scholarly journals Hypopigmented burn hypertrophic scar contains melanocytes that can be signaled to re-pigment by synthetic alpha-melanocyte stimulating hormone in vitro

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0248985
Author(s):  
Bonnie C. Carney ◽  
Taryn E. Travis ◽  
Lauren T. Moffatt ◽  
Laura S. Johnson ◽  
Melissa M. McLawhorn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Animal Model ◽  
Hypertrophic Scar ◽  
Hypertrophic Scars ◽  
Synthetic Genes ◽  
Melanocyte Stimulating Hormone ◽  
En Face ◽  
Initial Work ◽  
Skin Biopsies

There are limited treatments for dyschromia in burn hypertrophic scars (HTSs). Initial work in Duroc pig models showed that regions of scar that are light or dark have equal numbers of melanocytes. This study aims to confirm melanocyte presence in regions of hypo- and hyper-pigmentation in an animal model and patient samples. In a Duroc pig model, melanocyte presence was confirmed usingen facestaining. Patients with dyschromic HTSs had demographic, injury details, and melanin indices collected. Punch biopsies were taken of regions of hyper-, hypo-, or normally pigmented scar and skin. Biopsies were processed to obtain epidermal sheets (ESs). A subset of ESs wereen facestained with melanocyte marker, S100β. Melanocytes were isolated from a different subset. Melanocytes were treated with NDP α-MSH, a pigmentation stimulator. mRNA was isolated from cells, and was used to evaluate gene expression of melanin-synthetic genes. In patient and pig scars, regions of hyper-, hypo-, and normal pigmentation had significantly different melanin indices. S100βen facestaining showed that regions of hyper- and hypo-pigmentation contained the same number of melanocytes, but these cells had different dendricity/activity. Treatment of hypo-pigmented melanocytes with NDP α-MSH produced melanin by microscopy. Melanin-synthetic genes were upregulated in treated cells over controls. While traditionally it may be thought that hypopigmented regions of burn HTS display this phenotype because of the absence of pigment-producing cells, these data show that inactive melanocytes are present in these scar regions. By treating with a pigment stimulator, cells can be induced to re-pigment.

55 Melanocytes in Hypopigmented Burn Scar Can Be Stimulated to Produce Melanin

2020 ◽  
Vol 41 (Supplement_1) ◽  
pp. S36-S37
Author(s):  
Bonnie C Carney ◽  
Taryn E Travis ◽  
Lauren T Moffatt ◽  
Laura S Johnson ◽  
Melissa M McLawhorn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skin Color ◽  
Normal Skin ◽  
Burn Scar ◽  
Synthetic Genes ◽  
Dopachrome Tautomerase ◽  
En Face ◽  
Pilot Work ◽  
Bright Field Microscopy ◽  
Skin Biopsies

