scholarly journals Expression of pro-inflammatory markers by human dermal fibroblasts in a three-dimensional culture model is mediated by an autocrine interleukin-1 loop

2004 ◽  
Vol 379 (2) ◽  
pp. 351-358 ◽  
Author(s):  
Daniela KESSLER-BECKER ◽  
Thomas KRIEG ◽  
Beate ECKES
Keyword(s):  
Keratinocyte Growth Factor ◽  
Interleukin 1 ◽  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Inflammatory Processes ◽  
Cox 2 ◽  
Extracellular Matrix Interactions

In vivo, fibroblasts reside in connective tissues, with which they communicate in a reciprocal way. Such cell–extracellular matrix interactions can be studied in vitro by seeding fibroblasts in collagen lattices. Depending upon the mechanical properties of the system, fibroblasts are activated to assume defined phenotypes. In the present study, we examined a transcriptional profile of primary human dermal fibroblasts cultured in a relaxed collagen environment and found relative induction (>2-fold) of 393 out of approx. 7100 transcripts when compared with the same system under mechanical tension. Despite down-regulated proliferation and matrix synthesis, cells did not become generally quiescent, since they induced transcription of numerous other genes including matrix metalloproteinases (MMPs) and growth factors/cytokines. Of particular interest was the induction of gene transcripts encoding pro-inflammatory mediators, e.g. cyclo-oxygenase-2 (COX-2), and interleukins (ILs)-1 and -6. These are apparently regulated in a hierarchical fashion, since the addition of IL-1 receptor antagonist prevented induction of COX-2, IL-1 and IL-6, but not that of MMP-1 or keratinocyte growth factor (KGF). Our results suggest strongly that skin fibroblasts are versatile cells, which adapt to their extracellular environment by displaying specific phenotypes. One such phenotype, induced by a mechanically relaxed collagen environment, is the ‘pro-inflammatory’ fibroblast. We propose that fibroblasts that are embedded in a matrix environment can actively participate in the regulation of inflammatory processes.

scholarly journals Neuroprotective Effect of Optimized Yinxieling Formula in 6-OHDA-Induced Chronic Model of Parkinson’s Disease through the Inflammation Pathway

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Renrong Wei ◽  
Cuiping Rong ◽  
Qingfeng Xie ◽  
Shouhai Wu ◽  
Yuchao Feng ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Calcium Binding ◽  
Dopaminergic Neurons ◽  
Neuroprotective Effect ◽  
Interleukin 1 ◽  
Mrna Levels ◽  
Cox 2

Parkinson’s disease (PD) is characterized by progressive degeneration of dopaminergic neurons in the substantia nigra (SN)-striatum circuit, which is associated with glial activation and consequent chronic neuroinflammation. Optimized Yinxieling Formula (OYF) is a Chinese medicine that exerts therapeutical effect and antiinflammation property on psoriasis. Our previous study has proven that pretreatment with OYF could regulate glia-mediated inflammation in an acute mouse model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Given that PD is a chronic degeneration disorder, this study applied another PD animal model induced by striatal injection of 6-hydroxydopamine (6-OHDA) to mimic the progressive damage of the SN-striatum dopamine system in rats. The OYF was administrated in the manner of pretreatment plus treatment. The effects of the OYF on motor behaviors were assessed with the apomorphine-induced rotation test and adjusting steps test. To confirm the effect of OYF on dopaminergic neurons and glia activation in this model, we analyzed the expression of tyrosine hydroxylase (TH) and glia markers, ionized calcium-binding adapter molecule 1 (Iba-1), and glial fibrillary acidic protein (GFAP) in the SN region of the rat PD model. Inflammation-associated factors, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), were further evaluated in this model and in interferon-γ- (INF-γ-) induced murine macrophages RAW264.7 cells. The results from the in vivo study showed that OYF reversed the motor behavioral dysfunction in 6-OHDA-induced PD rats, upregulated the TH expression, decreased the immunoreactivity of Iba-1 and GFAP, and downregulated the mRNA levels of TNF-α and COX-2. The OYF also trended to decrease the mRNA levels of IL-1β and iNOS in vivo. The results from the in vitro study showed that OYF significantly decreased the mRNA levels of TNF-α, IL-1β, IL-6, iNOS, and COX-2. Therefore, this study suggests that OYF exerts antiinflammatory effects, which might be related to the protection of dopaminergic neurons in 6-OHDA-induced chronic neurotoxicity.

