Effect of MMP/TIMP Balancing of Cynoglossus semilaevis Shell Extracts on Skin Protection

Cynoglossus semilaevis shell is a by-product of the Cynoglossus semilaevis, a species of fish mainly distributed along the west coast of Korea. As its skin is very tough and difficult to process, it is not useful as food. For this reason, most of it is discarded except for a small amount that is used as feed, which results in environmental pollution. Considering this, there is a need for research on the development of functional materials using Cynoglossus semilaevis shell. This study focused on the mechanism of in vitro expression function of Cynoglossus semilaevis shell extract (CSE) for skin tissue in human dermal fibroblasts that induced or did not induce wrinkles by UV-B irradiation and aims to use it as a functional material for human skin beauty or wrinkle improvement through extraction and purification. According to the ELISA results using human dermal fibroblast cells, CSE reduced MMP-1 and elastase activity by up to 21.89% and 12.04%, respectively, in a concentration-dependent manner, and increased PIP synthesis by up to 62.24% in a concentration-dependent manner. The RT-PCR test results using mRNA showed the MMP-1, 2, and 3 expression levels were suppressed in the CSE-treated group compared to the UVB-induced group and caused a concentration-dependent increase in TIMP-1 in the CSE-treat group. These results suggest that CSE can maintain and improve skin tissue conditions through MMP/TIMP balancing in human dermal fibroblast cell lines and indicate its potential as a functional material for improving skin diseases and suppressing photo-aging.

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Evaluation of the efficacy of ethanol leaf extract of Helichrysum petiolare Hilliard and B.L. Burtt against skin aging

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i12.22 ◽

2021 ◽

Vol 19 (12) ◽

pp. 2631-2638

Author(s):

Idowu Jonas Sagbo ◽

Wilfred Otang-Mbeng

Keyword(s):

Leaf Extract ◽

Dermal Fibroblast ◽

Stress Protein ◽

Inhibitory Effect ◽

Human Dermal Fibroblast ◽

Skin Aging ◽

Protein Glycation ◽

No Production ◽

Dependent Manner ◽

Concentration Dependent Manner

Purpose: To determine the efficacy of Helichrysum petiolare ethanol leaf extract against skin aging.Methods: The cytotoxic potential of the plant extract towards human dermal fibroblast (MRHF) cells was determined by Hoechst 33342/Propidium iodide (PI) staining. Effect of H. petiolare extract on reactive oxygen species (ROS) levels in MRHF cells and NO (nitric oxide) production in RAW 246.7 cells activated by LPS (lipopolysaccharides) was investigated. The inhibitory effect of the extract againstcollagenase, elastase, tyrosinase and protein glycation was also evaluated.Results: The extract did not display cytotoxicity towards MRHF cells at the tested concentrations when compared to the trend seen with the untreated control (p < 0.05). The extract caused a significant decrease (p < 0.05) in ROS levels in MRHF cells in a concentration-dependent manner and also demonstrated a reduction in NO production in RAW cells with no toxicity. Furthermore, the extract produced a weak inhibition of collagenase, elastase and tyrosinase activities when compared to the corresponding positive controls, but effectively inhibited protein glycation at the tested concentrations.Conclusion: The findings suggest that the ethanol leaf extract from H. petiolare has the potential to mitigate skin aging and therefore needs to be further investigated for possible clinical applications. Keywords: Cytotoxicity, Efficacy, Helichrysum petiolare, MRHF cells, Oxidative stress, Protein glycation; skin aging

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Dendranthema zawadskii var. lucidum (Nakai) J.H. Park Extract Inhibits Cellular Senescence in Human Dermal Fibroblasts and Aging-Related Inflammation in Rats

Processes ◽

10.3390/pr9050801 ◽

2021 ◽

Vol 9 (5) ◽

pp. 801

Author(s):

Jehun Choi ◽

Gwi-Yeong Jang ◽

Jeonghoon Lee ◽

Hae-Young Chung ◽

Hyung-Jun Noh ◽

...