Abstract Introduction Currently, there are limited clinical treatments for dyschromia in burn hypertrophic scars (HTSs). Initial pilot work in a Duroc pig model showed that melanocyte quantity does not influence pigmentation. Regions of scar that are light or dark have equal numbers of melanocytes. This study aims to confirm melanocyte presence in regions of hypo- and hyper-pigmentation. If melanocytes are present, pigmentation stimulators may be a useful therapy. Methods Following IRB approval, patients with dyschromic HTSs were enrolled and demographic, injury and treatment details, and melanin indices by Skin Color Catch probes (SCC) were collected. Punch biopsies were taken of distinct regions of hyper-, hypo-, or normally pigmented scar and skin. Biopsies were processed to obtain epidermal sheets (ESs). A subset of ESs were stained using en face staining with melanocyte marker, S100b. Melanocytes were isolated from a different subset. Melanocytes were treated with NDP a-MSH, a pigmentation stimulator. mRNA was isolated from cells, and qRT-PCR was used to evaluate gene expression of melanin-synthetic genes, tyrosinase (TYR), tyrosinase-related protein-1 (TYRP1) and dopachrome tautomerase (DCT). Results 8 patients were enrolled and the cohort included patients with Fitzpatrick skin types 5 (n=6) and 3 (n=2). HTSs ranged from 4 to 395 months old and dyschromia developed mainly due to healing by re-epithelialization and was refractory to prior scar treatments. By SCC, regions of hyper-, hypo-, and normal pigmentation had significantly different melanin indices (864.9 ± 19.8 vs. 627.1 ± 30.3 vs. 786.5 ± 21.4, p< 0.01). S100b en face staining showed that regions of hyper- and hypo-pigmentation contained the same number of melanocytes (14.6 ± 3.1 vs. 14.5 ± 3.2 melanocytes/HPF (n=5, p=0.99), but these cells had different dendricity, or activity (3.9 ± 0.5 vs. 0.4 ± 0.2 dendrites/cell (n=5, p< 0.0001). When hypo-pigmented melanocytes were treated with NDP a-MSH, cells produced melanin visible by bright-field microscopy (n=3). TYR, TYRP1, and DCT gene expression were upregulated in treated cells compared to controls. Conclusions While traditionally, it may be thought that hypo-pigmented regions of burn HTS display this phenotype because of the absence of pigment-producing cells, these data show that melanocytes are present in these scar regions. The cells are present, but they are inactive in pigment production. By treating the cells with a pigment stimulator, cells can be induced to re-pigment. Applicability of Research to Practice Patients were asked, on a scale of 1–10 with 1 being “no, not at all”, and 10 being “yes, very much”, “Is the scar color different from the color of your normal skin at present?” All patients rated this question as 10/10, indicating that scar dyschromia is a pervasive problem. Patients’ quality of life may be improved with the development of treatments for hypopigmentation.

scholarly journals The serine proteases dipeptidyl-peptidase 4 and urokinase are key molecules in human and mouse scar formation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Vera Vorstandlechner ◽  
Maria Laggner ◽  
Dragan Copic ◽  
Katharina Klas ◽  
Martin Direder ◽  
...  
Keyword(s):  
Gene Expression ◽  
Serine Proteases ◽  
Hypertrophic Scar ◽  
Normal Skin ◽  
Dipeptidyl Peptidase ◽  
Scar Formation ◽  
Hypertrophic Scars ◽  
Dipeptidyl Peptidase 4

AbstractDespite recent advances in understanding skin scarring, mechanisms triggering hypertrophic scar formation are still poorly understood. In the present study, we investigate mature human hypertrophic scars and developing scars in mice at single cell resolution. Compared to normal skin, we find significant differences in gene expression in most cell types present in scar tissue. Fibroblasts show the most prominent alterations in gene expression, displaying a distinct fibrotic signature. By comparing genes upregulated in murine fibroblasts during scar development with genes highly expressed in mature human hypertrophic scars, we identify a group of serine proteases, tentatively involved in scar formation. Two of them, dipeptidyl-peptidase 4 (DPP4) and urokinase (PLAU), are further analyzed in functional assays, revealing a role in TGFβ1-mediated myofibroblast differentiation and over-production of components of the extracellular matrix in vitro. Topical treatment with inhibitors of DPP4 and PLAU during scar formation in vivo shows anti-fibrotic activity and improvement of scar quality, most prominently after application of the PLAU inhibitor BC-11. In this study, we delineate the genetic landscape of hypertrophic scars and present insights into mechanisms involved in hypertrophic scar formation. Our data suggest the use of serine protease inhibitors for the treatment of skin fibrosis.

Effect of Auraptene on angiogenesis in Xenograft model of breast cancer

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mohammad Reza Shiran ◽  
Elham Mahmoudian ◽  
Abolghasem Ajami ◽  
Seyed Mostafa Hosseini ◽  
Ayjamal Khojasteh ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Animal Model ◽  
Protein Expression ◽  
Western Blot ◽  
Xenograft Model ◽  
Dependent Manner ◽  
Concentration Dependent Manner

