Characterisation of structural changes in collagen with Raman spectroscopy

Raman spectroscopy can detect conformational changes in collagen structures and these findings are reviewed in this article. More specifically, some progressive diseases are mainly caused by alterations of collagen molecules but what is occurring at the biochemical level of this complex molecule usually remains unclear. While it may be true that a number of analytical techniques can analyze collagen, most of them have a series of limitations that limit their applicability to a wide range of samples. To understand in more detail the progression of a disease due to changes in the collagen structure, a technique that can detect subtle alterations at the biochemical level is needed. Raman spectroscopy is a label-free and noninvasive technique that can easily pick up on any conformational changes reflected primarily at the lipids, amides and proline and hydroxyproline regions. This review is the first compilation of studies of conformational changes in collagen molecules, providing help to understand changes in collagen biochemistry that can be of relevance to the human wound healing, ageing and pathologies.     

FTIR analysis of natural and synthetic collagen

Collagen is the most abundant protein in humans and animals, comprising of one third of the total proteins that accounts for three quarters of the dry weight skin in humans. Collagen containing a range of proteins has been reported for tissue engineering applications, but, only a small number of studies related to chemical structure evaluation of collagen are found in the literature. Collagen can be obtained from both the natural and synthetic sources and offers a wide range of biomedical applications due to its excellent biocompatibility and low immunogenicity. Hence, it is important to identify chemical structural properties of collagen and Fourier transform infrared (FTIR) appears to be a technique of choice to study their chemical structure. This review aims to highlight the use of FTIR to study collagen-based biomaterials, using it for characterization of collagen extracted from various sources. Characterization of collagen-based materials used in wound healing, skin substitutes, derma fillers, and aging of skin, collagen containing drug delivery agents, collagen-based materials used in tissue engineering, bone regeneration, and osteogenic differentiation is discussed in detail. FTIR analysis of collagen-containing materials used for dental applications, cleft-palate, and in alveolar-ridge preservation has also been highlighted.     

Synchrotron small angle X-ray scattering study on the conformation of polystyrene in compressed CO2-toluene mixture

Synchrotron small angle X-ray scattering (SAXS) was performed to investigate the effect of dissolved CO2 in toluene on the conformation of polystyrene (PS) in the solution. It has been found that the second virial coefficient A2 and the radius of gyration Rg decrease with the increasing antisolvent CO2 pressure. The scattering intensity of PS chain followed l(h)∞h-αunder different antisolvent pressures (0, 0.6, 1.5, 2.4, 3.3, and 4.2 MPa), suggesting that the PS chain has self-similar structure behavior or a fractal structure in the presence of antisolvent CO2.All this reveals a large effect of antisolvent pressure or the solubility of CO2 in the solution on PS structure. The fractal dimensions increase with the increasing antisolvent pressure, indicating that the polymer chain changes from a swollen coil into a rather dense globule in the course of adding antisolvent CO2.     

Application of a micro-Brillouin scattering technique to characterize bone in the GHz range

The evaluation of elastic properties of bone matrix has been investigated using several techniques such as nanoindentation and scanning acoustic microscopy (SAM). These techniques make use of good spatial resolution, which can prevent effects due to microstructures at the level of several hundreds of microns. In this paper, micro-Brillouin scattering (μ-BR) is introduced as another possible technique to characterize the elastic properties of bone. This technique is well known as a non-contact and non-destructive method to evaluate viscoelastic properties of transparent materials in the GHz range. Using thin, translucent bone specimens with thicknesses of around 100 μm, and the reflection induced optical geometry, ultrasonic wave velocities in the GHz range were obtained. Because this technique optically measures thermal phonons in the specimen, we can easily measure in-plane anisotropy of wave velocities by rotating the specimen. In a single trabecula, the site matched data between SAM and μ-BR showed good correlation, revealing the applicability of this technique to characterize material properties of bone. Some recent results on the anisotropy in a trabecula and the elasticity evaluation of newly and matured bones are also introduced.     

