Colorectal cancer is the second most common cancer and the second with the highest associated deaths in the world. Methods used in clinical practice for colon cancer diagnosis are fairly effective but quite unpleasant and not always applicable in situations where the patient has symptoms of colonic obstruction. This problem can be solved by the use of optical methods that can be applied less invasively.
This study presents the results of classification of cancerous and healthy colon tissue absorption coefficient spectra. The absorption coefficient was measured using direct calculations from the total reflectance and total transmittance spectra obtained ex vivo. Classification was performed using support vector machine, multilayer perceptron and linear discriminant analysis.
The interest of using light in clinical practice is increasing strongly and many applications work at various wavelengths from the ultraviolet to the infrared. Due to this great range of applications, the determination of the optical properties of biological tissues in a wide spectral range becomes of interest. The liver is an important organ, since it has a major role in the human body and various pathologies are known to develop within it. For these reasons, this study concerns the estimation of the optical properties of human normal and pathological (metastatic carcinoma) liver tissues between 200 and 1000 nm. The obtained optical properties present the expected wavelength dependencies for both tissues – the refractive index, the absorption and the scattering coefficients decrease with the wavelength and the anisotropy and light penetration depth increase with the wavelength. Although similar behavior was observed for the various properties between the normal and pathological tissues, evidence of smaller blood content in the pathological tissues was found. A possible explanation is that the cancer cells destroy liver’s vasculature and internal architecture, providing though a reduction in the blood content. For low wavelengths, it was observed a matching between the scattering and the reduced scattering coefficients, which implies a nearly zero anisotropy in that range. The scattering coefficient decreases from nearly 140 cm-1 (at 200 nm) to 80 cm-1 (at 1000 nm) for the normal liver and from nearly 140 cm-1 (at 200 nm) to 95 cm-1 (at 1000 nm) for the pathological tissue.
To characterize the optical clearing treatments in human colorectal tissues and possibly to differentiate between treatments of normal and pathological tissues, we have used a simple indirect method derived from Mie scattering theory to estimate the kinetics of the reduced scattering coefficient. A complementary method to estimate the kinetics of the scattering coefficient is also used so that the kinetics of the anisotropy factor and of the refractive index are also calculated. Both methods rely only on the thickness and collimated transmittance measurements made during treatment. The results indicate the expected time dependencies for the optical properties of both tissues: an increase in the refractive index and anisotropy factor and a decrease in the scattering coefficients. The similarity in the kinetics obtained for normal and pathological tissues indicates that optical clearing treatments can be applied also in pathological tissues to produce similar effects. The estimated time dependencies using experimental spectral data in the range from 400 to 1000 nm allowed us to compare the kinetics of the optical properties between different wavelengths.
Knowledge of the optical properties of tissues is necessary, since they change from tissue to tissue and can differ between normal and pathological conditions. These properties are used in light transport models with various areas of application. In general, tissues have significantly high scattering coefficient when compared to the absorption coefficient and such difference usually increases with decreasing wavelength. The study of the wavelength dependence of the optical properties has been already made for several animal and human tissues, but extensive research is still needed in this field. Considering that most of the Biophotonics techniques used in research and clinical practice use visible to NIR light, we have estimated the optical properties of colorectal muscle (muscularis propria) between 400 and 1000 nm. The samples used were collected from patients undergoing resection surgery for colorectal carcinoma. The estimated scattering coefficient for colorectal muscle decreases exponentially with wavelength from 122 cm-1 at 400 nm to 95 cm-1 at 650 nm and to 91 cm-1 at 1000 nm. The absorption coefficient shows a wavelength dependence according to the behavior seen for other tissues, since it decreases from 8 cm-1 at 400 nm to 2.6 cm-1 at 650 nm and to 1.3 cm-1 at 1000 nm. The estimated optical properties differ from the ones that we have previously obtained for normal and pathological colorectal mucosa. The data obtained in this study covers an extended spectral range and it can be used for planning optical clearing treatments for some wavelengths of interest.
The optical dispersion and water content of human liver were experimentally studied to estimate the optical dispersions of tissue scatterers and dry matter. Using temporal measurements of collimated transmittance [Tc(t)] of liver samples under treatment at different glycerol concentrations, free water and diffusion coefficient (Dgl) of glycerol in liver were found as 60.0% and 8.2×10−7 cm2/s, respectively. Bound water was calculated as the difference between the reported total water of 74.5% and found free water. The optical dispersion of liver was calculated from the measurements of refractive index (RI) of tissue samples made for different wavelengths between 400 and 1000 nm. Using liver and water optical dispersions at 20°C and the free and total water, the dispersions for liver scatterers and dry matter were calculated. The estimated dispersions present a decreasing behavior with wavelength. The dry matter dispersion shows higher RI values than liver scatterers, as expected. Considering 600 nm, dry matter has an RI of 1.508, whereas scatterers have an RI of 1.444. These dispersions are useful to characterize the RI matching mechanism in optical clearing treatments, provided that [Tc(t)] and thickness measurements are performed during treatment. The knowledge of Dgl is also important for living tissue cryoprotection applications.
Optical properties of biological tissues are unique and may be used for tissue identification, tissue discrimination or even to identify pathologies. Early stage colorectal cancer evolves from adenomatous polyps that arise in the inner layer of the colorectal tube — the mucosa. The identification of different optical properties between healthy and pathological colorectal tissues might be used to identify different tissue components and to develop an early stage diagnosis method using optical technologies. Since most of the biomedical optics techniques use light within the visible and near infrared wavelength ranges, we used the inverse adding-doubling method to make a fast estimation of the optical properties of colorectal mucosa and early stage adenocarcinoma between 400 and 1000 nm. The estimated wavelength dependencies have provided information about higher lipid content in healthy mucosa and higher blood content in pathological tissue. Such data has also indicated that the wavelength dependence of the scattering coefficient for healthy mucosa is dominated by Rayleigh scattering and for pathological mucosa it is dominated by Mie scattering. Such difference indicates smaller scatterer size in healthy mucosa tissue. Such information can now be used to develop new diagnosis or treatment methods for early cancer detection or removal. One possibility is to use optical clearing technique to improve tissue transparency and create localized and temporary tissue dehydration for image contrast improvement during diagnosis or polyp laser removal. Such techniques can now be developed based on the different results that we have found for healthy and pathological colorectal mucosa.
Colorectal carcinoma is a major health concern worldwide and its high incidence and mortality require accurate screening methods. Following endoscopic examination, polyps must be removed for histopathological characterization. Aiming to contribute to the improvement of current endoscopy methods of colorectal carcinoma screening or even for future development of laser treatment procedures, we studied the diffusion properties of glucose and water in colorectal healthy and pathological mucosa. These parameters characterize the tissue dehydration and the refractive index matching mechanisms of optical clearing (OC). We used ex vivo tissues to measure the collimated transmittance spectra and thickness during treatments with OC solutions containing glucose in different concentrations. These time dependencies allowed for estimating the diffusion time and diffusion coefficient values of glucose and water in both types of tissues. The measured diffusion times for glucose in healthy and pathological mucosa samples were 299.2±4.7 s and 320.6±10.6 s for 40% and 35% glucose concentrations, respectively. Such a difference indicates a slower glucose diffusion in cancer tissues, which originate from their ability to trap far more glucose than healthy tissues. We have also found a higher free water content in cancerous tissue that is estimated as 64.4% instead of 59.4% for healthy mucosa.
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