Staphylococcus epidermidis (S. epidermidis) is one of the leading nosocomial pathogens, particularly associated with periprosthetic infections of biomedical implants. Silicon nitride (Si3N4), a nonoxide biomaterial widely used in spinal implants, has shown bacteriostatic effects against both gram-positive and gram-negative bacteria; however, the physicochemical interactions between Si3N4 and bacteria yet remain conspicuously unexplored. In situ time-lapse Raman spectroscopic experiments were conducted by exposing S. epidermidis for 12, 24, and 48 h to Si3N4 substrates to understand the evolution of bacterial metabolism and to elucidate the ceramics antimicrobial behavior. The Raman probe captured an initial metabolic response of the bacteria to the adverse chemistry of the Si3N4 surface, which included peroxidation of membrane phospholipids and protein structural modifications to adjust for survivorship. However, beyond 24 h of exposure, the Raman signals representing DNA, lipids, proteins, and carbohydrates showed clear fingerprints of bacterial lysis. Bands related to biofilm formation completely disappeared or underwent drastically reduced intensity. Bacterial lysis was confirmed by conventional fluorescence microscopy methods. Spectroscopic experiments suggested that a pH change at the Si3N4’s surface induced variations in the membrane structure and D-alanylation of teichoic acids in its peptidoglycan layer. Concurrent stimulation of peptidoglycan hydrolase (i.e., an enzyme involved with autolysis) ultimately led to membrane degradation and cellular death. An additional finding was that modulating the Si3N4 surface by increasing the population of amine groups improved the efficiency of the substrate against S. epidermidis, thus suggesting that optimization of the near-surface (alkaline) conditions may be a viable approach to bacterial reduction.
The possibility of examining soft tissues by Raman spectroscopy is challenged in an attempt to probe human age for the changes in biochemical composition of skin that accompany aging. We present a proof-of-concept report for explicating the biophysical links between vibrational characteristics and the specific compositional and chemical changes associated with aging. The actual existence of such links is then phenomenologically proved. In an attempt to foster the basics for a quantitative use of Raman spectroscopy in assessing aging from human skin samples, a precise spectral deconvolution is performed as a function of donors’ ages on five cadaveric samples, which emphasizes the physical significance and the morphological modifications of the Raman bands. The outputs suggest the presence of spectral markers for age identification from skin samples. Some of them appeared as authentic “biological clocks” for the apparent exactness with which they are related to age. Our spectroscopic approach yields clear compositional information of protein folding and crystallization of lipid structures, which can lead to a precise identification of age from infants to adults. Once statistically validated, these parameters might be used to link vibrational aspects at the molecular scale for practical forensic purposes.
Environmental metastability of zirconia (ZrO 2 ) ceramic in the human body [represented by a tetragonal-to-monoclinic (t→m ) phase transformation] takes place on the surface of the artificial joint and proceeds with time toward its interior. Its quantitative characterization is mandatory for the safety of joint implants and consists of the assessment of the in-depth monoclinic profile fraction as compared to that of the initially untransformed material. We attempt to fully establish a characterization protocol and present two different nondestructive approaches for resolving highly graded phase-transformation profiles along the hip-joint subsurface by confocal Raman microprobe technique. A series of partially transformed tetragonal zirconia polycrystal and zirconia-toughened alumina ceramics are used as screening samples. Probe biases could be eliminated and the real transformation profiles retrieved through a deconvolution procedure of Raman experimental data collected as a function of pinhole aperture and focal depth, respectively. Confirmation of the confocal assessments was made by a destructive cross-sectional inspection by both laser optical microscope and Raman spectral line scans. This study unveils for the first time the real quantitative amount of surface phase-transformation fractions and the related subsurface profiles in zirconia-based retrieved medical samples.
Quantitative experimental assessments of stress/strain with sub-micrometer spatial resolution are shown in both
crystalline and amorphous structures of a metal oxide semiconductor (MOS), consisting of Si and SiOx, respectively. A
piezo-spectroscopic (PS) approach, based on the wavelength shift of a spectroscopic band in a solid in response to an
applied strain or stress, has been used throughout this investigation. Although the PS behavior in such different structures
obeyed completely different physical rules, its rationalization gave access to stress/strain information, regardless of the
particular spectroscopic transition involved. In this paper, we applied two complementary PS analytic procedures
suitable for MOS semiconductor devices: (i) using the LO Raman transition for rationalizing the stress state on the
crystalline-Si side of the device; and, (ii) using the electro-stimulated spectrum of oxygen point defects for analyzing the
amorphous SiOx side of the device. As a further step, in this study, we challenged a nanometer-scale spatial resolution in
stress assessments by applying spatially resolved PS procedures, which involved deconvolution of both laser and
electron probes in the respective materials.
Confocal spectroscopic techniques are applied to selected Raman bands to study the microscopic features of acetabular cups made of ultra-high molecular weight polyethylene (UHMWPE) before and after implantation in vivo. The micrometric lateral resolution of a laser beam focused on the polymeric surface (or subsurface) enables a highly resolved visualization of 2-D conformational population patterns, including crystalline, amorphous, orthorhombic phase fractions, and oxidation index. An optimized confocal probe configuration, aided by a computational deconvolution of the optical probe, allows minimization of the probe size along the in-depth direction and a nondestructive evaluation of microstructural properties along the material subsurface. Computational deconvolution is also attempted, based on an experimental assessment of the probe response function of the polyethylene Raman spectrum, according to a defocusing technique. A statistical set of high-resolution microstructural data are collected on a fully 3-D level on -ray irradiated UHMWPE acetabular cups both as-received from the maker and after retrieval from a human body. Microstructural properties reveal significant gradients along the immediate material subsurface and distinct differences are found due to the loading history in vivo, which cannot be revealed by conventional optical spectroscopy. The applicability of the confocal spectroscopic technique is valid beyond the particular retrieval cases examined in this study, and can be easily extended to evaluate in-vitro tested components or to quality control of new polyethylene brands. Confocal Raman spectroscopy may also contribute to rationalize the complex effects of -ray irradiation on the surface of medical grade UHMWPE for total joint replacement and, ultimately, to predict their actual lifetime in vivo.
We aim to establish a confocal spectroscopic technique able to study the features of fluorescence spectra arising from native Cr3+ impurity in polycrystalline alumina (Al2O3) as a biomaterial and to use their emission lines as microscopic probes for the characterization of residual stress fields stored in artificial hip prostheses during their implantation in vivo. As an application of the technique, we report for the first time concerning the evolution of microscopic (residual) stress fields stored on the surface and in the subsurface of N=7 retrieved Al2O3 hip joints after exposure in the human body from a few months to 19 yr. The micrometric diameter of the laser beam waist impinging on the joint surface (typically about 1 µm in lateral resolution) enables us to estimate the patterns and magnitude of residual stress with high spatial resolution, at least comparable with the grain size of the material. In addition, a selected confocal configuration for the optical probe enables minimization of the probe size along the in-depth direction. According to a statistical collection of data on the microscopic level for retrieved femoral heads in toto, a residual stress field arising from loading history in vivo during the lifetime of the Al2O3 femoral head can be revealed. Finally, an interpretation is given of microscopic wear mechanisms in Al2O3 artificial hip joints consistent with the observed evolution of surface residual stress fields on elapsed time in vivo.
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