Valter Sergo

Mechanical and chemical consequences of the residual stresses in plasma sprayed hydroxyapatite coatings

The residual stresses in thick hydroxyapatite coatings, deposited by plasma spraying, have been determined experimentally using Raman piezo-spectroscopy. The stress dependence of the centre position of the 980 cm 1 Raman band, owing to the symmetric stretching of the phosphate ion, PO3(4), has been established and found to be 2.47 cm 3 GPa 1. Using this calibration, the residual stresses in hydroxyapatite coatings deposited onto Ti-6A1-4V substrates in air have been found to be 100 MPa (tensile), whereas those deposited in a vacuum have been found to be 60 MPa (compressive). Although desirable from a mechanical point of view, it is shown that coating under residual compression are thermodynamically more stable and, hence, the dissolution of the ionic species, necessary in the exchange between bone and hydroxyapatite coating, can be impeded. It is calculated that for the coating under examination the stresses have an effect comparable with almost an order of magnitude change of the [OH] concentration. The analysis explains the dissolution behaviour of hydroxyapatite coatings subject to cyclic stress reported previously.

Biomedical applications of Raman and infrared spectroscopy to diagnose tissues

The objective of the article is to review biomedical applications which became possible after the development of sensitive and high throughput Raman and Fourier transform infrared spectrometers in the past decade. Technical aspects of the instrumentation are briefly described. Then the broad range of vibrational spectroscopic applications with the focus on imaging and fiber-optical methods are discussed to study mineralized tissue (bone, teeth), skin, brain, the gastrointestinal tract (mouth, pharynx, esophagus, colon), breast, arteries, cartilage, cervix uteri, the urinary tract (prostate, bladder), lung, ocular tissue, liver, heart and spleen. Experimental studies are summarized demonstrating the possibilities and prospects of these methods in various fields of biodiagnostics to detect and characterize diseases, tumors and other pathologies. Infrared (IR) and Raman spectroscopy have recently been applied to address various biomedical is- sues. The basis for these applications is that IR and Raman spectroscopy are vibrational spectroscopic techniques capable of providing details of the chemical composition and molecular structures in cells and tissues. In principle, diseases and other pathological anomalies lead to chemical and structural changes on the molecular level which also change the vibrational spectra and which can be used as sensitive, phenotypic markers of the disease. As these spectral changes are very specific and unique, they are also called fingerprint. The advantages of the methods include that they are non-destructive and do not require extrinsic contrast-enhancing agents. Early reports in the literature regarding the utility of IR and Raman spectroscopy to biomedical problems were based on macroscopic acquisition of spectral data only at sin- gle points which required an a priori knowledge of the location or a pre-selection of the probed position. Since the inhomogeneous nature of tissue was not considered in these early studies, an accurate corre- lation between the histopathology of the sampled area and the corresponding spectra was not possible. Therefore, many of the early results were spurious and they will not be presented here. Considerable progress was made in the past ten years because high throughput and more sensitive instruments be- came available for Raman and IR microspectroscopic imaging. They enable to microscopically collect larger number of spectra from larger sample populations in less time, improving statistical significance and spatial specificity. Simultaneously, fiber-optical probes have been developed for in vivo applications.

Effect of ZrO2 content on textural and structural properties of CeO2 / ZrO2 solid solutions made by citrate complexation route

Abstract Single phase homogeneous CeO 2 –ZrO 2 solid solutions with various compositions were synthesized using a citrate complexation route. Investigation of the sintering behaviour disclosed important modifications of the textural and, in particular, structural properties, which apparently create a strongly defective structure that could explain the unusual redox properties of these catalytic systems.

Raman and FTIR microscopic imaging of colon tissue: a comparative study.

Colon tissue constitutes a valid model for the comparative analysis of soft tissue by Raman and Fourier transform infrared (FTIR) imaging because it contains four major tissue types such as muscle tissue, connective tissue, epithelium and nerve cells. Raman microscopic images were recorded in the mapping mode using 785 nm laser excitation and a step size of 10 microm from three regions within a thin section that encompassed mucus, mucosa, submucosa, and longitudinal and circular muscle layers. FTIR microscopic images that were composed of 4, 8 and 9 individual images of 4096 spectra each were recorded from the same regions using a FTIR spectrometer coupled to a microscope with a focal plane array detector. Furthermore, Raman microscopic images were recorded at a step size of 2.5 microm from three ganglia that belong to the myenteric plexus. The results are discussed with respect to lateral resolution, spectral resolution, acquisition time and sensitivity of both modalities.

