Bone apatite anisotropic structure control via designing fibrous scaffolds

Bone tissue has an anisotropic structure, associated with the collagen fibrils'' orientation and the c-axis direction of the bone apatite crystal. The bone regeneration process comprises two main phases: bone mineral density restoration (bone quantity), and subsequent recovery of bone apatite c-axis orientation (bone quality). Bone quality is the determinant factor for mechanical properties of bone. Control of osteoblast alignment is one of the strategies for reconstructing bone quality since the collagen/apatite matrix orientation in calcified tissues is dependent on the osteoblast orientation. In this work, fibrous scaffolds designed for reconstruction of bone quality via cell alignment control was investigated. The fibrous scaffolds were fabricated using the electrospinning method with poly(lactic acid) at various fiber collecting speeds. The degree of fiber alignment in the prepared fibrous scaffolds increased with increasing fiber collecting speed, indicating that the fibers were oriented in a single direction. The alignment of osteoblasts on the fibrous scaffolds as well as the subsequent apatite c-axis orientation increased with increasing fiber collecting speed. We successfully controlled cell alignment and apatite c-axis orientation using the designed morphology of fibrous scaffolds. To the best of our knowledge, this is the first report demonstrating that adjusting the degree of fiber orientation for fibrous scaffolds can manipulate the regeneration of bone quality.

Osteoblast alignment on the fibrous scaffolds as well as the subsequent apatite c-axis orientation increased with increasing fiber collecting speed. We successfully controlled cell alignment and apatite c-axis orientation using the designed morphology of fibrous scaffolds.     

Microgrooved collagen-based corneal scaffold for promoting collective cell migration and antifibrosis

Collagen is a promising material for corneal tissue engineering. The surface topography of collagen is critical for cornea reconstruction and regeneration. Herein, we fabricated collagen films with microgrooved surface to investigate the effect of collagen film topography on corneal cell migration and antifibrosis. We found that the patterned films with microscale grooves could greatly affect the orientation, proliferation, migration, and gene expression of rabbit corneal epithelial cells and keratocytes. Compared with the natural cornea, the optimized collagen films with microgroove pattern show similar swelling performance, optical clarity, and biodegradability, which could efficiently promote the epithelial cell migration, accelerate wound healing process, and inhibit the fibrosis of keratocytes. These results suggest that collagen films with microgroove pattern are promising in corneal tissue engineering.

Microgrooved collagen membrane can effectively promote the epithelialization of corneal epithelial cells and inhibit the fibrosis of corneal stromal cells.     

Non-invasive evaluation of therapeutic response in keloid scar using diffuse reflectance spectroscopy

The pathogenesis and ideal treatment of keloid are still largely unknown, and it is essential to develop an objective assessment of keloid severity to evaluate the therapeutic response. We previously reported that our diffuse reflectance spectroscopy (DRS) system could assist clinicians in understanding the functional and structural condition of keloid scars. The purpose of this study was to understand clinical applicability of our DRS system on evaluating the scar severity and therapeutic response of keloid. We analyzed 228 spectral data from 71 subjects with keloid scars. The scars were classified into mild (0-3), moderate (4-7) and severe (8-11) according to the Vancouver scar scale. We found that as the severity of the scar increased, collagen concentration and water content increased, and the reduced scattering coefficient at 800 nm and oxygen saturation (SaO₂) decreased. Using the DRS system, we found that collagen bundles aligned in a specific direction in keloid scars, but not in normal scars. Water content and SaO₂ may be utilized as reliable parameters for evaluating the therapeutic response of keloid. In conclusion, the results obtained here suggest that the DRS has potential as an objective technique with which to evaluate keloid scar severity. In addition, it may be useful as a tool with which to track longitudinal response of scars in response to various therapeutic interventions.OCIS codes: (170.5280) Photon migration, (170.4580) Optical diagnostics for medicine, (170.6510) Spectroscopy, tissue diagnostics, (290.1990) Diffusion     

Tough and tissue-adhesive polyacrylamide/collagen hydrogel with dopamine-grafted oxidized sodium alginate as crosslinker for cutaneous wound healing

Natural collagen has good biocompatibility and ability to promote tissue regeneration; however, its low flexibility and easy degradation hinder its applications in wound repair. In this study, we synthesized a skin wound-repairing hydrogel with good bioactivity and high toughness and adhesion. Inspired by the good adhesion of natural mussels, dopamine was grafted onto oxidized sodium alginate to synthesize a new crosslinker (COA), which was introduced into the collagen/polyacrylamide (PAM-Col) double network to synthesize hydrogel. The morphological characterization of the hydrogel using scanning electron microscopy confirmed that the hydrogel formed a more chaotic interconnected structure after the introduction of COA. PAM-Col-COA hydrogel had good mechanical properties, skin tissue adhesion, water absorption, and sustained biological activity. In vivo wound healing experiments showed that hydrogel accelerates the wound healing process and has potential applications in wound dressings.

