FTIR Imaging Research Articles

Stable Liposome in Cosmetic Platforms for Transdermal Folic acid delivery for fortification and treatment of micronutrient deficiencies

Oral folate fortification has been successful in many developed nations, however, developing countries still face low compliance and high incidence of folate deficiency associated with low birth weight infants and preterm deliveries. We report safe and efficient approach for transdermal systemic folate delivery using fluidising liposomes (120 ± 4 nm) stabilised within 3D matrix of naturally occurring cosmetic bases: Fuller’s earth and henna with room temperature stability. The proof of stratum corneum fluidisation was established ex-vivo by Langmuir-Blodgett film, FTIR and confocal imaging in rat skin. In-vivo topical application in rats showed 11-fold increase in plasma folate within 2 hr, confirming systemic delivery through skin. Efficacy study in folate deficient rats over 4 weeks showed significantly higher plasma levels compared to oral delivery with significant skin depot. Sub-acute toxicity studies in rats at 750-fold higher doses showed safety after 4 weeks daily application. Primary irritation patch test on 25 healthy human volunteers proved non-irritant nature of the nutricosmetics. The technology is first demonstration of transdermal folate fortification with nanosized liposome incorporated in cosmetics, without synthetic surfactants/ethanol or need of external energy. The platform technology opens the possibility of delivering multiple nutrients systemically through skin and can be scaled for affordable community fortification.

Chemometric Models for Quantification of Carbamazepine Anhydrous and Dihydrate Forms in the Formulation

Carbamazepine (CBZ) exists in anhydrous and dihydrate forms. These forms differ in their solubility, dissolution rate, and subsequently in their oral bioavailability. The objective of this study is to develop multivariate chemometric models for estimation of the low level of carbamazepine dihydrate (CBZ-DH) in the CBZ formulations containing excipients of the commercial formulation. The selected excipients were mixed in proportions to make sample matrices ranging from 0% to 50% CBZ-DH. Fourier transform infrared (FTIR), near infrared (NIR), and hyperspectral imaging data were mathematically pretreated before the development of partial least square and principal component analysis regression models. The developed partial least squares regression and principal component analysis models demonstrated predictability of CBZ and CBZ-DH by multiple scattering correction and standard normal variate processing methods. Among the spectroscopic techniques used the model performance parameters such as root-mean-square error, standard error, and bias were found to be low for NIR compared to FTIR. The treated data have shown better model fitting than without treatment, which was demonstrated by correlation coefficient of 0.9778, 0.9824, and 0.9852 for FTIR, NIR, and hyperspectral imaging, respectively. Furthermore, the predicted values were found to be very close to the selected low level of independent samples having 5% CBZ-DH in tablet formulation.

Comparison of Raman and Fourier Transform Infrared Spectroscopy for the Quantification of Microplastics in the Aquatic Environment

Microplastics (MPs, <5 mm) have been reported as emerging environmental contaminants, but reliable data are still lacking. We compared the two most promising techniques for MP analysis, namely, Raman and Fourier transform infrared (FTIR) spectroscopy, by analyzing MPs extracted from North Sea surface waters. Microplastics >500 μm were visually sorted and manually analyzed by μ-Raman and attenuated total reflection (ATR)-FTIR spectroscopy. Microplastics ≤500 μm were concentrated on gold-coated filters and analyzed by automated single-particle exploration coupled to μ-Raman (ASPEx-μ-Raman) and FTIR imaging (reflection mode). The number of identified MPs >500 μm was slightly higher for μ-Raman (+23%) than ATR-FTIR analysis. Concerning MPs ≤500 μm, ASPEx-μ-Raman quantified two-times higher MP numbers but required a four-times higher analysis time compared to FTIR imaging. Because ASPEx-μ-Raman revealed far higher MP concentrations (38-2621 particles m-3) compared to the results of previous water studies (0-559 particles m-3), the environmental concentration of MPs ≤500 μm may have been underestimated until now. This may be attributed to the exceptional increase in concentration with decreasing MP size found in this work. Our results demonstrate the need for further research to enable time-efficient routine application of ASPEx-μ-Raman for reliable MP counting down to 1 μm.

