scholarly journals Discovery of Lactoferrin as A Stimulant for hADSC-Derived EV Secretion and Proof of Enhancement of Resulting EVs through Skin Model

2021 ◽  
Vol 22 (20) ◽  
pp. 10993
Author(s):  
Junho Kim ◽  
Ga Eun You ◽  
Minkyu Woo ◽  
Nicole Hyesoo Chang ◽  
Jungsun Lee
Keyword(s):  
Membrane Protein ◽  
Extracellular Vesicles ◽  
Collagen Synthesis ◽  
Dermal Fibroblasts ◽  
Human Lactoferrin ◽  
Protein Markers ◽  
Skin Model ◽  
Low Concentration ◽  
Low Concentrations ◽  
Enhanced Secretion

Extracellular vesicles (EVs) are secreted from hADSCs in low concentrations, which makes it difficult to utilize them for the development of therapeutic products. To overcome the problem associated with low concentration, we proposed human lactoferrin (hLF) as a stimulant for the secretion of hADSC-derived EVs. hLF has been reported to upregulate intracellular Ca2+, which is known to be capable of increasing EV secretion. We cultured hADSCs in hLF-supplemented media and analyzed the changes in intracellular Ca2+ concentration. The characteristics of hADSC-derived EVs secreted by hLF stimulation were analyzed through their number, membrane protein markers, and the presence of hLFs to EVs. The function of hADSC-derived EVs was investigated through their effects on dermal fibroblasts. We found that hLF helped hADSCs effectively uptake Ca2+, resulting in an increase of EVs secretion by more than a factor of 4. The resulting EVs had enhanced proliferation and collagen synthesis effect on dermal fibroblasts when compared to the same number of hADSC-derived EVs secreted without hLF stimulation. The enhanced secretion of hADSC-derived EVs increased collagen synthesis through enhanced epidermal penetration, which resulted from increased EV numbers. In summary, we propose hLF to be a useful stimulant in increasing the secretion rate of hADSC-derived EVs.

Protein biomarkers in breast cancer-derived extracellular vesicles for use in liquid biopsies

2021 ◽  
Author(s):  
Dan Li ◽  
Wenjia Lai ◽  
Di Fan ◽  
Qiaojun Fang
Keyword(s):  
Breast Cancer ◽  
Early Diagnosis ◽  
Extracellular Vesicles ◽  
Liquid Biopsy ◽  
Breast Cancer Diagnosis ◽  
Extracellular Vesicle ◽  
Detection Methods ◽  
Protein Markers ◽  
Liquid Biopsies ◽  
Diagnosis And Prognosis

Breast cancer is the most common malignant disease in women worldwide. Early diagnosis and treatment can greatly improve the management of breast cancer. Liquid biopsies are becoming convenient detection methods for diagnosing and monitoring breast cancer due to their non-invasiveness and ability to provide real-time feedback. A range of liquid biopsy markers, including circulating tumor proteins, circulating tumor cells, and circulating tumor nucleic acids, have been implemented for breast cancer diagnosis and prognosis, with each having its own advantages and limitations. Circulating extracellular vesicles are messengers of intercellular communication that are packed with information from mother cells and are found in a wide variety of bodily fluids; thus, they are emerging as ideal candidates for liquid biopsy biomarkers. In this review, we summarize extracellular vesicle protein markers that can be potentially used for the early diagnosis and prognosis of breast cancer or determining its specific subtypes.

scholarly journals Low Concentrations of Chlorhexidine Inhibit the Formation and Structural Integrity of Enzyme-Treated Multispecies Oral Biofilms

2021 ◽  
Vol 12 ◽  
Author(s):  
Kay Andrin Gränicher ◽  
Lamprini Karygianni ◽  
Thomas Attin ◽  
Thomas Thurnheer
Keyword(s):  
Structural Integrity ◽  
Fluorescent Dyes ◽  
Combined Treatment ◽  
Extracellular Proteins ◽  
Laser Scanning Microscopy ◽  
Extracellular Dna ◽  
Anaerobic Growth ◽  
Low Concentration ◽  
Low Concentrations ◽  
Dental Hard Tissues