Natural collagen has good biocompatibility and ability to promote tissue regeneration; however, its low flexibility and easy degradation hinder its applications in wound repair.     

A novel lamellar structural biomaterial and its effect on bone regeneration

To evaluate a novel lamellar structural biomaterial as a potential biomaterial for guided bone regeneration, we describe the preparation of a collagen membrane with high mechanical strength and anti-enzyme degradation ability by using the multi-level structure of Ctenopharyngodon idella scales. The physical and chemical properties, in vitro degradation, biocompatibility, and in vivo osteogenic activity were preliminarily evaluated. In conclusion, it was shown that the multi-layered collagen structure material had sufficient mechanical properties, biocompatibility, and osteogenic ability. Meanwhile, it is also shown that there is a gap in current clinical needs, between the guided tissue regeneration membrane and the one being used. Therefore, this study provides useful insights into the efforts being made to design and adjust the microstructure to balance its mechanical properties, degradation rate, and osteogenic activity.

To evaluate a novel lamellar structural biomaterial for guided bone regeneration, we describe the preparation of a collagen membrane with high mechanical strength and anti-enzyme degradation ability using Ctenopharyngodon idella scales.     

Comparative study of aqueous solution processed ZnO/GaAs and ZnO/porous GaAs films

In this paper, we investigate the structural and photoluminescence properties of aqueous solution-processed ZnO/GaAs and ZnO/porous GaAs films. According to X-ray diffraction (XRD) analysis, a ZnO film deposited on porous GaAs shows a monocrystalline structure with a-axis orientation, which is desirable for light emitting applications. The results obtained from atomic force microscopy (AFM) data confirm that a porous GaAs substrate is beneficial to deposit a uniform array of ZnO nanostructures with sizes down to 12 nm and a relatively low surface roughness (2.6 nm). Under excitation wavelength λ_exc = 375 nm, ZnO/GaAs and ZnO/porous GaAs films showed emissions in most of the visible spectral region (450–750 nm). Our study reveals that changing the wavelength of the excitation UV radiation makes it possible to control the photoluminescence (PL) properties of ZnO films. Enhancement of the PL intensity was noticed in the UV and visible spectral regions when ZnO is deposited on porous GaAs, which is promising for optoelectronic device applications.

In this paper, we investigate the structural and photoluminescence properties of aqueous solution-processed ZnO/GaAs and ZnO/porous GaAs films.     

Modifying collagen with alendronate sodium for bone regeneration applications

Phosphorylated materials are attractive candidates for bone regeneration because they may facilitate the construction of a phosphorylated bone extracellular matrix (ECM) to build a beneficial environment for bone formation. Here, we designed and synthesized a new phosphorylated material, collagen type I phosphorylated with alendronate sodium (Col-Aln), based on the biodegradable osteoconductive collagen backbone. Col-Aln can distinctly accelerate in vitro mineralization in simulated body fluid. Col-Aln showed good biocompatibility with bone marrow mesenchymal stem cells (BMSCs) and promoted their adhesion as well as the osteogenic differentiation of BMSCs more effectively than did pure collagen. Furthermore, collagen and Col-Aln scaffolds implanted into a critical-sized rat cranial defect for 4 and 8 weeks were shown to degrade in vivo and helped to facilitate bone growth in the defect, while the phosphate-containing Col-Aln scaffold significantly promoted new bone formation. Col-Aln provides a new strategy to integrate bioactive phosphate molecules via covalent grafting onto biopolymers and has promise for bone regeneration applications.

Efficient covalent bonding with phosphate-containing alendronate prompts the fast mineralization and osteoinduction of the collagen scaffold.     

