Smoking Induced Hemolysis: Spectral and microscopic investigations.

Tobacco smoke contains at least 3500 chemicals such as carcinogens, mutagens, free radicals, heavy metals, and even radioactive materials1,2. According to the American Cancer Society’s report, smoking accounts for 30% of all cancer deaths in the US; further, 80% of lung cancer deaths are exclusively due to smoking3. In addition, smokers are at three-times greater risk of cardiovascular diseases than the nonsmokers. Moreover, smoking causes more deaths than the combined mortality due to HIV and motor vehicle injuries2,3. In addition, smoking causes chronic nonfatal diseases such as cataract, arthritis, erectile dysfunction, etc. However, only a few studies have investigated smoking-induced damage to the blood4,5,6,7,8. Optical biopsy is a new technique where the light of UV or visible radiation is employed as a tool to probe the intrinsic conditions of tissues (normal, benign, or malignant) or body fluids (blood, urine, saliva, etc.). When the light of a particular wavelength falls on a tissue or body fluid, it undergoes scattering or absorption because light photons interact essentially with the biomolecules. Some of these biomolecules can also produce fluorescence or incoherent molecular scattering leading to Raman shifts. Therefore, the different spectra obtained from such biomolecules serve as biomarkers of different diseases. Many studies have focused on the fluorescence and Raman spectra of a variety of malignant tissues in vivo and in vitro9,10. Similar studies have focused on the detection of malignancy of lung, liver, etc.11,12 (and inherited blood disorders, such as thalassemia and sickle cell anemia)13 from the spectral features of blood and urine14. The fluorescent spectral technique can be used for molecular diagnosis, and the present study can be considered as a logical extension of the spectral technique of blood components for monitoring smoking-induced damage on erythrocytes. The atomic force microscopic (AFM) analysis is the morphological and structural analysis of erythrocyte membrane to confirm the finding of spectral investigations. The AFM is a scanning probe microscopy (SPM), with the resolution of the order of a few nanometers and 1000 times better than the optical diffraction limit. By using an AFM, it is possible to measure the roughness and hardness of a sample surface at a high resolution. In comparison to the scanning electron microscope (SEM), AFM can provide a three-dimensional surface profile. In short, AFM is one of the cutting-edge techniques for imaging and measuring at micro and nanometer scale15,16,17.