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Published on: August 16, 2014
Effect of TGFβ and PDGF-B blockade on corneal myofibroblast development in mice
V Singh1, M R Santhiago, F L Barbosa
1The Cole Eye Institute, The Cleveland Clinic, 9500 Euclid Ave., Cleveland, OH 44195, USA.
Abstract:
The purpose of this study was to investigate the role of transforming growth factor beta (TGFβ) and/or platelet-derived growth factor-B (PDGF-B) blockade on the differentiation of vimentin and alpha-smooth muscle actin (αSMA)-expressing myofibroblasts associated with haze in mice. Mouse corneas had haze-generating irregular PTK (phototherapeutic keratectomy) and topical treatment with the vectors. Six study groups of PTK treated corneas, with four corneas per group in each experiment, were Group 1) treated with TGFβ-KDEL vector interfering with TGFβ signaling through anomalous sorting of cytokine bound to the expressed altered receptor; Group 2) treated with PDGF-B-KDEL vector interfering with PDGF signaling through anomalous sorting of cytokine bound to the expressed altered receptor; Group 3) treated with both TGFβ-KDEL vector and PDGF-B-KDEL vector to interfere with signaling of both cytokines; Group 4) empty pGFPC1 vector; Group 5) empty pCMV vector; and Group 6) no vector treatment control. At one month after surgery, the corneas were analyzed by immunocytochemistry (IHC) for central stromal cells expressing myofibroblast markers vimentin and αSMA. The stroma of corneas treated with the TGFβ-KDEL vector alone (p < 0.05) or both the TGFβ-KDEL and PDGF-B-KDEL vectors (P < 0.05) had significantly lower density of vimentin-positive cells compared to the corresponding control group. The central stroma of corneas treated with the TGFβ-KDEL vector (p < 0.05) or the PDGF-B-KDEL vector (p < 0.05) had lower density of αSMA-positive cells compared to the corresponding control group. The density of αSMA-positive stromal cells was also significantly lower (p < 0.05) when both the TGFβ-KDEL and PDGF-B-KDEL and vectors were applied together compared to the corresponding control groups. This study provides in situ evidence that TGFβ and PDGF-B have important roles in modulating myofibroblast generation in the mouse cornea after haze-associated injury.
Insights
Blocking transforming growth factor beta (TGFβ) and platelet-derived growth factor-B (PDGF-B) reduced myofibroblast markers in mouse corneas after injury. This suggests TGFβ and PDGF-B are key in haze development following corneal procedures.
Area of Science:
- Ophthalmology
- Cell Biology
- Molecular Biology
Background:
- Corneal haze after phototherapeutic keratectomy (PTK) is associated with myofibroblast differentiation.
- Transforming growth factor beta (TGFβ) and platelet-derived growth factor-B (PDGF-B) are implicated in this process.
Purpose of the Study:
- To investigate the role of TGFβ and PDGF-B blockade in myofibroblast differentiation.
- To assess the impact of targeted vector treatments on corneal haze markers.
Main Methods:
- Mouse corneas underwent PTK and received topical vector treatments targeting TGFβ and/or PDGF-B signaling.
- Immunocytochemistry was used to analyze vimentin and alpha-smooth muscle actin (αSMA) expression in stromal cells one month post-surgery.
Main Results:
- Treatment with TGFβ-KDEL vector alone or combined with PDGF-B-KDEL vector significantly reduced vimentin-positive cells.
- Both TGFβ-KDEL and PDGF-B-KDEL vectors, individually or combined, decreased αSMA-positive cells compared to controls.
Conclusions:
- TGFβ and PDGF-B play significant roles in myofibroblast generation in the mouse cornea post-injury.
- Targeting these growth factors offers a potential strategy for managing corneal haze.