Abstract Objectives Angiogenesis is the most important challenge in breast cancer treatment. Recently, scientists become interesting in rare natural products and intensive researches was performed to identify their pharmacological profile. Auraptene shows helpful effects such as cancer chemo-preventive, anti-inflammatory, anti-oxidant, immuno-modulatory. In this regard, we investigated the anti-angiogenesis effect of Auraptene in in-vitro and in-vivo model of breast cancer. Methods In this study, 4T, MDA-MB-231 and HUVEC cell lines were used. The proliferation study was done by MTT assay. For tube formation assay, 250 matrigel, 1 × 104 HUVEC treated with Auraptene, 20 ng/mL EGF, 20 ng/mL bFGF and 20 ng/mL VEGF were used. Gene expression of important gene related to angiogenesis in animal model of breast cancer was investigated by Real-time PCR. Protein expression of VCAM-1 and TNFR-1 gene related to angiogenesis in animal model of breast cancer was investigated by western-blot. Results Auraptene treatment led to reduction in cell viability of MDA-MB-231 in a concentration-dependent manner. Also, we observed change in the number of tubes or branches formed by cells incubated with 40 and 80 μM Auraptene. Auraptene effect the gene expression of important gene related to angiogenesis (VEGF, VEGFR2, COX2, IFNɣ). Moreover, the western blot data exhibited that Auraptene effect the protein expression of VCAM-1 and TNFR-1. Conclusions Overall, this study shows that Auraptene significantly suppressed angiogenesis via down-regulation of VEGF, VEGFR2, VCAM-1, TNFR-1, COX-2 and up-regulation of IFNγ.

scholarly journals Condition Medium of Glioblastoma Cell Lines Decreases the Viability of Glioblastoma Cells by Modulating Gene Expression Profile

2021 ◽  
Author(s):  
Mervenur Yavuz ◽  
Siddika Akgul ◽  
Egemen Kaya ◽  
Turan Demircan
Keyword(s):  
Gene Expression ◽  
Conditioned Medium ◽  
Cell Lines ◽  
Animal Experiments ◽  
Gene Expression Pattern ◽  
Incurable Disease ◽  
Initial Work ◽  
Migration Capacity ◽  
U87 Cells

Grade IV neoplasm of the central nervous system, GBM, is associated with poor prognosis and relatively short overall survival. Due to the current limitations in treatment methods, GBM is characterized as an incurable disease, and research to advance therapeutic options is required. Conditioned medium is commonly used in in-vitro studies complementary to animal experiments to simulate tumor microenvironment and has the potential to challenge and expand our current understanding of secretome effect on tumor characteristics. This study aimed to investigate the effects of conditioned mediums of GBM cell lines on each other. Conditioned mediums' cellular and molecular effects were evaluated using commonly employed techniques such as MTT assay, colony formation assay, wound healing assay, EdU labeling-based flow cytometry, and qRT-PCR. Our study demonstrated that conditioned medium harvested from U87 or LN229 cells at 48th h exhibited an anti-growth activity on each other by changing the gene expression pattern. Furthermore, the conditioned medium of LN229 decreased the migration capacity of U87 cells, and the conditioned medium of U87 cells significantly suppressed the LN229 proliferation. We believe that this initial work provides new insights for a better understanding of GBM cell lines' secretome roles and highlights the necessity of further studies to unveil the secretome content.

Inhibitory Effect of Tetrandrine on the Proliferation of Human Dermal Fibroblasts Derived from Hypertrophic Scars

2011 ◽  
Vol 268-270 ◽  
pp. 838-840
Author(s):  
De Wu Liu ◽  
Xiang Hu ◽  
De Ming Liu ◽  
Ping Zou
Keyword(s):  
Hypertrophic Scar ◽  
Culture Medium ◽  
Inhibitory Effect ◽  
Dermal Fibroblasts ◽  
Dependent Manner ◽  
Human Dermal Fibroblasts ◽  
Hypertrophic Scars ◽  
Result Show ◽  
Dose Dependent

Tetrandrine can inhibit the proliferation and collagen synthesis of fibroblasts in lung and liver tissue confirmed by a series of clinical research. In this chapter, we investigated the effect of Tetrandrine on the proliferation of human dermal fibroblasts derived from hypertrophic scars. The dermal fibroblasts were isolated from human hypertrophic scar tissues and cultured in vitro. Tetrandrine with different concentration were added to culture medium respectively. The proliferative activities were determined. The result show that when the concentration of added Tetrandrine increased from 5μg/ml to 80μg/ml, the proliferative activities of cultured dermal fibroblasts were decreased gradually in dose-dependent manner. It conclusions that Tetrandrine can obviously inhibit the proliferation of human dermal fibroblasts derived from hypertrophic scars.