Degree of conversion of Filtek Silorane Adhesive System and Clearfil SE Bond within the hybrid and adhesive layer: An in situ Raman analysis

OBJECTIVES To examine the degree of conversion (DC) of the adhesive interfaces created by Filtek Silorane Adhesive and Clearfil SE Bond using micro-Raman spectroscopy. METHODS The adhesives were applied on human dentin in accordance with manufacturers instructions. Specimens were cut to expose the bonded interfaces to the micro-Raman beam (Ranishaw InVia; laser wl 785 nm). Raman spectra were collected along the dentin/self-etching primer/adhesive interface at 1 microm intervals. The relative intensities of bands associated with mineral (P-O functional group at 960 cm(-1)) and adhesive (C-C-O group at 605 cm(-1)) components within the bonded interface were used to detect monomer penetration into the dentin matrix and to calculate the degree of conversion (C=C at 1640 cm(-1) as reaction peak, C-C-O at 605 cm(-1) as reference peak). Data were statistically analyzed with two-way ANOVA. RESULTS DC of Filtek Silorane Adhesive was 69+/-7% in the adhesive layer, increasing (p<0.05) to 93+/-5% in the primer and 92+/-9% in the hybrid layer. Clearfil SE Bond showed a DC of 83+/-3% in the hybrid and 85+/-3% in the adhesive layer. Thus, Filtek Silorane Adhesive showed a higher DC than Clearfil SE Bond in the hybrid layer (p<0.05), but a lower DC in the adhesive (p<0.05). SIGNIFICANCE As high DC is a fundamental pre-requisite for the stability of the bond over time, this study supports the hypothesis that optimal stability of Filtek Silorane Adhesive can be obtained. However, further research is needed to investigate the mechanical properties of the hybrid layer created by Filtek Silorane Adhesive and its long-term stability.

Raman mapping and FTIR imaging of lung tissue: congenital cystic adenomatoid malformation

Congenital cystic adenomatoid malformations (CCAMs) are benign masses of non-functional lung tissue developing from an overgrowth of the terminal bronchioles with subsequent suppressing of alveolar growth. For the first time CCAMs are studied by Raman mapping and Fourier transform infrared (FTIR) imaging. Both vibrational spectroscopic methods are able to analyze the biochemical composition of tissues and their pathological changes at the molecular level. Cryosections were prepared on calcium fluoride substrates from CCAMs and from normal lung tissue of two infant patients who underwent surgery. Raman maps were collected at a step size of 100 microm in order to assess the whole tissue section and at a smaller step size of 10 microm in order to resolve details in selected areas. FTIR images were collected in the macroscopic and microscopic modes. Data sets were segmented by cluster analysis and the mean spectra of each cluster were compared. At low lateral resolution a lower red blood cell content and higher lipid content were found in CCAMs than in normal lung tissue. At higher lateral resolution, accumulations of lipids and glycogen were identified in CCAMs. The lipid aggregates contain a high concentration of phosphatidylcholine. It is discussed how the combined application of Raman mapping and FTIR imaging might improve the differential diagnosis of lung malformations and how both modalities might be applied to other bioanalytical and biomedical problems in the future.

