical biocompatibility of hydrophobic and hydrophilic polymers for intraocular and intrastromal implants and for soft contact lenses is considered with respect to the presence of monomeric reactive centres in hydrophobic materials and the water amount in hydrophilic ones. The progress of the multichannel Raman technique for 'in-vivo' measurements is also described.

Calcium phosphate materials containing alumina: Raman spectroscopical, histological, and ultrastructural study.

Variable alumina quantities were added to two types of calcium phosphate materials-hydroxyapatite ceramics with Ca/P = 1.67 and calcium metaphosphate glass with Ca/P = 1--in order to increase their mechanical properties. Raman spectroscopy shows that alumina interacts with the phosphate group of these materials, while thermomechanical analysis shows that their elastic modulus has a value similar to that of bone. Histological sections demonstrate that the surface in close contact with ceramic materials shows a good integration between bone and biomaterial. All ceramic specimens are penetrated by well-stained, presumably glycoprotein, matrix, that consistently forms a thin network in close contact with the implant. After 6 weeks bone growing on both ceramic and glass shows signs of maturation with a lamellar structure and an apparently normal mineralization. In the case of the glass, inside this newly formed bone, were often observed two layers, the internal one showing a well defined lamellar structure.

Structural properties and biocompatibility of polymers used in ophthalmology.

The chemico-physical and structural properties and biocompatibility of new polymers used for intraocular lens manufacture are studied. In particular, the vibrational infrared and Raman spectra of fluorinated polymethacrylates are presented and discussed in relation to those of polymethylmethacrylate (PMMA), in order to correlate their structure with their properties and biocompatibility. The thermal analysis points out that the polymers are more stable than polymethylmethacrylate and the biocompatibility tests show that they are as biocompatible as polymethylmethacrylate.

Raman spectroscopy for the study of porous composite alumina coating.

A new acetabular cup prosthesis made of alumina and bioactive glass was studied and evaluated using nondestructive and noninvasive Raman laser spectroscopy. The prosthesis consists of an alumina socket coated with a double layer of alumina beads bonded to the prosthesis with a layer of glass. Raman spectroscopy showed that both the socket and the beads are composed of alpha-Al2O3 (corundum), while the bioactive glass (alkaline and alkaline earth alumosilicate with a small quantity of zirconium oxide) showed this to have an intermediate structure between that of vitreous silica and that of the alkaline and alkaline earth disilicate glasses. The Raman spectra of the contact surfaces between the bioactive glass and the alumina of the assembled prosthesis and of an alumina disk coated with the same glass showed the presence of new bands which may be correlated with a chemical interaction between the components of the prosthesis.

Structural modifications and biological compatibility of doped bio-active glasses.

The Raman laser and infrared spectra of doped bio-active glasses of the 45S5 type are presented and discussed. The spectroscopic results show that the doping agents cause the destruction of the basic glass structure and the consequent formation of SiO4(4-) units in the glass network. When the doped glasses have been immersed in a physiological solution (199 medium), a film of calcite forms on the glass surface and this modification is related to the type of doping agent used, decisive for close linking between metal supports and the glass. The presence of doping agents does not prevent the normal growth of the bone onto the surface of doped bioactive glasses. Histological tests show that tissue response to very fine powders of doped glasses increases up to 15 days more or less according to the structural modifications revealed by spectroscopic measurements.

Molecular interactions between nucleic acids and antitumor substances by Raman and NMR spectroscopy.

1-Methyladenosine (1-MeAdo) and protonated 1-Methyladenosine (1-MeAdo.H+) were chosen as a model to study the molecular interactions between the carcinogen dimethylsulphate (DMS) and the base adenine, and the interactions between the antitumoral electrophilic cis-diamminedichlorophatinum (II) (cis-Pt) and the methylation product of the base, respectively. Raman and multinuclear NMR studies show the molecular perturbations caused by the carcinogen on the base and those of the antitumoral substance on the reaction product base-carcinogen.