Platinum-gold nanoraspberries as effective photosensitizer in anticancer photothermal therapy.

BACKGROUND: New nanophotosensitizers for photothermal cancer therapy (PTT) are still sought. In this paper we propose fancy shaped, non agglomerated core/shell PtAu NRs nanoraspberries (PtAu NRs) as potential nanophotosensitizers in PTT. RESULTS: Light microscopy images of two colon cancer cell lines (SW480, SW620) showed, that the laser irradiation combined with PtAu NRs caused visible changes in the cell morphology. Fourier Transform InfraRed (FTIR) and Raman spectroscopies showed chemical changes in the DNA, phospholipids, lipids and protein structures caused by laser irradiation in the presence of PtAu NRs. The MTS assay showed ~ 25% mortality of cancer cells due to the addition of PtAu NRs to the cell culture, while for laser irradiation combined with nanoparticles, the mortality of cancer cells increased to 65% for the 650 nm laser and to 60% for the 808 nm laser. The calculated photothermal conversion efficiency reached 62% and 51% for the 650 nm and 808 nm lasers, respectively. CONCLUSIONS: PtAu NRs could be applied as effective light-absorbers in the PTT anticancer therapy.

FTIR analysis of molecular composition changes in hazel pollen from unpolluted and urbanized areas.

ABSTRACT: In this study, the effect of urbanization and environmental pollution on qualitative (structural) and quantitative changes of the Corylus avellana (hazel) pollen was investigated using scanning electron microscopy, Fourier Transform Infrared (FTIR) spectroscopy and curve-fitting analysis of amide I profile. The obtained spectroscopic results show significant variations in the fraction of proteins in the hazel pollen, which probably depend on various degrees of anthropopression. Our results suggest that alterations in the chemical composition of pollen, induced by urbanization and air pollutants, may intensify the allergenic potential and may cause the increase in the incidence of allergies in people. Mutations in nucleic acids are accompanied by a number of molecular changes leading to the formation of allergenic proteins. It seems that the type of habitat, where the pollen grew, affects the individual differentiation. Indeed, it was found that in the site exhibiting low pollution, the hazel pollen contain a lower amount of proteins than to the ones from a site with high anthropopression. Hence, FTIR spectroscopy and curve-fitting analysis of amide I profile can be successfully applied as tools for identifying quantitative and qualitative changes of proteins in hazel pollen. GRAPHICAL ABSTRACT: Anthropogenic factors such as air pollution and urbanization lead to changes in structure and chemical composition of hazel pollen. Fourier Transform Infrared spectroscopy (FTIR) and Gaussian analysis showed structural changes in hazel pollen collected from sites with different absorbance values of individual chemical functional groups and changes in the secondary structure of proteins of the pollen.

Size-dependent theoretical and experimental photothermal conversion efficiency of spherical gold nanoparticles.

INTRODUCTION: Due to their biocompatible and plasmonic properties, gold nanoparticles (Au NPs) are good candidates to be photosensitizers in photothermal cancer therapy (PTT). MATERIALS AND METHODS: In this paper, the dependence of the NIR-light-to-heat energy on Au NPs size was investigated. Moreover, to determine the photosensitizing properties of gold nanoparticles, PTT was conducted on two colon cell lines: SW480 and SW620 by irradiating them with two lasers having different wavelengths. RESULTS: Transmission electron microscopy showed that the respective sizes of Au NPs were 10 nm, 12 nm and 16 nm. Moreover, local as well as global structural measurements showed that all synthesized Au NPs were crystalline and UV-Vis spectroscopy revealed that with increasing nanoparticles size the position of the surface Plasmon resonance (SPR) peaks is shifted to higher wavelengths. Decrease of cells viability was observed, when they were cultured with Au NPs and irradiated by 650 nm and 808 nm lasers. Moreover, FTIR and Raman spectra of cells, showed structural changes in DNA, phospholipids, proteins and cholesterol caused by the addition of nanoparticles and laser irradiation. The chemical changes were more pronounced in the cells cultured with Au NPs and irradiated by 650 nm lasers and these changes were dependent on the nanoparticle size. Moreover, the viability of cells investigated by the MTS assay showed, that the percentage of dead cells (∼40%) is the highest for cells cultured with 8 nm Au NPs and irradiated by the 650 nm laser. The photothermal conversion efficiency calculated from the experimental results showed a decrease of this parameter from 70% to 55% and from 61% to 48% with increasing particle size, for 650 nm and 808 nm lasers, respectively. CONCLUSIONS: The obtained results showed that the photothermal conversion efficiency of Au NPs is size-tunable, and can be correlated with the absorption/extinction ratios calculated by the Mie theory.