scholarly journals Validation of in vitro assays in three-dimensional human dermal constructs

2018 ◽  
Vol 41 (11) ◽  
pp. 779-788 ◽  
Author(s):  
Ayesha Idrees ◽  
Valeria Chiono ◽  
Gianluca Ciardelli ◽  
Siegfried Shah ◽  
Richard Viebahn ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Dermal Fibroblasts ◽  
In Vitro Assays ◽  
Dimensional System ◽  
Type I ◽  
Human Dermal Fibroblasts ◽  
Cell Viability Assay ◽  
Normal Human ◽  
Normal Human Dermal Fibroblasts

Three-dimensional cell culture systems are urgently needed for cytocompatibility testing of biomaterials. This work aimed at the development of three-dimensional in vitro dermal skin models and their optimization for cytocompatibility evaluation. Initially “murine in vitro dermal construct” based on L929 cells was generated, leading to the development of “human in vitro dermal construct” consisting of normal human dermal fibroblasts in rat tail tendon collagen type I. To assess the viability of the cells, different assays CellTiter-Blue®, RealTime-Glo™ MT, and CellTiter-Glo® (Promega) were evaluated to optimize the best-suited assay to the respective cell type and three-dimensional system. Z-stack imaging (Live/Dead and Phalloidin/DAPI-Promokine) was performed to visualize normal human dermal fibroblasts inside matrix revealing filopodia-like morphology and a uniform distribution of normal human dermal fibroblasts in matrix. CellTiter-Glo was found to be the optimal cell viability assay among those analyzed. CellTiter-Blue reagent affected the cell morphology of normal human dermal fibroblasts (unlike L929), suggesting an interference with cell biological activity, resulting in less reliable viability data. On the other hand, RealTime-Glo provided a linear signal only with a very low cell density, which made this assay unsuitable for this system. CellTiter-Glo adapted to three-dimensional dermal construct by optimizing the “shaking time” to enhance the reagent penetration and maximum adenosine triphosphate release, indicating 2.4 times higher viability value by shaking for 60 min than for 5 min. In addition, viability results showed that cells were viable inside the matrix. This model would be further advanced with more layers of skin to make a full thickness model.

scholarly journals The chemotactic attraction of human fibroblasts to a lymphocyte-derived factor.

1976 ◽  
Vol 144 (5) ◽  
pp. 1188-1203 ◽  
Author(s):  
A E Postlethwaite ◽  
R Snyderman ◽  
A H Kang
Keyword(s):  
Connective Tissue ◽  
Peripheral Blood ◽  
Human Fibroblasts ◽  
Peripheral Blood Lymphocytes ◽  
Dermal Fibroblasts ◽  
Chemotactic Factor ◽  
Human Dermal Fibroblasts ◽  
Heat Stable

A quantitative assay that measures fibroblast chemotaxis in vitro is described. Application of this technique has revealed that peripheral blood lymphocytes stimulated by antigen or mitogen in vitro produce a factor that is chemotactic for human dermal fibroblasts. This lymphocyte-derived chemotactic factor for fibroblasts (LDCF-F) is different from the lymphokine that is chemotactic for monocytes or macrophages. Macrophages are required for the generation of LDCF-F by T lymphocytes stimulated by phytohemagglutinin. The fibroblast chemotactic factor is heat stable (56 degrees C for 30 min), trypsin sensitive, and neuraminidase resistant. LDCF-F could function to attact connective tissue fibroblasts to sites at which cell-mediated immune reactions are occurring in vivo.

scholarly journals Induction of fibroblast chemotaxis by fibronectin. Localization of the chemotactic region to a 140,000-molecular weight non-gelatin-binding fragment.