Keyword(s):

Cellular Senescence ◽

Cell Cycle Progression ◽

Inflammatory Responses ◽

Dermal Fibroblast ◽

Dermal Fibroblasts ◽

Dependent Manner ◽

Aged Rats ◽

Age Related ◽

Dendranthema Zawadskii ◽

Beta Galactosidase

Senescence is the phenomenon by which physiological functions of organisms degenerate with time. Cellular senescence is marked by an inhibition of cell cycle progression. Beta-galactosidase accumulates in the lysosomes of aged cells. In this study, human dermal fibroblast cells (HDFs) were treated with 0.5 μM doxorubicin for 4 h to induce cellular senescence. Senescence-associated beta-galactosidase (SA-β-gal) activity was then measured 72 h after treatment with aerial parts of Dendranthema zawadskii var. lucidum (Nakai) J.H. Park (DZ) extract. Treatment with DZ extract significantly decreased SA-β-gal activity in a dose-dependent manner in HDFs. Additionally, DZ extract treatment reduced age-related oxidative stress and inflammation in the aortas of aged rats. The reactive oxygen species (ROS) levels in aortas of aged control rats were higher than those in young rats. However, DZ extract-fed aged rats showed significantly lower ROS levels than the aged control rats. When the aged rats were treated with DZ extract at either 0.2 or 1.0 mg∙kg−1∙day−1, NF-κB levels in aorta tissue decreased significantly compared to those in aorta tissue of the aged control rats without DZ treatment. In addition, DZ extract-fed aged rat aortas showed significant reductions in expression of iNOS and COX-2 induced by NF-κB translocation. Therefore, these results suggest that DZ effectively inhibited senescence-related NF-κB activation and inflammation. DZ extract may have a role in the prevention of the vascular inflammatory responses that occur during vascular aging.

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Abstract 189: HuR, RNA Stabilizing Protein, Regulation of Pluripotency and Differentiation in iPSCs

Circulation Research ◽

10.1161/res.121.suppl_1.189 ◽

2017 ◽

Vol 121 (suppl_1) ◽

Author(s):

Sahana Suresh Babu ◽

Johnson Rajasingh ◽

Wing Tak Wong ◽

Prasanna Krishnamurthy

Keyword(s):

Rna Binding ◽

Rna Binding Proteins ◽

Dermal Fibroblast ◽

Critical Role ◽

Human Dermal Fibroblast ◽

Dermal Fibroblasts ◽

General Observation ◽

Transcript Stability ◽

Differentiated Cells ◽

Induced Pluripotent

Background: The Hu family of RNA-binding proteins, HuR (also known as ELAVL1 or human embryonic lethal abnormal vision-like protein), binds to the 3’-untranslated region of mRNAs and regulates transcript stability and translation. Global deletion of HuR is embryonically lethal in mice and plays a critical role in progenitor cell survival and biology. Induced-pluripotent stem cells (iPSC) have distinct transcriptional machinery for the maintenance of pluripotency and achievement of differentiation. However, the exact role of HuR in pluripotency or differentiation of iPSC to cardiomyocytes (iCM) remains unclear. Methods: HuR knockdown in human dermal fibroblast-derived iPSCs was achieved by CRISPR/Cas9 or lentiviral shRNA transduction and subsequently differentiated into cardiomyocytes (iCM). Then, the expression of HuR, pluripotency and cardiomyocyte markers were evaluated on days 0, 1, 3, 6, 8 and 17 following the initiation of differentiation. Results: At basal level, HuR expression was higher in the iPSCs compared to dermal fibroblasts. Upon differentiation of iPSCs into iCM, HuR mRNA expression gradually reduced with significantly lower levels on day 17. As expected, pluripotency markers gradually reduced upon differentiation with significantly lower levels from day 6 onwards. We observed a corresponding increase in ISL1, MESP1 (mesoderm/cardiac progenitor markers) from day 3 through day 8 with a steep fall from day 8 to day 17. This was associated with Myosin light chain-2V and GATA4 expression increases from day 8 through day 17. Interestingly, knockdown of HuR resulted in clumps of colonies with differentiated cells and a corresponding increase in cardiac-troponin positive cells. However, as a general observation, HuR knockdown reduced beating intensity compared to wild type cells. Conclusions: Based on these data, we could speculate that HuR might be necessary for maintenance of pluripotency and loss of which renders cells to differentiate in culture. HuR knockdown yields higher number of c-troponin positive cells but its effect on functional maturity of iCM needs to be further evaluated.

