Preliminary study on preperation,characterization and anti-melanogenesis effect of exosomes derived from Schizochytrium limacinum dregs
[Objective]Exosomes are membranous vesicles secreted by biological cells,with diameters ranging from 30 to 150 nm.They contain a diverse array of proteins and RNA molecules,which are crucial carriers for intercellular transmission of biological information.Exosomes have become a focus point in scientific research and biomedicine industry in recent years due to their broad application prospects and high demands.In this study,we extracted exosomes from Schizochytrium limacinum dregs and explored their cosmetic functions for the first time.We aimed at providing information for the utilization of S.limacinum dregs waste products and researches of plant exosomes,while providing new perspective on the potential of exosomes as active substance for skin whitening.[Methods]By gradually increasing the centrifuge speed,an ultrafast centrifuge was used to separate other impurities from the S.limacinum dregs after the extraction of algal oil,and finally the exosomes were extracted from it.The morphology of exosomes was observed using transmission electron microscopy(TEM).The concentration and size of exosomes were analyzed with a flow nanoanalyzer(NanoFCM),and their protein concentrations were evaluated using the bicinchoninic acid method.In addition,the Oryzias melastigma model was creatively established to test the anti-melanogenesis effect of exosomes and explore the beauty clinic functions of exosomes.[Results]The exosomes isolated from the S.limacinum dregs by differential centrifugation were studied in detail,and the effects of melanin accumulation in O.melastigma after exposure to the exosomes of S.limacinum were investigated.The extracted exosomes showed a characteristic round or oval cup-like shape with a double-layer membrane and an intact membrane structure.The particle size,measured by TEM,was approximately 100 nm,aligning with the particle size of(90.8±19.9)nm obtained from the NanoFCM.The concentration of protein was 0.3 mg/g and that of exosomes was around 2.96×1012 mL-1.These data revealed that S.limacinum could produce exosomes at high concentrations,making it an excellent plant source for exosome extraction.The results suggested that the low,medium and high concentrations of exosomes from the residue of S.limacinum showed a clear inhibitory effect on O.melastigma model.Observed with the positive control group,the number of melanin spots formed on O.melastigma was 0,which reflected an obvious effect of N-phenylthiourea(PTU)on inhibiting melanin formation.The number of melanin spots formed by O.melastigma when exposed to high concentrations of exosomes(20 μg/mL)derived from S.limacinum dregs was also 0,and the number of melanin spots formed by O.melastigma when exposed to medium concentrations of exosomes(10 μg/mL)derived from S.limacinum dregs was 1.O.melastigma exposed to low concentrations of exosomes(1 μg/mL and 5 μg/mL)derived from S.limacinum dregs produced 3 melanin spots.Moreover,there was a positive correlation between the inhibition rate and concentration:as the concentration increased,the inhibition effect became more pronounced.The results of the study confirm that the exosomes from the dregs of S.limacinum significantly inhibit the production of melanin and can be further explored as a potential whitening active substance.[Conclusions]This study successfully prepared and identified exosomes derived from the S.limacinum dregs and established the model of O.melastigma,confirming that the anti-melanogenesis effect of exosomes derived the S.limacinum dregs.The results from above characterizations indicate that our research provides new insights into the high-value utilization of S.limacinum dregs after industrial extraction of algal oil.This work also serves as a theoretical basis for developing efficacy evaluation of whitening products with skin care effects.At the same time,due to its remarkable anti-melanogenesis effect,the derived exosomes can be used as new candidates for anti-melanin drugs in cosmeceutical applications.
exosomeSchizochytrium limacinumanti-melanogenesis effectOryzias melastigma model
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