1981 ◽  
Vol 153 (2) ◽  
pp. 494-499 ◽  
Author(s):  
A E Postlethwaite ◽  
J Keski-Oja ◽  
G Balian ◽  
A H Kang
Keyword(s):  
Molecular Weight ◽  
Tissue Injury ◽  
Dermal Fibroblasts ◽  
Human Dermal Fibroblasts ◽  
Human Monocytes ◽  
Directional Migration

Plasma and cell-derived fibronectin are potent chemoattractants for human dermal fibroblasts in vitro. The chemotactic property of fibronectin resides in a major 140,000-mol wt non-gelatin-binding fragment of the native molecule. Human monocytes and neutrophils do not recognize fibronectin as a chemotactic stimulus. These findings suggest that fibronectin and perhaps certain fragments of fibronectin may function in vivo as a specific chemoattractant for fibroblasts and could, therefore, induce directional migration of fibroblasts to sites of tissue injury, remodeling or morphogenesis.

In vivo E-selectin upregulation correlates early with infiltration of PMN, later with PBL entry: MAbs block both

1996 ◽  
Vol 270 (1) ◽  
pp. H183-H193 ◽  
Author(s):  
R. M. Binns ◽  
S. T. Licence ◽  
A. A. Harrison ◽  
E. T. Keelan ◽  
M. K. Robinson ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Interleukin 1 ◽  
Inflammatory Infiltration ◽  
Necrosis Factor Alpha ◽  
Inflammatory Processes ◽  
Factor Alpha ◽  
Kinetics Of ◽  
Necrosis Factor

The endothelial molecule E-selectin binds most leukocyte subsets in vitro. Yet its role in regulating the very different kinetics of inflammatory infiltration of different leukocyte subsets in vivo is unclear. The kinetics of E-selectin upregulation and polymorphonuclear leukocyte (PMN) and blood lymphocyte (PBL) localization in inflammation induced by interleukin-1 alpha (IL-1 alpha), tumor necrosis factor-alpha (TNF-alpha), phytohemagglutinin (PHA), and phorbol myristate acetate (PMA) were investigated in a well-established inbred pig trafficking model. They differed markedly both for these three labeled indicators of inflammation and in each of the four inflammatory processes. In each, E-selectin upregulation correlated with early PMN entry and later with PBL infiltration but was more protracted than both. The importance of E-selectin was confirmed by marked inhibition of PMN and PBL entry (up to > 60%) by F(ab')2 anti-E-selectin. Involvement of other molecules was illustrated by similar or greater inhibition with anti-CD18 F(ab')2. We conclude that, like CD18, E-selectin is necessary for most PMN and PBL infiltration but alone is insufficient, consistent with the involvement of several alternative multistep molecular mechanisms in this entry.

scholarly journals Synthesis, Characterization of Novel N'(2-Phenoxyacetyl)Nicotinohydrazide and N'(2-Phenoxyacetyl)Isonicotinohydrazide Derivatives as Anti-Inflammatory and Analgesic Agents

2021 ◽  
Author(s):  
H. M. Pallavi ◽  
Fares Hezam Al-Ostoot ◽  
Hamse Kameshwar Vivek ◽  
Shaukath Ara Khanum
Keyword(s):  
Acetic Acid ◽  
Three Dimensional ◽  
Thermal Pain ◽  
Elemental Analyses ◽  
Analgesic Agents ◽  
Ft Ir ◽  
Cox 2 ◽  
Anti Inflammatory

Abstract A new derivatives of isonicotinohydrazide 9 (a-e) and 10(a-e) were designed, synthesized, characterized based on the different spectroscopic analysis FT-IR, -1H, -13C-NMR, mass spectra, and also elemental-analyses, and identified as a remarkable anti-inflammatory along with analgesic effect. In-vitro and in-vivo anti-inflammatory and analgesic actions were performed using analgesic acetic acid-induced squirming and Hot plate latency tests using male Swiss albino mice. The study was supported by the in-silico and ADMET approaches. Among the series, compound (10e) showed the highest IC50 value for COX-1 inhibition, whereas compounds (9e) and (10e) exhibited the highest COX-2 SI. Further, in silico studies provide a putative bind site for the potent compound in a three-dimensional geometrical view. The results revealed that the title compound (10e) with bromo group exhibited potent COX-2 inhibitor at different times which is on par with the normal drug used in the assay system. Substitution of halogens at N’(2-phenoxyacetyl)nicotinohyrazide showed (10e) potent analgesic outcome on acetic acid-induced squirming response and thermal pain among the series 9(a-e) and 10(a-e). The overall pharmacological result suggests that the newly synthesized compounds possess good anti-inflammatory along with analgesic-activity which was evaluated through in-vitro, in-vivo, and --silico studies.

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