Surface-enhanced Raman spectroscopy of blood plasma and serum using Ag and Au nanoparticles: a systematic study

Surface-enhanced Raman spectroscopy (SERS) is a good candidate for the development of fast and easy-to-use diagnostic tools, possibly used on biofluids in point-of-care or screening tests. In particular, label-free SERS spectra of blood serum and plasma, two biofluids widely used in diagnostics, could be used as a metabolic fingerprinting approach for biomarker discovery. This study aims at a systematic evaluation of SERS spectra of blood serum and plasma, using various Ag and Au aqueous colloids, as SERS substrates, in combination with three excitation lasers of different wavelengths, ranging from the visible to the near-infrared. The analysis of the SERS spectra collected from 20 healthy subjects under a variety of experimental conditions revealed that intense and repeatable spectra are quickly obtained only if proteins are filtered out from samples, and an excitation in the near-infrared is used in combination with Ag colloids. Moreover, common plasma anticoagulants such as EDTA and citrate are found to interfere with SERS spectra; accordingly, filtered serum or heparin plasma are the samples of choice, having identical SERS spectra. Most bands observed in SERS spectra of these biofluids are assigned to uric acid, a metabolite whose blood concentration depends on factors such as sex, age, therapeutic treatments, and various pathological conditions, suggesting that, even when the right experimental conditions are chosen, great care must be taken in designing studies with the purpose of developing diagnostic tests.

Label-free surface-enhanced Raman spectroscopy of biofluids: fundamental aspects and diagnostic applications

In clinical practice, one objective is to obtain diagnostic information while minimizing the invasiveness of the tests and the pain for the patients. To this end, tests based on the interaction of light with readily available biofluids including blood, urine, or saliva are highly desirable. In this review we examine the state of the art regarding the use of surface-enhanced Raman spectroscopy (SERS) to investigate biofluids, focusing on diagnostic applications. First, a critical evaluation of the experimental aspects involved in the collection of SERS spectra is presented; different substrate types are introduced, with a clear distinction between colloidal and non-colloidal metal nanostructures. Then the effect of the excitation wavelength is discussed, along with anomalous bands and artifacts which might affect SERS spectra of biofluids. The central part of the review examines the literature available on the SERS spectra of blood, plasma, serum, urine, saliva, tears, and semen. Finally, diagnostic applications are critically discussed in the context of the published evidence; this section clearly reveals that SERS of biofluids is most promising as a rapid, cheap, and non-invasive tool for mass screening for cancer.

Degree of conversion and interfacial nanoleakage expression of three one-step self-etch adhesives.

Suboptimally polymerized monomers may be responsible for the reduced material properties of simplified adhesives and their inherent bonded interface instability. This study was performed to determine the degree of cure within the hybrid layers produced by three one-step self-etch adhesives in situ using Raman microspectroscopy and to investigate nanoleakage expression. Dentin disks were bonded with AdheSE One, Adper Prompt L-Pop, or iBond. Composite layers of 2-mm thickness were built up in bulk on the polymerized adhesive surfaces and then the adhesive-dentin interfaces were exposed to a micro-Raman beam. Adhesive penetration was calculated using the relative intensities of bands associated with mineral and adhesive, and the degree of conversion (DC) was evaluated. Interfacial nanoleakage expression was evaluated on the same specimens. The DC values for the tested adhesives were found to increase in the following order: AdheSE One (48 +/- 16%) < Adper Prompt L-Pop (83 +/- 2%) = iBond (90 +/- 6%; P < 0.05). AdheSE One showed greater nanoleakage expression than iBond or Adper Prompt L-Pop. Increased nanoleakage expression was associated with AdheSE One that showed the lowest DC. This suggests that a low DC may affect the quality and the long-term stability of the adhesive interface owing to the elution of unreacted monomers forming a porous and highly permeable hybrid layer.

Poly-l-lysine-Coated Silver Nanoparticles as Positively Charged Substrates for Surface-Enhanced Raman Scattering

Positively charged nanoparticles to be used as substrates for surface-enhanced Raman scattering (SERS) were prepared by coating citrate-reduced silver nanoparticles with the cationic polymer poly-l-lysine. The average diameter of the coated nanoparticles is 75 nm, and their zeta potential is +62.3 ± 1.7 mV. UV-vis spectrophotometry and dynamic light scattering measurements show that no aggregation occurs during the coating process. As an example of their application, the so-obtained positively charged coated particles were employed to detect nanomolar concentrations of the anionic chromophore bilirubin using SERS. Because of their opposite charge, bilirubin molecules interact with the coated nanoparticles, allowing SERS detection. The SERS intensity increases linearly with concentration in a range from 10 to 200 nM, allowing quantitative analysis of bilirubin aqueous solutions.