Comparison of spectral and spatial denoising techniques in the context of High Definition FT-IR imaging hyperspectral data

The recent emergence of High Definition (HD) FT-IR and Quantum Cascade Laser (QCL) Microscopes elevated the IR imaging field very close to clinical timescales. However, the speed of acquisition and data quality are still the critical factors in reaching the clinic. Denoising offers aide in both aspects if performed properly. However, there is a lack of a direct comparison of the efficiency of denoising techniques in IR imaging in general. To achieve such comparison within a rigorous framework and obtaining the critical information about signal loss, a simulated dataset strongly bound by experimental parameters was created. Using experimental structural and spectral information and experimental noise levels data as an input for the simulation, a direct comparison of spatial (Fourier transform, Mean Filter, Weighted Mean Filter, Gauss Filter, Median Filter, spatial Wavelets and Deep Neural Networks) and spectral (Savitzky-Golay, Fourier transform, Principal Component Analysis, Minimum Noise Fraction and spectral Wavelets) denoising schemes was enabled. All of these techniques were compared on the simulated dataset, taking into account SNR gain, signal distortion and sensitivity to tuning parameters as comparison metrics. Later, the best techniques were applied to experimental data for validation. The results presented here clearly show the benefit of using hyperspectral denoising schemes such as PCA and MNF which outperform other methods.

Fourier-Transform Infrared Imaging Spectroscopy and Laser Ablation -ICPMS New Vistas for Biochemical Analyses of Ischemic Stroke in Rat Brain.

Objective: Stroke is the main cause of adult disability in the world, leaving more than half of the patients dependent on daily assistance. Understanding the post-stroke biochemical and molecular changes are critical for patient survival and stroke management. The aim of this work was to investigate the photo-thrombotic ischemic stroke in male rats with particular focus on biochemical and elemental changes in the primary stroke lesion in the somatosensory cortex and surrounding areas, including the corpus callosum.Materials and Methods: FT-IR imaging spectroscopy and LA-ICPMS techniques examined stroke brain samples, which were compared with standard immunohistochemistry studies.Results: The FTIR results revealed that in the lesioned gray matter the relative distribution of lipid, lipid acyl and protein contents decreased significantly. Also at this locus, there was a significant increase in aggregated protein as detected by high-levels Aβ1-42. Areas close to the stroke focus experienced decrease in the lipid and lipid acyl contents associated with an increase in lipid ester, olefin, and methyl bio-contents with a novel finding of Aβ1-42 in the PL-GM and L-WM. Elemental analyses realized major changes in the different brain structures that may underscore functionality.Conclusion: In conclusion, FTIR bio-spectroscopy is a non-destructive, rapid, and a refined technique to characterize oxidative stress markers associated with lipid degradation and protein denaturation not characterized by routine approaches. This technique may expedite research into stroke and offer new approaches for neurodegenerative disorders. The results suggest that a good therapeutic strategy should include a mechanism that provides protective effect from brain swelling (edema) and neurotoxicity by scavenging the lipid peroxidation end products.

Dynamic structure and composition of bone investigated by nanoscale infrared spectroscopy.

Bone is a highly organized tissue in which each structural level influences the macroscopic and microscopic mechanical behavior. In particular, the quantity, quality, and distribution of the different bone components, i.e. collagen matrix and hydroxyapatite crystals, are associated with bone strength or fragility. Common spectroscopic techniques used to assess bone composition have resolutions limited to the micrometer range. In this study, our aims were two-fold: i) to develop and validate the AFM-IR methodology for skeletal tissues and ii) to apply the methodology to sheep cancellous bone with the objective to obtain novel findings on the composition and structure of trabecular packets.To develop the methodology, we assessed spatial and temporal reproducibility using a known homogeneous material (polymethylmethacrylate, PMMA). We verified that the major peak positions were similar and not shifted when compared to traditional Fourier Transform Infrared imaging (FTIRI). When AFM-IR was applied to sheep cancellous bone, the mineral-to-matrix ratio increased and the acid phosphate substitution ratio decreased as a function of tissue maturity. The resolution of the technique enabled visualization of different stages of the bone maturation process, particularly newly-formed osteoid prior to mineralization. We also observed alternating patterns of IR parameters in line and imaging measurements, suggesting the apposition of layers of alternating structure and / or composition that were not visible with traditional spectroscopic methods. In conclusion, nanoscale IR spectroscopy demonstrates novel compositional and structural changes within trabecular packets in cancellous bone. Based on these results, AFM-IR is a valuable tool to investigate cancellous bone at the nanoscale and, more generally, to analyze small dynamic areas that are invisible to traditional spectroscopic methods.