Synchrotron small angle X-ray scattering study on the conformation of polystyrene in compressed CO<Subscript>2</Subscript>-toluene mixture

Synchrotron small angle X-ray scattering (SAXS) was performed to investigate the effect of dissolved CO₂ in toluene on the conformation of polystyrene (PS) in the solution. It has been found that the second virial coefficientA ₂ and the radius of gyrationR _g decrease with the increasing antisolvent CO₂ pressure. The scattering intensity of PS chain followedI(h) ∞h ^−α under different antisolvent pressures (0, 0.6, 1.5, 2.4, 3.3, and 4.2 MPa), suggesting that the PS chain has self-similar structure behavior or a fractal structure in the presence of antisolvent CO₂. All this reveals a large effect of antisolvent pressure or the solubility of CO₂ in the solution on PS structure. The fractal dimensions increase with the increasing antisolvent pressure, indicating that the polymer chain changes from a swollen coil into a rather dense globule in the course of adding antisolvent CO₂.     

Concentration dependence of F nuclear magnetic resonance decay shapes and second moments in bone mineral

¹⁹F nuclear magnetic resonance (NMR) spin-echoes and free induction decays (FIDs) have been observed from samples of fluoridated trabecular canine bone powder, with fluoride concentrations ([F]) ranging from approximately 10 to 33 mg F/g Ca. Curve fitting of echo envelopes and FIDs was performed using a two-component model function, where one of the components incorporates the effects of one-dimensional dipolar coupling. This function provides a good match for both echo envelopes and FIDs. Estimates of the total second moment and its homonuclear (F–F coupling) component were obtained from the fitting procedure. Based on the second moment measurements, it is argued that ¹⁹F spins in bone mineral typically experience weaker heteronuclear dipolar coupling than those in the mineral hydroxyapatite (HAP), which is often considered to be a prototype for bone mineral.     

生物组织中光子迁移的统计性质研究

研究了生物组织中散射光子程长的概率分布函数Ｐ（ι）及其一阶统计性质,并由Ｐ（ι）导出了阶统计量－－漫射光时间自相关函数ｇ１（τ）的表达式,讨论了影响ｇ１（τ）衰减快慢的因素。     

Small-x behavior of the structure function and its slope for “frozen” and analytic strong-coupling constants

Using the leading-twist approximation of the Wilson operator product expansion with “frozen” and analytic versions of the strong-coupling constant, we show that the Bessel-inspired behavior of the structure function F₂ and its slope ∂lnF₂/∂ln(1/x) at small values of x, obtained for a flat initial condition in the DGLAP evolution equations, leads to good agreement with experimental data of deep-inelastic scattering at DESY HERA.     

Phosphorus-31 spin-lattice NMR relaxation in bone apatite and its mineral standards

Phosphorus-31 spin-lattice relaxation, both in the laboratory (B(0)=4.7 T) and rotating frame (B(1)=2.2 mT), was studied in the following samples: mineral of whole human bone (samples B1-B6), apatite prepared from bone (BHA), natural brushite (BRU), synthetic hydroxyapatite hydrated (HAh) and calcined (HAc), and synthetic carbonatoapatite of type B (CHA-B) with 9 wt% of CO(3)(2-). The T(1)(P) relaxation time was determined directly using the saturation recovery technique, while the T(1 rho)(P) relaxation time was measured via (1)H-->(31)P CP by incrementing the (31)P spin-lock. In order to avoid an effect of magic-angle spinning (MAS) on CP and relaxation, the experiments were carried out on static samples. The (31)P spin-lattice relaxation was discussed for trabecular and cortical bone tissue from adult subjects in comparison to the synthetic mineral standards. None of the reference materials has matched accurately the relaxation behaviour of the bone mineral. The most striking differences between the examined substances were observed for T(1)(P), which for human bone was sample dependent and appeared in the range 55-100 s, while for HAh, HAc, and CHA-B was 7.2, 10.0, and 25.8 s, respectively. Possible reasons of so large relaxation diversity were discussed. It has been suggested that T(1)(P) of apatites is to some extent dependent on the concentration of the structural hydroxyl groups, and this in turn is controlled by the material crystallinity. It was also found that T(1)(P) decreased on hydration by ca. 30%. For T(1rho)(P), both its magnitude and dependence on the CP contact time gave useful structural information. The dehydrated samples (HAc and BHA) had long T(1 rho)(P) over 250 ms. Those, which contained water, either structural (BRU) or adsorbed on the crystal surface (HAh, CHA-B, and B1-B6), had shorter T(1 rho)(P) below 120 ms. It was concluded that the effect of water on T(1 rho)(P) is much more pronounced than on T(1)(P). The interpretation has involved P-OH groups and adsorbed water, which cover the apatite crystal surface.     

