POTENTIALS OF PLANT-DERIVED EXOSOME-LIKE NANOPARTICLES FROM PHYSALIS PERUVIANA FRUIT FOR HUMAN DERMAL FIBROBLAST (HDF) REGENERATION AND REMODELING

• Main Author: Natania, Filia
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/76741
• Institution: Institut Teknologi Bandung

Skin is the protector of the human body and prone to wounds. Wounds must heal quickly and restore skin to its normal condition. Plant-derived exosome-like nanoparticles (PDEN) has now been widely researched for its function, such as for cell-free wound healing treatment, because they are nano-sized, nonimmunogenic, nontoxic, contains important cargos, relatively cheap, and easily gained. This research will study how PDEN from Physalis peruviana fruit (PENC) and their effects on human dermal fibroblast (HDF) regeneration and remodeling. PENC are isolated using gradual filtration and centrifugation, followed by sedimentation using PEG6000. PENC are characterized by PSA (particle size analyzer), ZPA (zeta potential analyzer), TEM (for morphology), BCA (for protein concentration), and their internalization using PKH67. Functionality tests are done using MTT assay (for cytotoxicity and proliferation), scratch assay (for migration), immunocytochemistry (ICC) for scratch and collagen-1, ELISA for MMP-1, and ROS assay. Isolated PENC have size around 190-220 nm, spherical, have membrane, and their zeta potential is -6.7 mV, also can be stored in 4°C up to 2 weeks in aqua bides. Protein concentration range between 170-1,395 ?g/mL. Using PKH67, PENC can enter HDF within 6 hours. MTT assay showed PENC concentration of 2.5 until 500 ?g/mL are not toxic to cell. PENC induce cell proliferation in 1, 3, 5, and 7 days. PENC, concentration 2.5, 5, and 7.5 ?g/mL, also accelerate HDF migration using scratch assay in two days. In remodeling, PENC can upregulate collagen-1 expression from day-7 to 14 compared to control. MMP-1 declines from day-2 to 7 in every PENC concentration and increases in day-14. Meanwhile ROS declines from day-2 to 7 and 14, with PENC give rise to ROS level compared to control. Overall, PENC possess characteristics of PDEN, PENC with concentration of 2.5, 5, and 7.5 ?g/mL are potential to induce HDF proliferation and migration, upregulate collagen I production, decrease MMP-1 with PENC concentration 2.5, 5, and 7.5 ?g/mL , and upregulate ROS formation in day 7 with PENC 5 and 7.5 ?g/mL. PENC has the potential for wound healing and this research serves as fundamental knowledge for future more detailed and extensive studies.