Label-free FTIR spectroscopy detects and visualizes the early stage of pulmonary micrometastasis seeded from breast carcinoma

An application of FTIR spectroscopic imaging for the identification and visualization of early micrometastasis from breast cancer to lungs in a murine model is shown. Spectroscopic and histological examination is focused on lung cross-sections derived from animals at the early phase of metastasis (early micrometastasis, EM) as compared to healthy control (HC) and late phase of metastasis (advanced macrometastasis, AM) using murine model of metastatic breast cancer with 4T1 cells orthotopically inoculated. FTIR imaging allows for a detailed, objective and label-free differentiation and visualization of EM foci including large and small micrometastases as well as single cancer cells grouped in clusters. An effect of the EM phase on the entire lung tissue matrix as well as characteristic biochemical profiles for HC and advanced macrometastasis were determined from morphological and spectroscopic points of view. The extraordinary sensitivity of FTIR imaging toward EM detection and discrimination of AM borders confirms its applicability as a complementary tool for the histopathological assessment of the metastatic cancer progression.

Preparation and characterization of engineered stones based geopolymer composites

The objective of the present paper is to study the effect of combination between the low liquid/solid (l/s) ratio and pressure compaction on the properties of the hardened engineered stone alkaline hybrid composites of water cooled slag/metakaolin using granite as filler and metakaolin using fine sand as filler in the ratio of 1:3, while the used activator was 5% sodium hydroxide with liquid to solid ratio of 10% to ensure low water content for better organization of the binding materials upon pressure compaction. It is well known that these combinations increase the possibility of preparing engineered stones composites with excellent engineering properties. The results showed a high dense nano- or near-nano-pore structure with high degree of homogeneity and the high strength of alkaline hardened paste compacted with low l/s ratio.The properties of the produced engineered stone alkaline hybrid composites have been studied through measurement of compressive strength, water absorption, bulk density, FTIR, petrographic examination and SEM-BSE imaging. The results exposed the possibility of forming a compact and dense composite on compacting with pressures ranging from 30 to 70 MPa.

Low numbers of microplastics detected in drinking water from ground water sources

Microplastic particles have been detected in various natural habitats and the digestive tracts of several species. These particles have also been reported in commercially available seafood, salt or bottled water starting discussions on potential implications for human health. To be able to assess the related risks, exposure concentrations and pathways need to be known. Here, we analysed ground water and drinking water for the presence of microplastics (>20 μm) using FTIR imaging. Samples were taken at different positions within the drinking water supply chain. Determined concentrations ranged from 0 to 7 microplastics m−3 raw water or drinking water with an overall mean of 0.7 microplastics m−3. These particles were identified as polyethylene, polyamide, polyester, polyvinylchloride or epoxy resin and between 50 and 150 μm in size. Plastic is a resistant and durable material, still, the abrasion of plastic equipment used during water purification or transport is a likely explanation for the plastic particles detected in water samples.