Electromagnetic field of extremely low frequency has an impact on selected chemical components of the honeybee.

The electromagnetic field (EMF) is an environmental factor affecting living organisms. The aim of this study was to demonstrate the effect of an extremely low frequency electromagnetic field (ELF-EMF) on selected chemical components of the honeybee (Apis mellifera L.) using Fourier Transform Infrared (FTIR) spectroscopy. The FTIR method provides information on the chemical structure of compounds through identification and analysis of functional groups. The honeybees were treated with EMF at a frequency of 50 Hz and magnetic induction of 1.6 mT for 2, 6, 12, 24 and 48 hours. Analysis of FTIR spectra showed that EMF exposure longer than 2 hours induced changes in the structure of chemical compounds, especially in the IR region corresponding to DNA, RNA, phospholipids and protein vibrations, compared to control samples (bees not EMF treated). The results confirm the effect of EMF on bees depending on the duration of exposure.

From spherical to bone-shaped gold nanoparticles-Time factor in the formation of Au NPs, their optical and photothermal properties.

Using the same synthesis method, which was stopped at different time intervals, gold nanoparticles (Au NPs) with different shapes, from spherical to bone-shaped, were obtained. The physical properties of the synthesized Au NPs were investigated using transmission electron microscopy (TEM) combined with selected area electron diffraction patterns (SAED) and ultraviolet-visible spectroscopy (UV-vis). The TEM images showed, that stopping the synthesis after one minute lead to the formation of small spherical Au NPs, which evolved to the cubic shape, rods and bone-shaped Au NPs after 15 min, 30 min, 2 h, respectively. SAED patterns showed, that all the obtained Au NPs were crystalline. UV-vis spectra revealed, that the light absorbance depends on the shape of the Au NPs. Moreover, the effects of the time factor in the formation of Au NPs on the effective conversion of electromagnetic energy into thermal energy, was studied. Furthermore, simulated photothermal therapy (PTT) in combination with the obtained NPs, was done for two cancer cell lines SW480 and SW620. The mortality of cells after using the differently shaped Au NPs as photosensitizers is between 18 % and 52 % and increases with the decrease of the synthesis time.

Size effect of platinum nanoparticles in simulated anticancer photothermal therapy.

Platinum nanoparticles (Pt NPs) with properties including damage of DNA, enzmatic activity and possibility of light absorption in the biological range could find application as effective photosensitizers in photothermal anticancer therapy (PTT). The photothermal properties of Pt NPs depend on their shape, size and crystalline structure. Therefore, in this paper the effect of Pt NPs size on photothermal efficiency is determined. For this purpose, spherical, crystalline 80 nm PtI NPs and 2 nm PtII NPs were synthesized and characterized by transmission electron microscopy (TEM), selected area diffraction patterns (SAED) and X-ray diffraction (XRD). The possibilities of using Pt NPs in PTT were investigated using two colon cancer cell lines: SW480 and SW620, which were cultured with both Pt NPs and irradiated by two, 650 nm and 808 nm lasers. Microscopy images of cells and MTS assay showed, that the PTT is the most effective when 2 nm nanoparticles and the 650 nm laser were used. The mortality of cells was around 62% for SW480 and 70% for SW620. Furthermore, higher temperatures after irradiation of Pt NPs by lasers were observed for the 2 nm Pt NPs for both wavelengths. Consequently, the values of photothermal efficiency are higher by approximately 5% and 6% for 2 nm Pt NPs, than for 80 nm ones, which were irradiated by 650 and 808 nm lasers, respectively. Moreover, the results obtained from experimental data corresponded with Mie theory.