Terminal aspartic acids promote the self-assembly of collagen mimic peptides into nanospheres

The development of novel strategies to construct collagen mimetic peptides capable of self-assembling into higher-order structures plays a critical role in the discovery of functional biomaterials. We herein report the construction of a novel type of amphiphile-like peptide conjugating the repetitive triple helical (GPO)_m sequences characteristic of collagen with terminal hydrophilic aspartic acids. The amphiphile-like collagen mimic peptides containing a variable length of (Gly-Pro-Hyp)_m sequences consistently generate well-ordered nanospherical supramolecular structures. The C-terminal aspartic acids have been revealed to play a determinant role in the appropriate self-assembly of amphiphile-like collagen mimic peptides. Their presence is a prerequisite for self-assembly, and their lengths could modulate the morphology of final assemblies. We have demonstrated for the first time that amphiphile-like collagen mimic peptides with terminal aspartic acids may provide a general and convenient strategy to create well-defined nanostructures in addition to amphiphile-like peptides utilizing β-sheet or α-helical coiled-coil motifs. The newly developed assembly strategy together with the ubiquitous natural function of collagen may lead to the generation of novel improved biomaterials.

Amphiphile-like collagen mimic peptides with terminal aspartic acids may provide a general and convenient strategy to create well-defined nanostructures.     

The enhanced osteogenesis and osteointegration of 3-DP PCL scaffolds via structural and functional optimization using collagen networks

Optimal balance between biological activity and mechanical stability should be meticulously considered during scaffold design for bone tissue engineering applications. To fabricate an individualized construct with biomechanical and biological functionality for bone tissue regeneration, a polycaprolactone–collagen (PCL–COL) composite construct was developed through the combination of three-dimensional printing (3-DP) technology and biomimetic collagen matrix incorporation, with a 3-DP PCL framework maintaining the mechanical stability and a porous collagen matrix improving the biological activity. The results indicate that the compressive modulus of the composite constructs increased synergistically (over 40 MPa), providing sufficient mechanical support during new bone formation. On the other hand, the collagen matrix with a micro-porous architecture structurally increased scaffold areas and provided cellular adhesion sites, allowing for the functional construction of a favorable 3D microenvironment for BMSC adhesion, proliferation and extracellular matrix production. Moreover, critical-sized long bone defect (CSD) implantation demonstrated that the optimized composite constructs could promote bone tissue regeneration (5.5-fold) and bone-material osteointegration (4.7-fold), and decrease fibrosis encapsulation, compared to pristine PCL. The results indicate that these biomimetically ornamented PCL–COL constructs exhibit favorable mechanical properties and biological functionality, demonstrating great potential as an effective bone graft substitute for bone defect treatment. Meanwhile, they can also harness the advantages of 3-DP technology and a collagen-based functionalized strategy, facilitating the creation of customized and functional PCL–COL constructs for clinical translation.

Optimal balance between biological activity and mechanical stability should be meticulously considered during scaffold design for bone tissue engineering applications.     

Enzymatic synthesis and self-assembly of glycolipids: robust self-healing and wound closure performance of assembled soft materials

In developing countries, wounds are a major health concern and pose a significant problem. Hence, the development of new materials that can act as scaffolds for in situ tissue regeneration and regrowth is necessary. In this report, we present a new class of injectable oleogel and composite gel derived from glycolipids that provide reversible interlinked 3D fiberous network architecture for effective wound closure by tissue regrowth and regeneration. Glycolipids were derived from α-chloralose and various vinyl esters using Novozyme 435, an immobilized lipase B from Candida antarctica as a catalyst, in good yield. These glycolipids undergo spontaneous self-assembly in paraffin oil to form an oleogel, in which curcumin was successfully incorporated to generate a composite gel. Morphological analysis of the oleogel and composite gel clearly revealed the formation of a 3D fiberous network. Rheological investigation revealed the thermal and mechanical processability of the oleogel and composite gel under various experimental conditions. Interestingly, the developed injectable oleogel and composite gel are able to accelerate the wound healing process by regulating the overlapping phases of inflammation, cell proliferation and extracellular matrix remodelling. Since chloralose displays anesthetic properties, this study will establish a new strategy to develop anesthetic wound healing oleogels in the future.

In this report, we present a new class of injectable oleogels and a composite gel derived from glycolipids that provide a reversible interlinked 3D fiberous network architecture for effective wound closure by tissue regrowth and regeneration.     