scholarly journals Single cell landscape of hypertrophic scars identifies serine proteases as key regulators of myofibroblast differentiation

2020 ◽  
Author(s):  
Vera Vorstandlechner ◽  
Maria Laggner ◽  
Dragan Copic ◽  
Yiyan Chen ◽  
Bahar Golabi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Serine Proteases ◽  
Hypertrophic Scar ◽  
Normal Skin ◽  
Cell Types ◽  
Scar Formation ◽  
Myofibroblast Differentiation ◽  
Hypertrophic Scars ◽  
Dipeptidyl Peptidase 4

AbstractDespite recent advances in understanding skin scarring, mechanisms triggering hypertrophic scar formation are still poorly understood. In the present study we performed single-cell sequencing of mature human hypertrophic scars and developing scars in mice.Compared to normal skin, we found significant differences in gene expression in most cell types present in scar tissue. Fibroblasts (FBs) showed the most prominent alterations in gene expression, displaying a distinct fibrotic signature. By comparing genes upregulated in murine FBs during scar development with genes highly expressed in mature human hypertrophic scars, we identified a group of serine proteases, tentatively involved in scar formation. Two of them, dipeptidyl-peptidase 4 (DPP4) and urokinase (PLAU), were further analyzed in functional assays, revealing a role in TGFβ1-mediated myofibroblast differentiation and over-production of components of the extracellular matrix (ECM) without interfering with the canonical TGFβ1-signaling pathway.In this study, we delineate the genetic landscape of hypertrophic scars and present new insights into mechanisms involved in hypertrophic scar formation. Our data suggest the use of serine protease inhibitors for the treatment of skin fibrosis.

scholarly journals Overexpression of ubiquitin-specific peptidase 15 in systemic sclerosis fibroblasts increases response to transforming growth factor β

Rheumatology ◽  
2019 ◽  
Vol 58 (4) ◽  
pp. 708-718
Author(s):  
Christine Galant ◽  
Joel Marchandise ◽  
Maria S Stoenoiu ◽  
Julie Ducreux ◽  
Aurélie De Groof ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Transforming Growth Factor ◽  
Cell Metabolism ◽  
Fibroblast Cell ◽  
Transforming Growth Factor Β ◽  
Primary Fibroblast ◽  
Skin Biopsies ◽  
Fibroblast Cell Lines

Abstract Objective Ubiquitination of proteins leads to their degradation by the proteasome, and is regulated by ubiquitin ligases and substrate-specific ubiquitin-specific peptidases (USPs). The ubiquitination process also plays important roles in the regulation of cell metabolism and cell cycle. Here, we found that the expression of several USPs is increased in SSc tenosynovial and skin biopsies, and we demonstrated that USP inhibition decreases TGF-β signalling in primary fibroblast cell lines. Methods High-density transcriptomic studies were performed using total RNA obtained from SSc tenosynovial samples. Confirmatory immunostaining experiments were performed on tenosynovial and skin samples. In vitro experiments were conducted in order to study the influence of USP modulation on responses to TGF-β stimulation. Results Tenosynovial biopsies from SSc patients overexpressed known disease-associated gene pathways: fibrosis, cytokines and chemokines, and Wnt/TGF-β signalling, but also several USPs. Immunohistochemistry experiments confirmed the detection of USPs in the same samples, and in SSc skin biopsies. Exposure of primary fibroblast cell lines to TGF-β induced USP gene expression. The use of a pan-USP inhibitor decreased SMAD3 phosphorylation, and expression of COL1A1, COL3A1 and fibronectin gene expression in TGF-β-stimulated fibroblasts. The effect of the USP inhibitor resulted in increased SMAD3 ubiquitination, and was blocked by a proteasome inhibitor, thereby confirming the specificity of its action. Conclusion Overexpression of several USPs, including USP15, amplifies fibrotic responses induced by TGF-β, and is a potential therapeutic target in SSc.

