Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail
Abstract Introduction Fibrotic cocktail (FC) is a combination of pro-fibrotic and pro-inflammatory mediators that induces early fibrotic changes in organotypic lung models. We hypothesised that transforming growth factor beta 1 (TGF-β1) alone induces a pro-fibrotic effect similar to FC. Our aim was...
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BMC
2025-01-01
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| Series: | Respiratory Research |
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| Online Access: | https://doi.org/10.1186/s12931-025-03110-2 |
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| author | Carlos Machahua Thomas M. Marti Patrick Dorn Manuela Funke-Chambour |
| author_facet | Carlos Machahua Thomas M. Marti Patrick Dorn Manuela Funke-Chambour |
| author_sort | Carlos Machahua |
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| description | Abstract Introduction Fibrotic cocktail (FC) is a combination of pro-fibrotic and pro-inflammatory mediators that induces early fibrotic changes in organotypic lung models. We hypothesised that transforming growth factor beta 1 (TGF-β1) alone induces a pro-fibrotic effect similar to FC. Our aim was to compare the pro-fibrotic effects of TGF-β1 with FC in human precision-cut lung slices (PCLS). Methods PCLS from “healthy” lung tissue of cancer patients undergoing surgery (n = 7) were incubated with TGF-β1, FC or control for 72 h. Gene expression markers for myofibroblasts differentiation, extracellular matrix (ECM), as well as TGF-β receptors were assessed (RT-qPCR). ECM proteins expression in lysates and supernatant was assessed by ELISA and immunofluorescence. Results We found that TGF-β1 significantly increased gene expression of ACTA2, COL1A1, CCN2, and VIM compared to control but also compared to FC. FC showed a significant increase of matrix metalloproteinase (MMP) 7 and 1 compared to control, while TGF-β receptor 2 was lower after FC compared to TGF-β1 or control. FC or TGF-β1 showed similar fibronectin protein expression in lysates and supernatants, while type I collagen protein expression in lysates was significantly greater with TGF-β1 compared to control. Conclusions Our findings show that TGF-β1 induces consistent pro-fibrotic changes in PCLS after 72 h. Compared to TGF-β1, FC treatment resulted in reduced gene expression of TGF-β receptor 2 and increased MMPs expression, potentially mitigating the early pro-fibrotic effects. Selecting specific pro-fibrotic stimuli may be preferable depending on the research question and time point of interest in lung fibrosis studies using PCLS. |
| format | Article |
| id | doaj-art-0eae888aea6b4fb990190ba66618870b |
| institution | Kabale University |
| issn | 1465-993X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Respiratory Research |
| spelling | doaj-art-0eae888aea6b4fb990190ba66618870b2025-02-02T12:37:58ZengBMCRespiratory Research1465-993X2025-01-012611710.1186/s12931-025-03110-2Fibrosis in PCLS: comparing TGF-β and fibrotic cocktailCarlos Machahua0Thomas M. Marti1Patrick Dorn2Manuela Funke-Chambour3Department for Pulmonary Medicine, Allergology and Clinical Immunology, Inselspital, Bern University Hospital, University of BernDepartment of General Thoracic Surgery, Inselspital, Bern University HospitalDepartment of General Thoracic Surgery, Inselspital, Bern University HospitalDepartment for Pulmonary Medicine, Allergology and Clinical Immunology, Inselspital, Bern University Hospital, University of BernAbstract Introduction Fibrotic cocktail (FC) is a combination of pro-fibrotic and pro-inflammatory mediators that induces early fibrotic changes in organotypic lung models. We hypothesised that transforming growth factor beta 1 (TGF-β1) alone induces a pro-fibrotic effect similar to FC. Our aim was to compare the pro-fibrotic effects of TGF-β1 with FC in human precision-cut lung slices (PCLS). Methods PCLS from “healthy” lung tissue of cancer patients undergoing surgery (n = 7) were incubated with TGF-β1, FC or control for 72 h. Gene expression markers for myofibroblasts differentiation, extracellular matrix (ECM), as well as TGF-β receptors were assessed (RT-qPCR). ECM proteins expression in lysates and supernatant was assessed by ELISA and immunofluorescence. Results We found that TGF-β1 significantly increased gene expression of ACTA2, COL1A1, CCN2, and VIM compared to control but also compared to FC. FC showed a significant increase of matrix metalloproteinase (MMP) 7 and 1 compared to control, while TGF-β receptor 2 was lower after FC compared to TGF-β1 or control. FC or TGF-β1 showed similar fibronectin protein expression in lysates and supernatants, while type I collagen protein expression in lysates was significantly greater with TGF-β1 compared to control. Conclusions Our findings show that TGF-β1 induces consistent pro-fibrotic changes in PCLS after 72 h. Compared to TGF-β1, FC treatment resulted in reduced gene expression of TGF-β receptor 2 and increased MMPs expression, potentially mitigating the early pro-fibrotic effects. Selecting specific pro-fibrotic stimuli may be preferable depending on the research question and time point of interest in lung fibrosis studies using PCLS.https://doi.org/10.1186/s12931-025-03110-2Precision cut lung slicesFibrotic cocktailTransforming growth factor beta 1Ex vivo modelFibrotic markersLung fibrosis |
| spellingShingle | Carlos Machahua Thomas M. Marti Patrick Dorn Manuela Funke-Chambour Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail Respiratory Research Precision cut lung slices Fibrotic cocktail Transforming growth factor beta 1 Ex vivo model Fibrotic markers Lung fibrosis |
| title | Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail |
| title_full | Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail |
| title_fullStr | Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail |
| title_full_unstemmed | Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail |
| title_short | Fibrosis in PCLS: comparing TGF-β and fibrotic cocktail |
| title_sort | fibrosis in pcls comparing tgf β and fibrotic cocktail |
| topic | Precision cut lung slices Fibrotic cocktail Transforming growth factor beta 1 Ex vivo model Fibrotic markers Lung fibrosis |
| url | https://doi.org/10.1186/s12931-025-03110-2 |
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