Polymer derived silicon oxycarbide ceramic monoliths: Microstructure development and associated materials properties

Polymer derived SiOC and SiCN ceramics (PDCs) are interesting candidates for additive manufacturing techniques to develop micro sized ceramics with the highest precision. PDCs are obtained by the pyrolysis of crosslinked polymer precursors at elevated temperatures. Within this work, we are investigating PDC SiOC ceramic monoliths synthesized from liquid polysiloxane precursor crosslinked with divinylbenzene for fabrication of conductive electromechanical devices. Microstructure of the final ceramics was found to be greatly influenced by the pyrolysis temperature. Crystallization in SiOC ceramics starts above 1200 °C due to the onset of carbothermal reduction leading to the formation of SiC and SiO2 rich phases. Microstructural characterisation using ex-situ X-ray diffraction, FTIR, Raman spectra and microscopy imaging confirms the formation of nano crystalline SiC ceramics at 1400 °C. The electrical and mechanical properties of the ceramics are found to be significantly influenced by the phase separation with samples becoming more electrically conducting but with reduced strength at 1400 °C. A maximum electrical conductivity of 101 S cm−1 is observed for the 1400 °C samples due to enhancement in the ordering of the free carbon network. Mechanical testing using the ball on 3 balls (B3B) method revealed a characteristic flexural strength of 922 MPa for 1000 °C amorphous samples and at a higher pyrolysis temperature, materials become weaker with reduced strength.

New insights on the macromolecular building of rainbow trout (O. mykiss) intestine: FTIR Imaging and histological correlative study

Fish intestine plays various physiological functions and its integrity is essential to guarantee fish growth and welfare. Several laboratory techniques have been proposed for studying morpho-physiology of fish intestine, but classical histology still represents one of the gold standards. Nevertheless, this technique is time consuming and does not provide comprehensive information on the biochemical composition of tissues. The present study, introduces a new approach for studying fish intestine, the Fourier Transform Infrared (FTIR) spectroscopy. This is a fast, label-free analytical technique, which analyses the vibrational transitions induced on tissues by the interaction with the electromagnetic radiation. On the base of vibrational features of some functional groups classes and molecular bonds, FTIR allows the analysis of the macromolecular composition of the sample. Rainbow trout (O. mykiss) represents an important species for modern aquaculture and deeper knowledge about its intestine anatomy and macromolecular composition is desirable. In the present study, besides the traditional histological analysis, the biochemical building of the intestine mucosa was analyzed for the first time by FTIR. Spectral data can be considered a complement to traditional histological analysis.

Subcellular mapping of living cells via synchrotron microFTIR and ZnS hemispheres

FTIR imaging is a label-free, non-destructive method valuably exploited in the study of the biological process in living cells. However, the long wavelength/low spatial resolution and the strong absorbance of water are still key constrains in the application of IR microscopy ex vivo. In this work, a new retrofit approach based on the use of ZnS hemispheres is introduced to significantly improve the spatial resolution on live cell FTIR imaging. By means of two high refractive index domes sandwiching the sample, a lateral resolution close to 2.2 μm at 6 μm wavelength has been achieved, i.e. below the theoretical diffraction limit in air and more than twice the improvement (to ~λ/2.7) from our previous attempt using CaF2 lenses. The ZnS domes also allowed an extended spectral range to 950 cm−1, in contrast to the cut-off at 1050 cm−1 using CaF2. In combination with synchrotron radiation source, microFTIR provides an improved signal-to-noise ratio through the circa 12 μm thin layer of medium, thus allowing detailed distribution of lipids, protein and nucleic acid in the surround of the nucleus of single living cells. Endoplasmic reticula were clearly shown based on the lipid ν(CH) and ν(C=O) bands, while the DNA was imaged based on the ν(PO2−) band highlighting the nucleus region. This work has also included a demonstration of drug (doxorubicin) in cell measurement to highlight the potential of this approach.Graphical abstract

Effect of processing parameters and controlled environment storage on the disproportionation and dissolution of extended-release capsule of phenytoin sodium