Counterion condensation in ionic micelles as studied by a combined use of SANS and SAXS

We report a combined use of small-angle neutron scattering (SANS) and small-angle X-ray scattering (SAXS) to the study of counterion condensation in ionic micelles. Small-angle neutron and X-ray scattering measurements have been carried out on two surfactants cetyltrimethylammonium bromide (CTABr) and cetyltrimethylammonium chloride (CTACl), which are similar but having different counterions. SANS measurements show that CTABr surfactant forms much larger micelles than CTACl. This is explained in terms of higher condensation of Br⁻ counterions than Cl⁻ counterions. SAXS data on these systems suggest that the Br⁻ counterions are condensed around the micelles over smaller thickness than those of Cl⁻ counterions.     

Review of recent results in deep inelastic scattering

This talk gives a summary of recent results in deep inelastic lepton hadron scattering. This includes structure functions from inclusive measurements as well as fragmentation in semi-inclusive processes, mainly with respect to data from colliders such as HERA at DESY, and their associated phenomenology.     

The role of collagen in bone structure: an image processing approach

Bone collagen structure in normal and pathological tissues is illustrated using techniques of thin section transmission electron microscopy and computer-assisted analysis. The normal bone collagen types, fibril architecture and diameter are described. In pathological tissue, deviations from normal fine structure are reflected in abnormal arrangements of collagen fibrils and abnormalities in fibril diameter. Computer analyses of normal fibril positive staining patterns are presented in order to provide a basis for comparing such patterns with pathological ones.     

Efficiency of 1H --> 31P NMR cross-polarization in bone apatite and its mineral standards

Human bone mineral was studied using solid-state ³¹P NMR with cross-polarization (CP) from protons. The CP efficiency was determined for trabecular and cortical bone tissue from human adults and compared with synthetic mineral standards. The study shows the similarity between carbonatoapatite of type B and bone mineral as shown by their CP behaviour. The method can be used for the characterization of synthetic apatite-based implant materials.     

Step-addition Method for Enhancing the Yield of Gold Nanoparticles in Block Copolymer Solution

Triblock copolymer poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) has been used to synthesize gold nanoparticles from hydrogen tetrachloroaureate (III) hydrate (HAuCl₄·3H₂O) salt in aqueous solution at room temperature. Measurements were performed using the triblock copolymer Pluronic P85 (EO₂₆PO₃₉EO₂₆) at a fixed concentration (1 wt%) mixed with varying HAuCl₄·3H₂O concentration in the range of 0.001 to 0.1 wt%. The surface plasmon resonance (SPR) band in UV-visible absorption spectra confirmed the formation of the gold nanoparticles. The maximum yield of the nanoparticles was found at 0.005 wt% of the salt solution. Small-angle neutron scattering (SANS) does not show any significant change in the scattering profile in these suspensions of the nanoparticles. A similar behavior was also observed in dynamic light scattering (DLS) experiments where autocorrelation function was found to be independent of the salt concentration. This can be understood since a high-block copolymer-to-gold ion ratio (r ～ 22) is required in the reduction reaction to produce gold particles. As a result, a very small fraction of the block copolymers were associated with the gold nanoparticles, and hence lead to a very low yield. Both SANS and DLS basically see the micelles of most of these block copolymers, which are not associated with nanoparticles. Based on this explanation, a step-addition method was used to enhance the yield of gold nanoparticles by manifold, where the gold salt is added in small steps to maintain higher value of r (>22), and therefore continuous formation of nanoparticles.     

