Using biospeckle and LIBS techniques with artificial intelligence to monitor phthalocyanine-gold nanoconjugates as a new drug delivery mediator for in vivo PDT

Photodynamic therapy (PDT) is a promising treatment method that generates reactive oxygen species by exciting a photosensitizer in the presence of light. The positive treatment outcome of PDT is enhanced by the photosensitizer's ability to get out quickly from normal tissue. This study non-destructively evaluated the biocompatibility of novel synthesized phthalocyanine-gold nanoconjugates (Pc-Au NCs) as a drug delivery mediator for in vivo PDT. We first proposed a novel noncontact and safe biospeckle optical imaging technique to visualize the quantitative changes in the internal dynamics of biological tissue in response to PDT treatment. Second, laserinduced breakdown spectroscopy (LIBS) was utilized to emphasize the accumulation of gold nanoparticles (Au NPs) within the normal and cancerous tissue, which in turn verifies the cytotoxic and treatment effects of the PcAu NCs. The biospeckle images and LIBS spectra were obtained from tumor and normal tissues of male BALB/c malignancy-bearing mice before and after exposure to PDT. Then, they were statistically treated via different artificial intelligence techniques. For biospeckle analysis, the proposed method was established from the combination of second and higher-order statistical features extracted from the gray-level co-occurrence matrix (GLCM) and local contrast analysis (LCA), respectively, through principal component analysis (PCA). Regarding LIBS analysis, the powerful, most popular ensemble sequential learning algorithm Adaptive Boost was implemented on highly uncorrelated variables extracted from the raw spectra. The results, confirmed by the histological investigation, indicated the potential of the biospeckle and LIBS statistical properties in demonstrating the efficiency of the Pc-Au NCs as a drug delivery mediator for in vivo PDT.