Phenytoin sodium (PS) is a narrow therapeutic index drug that dictates maintenance of narrow plasma concentration of the drug. This requires a good quality product that meets regulatory specifications throughout shelf-life as well as during the usage period. Quality of the drug product may change if not develop in a scientific fashion which may have ramifications on clinical outcome. The focus of the study was to understand the effect of process variables and storage conditions on the disproportionation and phase transformation of the drug, and dissolution of extended-release capsules of PS. Four formulations were prepared to contain either powder blend or granules of PS with commonly used excipients. The granules were prepared in a high shear mixture by granulating with water, ethanol or mixture thereof. The capsules were stored in pharmacy vials for 4-weeks at 40 °C/75% RH and 30 °C/65% RH. Formulations were characterized by spectroscopies (FTIR, NIR, and chemical imaging), X-ray powder diffraction (XRPD), dissolution and assay. Granules filled capsule did not meet USP dissolution specifications and there was a significant decrease in dissolution on exposure to stability conditions. Dissolution decreased from 77.56 ± 3.83% to 42.34 ± 4.31%, and 63.96 ± 6.12 to 46.53 ± 2.91 in physical mixture and water granulated formulations, respectively, after four weeks exposure at 40 °C/75% RH. Spectroscopic (NIR, FTIR, and chemical imaging) data indicated disproportionation of the drug during the storage period. Moreover, XRPD data explained a decrease in dissolution in the initial granules filled capsules as well as temperature and humidity exposed samples. It indicated the phase transformation of the drug and/or excipients, disproportionation and/or interactions. These changes further accelerated on exposure to stability conditions. In conclusion, our studies indicated the significant effect of process variables and stability conditions on the critical quality attributes of PS extended-release capsule that has the potential impact on the clinical performance of the product.

Hyperspectral FTIR imaging of olive fruit for understanding ripening processes

A combination of micro-FTIR hyperspectral imaging and chemometrics is proposed to better understand biochemical changes in olive fruit during the ripening process without the need for fractionation and related analytical methods. Principal component analysis was used for the study of the spatial distribution of vibrational spectroscopic signatures in flesh, hypodermis and epidermis tissues. Partial least squares discriminant analysis was then employed to highlight the main spectral changes related to ripening. Modifications in cell wall polysaccharides were identified as the major processes taking place during ripening in both the flesh and the hypodermis whereas olive oil accumulation was characteristic in the flesh of the fruit. Evidence of solubilization and depolymerization of cell wall polysaccharides as well as demethylation with the subsequent formation of carboxylate groups were found. Spectral features attributed to polyphenols and sugars also change during the ripening process. No clear temporal trends were observed in the epidermal tissues.

Early changes in the extracellular matrix of the degenerating intervertebral disc, assessed by Fourier transform infrared imaging

Mechanical overloading induces a degenerative cell response in the intervertebral disc. However, early changes in the extracellular matrix (ECM) are challenging to assess with conventional techniques. Fourier Transform Infrared (FTIR) imaging allows visualization and quantification of the ECM. We aim to identify markers for disc degeneration and apply these to investigate early degenerative changes due to overloading and katabolic cell activity. Three experiments were conducted; Exp 1.: Invivo, lumbar spines of seven goats were operated: one disc was injected with chondroitinase ABC [cABC (mild degeneration)] and compared to the adjacent disc (control) after 24 weeks. Exp 2a: Exvivo, caprine discs received physiological loading (n=10) or overloading (n=10) in a bioreactor. Exp 2b: Cell activity was diminished prior to testing by freeze-thaw cycles, 18 discs were then tested as in Exp 2a. In all experiments, FTIR images (spectral region: 1000-1300cm-1) of mid-sagittal slices were analyzed using multivariate curve resolution. Invivo, FTIR was more sensitive than biochemical and histological analysis in identifying reduced proteoglycan content (P=0.046) and increased collagen content in degenerated discs (P<0.01). Notably, FTIR analysis additionally showed disorganization of the ECM, indicated by increased collagen entropy (P=0.011). Exvivo, the proteoglycan/collagen ratio decreased due to overloading (P=0.047) and collagen entropy increased (P=0.047). Cell activity affected collagen content only (P=0.044). FTIR imaging allows a more detailed investigation of early disc degeneration than traditional measures. Changes due to mild overloading could be assessed and quantified. Matrix remodeling is the first detectable step towards intervertebral disc degeneration.

