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Phenotypic responses to mechanical stress in fibroblasts from tendon, cornea and skin.

ABSTRACT: Primary fibroblasts isolated from foetal mouse cornea, skin and tendon were subjected to linear shear stress and analysed for morphological parameters and by microarray, as compared with unstimulated controls. Approx. 350 genes were either up- or down-regulated by a significant amount, with 51 of these being common to all three cell types. Approx. 50% of altered genes in tendon and cornea fibroblasts were changed in common with one of the other cell types, with the remaining approx. 50% being specific to tendon or cornea. In skin fibroblasts, however, less than 25% of genes whose transcription was altered were specific only to skin. The functional spectrum of genes that were up- or down-regulated was diverse, with apparent house-keeping genes forming the major category of up-regulated genes. However, a significant number of genes associated with cell adhesion, extracellular matrix and matrix remodelling, as well as cytokines and other signalling factors, were also affected. Somewhat surprisingly, in these latter categories the trend was towards a reduction in mRNA levels. Verification of the mRNA quantity of a subset of these genes was performed by reverse transcriptase PCR and was found to be in agreement with the microarray analysis. These findings provide the first in-depth analysis of phenotypic differences between fibroblast cells from different tissue sources and reveal the responses of these cells to mechanical stress.

SUBMITTER: Mackley JR

PROVIDER: S-EPMC1462727 | biostudies-literature | 2006 Jun

REPOSITORIES: biostudies-literature

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Phenotypic responses to mechanical stress in fibroblasts from tendon, cornea and skin.

Mackley Jennifer R JR Ando Joji J Herzyk Pawel P Winder Steven J SJ

The Biochemical journal 20060601 2

Primary fibroblasts isolated from foetal mouse cornea, skin and tendon were subjected to linear shear stress and analysed for morphological parameters and by microarray, as compared with unstimulated controls. Approx. 350 genes were either up- or down-regulated by a significant amount, with 51 of these being common to all three cell types. Approx. 50% of altered genes in tendon and cornea fibroblasts were changed in common with one of the other cell types, with the remaining approx. 50% being sp ...[more]

PMID: 16492137

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Project description:BackgroundTendon injuries are one of the most common musculoskeletal conditions in active patients. Platelet-rich plasma (PRP) has shown some promise in the treatment of tendon disorders, but little is known as to the mechanisms by which PRP can improve tendon regeneration. PRP contains numerous different growth factors and cytokines that activate various cellular signaling cascades, but it has been difficult to determine precisely which signaling pathways and cellular responses are activated after PRP treatment. Additionally, macrophages play an important role in modulating tendon regeneration, but the influence of PRP on determining whether macrophages assume a proinflammatory or anti-inflammatory phenotype remains unknown.PurposeTo use genome-wide expression profiling, bioinformatics, and protein analysis to determine the cellular pathways activated in fibroblasts treated with PRP. The effect of PRP on macrophage polarization was also evaluated.Study designControlled laboratory study.MethodsTendon fibroblasts or macrophages from rats were cultured and treated with either platelet-poor plasma (PPP) or PRP. RNA or protein was isolated from cells and analyzed using microarrays, quantitative polymerase chain reaction, immunoblotting, or bioinformatics techniques.ResultsPathway analysis determined that the most highly induced signaling pathways in PRP-treated tendon fibroblasts were TNFα and NFκB pathways. PRP also downregulated the expression of extracellular matrix genes and induced the expression of autophagy-related genes and reactive oxygen species (ROS) genes and protein markers in tendon fibroblasts. PRP failed to have a major effect on markers of macrophage polarization.ConclusionPRP induces an inflammatory response in tendon fibroblasts, which leads to the formation of ROS and the activation of oxidative stress pathways. PRP does not appear to significantly modulate macrophage polarization.Clinical relevancePRP might act by inducing a transient inflammatory event, which could then trigger a tissue regeneration response.

Dataset Information

Phenotypic responses to mechanical stress in fibroblasts from tendon, cornea and skin.

Publications

Phenotypic responses to mechanical stress in fibroblasts from tendon, cornea and skin.

Similar Datasets

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