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GPR43 REGULATES SODIUM BUTYRATE-INDUCED ANGIOGENESIS AND MATRIX REMODELING

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00515.2019 ◽

2020 ◽

Author(s):

Pollyana Ribeiro Castro ◽

Lucas Felipe Fernandes Bittencourt ◽

Sébastien Larochelle ◽

Silvia Passos Andrade ◽

Charles Reay Mackay ◽

...

Keyword(s):

Sodium Butyrate ◽

Granulation Tissue ◽

Local Treatment ◽

Dermal Fibroblasts ◽

Matrix Remodeling ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Fibrovascular Tissue

Butyrate is a short-chain fatty acid (SCFA) derived from microbiota and is involved in a range of cell processes in a concentration-dependent manner. Low concentrations of sodium butyrate (NaBu) was shown to be proangiogenic. However, the mechanisms associated with these effects are not yet fully known. Here, we investigated the contribution of the SCFA receptor GPR43 in the proangiogenic effects of local treatment with NaBu and its effects on matrix remodeling using the sponge-induced fibrovascular tissue model in mice lacking the GPR43 gene (GPR43-KO) and the wild-type (WT). We demonstrated that NaBu (0.2 mM intraimplant) treatment enhanced the neovascularization process, blood flow, and VEGF levels in a GPR43-dependent manner in the implants. Moreover, NaBu was able to modulate matrix remodeling aspects of the granulation tissue such as proteoglycans production, collagen deposition and α-SMA expression in vivo, besides to increase TGF-b1 levels in the fibrovascular tissue, in a GPR43-dependent manner. Interestingly, NaBu directly stimulated L929 murine fibroblasts migration, and TGF-β1 and collagen production in vitro. GPR43 was found to be expressed in human dermal fibroblasts, myofibroblasts and endothelial cells. Overall, our findings evidence that the metabolite-sensing receptor GPR43 contributes to the effects of low dose of NaBu in inducing angiogenesis and matrix remodeling during granulation tissue formation. These data provide important insights for the proposition of new therapeutic approaches based on NaBu, beyond the highly explored intestinal, anti-inflammatory, and anti-cancer purposes, as a local treatment to improve tissue repair, particularly, by modulating granulation tissue components.

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Wound Healing Potential of Spirulina Protein on CCD-986sk Cells

Marine Drugs ◽

10.3390/md17020130 ◽

2019 ◽

Vol 17 (2) ◽

pp. 130

Author(s):

Ping Liu ◽

Jeong-Wook Choi ◽

Min-Kyeong Lee ◽

Youn-Hee Choi ◽

Taek-Jeong Nam

Keyword(s):