Raman and FTIR spectroscopy in determining the chemical changes in healthy brain tissues and glioblastoma tumor tissues.

Glioblastoma, also called glioblastoma multiforme (GBM), is a particularly malignant form of primary brain tumor. This cancer accounts for 12-15% of all brain tumors. Despite the advances in neurosurgery, radio and chemotherapy the average survival rate is only 12.1-16.6 months. This is due not only to the late diagnosis of the disease, but also to ineffective treatment methods which result from the still low knowledge about the causes of glioblastoma development. Therefore, it is very important to look for new diagnostic methods of detection of the smallest features of cancer. Raman and infrared spectroscopy (FTIR) can be such methods. In this paper we discuss the chemical composition of sample glioblastoma brain tissues and marginal brain tissues using these two spectroscopy methods. Raman and FTIR spectra of cancer brain tissues showed that the highest differences in the chemical composition, compared to the control brain tissue, occur in the areas corresponding to lipids, collagen and proteins. Moreover, Raman spectroscopy also showed significant changes in the cancer tissues in the phosphatidylcholine and sphingomyelin. Interestingly, FTIR spectra after Kramers-Kronig transformations showed signals only for three peaks which corresponded to the vibrations of lipid function groups. Adjustment of the Lorenz function for these three peaks showed that only in the case of cancerous tissues the number of matching lines is different, compared to the control and marginal tissues. Therefore, we assume that lipids could be a spectroscopic marker for brain tumor. Furthermore, principal component analysis (PCA) showed that chemical changes seen between cancer and control tissues are significant and it is possible to differentiate the infected tissue from the healthy one. Interestingly, the PCA analysis also showed that adjacent brain tissues have different chemical composition than the control tissues.

Comparing dried and liquid blood serum samples of depressed patients: An analysis by Raman and infrared spectroscopy methods.

Depression is a serious mental illness. To study the mechanism of depression and search for new, more effective therapies, animal models are often used. Unfortunately, none of the available models reflects all the symptoms of depression. Therefore researchers are looking for new tools to diagnose depression. Unfortunately, the nowadays-available depression diagnosis methods are only psychological tests. However, it is known, that the amount of phospholipids, proteins and lipids decreases during depression. Raman and FTIR (Fourier Transform Infra Red) spectroscopies provide information on the chemical compounds in the measured sample e.g. blood serum. These spectroscopic techniques may thus become reliable and accurate tools for evaluating changes in the amount of phospholipids and proteins in depression disease. In this study differences between dried and liquid blood serum samples of healthy and depressed individuals measured by Raman (range 0-3000cm-1) and FTIR (Fourier Transform Infrared) (range 900-3000cm-1) spectroscopy were evaluated. The resulting spectra and accurate analysis led to the conclusion that an appropriate measurement of the background and the elimination of peaks from water had the greatest impact on the reliability of the results. Furthermore, after detailed studies of FTIR and Raman spectra of dried and liquid blood serum samples, including a complete analysis of peaks after Kramers-Kröning (KK) transformation, it was found that the sample preparation did not affect the results obtained by Raman spectroscopy. In FTIR measurements only a minimal effect on peak intensity was observed.

Qualitative and quantitative changes in phospholipids and proteins investigated by spectroscopic techniques in olfactory bulbectomy animal depression model.