Studies on interstitial carbon doping from a Ti precursor in a hierarchical TiO2 nanostructured photoanode by a single step hydrothermal route

Carbon doping from a Ti precursor in TiO₂ synthesized by a hydrothermal method was studied. The structural, optical and morphological study of the deposited material was carried out using X-ray diffraction, UV-vis spectroscopy and scanning electron microscopy characterization techniques. The elemental composition of the TiO₂ deposited with different precursor concentrations was studied using X-ray photoelectron spectroscopy and electron dispersive X-ray spectroscopy. The amount of elemental carbon in the TiO₂ matrix is found to be increased as the Ti precursor concentration is increased, which strengthens the proposed idea of carbon doping via a Ti precursor.

Interstitial carbon doping is possible directly from a Ti precursor (titanium(iv) isopropoxide) without using any other carbon source.     

Anisotropic architecture and electrical stimulation enhance neuron cell behaviour on a tough graphene embedded PVA: alginate fibrous scaffold

Tough scaffolds comprised of aligned and conductive fibers are promising for peripheral nerve regeneration due to their unique mechanical and electrical properties. Several studies have confirmed that electrical stimulation can control the axonal extension in vitro. However, the stimulatory effects of scaffold architecture and electrical stimulation have not yet been investigated in detail. Here, we assessed a comparison between aligned and random fibers made of graphene (Gr) embedded sodium alginate (SA) polyvinyl alcohol (PVA) (Gr-AP scaffolds) for peripheral nerve engineering. The effects of applied electrical stimulation and orientation of the fabricated fibers on the in vitro attachment, alignment, and proliferation of PC12 cells (a rat neuronal cell line) were investigated. The results revealed that the aligned fibrous Gr-AP scaffolds closely mimicked the anisotropic structure of the native sciatic nerve. Aligned fibrous Gr-AP scaffolds significantly improved mechanical properties as well as cell-scaffold integration compared to random fibrous scaffolds. In addition, electrical stimulation significantly improved PC12 cell proliferation. In summary, our findings revealed that aligned fibrous Gr-AP scaffolds offered superior mechanical characteristics and structural properties that enhanced neural cell–substrate interactions, resulting in a promising construct for nerve tissue regeneration.

Tough scaffolds comprised of aligned and conductive fibers are promising for peripheral nerve regeneration due to their unique mechanical and electrical properties.     

Impact of type-1 collagen hydrogel density on integrin-linked morphogenic response of SH-SY5Y neuronal cells

Cellular metabolism and behaviour is closely linked to cytoskeletal tension and scaffold mechanics. In the developing nervous system functional connectivity is controlled by the interplay between chemical and mechanical cues that initiate programs of cell behaviour. Replication of functional connectivity in neuronal populations in vitro has proven a technical challenge due to the absence of many systems of biomechanical regulation that control directional outgrowth in vivo. Here, a 3D culture system is explored by dilution of a type I collagen hydrogel to produce variation in gel stiffness. Hydrogel scaffold remodelling was found to be linked to gel collagen concentration, with a greater degree of gel contraction occurring at lower concentrations. Gel mechanics were found to evolve over the culture period according to collagen concentration. Less concentrated gels reduced in stiffness, whilst a biphasic pattern of increasing and then decreasing stiffness was observed at higher concentrations. Analysis of these cultures by PCR revealed a program of shifting integrin expression and highly variable activity in key morphogenic signal pathways, such as mitogen-associated protein kinase, indicating genetic impact of biomaterial interactions via mechano-regulation. Gel contraction at lower concentrations was also found to be accompanied by an increase in average collagen fibre diameter. Minor changes in biomaterial mechanics result in significant changes in programmed cell behaviour, resulting in adoption of markedly different cell morphologies and ability to remodel the scaffold. Advanced understanding of cell–biomaterial interactions, over short and long-term culture, is of critical importance in the development of novel tissue engineering strategies for the fabrication of biomimetic 3D neuro-tissue constructs. Simple methods of tailoring the initial mechanical environment presented to SH-SY5Y populations in 3D can lead to significantly different programs of network development over time.

Cellular metabolism and behaviour is closely linked to cytoskeletal tension and scaffold mechanics.     