Wound Healing ◽

Dermal Fibroblast ◽

Fibroblast Cell ◽

Underlying Mechanism ◽

Human Dermal Fibroblast ◽

Dermal Fibroblasts ◽

Cyclin Dependent Kinase ◽

Pi3k Inhibitor Ly294002 ◽

And Migration ◽

Proliferation And Migration

Wound healing is a dynamic and complex process. The proliferation and migration of dermal fibroblasts are crucial for wound healing. Recent studies have indicated that the extracts from Spirulina platensis have a positive potential for wound healing. However, its underlying mechanism is not fully understood. Our previous study showed that spirulina crude protein (SPCP) promoted the viability of human dermal fibroblast cell line (CCD-986sk cells). In this study, we further investigated the wound healing effect and corresponding mechanisms of SPCP on CCD-986sk cells. Bromodeoxyuridine (BrdU) assay showed that SPCP promoted the proliferation of CCD-986sk cells. The wound healing assay showed that SPCP promoted the migration of CCD-986sk cells. Furthermore, cell cycle analysis demonstrated that SPCP promoted CCD-986sk cells to enter S and G2/M phases from G0/G1 phase. Western blot results showed that SPCP significantly upregulated the expression of cyclin D1, cyclin E, cyclin-dependent kinase 2 (Cdk2), cyclin-dependent kinase 4 (Cdk4), and cyclin-dependent kinase 6 (Cdk6), as well as inhibited the expression of CDK inhibitors p21 and p27 in CCD-986sk cells. In the meanwhile, SPCP promoted the phosphorylation and activation of phosphoinositide 3-kinase (PI3K) and protein kinase B (Akt). However, the phosphorylation of Akt was significantly blocked by PI3K inhibitor (LY294002), which in turn reduced the SPCP-induced proliferation and migration of CCD-986sk cells. Therefore, the results presenting in this study suggested that SPCP can promote the proliferation and migration of CCD-986sk cells; the PI3K/Akt signaling pathway play a positive and important role in these processes.

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Dermal Olfactory Receptor OR51B5 Is Essential for Survival and Collagen Synthesis in Human Dermal Fibroblast (Hs68 Cells)

International Journal of Molecular Sciences ◽

10.3390/ijms22179273 ◽

2021 ◽

Vol 22 (17) ◽

pp. 9273

Author(s):

Bomin Son ◽

Wesuk Kang ◽

Soyoon Park ◽

Dabin Choi ◽

Taesun Park

Keyword(s):

Cell Survival ◽

Collagen Synthesis ◽

Dermal Fibroblast ◽

Olfactory Receptors ◽

Camp Response Element Binding ◽

Human Dermal Fibroblast ◽

Skin Aging ◽

Dermal Fibroblasts ◽

Nasal Tissue ◽

Extracellular Matrix Components

Skin dermis comprises extracellular matrix components, mainly collagen fibers. A decrease in collagen synthesis caused by several factors, including ultraviolet (UV) irradiation and stress, eventually causes extrinsic skin aging. Olfactory receptors (ORs) were initially considered to be specifically expressed in nasal tissue, but several ORs have been reported to be present in other tissues, and their biological roles have recently received increasing attention. In this study, we aimed to characterize the role of ORs in cell survival and collagen synthesis in dermal fibroblasts. We confirmed that UVB irradiation and dexamethasone exposure significantly decreased cell survival and collagen synthesis in Hs68 dermal fibroblasts. Moreover, we demonstrated that the mRNA expression of 10 ORs detectable in Hs68 cells was significantly downregulated in aged conditions compared with that in normal conditions. Thereafter, by individual knockdown of the 10 candidate ORs, we identified that only OR51B5 knockdown leads to a reduction of cell survival and collagen synthesis. OR51B5 knockdown decreased cAMP levels and dampened the downstream protein kinase A/cAMP-response element binding protein pathway, downregulating the survival- and collagen synthesis-related genes in the dermal fibroblasts. Therefore, OR51B5 may be an interesting candidate that plays a role in cell survival and collagen synthesis.

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Enhanced antioxidant properties of sericin-cecropin fusion protein against oxidative stress in human adult dermal fibroblasts

Journal of Bioactive and Compatible Polymers ◽

10.1177/0883911520973238 ◽

2020 ◽

pp. 088391152097323

Author(s):

Dyna Susan Thomas ◽

Chitra Manoharan ◽

Sandhya Rasalkar ◽

Rakesh Kumar Mishra ◽

Ravikumar Gopalapillai

Keyword(s):

