Understanding the Uptake Mechanism and Interaction Potential of the Designed Peptide and Preparation of Composite Fiber Matrix for Fungal Keratitis

Abstract

The conventional use of antibiotics for the treatment of infectious keratitis currently faces two major challenges: poor drug penetration and the emergence of antibiotic resistance in microbial strains. Cell-penetrating peptides (CPPs) with antimicrobial properties have the potential to address these challenges. However, their mode of action, mechanism of uptake, and interaction potential have not been explored in detail. In this study, we probed the mechanism of uptake and interaction potential of our previously designed peptides (VRF005 and VRF007). Our results showed that VRF005 undergoes direct translocation and induces a rough membrane surface, whereas VRF007 undergoes clathrin-mediated endocytic uptake. The gel shift assay showed that VRF005 is bound to genomic DNA, whereas VRF007 is bound to chitin and β-D-glucan. Gene expression studies revealed the effect of peptide VRF005 on Candida albicans transcription. Molecular docking and simulations showed that VRF005 forms noncovalent interactions (such as H-bonding and water bridges) with natamycin. It exhibited synergistic antifungal activity in the colony-forming assay. VRF005, functionalized in the polycaprolactone fiber matrix, showed sustained delivery and antifungal activity.

Understanding the cellular response of human tenon fibroblast on polycaprolactone-Aloe vera blend fiber

“…Polymeric solution of both 5% PCL and PCL-AV (9:1 ratio) were loaded in a syringe of 5 mL with 21 gauge needle and spun using Espin Nanotech, India. The fibers were spun with optimized 25 parameters of voltage at 12 kV, the flow rate of 15 μL/min, and a distance of 15 cm between the tip of the needle and collector. Coverslip coated with Silane was used as a collector.…”

Elucidating the Therapeutic Potential of Cell-Penetrating Peptides in Human Tenon Fibroblast Cells

“… 1 They exhibit therapeutic functions like antifungal activity, in addition to being a delivery vehicle. 2 , 3 Several CPPs such as pexiganan, omiganan, PXLO1, and PAC-113 are in clinical trials for antimicrobial activity. 4 , 5 The outcome of the clinical trials depends on their stability, metabolic activity, bioavailability, enzymatic degradation, and the environment regulating the activity.…”

“…Cell-penetrating peptides (CPPs) like transactivator of transcription (TAT), penetratin, polyarginine, and MPG are being explored for delivering drug molecules to the target site in various ways . They exhibit therapeutic functions like antifungal activity, in addition to being a delivery vehicle. , Several CPPs such as pexiganan, omiganan, PXLO1, and PAC-113 are in clinical trials for antimicrobial activity. , The outcome of the clinical trials depends on their stability, metabolic activity, bioavailability, enzymatic degradation, and the environment regulating the activity . Delivery systems such as hydrogels, fibers, nanoparticles, etc .…”