This research delves into the realm of image processing as a pivotal component in the evaluation of fibroblast stimulation for collagen... Show moreThis research delves into the realm of image processing as a pivotal component in the evaluation of fibroblast stimulation for collagen remodeling. The study focuses on unraveling the intricate synergy between electrospun silk fibroin-carbon nanotube (SF-CNT) fibers and electrical stimulation, working in harmony to enhance tissue regeneration. Building upon our previous work, we successfully engineered SF-CNT fibers through the electrospinning process, yielding highly aligned structures reminiscent of natural extracellular matrix proteins. These fibers were fortified with water stability through post-treatment with ethanol vapor, while subtle additions of carbon nanotubes (CNTs) significantly improved fiber alignment, strength, and conductivity without compromising biocompatibility. This innovative platform served as a cell culture matrix for fibroblasts harvested from pelvic organ prolapse (POP) patients, facilitating electrical stimulation that triggered a substantial increase in collagen production. In this study, we harnessed the power of various image-processing software tools, including ImageJ and Python, to analyze immunostained images of fibroblasts obtained from POP patients. Under carefully tailored electrical stimulation conditions, the stimulated cells exhibited an astonishing up to 11.97-fold increase in alpha-smooth muscle actin (α-SMA) expression, unequivocally signifying the successful activation of myofibroblasts. Additionally, in an animal model employing LOX-knockout mice to mimic collagen disorders associated with POP, the application of optimized electrical stimulation conditions for patient 003 led to a remarkable surge in collagen production and structural enhancement, underlining the potential of electrical stimulation in expediting tissue remodeling. Intriguingly, fibroblasts from patient 005 and patient 006 exhibited a distinct response, shedding light on the influence of POP severity on cellular behavior. This study firmly reinforces the imperative of personalized therapeutic approaches, emphasizing the need to customize treatment strategies to align with individual patient characteristics through innovative biological image analysis techniques. Show less