Oxidative Stress ◽

Fusion Protein ◽

Skin Diseases ◽

Dermal Fibroblast ◽

Antioxidant Properties ◽

Dermal Fibroblasts ◽

Protective Effects ◽

Uvb Radiation ◽

Silk Sericin ◽

Cecropin B

Chronic exposure to UVB radiation causes photoaging, immunosuppression, and ultimately photocarcinogenisis through the generation of reactive oxygen species (ROS). The ability of natural compounds in neutralizing the effects of oxidative stress is being explored with increased interest. Silk sericin, a biopolymer is reported to have diverse biological properties. In an effort to make the silk sericin pure, more effective and multifunctional, we have recombinantly expressed both functional sericin as well as sericin-cecropin B fusion proteins. Herein, we studied the antioxidant and anti-UVB potential of recombinant sericin and sericin-cecropin B proteins against oxidative stress using human primary dermal fibroblast cells. Treating the cells with recombinant sericin (RS) or sericin-cecropin B (RSC) prior to exposure to UVB and H2O2, effectively increased the cell viability by approximately 30% and 50%, respectively, in comparison to non-treated control. The protective effects were further evident in terms of significant reduction of LDH in oxidatively challenged cells treated with RS and RSC. A reduction in LDH release of at least 16 and 33% was observed with RS and RSC treatments, respectively, in comparison to exposed control. Further, elevated levels of catalase and superoxide dismutase (SOD) activity were observed. Importantly, the RSC fusion protein exhibited enhanced protective effects than cells treated with RS alone. Our results demonstrate that the functional attributes of cecropin B along with sericin activity in the fusion protein conferred enhanced protection against UVB- and H2O2-induced oxidative damage in human dermal fibroblasts. The improved antioxidant activity of recombinant sericin fusion biopolymer has great potential as a promising therapeutic agent for ROS-induced skin diseases.

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Anti-Melanogenesis, Antioxidant and Anti-Tyrosinase Activities of Scabiosa columbaria L.

Processes ◽

10.3390/pr8020236 ◽

2020 ◽

Vol 8 (2) ◽

pp. 236

Author(s):

Wilfred Otang-Mbeng ◽

Idowu Jonas Sagbo

Keyword(s):

Plant Extract ◽

Dermal Fibroblast ◽

Reducing Power ◽

Human Dermal Fibroblast ◽

Collagen Content ◽

Dependent Manner ◽

Tyrosinase Enzyme ◽

Fluorescence Imaging Microscopy ◽

Dose Dependent

Scabiosa columbaria is a plant traditionally used to treat skin ailments, such as scabies, wound bruises, sores and hyperpigmentation. To find a novel skin depigmenting agent, the present study was investigated to determine the possible anti-melanogenesis, antioxidant and anti-tyrosinase effects of methanol extract of S. columbaria leaves. Cytotoxicity towards human dermal fibroblast (MRHF) cells was assessed using the live-cell fluorescence imaging microscopy. The inhibitory effects of the extract on tyrosinase, collagenase and melanin synthesis were also investigated using standard in vitro method, while ferric reducing power (FRAP) was used to determine the antioxidant potential of the plant extract. The effect of the extract on collagen content in MRHF cells was also investigated. The plant extract displayed no meaningful cytotoxicity towards MRHF cells and no significant cell death was recorded at all the tested concentrations. The extract (25–100 µg/mL) effectively decreased melanin content in B16F10 (mouse melanoma) cells with moderate inhibition of tyrosinase enzyme in a dose-dependent manner. However, the extract also demonstrated no significant effect on collagenase and collagen content in MRHF cells, but showed strong antioxidant activity at the concentrations tested. The results suggest that S. columbaria could be a promising candidate in the treatment of skin hyperpigmentation disorders

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Attachment and Proliferation of Human Dermal Fibroblasts onto ECM-Immobilized PLGA Films

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.288-289.291 ◽

2005 ◽

Vol 288-289 ◽

pp. 291-294

Author(s):

Hyun Joo Son ◽

Dong Wook Han ◽

H.H. Kim ◽

Hee Joong Kim ◽

In Seop Lee ◽

...

