Characterizing Nanoparticle Isolated by Yam Bean (Pachyrhizus erosus) as a Potential Agent for Nanocosmetics: An in vitro and in vivo Approaches

Background: This study investigated the potential of Plant-Derived Exosome-Like Nanoparticles (PDENs) as cosmeceutical nanocarriers for treating skin problems, such as scar removal, face rejuvenation, anti-aging, and anti-pigmentation. background: Background. Depigmentation and skin-whitening products have long been widely used in Asian countries. Highly hazardous active substances, including hydroquinone, corticosteroids, and derivatives of mercury, are commonly included in skin-whitening treatments that may be easily found on the market. The use of these depigmenting agents can be followed by a variety of adverse effects, with very serious and fatal complications. Objective: Researchers isolated PDENs from Yam Bean (Pachyrhizus erosus) using PEG-based precipitation, gradual filtration, and various centrifugations at low temperatures. Followed by in vitro and in vivo studies using HDF cells and Zebrafish. Methods: The morphology of the YB-PDENs was determined using TEM analysis, they had a spherical shape with diameters of 236,83 ± 9,27nm according to PSA. The study found that YB-PDENs were stable in aquabidest at 4°C for one month of storage and had ~-26,5 mV of Zeta Potential. The concentration of YB-PDENs was measured using the BCA Assay, and internalization of YB-PDENs to HDF cells was observed using a Confocal Laser Scanning Microscope labelled with PKH67. method: Method and Result. We successfully isolated PDENs from Yam Bean (Pachyrhizus erosus) using PEG-based precipitation, with gradual filtration followed by various centrifugations at low temperatures. The morphology of Yam Bean PDENs (YB-PDENs) was determined using TEM analysis. It has a spherical shape, with diameters ranging from 100-150 nm and its size distribution and storage stability was determined using DLS analysis with an average size of 236,83nm ± 9,27nm. Within aquabidest at a temperature of 4C, it stable for one month storage according to DLS analysis and has -26.5mV of Zeta Potential. The concentration of YB-PDENs was measured using the BCA Assay had a concentration range 101,15 – 266,70 μg/mL. Internalization of YB-PDENs to Human Dermal Fibroblast (HDF) cells was seen using a Confocal Laser Scanning Microscope that had previously been labeled with PKH67, within one hour of incubation, YB-PDENs could also be absorbed by HDF cells. Using MTT Assay we see the viability of the HDF cells after incubated with YB-PDENs. It shows that within 24 and 72 hours of incubation with YB-PDENs, the viability of HDF remain more than 80%. To examining cell migration, we performed Scratch Assay using MuviCyteTM, the measurements are made by determining the length of the covered area. At the 2.5 μg/mL YB-PDENs have better migration results than other concentrations with p=0.006. Immunocytochemistry was performed after 14 days of incubation HDF with YB-PDENs using Col-1 and the collagen expressed well. In vivo study was conduct using Zebrafish as animal model to examine the anti-melanogenic effect of the YB-PDENs. The melanocytes in Zebrafish were observed to be decrease at each concentration compared to controls and the melanocytes appeared significantly after 48 hours treatment is not given. Result: As for cytotoxicity, after 24 and 72 hours of incubation with YB-PDENs, the viability of HDF cells remained more than 80%. The study also examined cell migration using the Scratch Assay and found that at 2,5 μg/mL, YB-PDENs had better migration results than other concentrations. Immunocytochemistry showed that collagen expression was higher after 14 days of incubation with YB-PDENs, and melanocytes in zebrafish decreased at each concentration compared with controls. Conclusion: In conclusion, this study is the first to extract and describe PDENs from Yam Bean (Pachyrhizus erosus), with YB-PDENs having a promising anti-melanogenic effect in skin treatment. This study highlights the potential of YB-PDENs as a promising alternative to depigmentation and skin whitening treatments.