SERS-Assisted Multivariate Data Analysis of Spectinomycin-Cu(II) Complex Released in Blood Serum from Stimuli-Responsive Drug Carrier: In-Vitro Kinetics Modeling and Pharmacodynamic Analysis

Abstract

The controlled drug delivery from stimuli-responsive drug carrier offers effective therapeutic potential, however, precise assessment of release kinetics and pharmacodynamic (PD) behavior remains challenging. In this research work, the surface-enhanced Raman spectroscopy (SERS) is used for the quantification of the Spectinomycin-Copper (SPM-Cu (II)) complex released in human blood serum. The silver nanoparticles (AgNPs) are prepared by the chemical reduction method for the active SERS substrate. The SPM-Cu(II) complex loaded on the stimuli responsive PVA/AgO hydrogel is released in blood serum, 36 hours release kinetics and in-vitro pharmacodynamics study is carried at different intervals as (1h, 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h and 36h).The multivariate data analysis techniques, Principal Component Analysis (PCA) is used for the qualitative analysis to study the intensity-based variability, Partial Least Squares Regression (PLSR) is used for the quantification of SPM-Cu Complex release kinetics while Hierarchical Cluster Analysis (HCA) is used for spectral variability and classify temporal release profiles. The release kinetics is additionally validated by UV-Vis spectroscopy quantification compared with SPM-Cu(II) Complex released in phosphate buffer solution of pH 7.4 at 37oC. The in-vitro release kinetics is determined using four kinetic models that included Zero-order, First-order, Higuchi, and Korsmeyer-Peppas. It is shown by mathematical fitting that the release is dominated by Higuchi (R2 = 0.934 in serum, 0.944 in PBS) and Korsmeyer Peppas models (n = 0.463 which is Fickian diffusion), indicating that the release is diffusion-controlled. Pharmacodynamic potential of released SPM-Cu(II) complex in blood serum is evaluated systematically by antibacterial activity, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and biofilm assay against Pseudomonas aeruginosa and Enterococcus faecalis. The Cytotoxicity against human liver cancer cell line (HepG2), and hemolytic analysis, are also performed to validate the release kinetics in blood serum. The results indicate the improved antibacterial and biocompatible properties of the SPM-Cu(II) released in blood serum from PVA/AgO hydrogel.