Application of FTIR imaging to detect dietary induced biochemical changes in brown and white adipocytes

Fourier Transform Infrared microspectroscopy Imaging (FTIR imaging) has powerful advantages when used as a complementary technique to histopathological investigations. The common histopathological investigations use a number of chemical fixatives and deparaffinizing agents that alter lipid profiles of biological tissues, particularly in lipid-rich adipose tissues. An infrared image is made of a multitude of pixels. For each pixel, identified by its (x,y) coordinates, corresponds an FTIR spectrum that reveals the chemical composition at this specific location. As a result, it allows biochemical analysis in selective locations within samples. Moreover, functional group distribution maps can be used as a staining free technique for spatial characterization of different biomolecules. Though FTIR imaging has been reported as a powerful approach to characterize adipose tissues, the biochemical heterogeneity within adipose tissues has not been acknowledged. This study shows relative changes in brown and white adipose tissues of mice due to consumption of high-fat diet. In particular, we report cryosectioning of adipose tissues, image acquisition, and different image analysis methods to evaluate dietary induced changes in lipids stored in brown and white adipocytes. Since biochemical changes in adipocytes constitute an important component in obesity investigations, this study shows the potential use of FTIR imaging to compare relative biochemical changes associated with dietary interventions.

Simultaneous cancer and tumor microenvironment subtyping using confocal infrared microscopy for all-digital molecular histopathology

Histopathology based on spatial patterns of epithelial cells is the gold standard for clinical diagnoses and research in carcinomas; although known to be important, the tissue microenvironment is not readily used due to complex and subjective interpretation with existing tools. Here, we demonstrate accurate subtyping from molecular properties of epithelial cells using emerging high-definition Fourier transform infrared (HD FT-IR) spectroscopic imaging combined with machine learning algorithms. In addition to detecting four epithelial subtypes, we simultaneously delineate three stromal subtypes that characterize breast tumors. While FT-IR imaging data enable fully digital pathology with rich information content, the long spectral scanning times required for signal averaging and processing make the technology impractical for routine research or clinical use. Hence, we developed a confocal design in which refractive IR optics are designed to provide high-definition, rapid spatial scanning and discrete spectral tuning using a quantum cascade laser (QCL) source. This instrument provides simultaneously high resolving power (2-μm pixel size) and high signal-to-noise ratio (SNR) (>1,300), providing a speed increase of ∼50-fold for obtaining classified results compared with present imaging spectrometers. We demonstrate spectral fidelity and interinstrument operability of our developed instrument by accurate analysis of a 100-case breast tissue set that was analyzed in a day, considerably speeding research. Clinical breast biopsies typical of a patients' caseload are analyzed in ∼1 hour. This study paves the way for comprehensive tumor-microenvironment analyses in feasible time periods, presenting a critical step in practical label-free molecular histopathology.

Comparative behavior between sunscreens based on free or encapsulated UV filters in term of skin penetration, retention and photo-stability

BackgroundThe growing incidence of photodamaging effects caused by UV radiation (e.g. sunburn, skin cancer) has increased the attention from health authorities which recommend the topical application of sunscreens to prevent these skin damages. The economic stakes for those companies involved in this international market are to develop new UV filters and innovative technologies to provide the most efficient, flexible and robust sunscreen products. Today the development of innovative and competitive sunscreen products is a complex formulation challenge. Indeed, the current sunscreens must protect against skin damages, while also being safe for the skin and being sensory and visually pleasant for the customers when applied on the skin. Organic UV filters, while proposing great advantages, also present the risk to penetrate the stratum corneum and diffuse into underlying structures with unknown consequences; moreover, their photo-stability are noted thorny outcomes in sunscreen development and subsequent performance. In recent years, the evaluation of the interaction between skin and sunscreen in terms of penetration after topical application has been considered from European authority but still its testing as their photo-stability assessment are not mandatory in most countries. ObjectiveThis study, based on in-vitro approaches, was performed to evaluate and compare the retention and the penetration of organic UV filters in free or encapsulated form inside the skin as well as their respective photo-stability. MethodsSunscreen formulation with a combination of Avobenzone and Octocrylene in “free form” and a formulation using the same UV filters but encapsulated in a sol-gel silica capsule, were analyzed and compared by FTIR Imaging Spectroscopy. Tape stripping method was used to investigate the penetration of these UV filters inside the stratum corneum. Their photo-stabilities were evaluated by spectroscopic measurements (FTIR, UV/Vis) and standard measurements were calculated: AUC (Area Under the Curve) and SPF (Sun Protection Factor). ResultWith traditional formulation, the organic UV filters penetrated significantly into the stratum corneum while the same UV filters combined with encapsulation technology remained on the skin surface. The encapsulation technology also improved significantly their stability. ConclusionEncapsulation technology is a promising strategy to improve the efficacy of sunscreen product using organic UV filters and to reduce safety problem. On the other hand, this study highlighted the pertinence of the FTIR Spectroscopy to test, compare and investigate sunscreen formulations.

