ATR-FTIR spectral monitoring of burn wound healing in skin by hierarchical cluster

Abstract

Efficient biochemical characterization of burn wound healing stages can improve clinical routine to adjust the patient treatment. The Fourier Transform Infrared spectroscopy coupled with Attenuated Total Reflectance (ATR-FTIR) is an analytical technique that has potential capability to provide spectral biomarkers in biological material. This study aims to evaluate the feasibility of using ATR-FTIR to classify burned skin, to be able, in the future, to follow the regenerative process in patients. Wistar rat burn tissues were evaluated by ATRFTIR spectroscopy at 3, 7, 14, 21 days after burn and compared with the healthy group samples (H). For the acquisition of one spectrum, 150 scans were averaged with a resolution of 4 cm-1 and wavenumbers ranging from 4000 to 400 cm-1. Analysis of the spectra was performed using MatLab R2017a (MathWorks, EUA) software. The fingerprint region between 900 to 1800 cm-1 was separated and normalized by amide I band area. For smoothing purpose, spectra were submitted to Savitzky-Golay filter with a polynomial of second order in a fifteen points window. All spectra data were submitted to the hierarchical cluster using the single method and standardized Euclidean distance. The classification results demonstrated separation to non-wounded groups with an accuracy of 91,8% (H vs. 3D), 87,83% (H vs. 7D) and 97,96% (H vs. 14D). Interestingly, when the results from the healthy group were compared to the 21D group, the accuracy dissimilarity was 26%, which suggests that they are chemically similar implying that after 21 days the burned lesions are entirely recovered. These findings indicate that when an adequate multivariate method is applied, ATR-FTIR is suitable to detect the wounded and non-wounded skin as well as its healing stages. In this way, it is possible to conclude that ATR-FTIR can be an auxiliary analysis for the clinical routine for skin burn wound healing.