The self-produced matrix of biofilms, consisting of extracellular polymeric substances, plays an important role in biofilm adhesion to surfaces and the structural integrity of biofilms. In dentistry, biofilms cause multiple diseases such as caries, periodontitis, and pulpitis. Disruption of these biofilms adhering to dental hard tissues may pose a major challenge since biofilms show higher tolerance to antimicrobials and antibiotics than planktonic cells. In this study, the effect of low concentrations of chlorhexidine (CHX) on enzyme-treated multispecies oral biofilm was investigated in an in vitro model. Six-species biofilms were enzymatically treated by anaerobic growth in a medium containing DNase I and proteinase K. Biofilms were exposed to a low concentration of CHX at defined time points. After 64h, biofilms were either harvested and quantified by cultural analyses or stained for confocal laser scanning microscopy (CLSM) analyses using either Live/Dead kit or different fluorescent dyes. A mixture of YoPro1 and SYTOX™ Green, Fluorescent Brightener 28 (Calcofluor), and SYPRO™ Ruby Protein Gel Stain was used to stain total DNA, exopolysaccharides, and extracellular proteins, respectively. Extracellular DNA (eDNA) was visualized via an indirect immunofluorescence assay (Mouse anti-DNA IgG, Goat anti-Mouse IgG, Streptavidin-Cy3). Overall, the total colony-forming units significantly decreased after combined treatment with a low concentration of CHX and enzymes compared to the group treated with CHX alone (p<0.001). These findings also apply to five species individually (Streptococcus mutans, Streptococcus oralis, Actinomyces oris, Veillonella dispar, and Candida albicans) occurring in the biofilms, with Fusobacterium nucleatum being the only exception. Furthermore, CLSM images showed less dense biofilms and a reduction in cell numbers after combined treatment compared to the group without enzymes. The combination of enzymes capable of disturbing the matrix integrity with antimicrobial agents thus appears to be a promising approach for biofilm disruption and killing.

scholarly journals Isolation and Characterization of Extracellular Vesicles from the Fungal Phytopathogen Colletotrichum higginsianum

2022 ◽  
Author(s):  
Brian D Rutter ◽  
Thi-Thu-Huyen Chu ◽  
Kamil K Zajt ◽  
Jean-Felix Dallery ◽  
Richard J O'Connell ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Experimental System ◽  
Protein Markers ◽  
Cell Wall Biogenesis ◽  
Colletotrichum Higginsianum ◽  
Isolation And Characterization ◽  
Large Numbers ◽  
Transgenic Lines ◽  
Fungal Phytopathogen ◽  
Fungal Phytopathogens

Fungal phytopathogens secrete extracellular vesicles (EVs) associated with enzymes and phytotoxic metabolites. While these vesicles are thought to promote infection, defining the true contents and functions of fungal EVs, as well as suitable protein markers, is an ongoing process. To expand our understanding of fungal EVs and their possible roles during infection, we purified EVs from the hemibiotrophic phytopathogen Colletotrichum higginsianum, the causative agent of anthracnose disease in multiple plant species, including Arabidopsis thaliana. EVs were purified in large numbers from the supernatant of protoplasts but not the supernatant of intact mycelial cultures. We purified two separate populations of EVs, each associated with over 700 detected proteins, including proteins involved in vesicle transport, cell wall biogenesis and the synthesis of secondary metabolites. We selected two SNARE proteins (Snc1 and Sso2) and one 14-3-3 protein (Bmh1) as potential EV markers and generated transgenic lines expressing fluorescent fusions. Each marker was confirmed to be protected inside EVs. Fluorescence microscopy was used to examine the localization of each marker during infection on Arabidopsis leaves. These findings further our understanding of EVs in fungal phytopathogens and will help build an experimental system to study EV inter-kingdom communication between plants and fungi.

scholarly journals Asiaticoside induces cell proliferation and collagen synthesis in human dermal fibroblasts

2015 ◽  
Vol 34 (2) ◽  
pp. 96
Author(s):  
Linda Yuliati ◽  
Etik Mardliyati ◽  
Kusmarinah Bramono ◽  
Hans Joachim Freisleben
Keyword(s):  
Cell Proliferation ◽  
Wound Healing ◽  
Retinoic Acid ◽  
Collagen Synthesis ◽  
Active Agent ◽  
Dermal Fibroblasts ◽  
Type Iii Collagen ◽  
Type I ◽  
Human Dermal Fibroblasts ◽  
Type Iii