Keyword(s):

Cell Attachment ◽

Dermal Fibroblast ◽

Supporting Cell ◽

Human Dermal Fibroblast ◽

Dermal Fibroblasts ◽

Water Soluble ◽

Seeding Density ◽

Tetrazolium Salt ◽

Cell Seeding ◽

Plga Film

In this study, human dermal fibroblast behaviors onto non-porous PLGA (75:25) films immobilized with 1, 10 and 100 µg/ml collagen (CN) or fibronectin (FN) were investigated according to different cell-seeding densities (1,000, 10,000 and 100,000 cells/ml). Cell attachment and proliferation were assessed using water soluble tetrazolium salt. The results indicated that 1 µg/ml of FN-immobilized PLGA film demonstrated significantly (p < 0.05) superior cellular attachment to the intact PLGA film after 4 hr of incubation. Moreover, the number of attached cells was shown to be directly proportional to that of initially seeded cells. After 48 hr, the cells showed significantly (p < 0.05) higher proliferation onto 1 or 10 µg/ml of FN-immobilized PLGA films than onto other PLGA films, regardless of the initial cell-seeding density. In terms of CN-immobilization, cell proliferation was appreciably increased but it was relatively lower than FN-immobilization. These results suggested that ECM-immobilization can enhance the cell affinity of hydrophobic scaffolds and be used to potential applications for tissue engineering by supporting cell growth.

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Dynamic transcriptome analysis reveals signatures of paradoxical effect of vemurafenib on human dermal fibroblasts

Cell Communication and Signaling ◽

10.1186/s12964-021-00801-3 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Eyleen Corrales ◽

Ella Levit-Zerdoun ◽

Patrick Metzger ◽

Silke Kowar ◽

Manching Ku ◽

...

Keyword(s):

Cancer Progression ◽

Dermal Fibroblast ◽

Acquired Resistance ◽

Braf Inhibitor ◽

Mek Inhibitor ◽

Human Dermal Fibroblast ◽

Dermal Fibroblasts ◽

Paradoxical Effect ◽

Combination Therapies ◽

Stromal Component

Abstract Background Vemurafenib (PLX4032) is one of the most frequently used treatments for late-stage melanoma patients with the BRAFV600E mutation; however, acquired resistance to the drug poses as a major challenge. It remains to be determined whether off-target effects of vemurafenib on normal stroma components could reshape the tumor microenvironment in a way that contributes to cancer progression and drug resistance. Methods By using temporally-resolved RNA- and ATAC-seq, we studied the early molecular changes induced by vemurafenib in human dermal fibroblast (HDF), a main stromal component in melanoma and other tumors with high prevalence of BRAFV600 mutations. Results Transcriptomics analyses revealed a stepwise up-regulation of proliferation signatures, together with a down-regulation of autophagy and proteolytic processes. The gene expression changes in HDF strongly correlated in an inverse way with those in BRAFV600E mutant malignant melanoma (MaMel) cell lines, consistent with the observation of a paradoxical effect of vemurafenib, leading to hyperphosphorylation of MEK1/2 and ERK1/2. The transcriptional changes in HDF were not strongly determined by alterations in chromatin accessibility; rather, an already permissive chromatin landscape seemed to facilitate the early accessibility to MAPK/ERK-regulated transcription factor binding sites. Combinatorial treatment with the MEK inhibitor trametinib did not preclude the paradoxical activation of MAPK/ERK signaling in HDF. When administered together, vemurafenib partially compensated for the reduction of cell viability and proliferation induced by trametinib. These paradoxical changes were restrained by using the third generation BRAF inhibitor PLX8394, a so-called paradox breaker compound. However, the advantageous effects on HDF during combination therapies were also lost. Conclusions Vemurafenib induces paradoxical changes in HDF, enabled by a permissive chromatin landscape. These changes might provide an advantage during combination therapies, by compensating for the toxicity induced in stromal cells by less specific MAPK/ERK inhibitors. Our results highlight the relevance of evaluating the effects of the drugs on non-transformed stromal components, carefully considering the implications of their administration either as mono- or combination therapies.

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