Depression becomes nowadays a high mortality civilization disease with one of the potential causes being impaired smell. In this study Raman, Fourier Transform Infra Red (FTIR) and Ultraviolet-Visible (UV-vis) spectroscopies were used to determine the changes in the quantity and structure of phospholipids and proteins in the blood serum of bulbectomized rats (OB_NaCl), which is a common animal depression model. The efficiency of amitriptyline (AMI) treatment was also evaluated. The obtained results show a significant decrease in the phospholipid and protein fractions (as well as changes in their secondary structures) in blood serum of bulbectomized rats. AMI treatment in bulbectomized rats increased protein level and did not affect the level of phospholipids. Structural information from phospholipids and proteins was obtained from UV-vis spectroscopy combined with the second derivative of the FTIR spectra. Indeed, the structure of proteins in blood serum of bulbectomized rats was normalized after amitriptyline therapy, while the damaged structure of phospholipids remained unaffected. These findings strongly suggest that impaired smell could be one of the causes of depression and may induce permanent (irreversible) damages into the phospholipid structure identified as shortened carbon chains. This study shows a possible new application of spectroscopic techniques in the diagnosis and therapy monitoring of depression.

The classification of lung cancers and their degree of malignancy by FTIR, PCA-LDA analysis, and a physics-based computational model.

Lung cancer has the highest mortality rate of all malignant tumours. The current effects of cancer treatment, as well as its diagnostics, are unsatisfactory. Therefore it is very important to introduce modern diagnostic tools, which will allow for rapid classification of lung cancers and their degree of malignancy. For this purpose, the authors propose the use of Fourier Transform InfraRed (FTIR) spectroscopy combined with Principal Component Analysis-Linear Discriminant Analysis (PCA-LDA) and a physics-based computational model. The results obtained for lung cancer tissues, adenocarcinoma and squamous cell carcinoma FTIR spectra, show a shift in wavenumbers compared to control tissue FTIR spectra. Furthermore, in the FTIR spectra of adenocarcinoma there are no peaks corresponding to glutamate or phospholipid functional groups. Moreover, in the case of G2 and G3 malignancy of adenocarcinoma lung cancer, the absence of an OH groups peak was noticed. Thus, it seems that FTIR spectroscopy is a valuable tool to classify lung cancer and to determine the degree of its malignancy.

Use of FTIR spectroscopy and PCA-LDC analysis to identify cancerous lesions within the human colon.

Colorectal cancer constitutes 33% of all cancer morbidity, so the research of the new methods for colorectal cancer diagnosis and chemotherapy monitoring is gaining its momentum. Diagnostic instruments are being sought, which enable the detection of single malignant cells based on the analysis of tissue material potentially reusable at further stages of diagnostic management. The most common approach to tissue specimen processing is paraffin-embedding. Yet, paraffin may cause background noise in spectroscopic measurements with the wavenumber ranging between 900cm-1 and 3500cm-1. However, the study by Depciuch et al. (2016) proved that appropriate specimen processing and paraffin-embedding technique as well as a strict measurement methodology may eliminate paraffin vibrations. As a result, spectroscopic measurements may become a reliable and precise method for the diagnosis and treatment monitoring in patients with colorectal cancer as long as the high standards of specimen processing are maintained. Chemotherapy is the main medical treatment in colorectal cancer. Unfortunately, the absence of tools which enable monitoring its efficacy leads to the partial response or non-response frequently seen in affected patients. Hence, diagnostic instruments are also being sought capable of monitoring treatment efficacy so as to enable early changes of chemotherapy regimen thus increasing the chance of cure. The paper aims at comparing the results of FTIR (Fourier Transform Infrared) spectroscopy in several types of colon tissue: healthy colon, cancerous colon, post-chemotherapy colon and healthy surgical margin of colon cancer sample. The obtained FTIR spectra along with the Principal Component Analysis-Linear Discriminant Analysis (PCA-LDC) as well as bandwidth analysis of the primary amide region revealed some differences between the spectra of healthy tissues as compared to cancerous tissues (pre- or post-chemotherapy). Apart from confirming that FTIR spectroscopy is a good source of information on the composition of analysed samples, this fact supports its application as a tool to facilitate understanding the pathophysiology of various conditions and to monitor efficacy of chemotherapy in cancer patients.