scholarly journals Single cell sequencing identifies serine proteases as regulators of myofibroblast differentiation

2020 ◽  
Author(s):  
Vera Vorstandlechner ◽  
Maria Laggner ◽  
Dragan Copic ◽  
Yiyan Chen ◽  
Bahar Golabi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Serine Proteases ◽  
Hypertrophic Scar ◽  
Normal Skin ◽  
Scar Formation ◽  
Myofibroblast Differentiation ◽  
Hypertrophic Scars ◽  
Dipeptidyl Peptidase 4 ◽  
Single Cell Sequencing

Abstract Despite recent advances in understanding skin scarring, mechanisms triggering hypertrophic scar formation are still poorly understood. In the present study we performed single-cell sequencing of mature human hypertrophic scars and developing scars in mice. Compared to normal skin, we found significant differences in gene expression in most cell types present in scar tissue. Fibroblasts (FBs) showed the most prominent alterations in gene expression, displaying a distinct fibrotic signature. By comparing genes upregulated in murine FBs during scar development with genes highly expressed in mature human hypertrophic scars, we identified a group of serine proteases, tentatively involved in scar formation. Two of them, dipeptidyl-peptidase 4 (DPP4) and urokinase (PLAU), were further analyzed in functional assays, revealing a role in TGFβ1-mediated myofibroblast differentiation and over-production of components of the extracellular matrix (ECM) without interfering with the canonical TGFbeta1-signaling pathway. In this study, we delineate the genetic landscape of hypertrophic scars and present new insights into mechanisms involved in hypertrophic scar formation. Our data suggest the use of serine protease inhibitors for the treatment of skin fibrosis.

Fibroblasts During Hypertrophic Scar Formation and Resolution

1973 ◽  
Vol 31 ◽  
pp. 428-429
Author(s):  
C. W. Kischer
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Proliferation ◽  
Fine Structure ◽  
Metabolic Activity ◽  
Hypertrophic Scar ◽  
Normal Skin ◽  
Ground Substance ◽  
Hypertrophic Scars ◽  
Scanning Electron

The morphology of the fibroblasts changes markedly as the healing period from burn wounds progresses, through development of the hypertrophic scar, to resolution of the scar by a self-limiting process of maturation or therapeutic resolution. In addition, hypertrophic scars contain an increased cell proliferation largely made up of fibroblasts. This tremendous population of fibroblasts seems congruous with the abundance of collagen and ground substance. The fine structure of these cells should reflect some aspects of the metabolic activity necessary for production of the scar, and might presage the stage of maturation.A comparison of the fine structure of the fibroblasts from normal skin, different scar types, and granulation tissue has been made by transmission (TEM) and scanning electron microscopy (SEM).

Neurofilaments and Paired Helical Filaments

1985 ◽  
Vol 43 ◽  
pp. 740-743
Author(s):  
J. Metuzals
Keyword(s):  
Animal Model ◽  
Posttranslational Modifications ◽  
Posttranslational Modification ◽  
Giant Axon ◽  
Paired Helical Filaments ◽  
Squid Giant Axon ◽  
In Vitro Experiments ◽  
Thin Sections ◽  
Structural Relationships

It has been demonstrated that the neurofibrillary tangles in biopsies of Alzheimer patients, composed of typical paired helical filaments (PHF), consist also of typical neurofilaments (NF) and 15nm wide filaments. Close structural relationships, and even continuity between NF and PHF, have been observed. In this paper, such relationships are investigated from the standpoint that the PHF are formed through posttranslational modifications of NF. To investigate the validity of the posttranslational modification hypothesis of PHF formation, we have identified in thin sections from frontal lobe biopsies of Alzheimer patients all existing conformations of NF and PHF and ordered these conformations in a hypothetical sequence. However, only experiments with animal model preparations will prove or disprove the validity of the interpretations of static structural observations made on patients. For this purpose, the results of in vitro experiments with the squid giant axon preparations are compared with those obtained from human patients. This approach is essential in discovering etiological factors of Alzheimer's disease and its early diagnosis.

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