FABRICATION OF PEO-CHITOSAN NANOFIBER MEMBRANE CONTAINING PROTEIN TARO TUBER (COLOCASIA ESCULENTA) USING ELECTROSPINNING FOR ACUTE WOUND DRESSING APPLICATION

• Main Author: Ayu Kusuma Wardhani, Riesca
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/51477
• Institution: Institut Teknologi Bandung

Wound healing is a complex regeneration process that involves several biological and molecular systems, such as coagulation, inflammation, proliferationmigration, and remodeling, to restore normal biological function. The bioactive wound dressing is applied to the wound site to accelerate the healing process while preventing infections. This wound dressing is not only permeable to water vapor and oxygen and provides an effective barrier against bacteria or other microorganisms from the outside environment but also has a bioactive molecule. Bioactive molecules such as proteins can support the attachment and growth of fibroblast cells that will enhance the wound healing process. Protein is one of the essential roles in the wound healing process because it is needed for enzyme and collagen synthesis, the proliferation of fibroblast cells, and the formation of connective tissue. Nanofiber membranes are included in the biological bandages of the bioactive category because they are synthesized chemically from natural materials. Natural polymers are the most widely used polymers for the treatment of wounds because of their ability to interact with biomolecules involved in the healing process. In this work, three biocompatible, biodegradable, and nontoxic polymers selected were two of natural polymers (CET’s protein and CS) and one of synthetic polymer (poly(ethylene) oxide (PEO)). These choices were encouraged by the promising properties of these polymers, which similarity toward structural components of ECM. All three polymers are fabricated in a three-dimensional structure using electrospinning techniques. Electrospinning is the most efficient processing method for producing continuous nanofiber for large scale from various polymers with small pore size, high porosity, and the sizeable surface-area-to-volume ratio. In this study, we aimed to invent a biomimetic system in the form of a nanofiber membrane with excellent material stability. Suitability of the use of crosslinking, it is expected that the PEO–CS–CET’s protein nanofibers can effectively function as potential candidates for wound dressing. CET’s protein containing 6.6% protein with molecular weight 53.5, 42, 24.4, 11.8, and 9.8 kDa shows no cytotoxicity on NIH-3T3 fibroblast cell. The protein profiles of the electrospun PEO–CS–CET’s protein nanofibers before and after HT crosslinking contained one major bioactive protein with a molecular weight of 14.4 kDa. The highest amino acids in the CET’s protein are arginine, aspartic acid, and glutamic acid. The electrical conductivity, surface tension, and viscosity of the blended solutions were increased with the increased of CET’s protein and chitosan content due to its ability to carry positive charges on the polymer chains. The scanning electron microscope (SEM) shows that the average diameter of the PEO– CS–CET’s protein nanofibers decreased with the increased of chitosan and CET’s protein content. The presence of 1% (w/v) chitosan resulted in better preservative of the nanofibrous structure after the stabilization process, so that the fiber structure is not destroyed. The results of the thermal gravimetric analyzer (TGA) confirm that the S4-H and S5-H fiber membranes cannot be tested for mechanical properties, swelling ratio, and degradation due to loss of fiber structure when stabilizing heat treatment. The ultimate tensile strength increased, and the ultimate strain decreased on the crosslinked nanofiber membranes as compared to noncrosslinked nanofiber membranes. GA vapor crosslinking shown higher water stability compared to HT crosslinking in 1% (w/v) CS and 1% (w/v) CET’s protein and 1% (w/v) CS and 2% (w/v) CET’s protein. Scanning electron microscopy of the crosslinked nanofibers indicated preservation of the structure after immersion in phosphate buffered saline. The in vitro antibacterial activity of the crosslinked nanofibers showed stronger bacteriostatic effect with Gram-positive bacteria than Gram-negative bacteria in the CS concentration of 0.25% and 1% (w/v). Human skin fibroblast cell proliferation on both crosslinked GA vapor and HT nanofibers with 1% (w/v) CS and 2% (w/v) CET’s protein was obtained the highest among all the other crosslinked nanofibers after seven days of cell culture. Our study not only present insights on the design of crosslinked electrospun CET’s protein–CS–PEO nanofibers, but also introduce other bioactive materials from plant-derived proteins for wound healing. This fiber membrane facilitates the growth of fibroblast cells so that its use is promising as a bioactive wound dressing.