Effect of porosity and pore morphology on the low-frequency dielectric response in sintered ZrO<Subscript>2</Subscript>—8 mol% Y<Subscript>2</Subscript>O<Subscript>3</Subscript> ceramic compact

Effect of porosity and pore size distribution on the low-frequency dielectric response, in the range 0.01–100 kHz, in sintered ZrO₂—8 mol% Y₂O₃ ceramic compacts have been investigated. Small-angle neutron scattering (SANS) technique has been employed to obtain the pore characteristics like pore size distribution, specific surface area etc. It has been observed that the real and the imaginary parts of the complex dielectric permittivity, for the specimens, depend not only on the porosity but also on the pore size distribution and pore morphology significantly. Unlike normal Debye relaxation process, where the loss tangent vis-à-vis the imaginary part of the dielectric constant shows a pronounced peak, in the present case the same increases at lower frequency region and an anomalous non-Debye type relaxation process manifests.     

Statistical Properties of Stimulated Raman Scattering in a Nonlinear Regime

Statistical properties of the Raman scattering of an intense step pulse are investigated by numerical solution of the equations of a semi-classical theory with assignment of a random Gaussian distribution of the initial polarization over the sample. The probability density functions of the intensity, energy, delay time, and width of a Stokes pulse and also of the position, height, and width of the spectral peaks of the first incidental Stokes component with the preservation of the phase memory of scattering centers are calculated. The influence of collisional dephasing on the fluctuation of these characteristics is considered.     

An Alternative Approach to the Extraction of Structure Functions in Deep Inelastic e-p Scattering at 5 to 20 GeV

Two structure functions W1(x,Q2) and W2(x,Q2) are determined by using the cross sections measured in the deep inelastic electron-proton scattering experiments at Stanford Linac in the energy range of 5 to 20 GeV. In this paper an alternative mathematical approach have been used in such determination, resulting in a larger number of points in the graphs of the structure functions.     

Calculation of the Gluon Distribution Function Using Alternative Method for the Proton Structure Function

A calculation of the proton structure function F2(x,Q2) is reported with an approximation method that relates the reduced cross section derivative and the F2(x, Q2) scaling violation at low x by using quadratic form for the structure function. This quadratic form approximation method can be used to determine the structure function F2 (x, Q2)from the HERA reduced cross section data taken at low x. This new approach can determine the structure functions F2(x,Q2) with reasonable precision even for low x values which have not been investigated. We observe that the Q2 dependence is quadratic over the full kinematic covered range. To test the validity of our new determined structure functions, wefind the gluon distribution function in the leading order approximation with our new calculation for the structure functions and compare them with the QCD parton distribution functions.     

Effect of interface/surface stress on the elastic wave band structure of two-dimensional phononic crystals

In the present Letter, the multiple scattering theory (MST) for calculating the elastic wave band structure of two-dimensional phononic crystals (PCs) is extended to include the interface/surface stress effect at the nanoscale. The interface/surface elasticity theory is employed to describe the nonclassical boundary conditions at the interface/surface and the elastic Mie scattering matrix embodying the interface/surface stress effect is derived. Using this extended MST, the authors investigate the interface/surface stress effect on the elastic wave band structure of two-dimensional PCs, which is demonstrated to be significant when the characteristic size reduces to nanometers.