Verification of the effectiveness of the Fourier transform infrared spectroscopy computational model for colorectal cancer.

Colorectal cancer is one of the most common cancers. Its formation is influenced by genetic and environmental factors. Despite the continuous development of diagnostic tools and cancer therapies, there are no methods that allow a real-time estimation of treatment efficiency. This method can be a vibrational spectroscopy. The resulting infrared spectrum (FTIR) of the tissue gives us information about the chemical composition and the content of the individual components. We have noticed that tumor tissues, healthy and after chemotherapy tissues, have different vibrational spectra. It was also shown that spectra acquired from normal (benign) tissues were similar to those derived from tissues post-chemotherapy. The similarity was greater, when the effectiveness of chemotherapy, confirmed by medical documentation, was better. Therefore, we decided to use the physical model proposed in our earlier paper to verify its correctness and to show whether a particular type of chemotherapy was effective or not. Comparison of the results obtained from the physical model with patients data have been found as close to the physical condition.

Qualitative and quantitative changes in phospholipids and proteins investigated by spectroscopic techniques in animal depression model.

Depression becomes nowadays a high mortality civilization disease with one of the major causes being chronic stress. Raman, Fourier Transform Infra Red (FTIR) and Ultraviolet-Visible (UV-vis) spectroscopies were used to determine the changes in the quantity and structure of phospholipids and proteins in the blood serum of rats subjected to chronic mild stress, which is a common animal depression model. Moreover, the efficiency of the imipramine treatment was evaluated. It was found that chronic mild stress not only damages the structure of the phospholipids and proteins, but also decreases their level in the blood serum. A 5weeks imipramine treatment did increase slightly the quantity of proteins, leaving the damaged phospholipids unchanged. Structural information from phospholipids and proteins was obtained by UV-vis spectroscopy combined with the second derivative of the FTIR spectra. Indeed, the structure of proteins in blood serum of stressed rats was normalized after imipramine therapy, while the impaired structure of phospholipids remained unaffected. These findings strongly suggest that the depression factor, which is chronic mild stress, may induce permanent (irreversible) damages into the phospholipid structure identified as shortened carbon chains. This study shows a possible new application of spectroscopic techniques in the diagnosis and therapy monitoring of depression.

Monitoring breast cancer treatment using a Fourier transform infrared spectroscopy-based computational model.

Breast cancer affects one in four women, therefore, the search for new diagnostic technologies and therapeutic approaches is of critical importance. This involves the development of diagnostic tools to facilitate the detection of cancer cells, which is useful for assessing the efficacy of cancer therapies. One of the major challenges for chemotherapy is the lack of tools to monitor efficacy during the course of treatment. Vibrational spectroscopy appears to be a promising tool for such a purpose, as it yields Fourier transformation infrared (FTIR) spectra which can be used to provide information on the chemical composition of the tissue. Previous research by our group has demonstrated significant differences between the infrared spectra of healthy, cancerous and post-chemotherapy breast tissue. Furthermore, the results obtained for three extreme patient cases revealed that the infrared spectra of post-chemotherapy breast tissue closely resembles that of healthy breast tissue when chemotherapy is effective (i.e., a good therapeutic response is achieved), or that of cancerous breast tissue when chemotherapy is ineffective. In the current study, we compared the infrared spectra of healthy, cancerous and post-chemotherapy breast tissue. Characteristic parameters were designated for the obtained spectra, spreading the function of absorbance using the Kramers-Kronig transformation and the best fit procedure to obtain Lorentz functions, which represent components of the bands. The Lorentz function parameters were used to develop a physics-based computational model to verify the efficacy of a given chemotherapy protocol in a given case. The results obtained using this model reflected the actual patient data retrieved from medical records (health improvement or no improvement). Therefore, we propose this model as a useful tool for monitoring the efficacy of chemotherapy in patients with breast cancer.