Collagen Biosynthesis in Normal Human Skin, Normal and Hypertrophic Scar and Keloid

Summary. A comparison of the rates of synthesis of collagen in normal skin, normal and hypertrophic scars, and keloids has been made by measuring the rate of incorporation of [¹⁴C]-proline into peptide-bound [¹⁴C]-hydroxyproline by tissue minces in vitro. The rate of synthesis of collagen, as measured by this technique, was significantly higher in skin than in normal scars whether the incorporation of radioactivity into hydroxyproline were expressed in terms of wet weight of tissue, weight of tissue DNA or weight of tissue hydroxyproline. The abnormal scar types exhibited similar rates of collagen synthesis, which were significantly higher than the rate in normal scars. Although the rates in both abnormal scar types appeared to be similar to that in normal skin when expressed in terms of wet weight of tissue, and weight of tissue hydroxyproline, they were seen to be lower than in skin in terms of weight of tissue DNA. The rate of synthesis of proteins generally, as measured by total radioactivity in non-diffusible peptides, was highest in normal skin and hypertrophic scar and lowest in keloid. The ratio of radioactivity in non-diffusible hydroxyproline to total non-diffusible radioactivity was almost twice as high in keloid as in normal scar, with intermediate values being observed in hypertrophic scar and normal skin. This indicated that collagen accounted for a higher proportion of the proteins being synthesised in keloid than in normal scar. The results confirm previous conclusions, from determination of the activity of the enzyme collagen proline hydroxylase, that the excessive accumulation of collagen in hypertrophic scars and keloids may, at least in part, be due to abnormally high rates of collagen synthesis in comparison to normal scars.     

Effect of nanoclay orientation on oxygen barrier properties of LbL nanocomposite coated films

Layer by layer (LbL) film deposition is an efficient technique used to produce thin coatings with high gas barrier properties. In this study, multilayer composite coatings with hydrogen bonding inter-layer interactions were deposited by LbL on a PET substrate, with an alternate deposition of a nanoclay layer and different intercalating polymers layers, namely chitosan (CS), polyethylene oxide (PEO), polyvinylpyrrolidone (PVP) and polyvinyl alcohol (PVA). The investigated coatings had two different structures, quadlayers and bilayers which are different in the number of layers in the repetitive unit (four and two respectively). The alignment of nanoclay platelets and the extent of their intercalation were studied using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD). The results showed that the dispersion level and the orientation of nanoclay particles depend considerably on the molecular structure of intercalating polymers and their interactions with nanoclay. An oxygen permeability model, specific to high filler loading composites, was then developed by considering only the aspect ratio and the volume fraction of the nanoparticles.

Quantify clay orientation in a layer by layer deposited films. Demonstrate the importance of interfacial region in a high clay loading nanocomposite film, in decreasing permeability.     

Bimodal spectroscopy for in vivo characterization of hypertrophic skin tissue : pre-clinical experimentation,data selection and classification

This study aims at investigating the efficiency of bimodal spectroscopy in detection of hypertrophic scar tissue on a preclinical model. Fluorescence and Diffuse Reflectance spectra were collected from 55 scars deliberately created on ears of 20 rabbits, amongst which some received tacrolimus injection to provide non-hypertrophic scar tissue. The spectroscopic data measured on hypertrophic and non-hypertrophic scar tissues were used for developing our classification algorithm. Spectral features were extracted from corrected data and analyzed to classify the scar tissues into hypertrophic or non-hypertrophic. The Algorithm was developed using k-NN classifier and validated by comparing to histological classification result with Leave-One-Out cross validation. Bimodal spectroscopy showed promising results in detecting hypertrophic tissue (sensibility 90.5%, specificity 94.4%). The features used for classification were extracted from the autofluorescence spectra collected at 4 CEFS with excitations at 360, 410, and 420 nm. This indicates the hypertrophic process may involve change in concentration of several fluorophores (collagen, elastin and NADH) excited in this range, or modification in volume of explored tissue layers (epidermis and dermis) due to tissue thickening.OCIS codes: (120.6200) Spectrometers and spectroscopic instrumentation, (300.6550) Spectroscopy, visible, (070.4790) Spectrum analysis     

The effect of solvent on Al-doped ZnO thin films deposited via aerosol assisted CVD

Aluminium-doped zinc oxide (AZO) thin films were deposited via aerosol assisted chemical vapour deposition (AACVD) from zinc acetylacetonate and aluminium chloride at 450 °C. The precursor solutions consisted of methanol in a mixture with one other secondary solvent, including toluene, tetrahydrofuran, n-hexane, cyclohexane, and ethyl acetate. The crystal structures, elemental compositions and surface morphologies of the resulting AZO films were determined, as well as the optoelectronic properties. It was found that the more polar solvents enhanced growth in the (002) plane of the wurtzite crystal structure, and that solutions with low viscosities resulted in superior grain growth. The film deposited from a solution consisting of methanol and ethyl acetate displayed the lowest visible transmittance, due to carbon contamination. However, it also exhibited 60% lower resistivity, in comparison to the film deposited using methanol only. This suggests that optoelectronic properties can be tuned for specific photovoltaic devices.