Relating Post-yield Mechanical Behavior in Polyethylenes to Spatially Varying Molecular Deformation Using Infrared Spectroscopic Imaging: Homopolymers

Stress–strain curves derived from tensile specimens are the primary characteristic of bulk polymers’ mechanical properties. Current tools, however, cannot provide molecular insights from this single bulk measurement. Hence, we use Fourier transform infrared (FT-IR) spectroscopic imaging to optically and nondestructively measure molecular structure and its spatial dependence in tensile specimens in high density polyethylene homopolymers. To overcome the limitations of FT-IR imaging, we use an emerging approach involving the use of tunable quantum cascade lasers that allows imaging through thick samples and facile polarized light imaging. Crystal structure and orientation are obtained from spatially varying measurements of molecular properties in the necking region. Local molecular (re)arrangements to characterize mechanical properties of drawn samples are deduced from spectral data. A modified Eyring model was developed to quantitatively understand spatial dependence in terms of a conformational volume. We...

Molecular docking of immunogenic peptide of Toxoplasma gondii and encapsulation with polymer as vaccine candidate

Toxoplasma gondii is one of the most widely spread parasitic organisms in the world. T. gondii causes primary, chronic infection and mortality. Major surface antigen 1 is the most abundant tachyzoite surface protein and highly conserved between species and causes strong humoural response. Some studies showed that the peptide sequence of surface antigen has immunity. Therefore, tachyzoite surface antigenic peptide sequence is one of the good candidates for vaccine development. However, conformational information and delivery systems are very important parameters for vaccine development. Computational chemistry which is used as an effective method to perform drug or vaccine design provides important information on structure–activity relationship, biological effects of functional groups, molecular geometry, design of enzyme inhibitors and antagonists. The interaction of immunological peptides with protein systems was carried out by means of computing the free energy of binding using the molecular docking technique. Due to the major histocompatibility complex (MHC), proteins play a substantial role for adaptive immunity, the crystal structure of a MHC class I, which plays a pivotal role in the adaptive branch of the immune system, was preferred for docking calculations. A delivery system based on poly(lactic-co-glycolic acid) (PLGA) nanoparticles and peptide loaded PLGA nanoparticles was prepared in this study to improve the bioavailability of tachyzoite surface antigenic peptide sequence. Double emulsion method (water-in-oil-in-water or w/o/w) was used for synthesis of PLGA and peptide loaded PLGA nanoparticles. The average particle size, polydispersity index and zeta potential values of PLGA and peptide loaded PLGA nanoparticles were measured with zeta-sizer by using dynamic light scattering (DLS) technique. The scanning electron microscope (SEM) (Zeiss Supra 50 V) was used for imagining the peptide loaded PLGA nanoparticles. Cell toxicity of nanoparticles was assayed on AGS (gastric adenocarcinoma) cell line. To evaluate mitochondrial activity of cells and toxicity studies, XTT methods were carried out. In this study, we aimed to obtain specific immunological peptide loaded PLGA nanoparticles and characterize the formation with FTIR, zeta sizer and SEM imaging, and evaluate cytotoxicity and carry out molecular docking calculations of peptide–MHC protein in order to enlight in vivo events as vaccine candidate against T. gondii.