The role of zinc deficiency-induced changes in the phospholipid-protein balance of blood serum in animal depression model by Raman, FTIR and UV-vis spectroscopy.

Depression is a serious mental illness. To study the mechanisms of diseases and search for new, more effective therapies, animal models are used. Unfortunately, none of the available models does reflect all symptoms of depression. Zinc deficiency is proposed as a new animal model of depression. However, it has not been yet validated in a detailed manner. Recently, spectroscopic techniques are increasingly being used both in clinical and preclinical studies. Here we examined the effect of zinc deficiency and amitryptyline treatment on the phospholipid - protein balance in the blood serum of rats using Raman, Fourier Transform Infra Red (FTIR) and UV-vis technique. Male Sprague Dawley rats were fed with a zinc ample diet (ZnA, 50mg Zn/kg) or a zinc deficient diet (ZnD, 3mg Zn/kg) for 4 weeks. Then amitriptyline administration (AMI, 10mg/kg, i.p.) was started. After injecting the drug for 2-weeks, blood samples were collected and analyzed. It was found that zinc deficiency decreases both the level of phospholipids and proteins and also causes structural changes in their structures. In the ZnD group amitriptyline treatment influenced the protein level and structure. UV-vis spectroscopy combined with the second derivative calculated from the FTIR spectra provided information that the proteins in blood serum of rat fed with a low Zn diet regain their intact structure after amitriptyline medication. Simultaneously, the antidepressant therapy did not have any effect on the level of phospholipids in this group of rats. Additionally, our results show, that amitriptyline administration can change the structure of phospholipids in rats subjected to zinc ample diet. This altered structure of phospholipids was identified as shortening of carbon chains. Our findings indicate that the decreased level of zinc may be the cause of depressive disorders, as it leads to changes in the phospholipid-protein balance necessary for the proper functioning of the body. This study also shows possible new applications of spectroscopic techniques in the diagnosis of affective disorders, and maybe even identifies markers of depressive disorders.

Analysis of morphological and molecular composition changes in allergenic Artemisia vulgaris L. pollen under traffic pollution using SEM and FTIR spectroscopy.

Nowadays, pollen allergy becomes an increasing problem for human population. Common mugwort (Artemisia vulgaris L.) is one of the major allergenic plants in Europe. In this study, the influence of air pollution caused by traffic on the structure and chemical composition of common mugwort pollen was investigated. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and curve-fitting analysis of amide I profile was applied to assess the morphological and structural changes of mugwort pollen grains collected from sites with different vehicle pollution levels. Microscopic observations support the conclusion, that the higher the car traffic, the smaller the pollen grains. The obtained results clearly show that air pollution had an impact on different maximum absorbance values of individual functional groups composing the chemical structure of pollen. Moreover, air pollution induced structural changes in macromolecules of mugwort pollen. In pollen collected from the unpolluted site, the content of sporopollenin (850 cm-1) was the highest, whereas polysaccharide concentration (1032 cm-1) was the lowest. Significant differences were observed in lipids. Pollen collected from the site with heavy traffic had the lowest content of lipids at 1709, 2071, and 2930 cm-1. The largest differences were observed in the spectra regions corresponding to proteins. In pollen collected from unpolluted site, the highest level of ß-sheet (1600 cm-1) and α-helix (1650 cm-1) was detected. The structural changes in proteins, observed in the second derivative of the FTIR spectrum and in the curve-fitting analysis of amide I profile, could be caused inter alia by air pollutants. Alterations in protein structure and in their content in the pollen may increase the sensitization and subsequent risk of allergy in predisposed people. The obtained results suggest that the changes in chemical composition of pollen may be a good indicator of air quality and that FTIR may be successfully applied in biomonitoring.