Aluminium-doped zinc oxide (AZO) thin films were deposited via aerosol assisted chemical vapour deposition (AACVD) from zinc acetylacetonate at 450 °C.     

Biofunctionalization of decellularized porcine aortic valve with OPG-loaded PCL nanoparticles for anti-calcification

Decellularized valve stents are widely used in tissue-engineered heart valves because they maintain the morphological structure of natural valves, have good histocompatibility and low immunogenicity. However, the surface of the cell valve loses the original endothelial cell coverage, exposing collagen and causing calcification and decay of the valve in advance. In this study, poly ε-caprolactone (PCL) nanoparticles loaded with osteoprotegerin (OPG) were bridged to a decellularized valve using a nanoparticle drug delivery system and tissue engineering technology to construct a new anti-calcification composite valve with sustained release function. The PCL nanoparticles loaded with OPG were prepared via an emulsion solvent evaporation method, which had a particle size of 133 nm and zeta potential of −27.8 mV. Transmission electron microscopy demonstrated that the prepared nanoparticles were round in shape, regular in size, and uniformly distributed, with an encapsulation efficiency of 75%, slow release in vitro, no burst release, no cytotoxicity to BMSCs, and contained OPG nanoparticles in vitro. There was a delay in the differentiation of BMSCs into osteoblasts. The decellularized valve modified by nanoparticles remained intact and its collagen fibers were continuous. After 8 weeks of subcutaneous implantation in rats, the morphological structure of the valve was almost complete, and the composite valve showed anti-calcification ability to a certain extent.

A novel composite valve with controlled release OPG was prepared by introducing tissue engineering technology and nano drug-loading system to introduce anti-calcification biological factor OPG on the decellularized valve.     

Comparison of NiOx thin film deposited by spin-coating or thermal evaporation for application as a hole transport layer of perovskite solar cells

We compared nickel oxide (NiO_x) deposited by thermal evaporation and that deposited by the spin-coating process, for use in the hole transport layers of inverted planar perovskite solar cells (PSCs). Spin-coating deposition for NiO_x HTL has been widely used, owing to its simplicity, low cost, and high efficiency. However, the spin-coating process has a technical limit to depositing a large-area uniformly. In contrast, thermal evaporation fabrication has a low price and is able to produce uniform and reproducible thin film. Hence, the chemical states, energy band alignment, surface morphologies, and microstructures of NiO_x deposited by spin coating and thermal evaporation were analyzed. The PSC with NiO_x HTL deposited by thermal evaporation showed a higher power conversion efficiency of 16.64% with open circuit voltage 1.07 V, short circuit current density of 20.68 mA cm⁻², and a fill factor of 75.51% compared to that of PSC with spin-coated NiO_x. We confirmed that thermal evaporation can deposit NiO_x to give a better performance as a HTL with higher reproducibility than spin-coating.

We compared nickel oxide (NiO_x) deposited by thermal evaporation and that deposited by the spin-coating process, for use in the hole transport layers of inverted planar perovskite solar cells (PSCs).     

Raman spectroscopic detection of carotenoids in cattle skin

Carotenoids, powerful anti-oxidants, play a significant role in protecting the skin from oxidation and help in balancing the redox status of skin. This study was aimed at investigating cattle skin to identify carotenoids in the lower epidermis (grain) and dermis (corium) layers for classification using Raman spectroscopy which is a powerful technique for the detection of carotenoids in cattle skin due to the strong resonance enhancement with 532 nm laser excitation. The spectral differences identified between these two layers were quantified by the univariate analysis of Raman peak heights and partial least squares (PLS) analysis. We compared the performance of the Raman spectroscopy method with the standard method, high performance liquid chromatography. The univariate analysis results demonstrated that the lower epidermis of the skin has a higher concentration of carotenoid than dermis using the carotenoid Raman peaks at 1151 cm⁻¹ and 1518 cm⁻¹. The carotenoid Raman intensity was linearly correlated with the total carotenoid concentration determined by standard HPLC methods. Partial Least Squares Regression analysis gives excellent results with R² = 0.99. Our results indicate that Raman spectroscopy is a potential tool to determine carotenoids in cattle skin with high precision.

The lower epidermis and dermis of cattle skin was investigated for the detection of carotenoids using Raman spectroscopy.