Background<br />Asiatiocoside, a saponin component isolated from Centella asiatica can improve wound healing by promoting the proliferation of human dermal fibroblasts (HDF) and synthesis of collagen. The skin-renewing cells and type I and III collagen synthesis decrease with aging, resulting in the reduction of skin elasticity and delayed wound healing. Usage of natural active compounds from plants in wound healing should be evaluated and compared to retinoic acid as an active agent that regulates wound healing. The aim of this study was to compare and evaluate the effect of asiaticoside and retinoic acid to induce greater cell proliferation and type I and III collagen synthesis in human dermal fibroblast.<br /><br />Methods<br />Laboratory experiments were conducted using human dermal fibroblasts (HDF) isolated from human foreskin explants. Seven passages of HDF were treated with asiaticoside and retinoic acid at several doses and incubated for 24 and 48 hours. Cell viability in all groups was tested with the MTT assay to assess HDF proliferation. Type I and III collagen synthesis was examined using the respective ELISA kits. Analysis of variance was performed to compare the treatment groups. <br /><br />Results<br />Asiaticoside had significantly stronger effects on HDF proliferation than retinoic acid (p&lt;0.05). The type III collagen production was significantly greater induction with asiaticoside compared to retinoic acid (p&lt;0.05). <br /><br />Conclusion<br />Asiaticoside induces HDF proliferation and type I and III collagen synthesis in a time- and dose-dependent pattern. Asiaticoside has a similar effect as retinoic acid on type I and type III collagen synthesis.

Transepithelial movement of calcium in crustaceans

1993 ◽  
Vol 184 (1) ◽  
pp. 1-16 ◽  
Author(s):  
D. S. Neufeld ◽  
J. N. Cameron
Keyword(s):  
Active Transport ◽  
Calcium Concentration ◽  
Cellular Mechanism ◽  
The Body ◽  
High Rate ◽  
Calcium Flux ◽  
Moult Cycle ◽  
Electrochemical Gradient ◽  
Low Concentration ◽  
Low Concentrations

The regulation of calcium in most crustaceans is especially challenging owing to the highly mineralized cuticle that must be recalcified after each moult, a process that often occurs in environments with low concentrations of calcium. The gill and carapace epithelia separate the major calcium-containing compartments of the body and therefore see large changes in the rate of calcium flux through the moult cycle. Large changes in the ultrastructure of these cells do not, however, correlate well with the periods of calcium movement and probably reflect other physiological events. Despite the challenges to regulating calcium levels at various acclimation salinities and moult stages, the calcium concentration in the blood is maintained relatively constant. There is a rapid increase to a high rate of calcium flux across both the gill and carapace epithelium shortly after the moult; on an area-specific basis these fluxes are among the highest reported for calcium-transporting epithelia. When in water with a very low concentration of calcium, the electrochemical gradient for calcium is directed outwards and net influx must occur by active transport. Evidence suggests that changes in the electrochemical gradient, permeability and active transport are all important in the ability of crustaceans to take up calcium from water with a low concentration of this ion. Although an enzyme transporter is presumably involved in the active transport of calcium across epithelia, very little is known about the cellular mechanism of the transepithelial movement of calcium in crustaceans.

scholarly journals Antiaging Activity of Peptide Identified from Fermented Trapa Japonica Fruit Extract in Human Dermal Fibroblasts

2020 ◽  
Vol 2020 ◽  
pp. 1-8 ◽  
Author(s):  
Jin Dong Jang ◽  
Minkyung Kim ◽  
Gun-He Nam ◽  
Young-Min Kim ◽  
Sang Moon Kang ◽  
...  
Keyword(s):  
Clinical Study ◽  
Mrna Expression ◽  
Collagen Synthesis ◽  
Dermal Fibroblasts ◽  
Dependent Manner ◽  
Human Dermal Fibroblasts ◽  
Fruit Extract ◽  
Necrosis Factor Alpha ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell

We have previously shown that Trapa japonica fruit extract (TJE) as well as its fermented extract (FTJ) can be potentially used to treat alopecia. In the current study, a newly synthesized peptide (PEP) was detected in an active compound isolated from FTJ. Several biological assays were conducted to verify the antiaging effects of TJE, FTJ, and PEP on the skin. We examined the effects of TJE, FTJ, and PEP on cell viability, collagen synthesis, and inhibition of mRNA expression of matrix metalloproteinases (MMPs), induced by tumor necrosis factor alpha (TNF-α), in human dermal fibroblasts (HDFs). In addition, a wound-healing assay of the human keratinocyte cell line (HaCaT) and a clinical study of antiaging activity were conducted. The findings confirmed that PEP exerted an effect on cell proliferation in a dose-dependent manner. Treatment with TJE, FTJ, and PEP increased collagen synthesis but inhibited TNF-α-induced mRNA expression of MMPs. Compared with TJE and FTJ, PEP promoted a significant level of wound recovery in HaCaT cells and also exhibited antiaging effect, as demonstrated by a clinical study. These results suggest that PEP shows potential as a skin antiaging cosmetic product.

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