RESUMO
Platelet derived growth factor (PDGF) is involved in wound healing in various organ systems. Its potential role in the context of peritoneal injury following long-term peritoneal dialysis is unclear. We used an adenovirus expressing the B chain of PDGF (AdPDGF-B) to assess its effect on pro-fibrotic pathways in the peritoneal membrane. To assess the transforming growth factor (TGF) beta independent effects of PDGF, we over-expressed PDGF-B in the peritoneum of either wild-type mice (Smad3+/+) or those with a deletion of the TGFbeta signaling protein Smad3 (Smad3(-/-)). PDGF-B induced sustained angiogenesis in both Smad3+/+ and Smad3(-/-) mice. Despite increased collagen gene expression, collagen accumulation was transient and fibrogenesis was associated with induction of collagenase activity. We observed epithelial to mesenchymal transition (EMT) involving the peritoneal mesothelial cells, as shown by increased SNAIL and decreased E-Cadherin expression with evidence of mesothelial cells expressing both epithelial and mesenchymal markers. Unlike TGFbeta-induced EMT, PDGF-B exposure did not lead to mobilization of the mesothelial cells; they remained as a single monolayer throughout the observation period. This "non-invasive" EMT phenomenon is a novel finding and may have implications concerning the role of EMT in peritoneal fibrosis and injury to other organ systems. The observed effects were similar in Smad3(-/-) and Smad3+/+ animals, suggesting that the PDGF-B effects were independent of TGFbeta or Smad signaling.
Assuntos
Epitélio/fisiologia , Mesoderma/fisiologia , Peritônio/citologia , Proteínas Proto-Oncogênicas c-sis/metabolismo , Proteína Smad3/metabolismo , Adenoviridae/genética , Adenoviridae/metabolismo , Animais , Caderinas/genética , Caderinas/metabolismo , Diferenciação Celular , Células Epiteliais/citologia , Células Epiteliais/fisiologia , Fibrose/patologia , Gelatinases/metabolismo , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Metaloproteinases da Matriz/genética , Metaloproteinases da Matriz/metabolismo , Mesoderma/citologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neovascularização Fisiológica/fisiologia , Peritônio/metabolismo , Peritônio/patologia , Fenótipo , Proteínas Proto-Oncogênicas c-sis/genética , Proteína Smad3/genética , Fator de Crescimento Transformador beta/metabolismoRESUMO
Posterior capsule opacification (PCO) or secondary cataract formation, following intraocular lens implantation, is a significant complication affecting an estimated 28% of cataract patients. Matrix metalloproteinases (MMPs) have been demonstrated to play a role in the formation of anterior subcapsular cataracts and it has been shown that the presence of MMP inhibitors (MMPI) decreases subcapsular cataract formation ex vivo. Since the mechanisms responsible for anterior subcapsular cataract formation and posterior capsule opacification are similar, it is reasonable to suggest that MMP inhibitors may also mitigate PCO. One of the most effective ways of delivering the inhibitors may be from the implanted intraocular lens (IOL) material itself. In the current work, delivery of three different MMP inhibitors from silicone rubber as a model IOL material was examined. Loading methods were developed which allowed continuous release of active MMPI for periods of over 5 months in some cases. Reduced migration rates were observed in human lens epithelial cells in vitro, suggesting that an effect on PCO may be possible. While further studies are necessary to tune the systems to achieve the desired rates of release, this work demonstrates that delivery of MMPI from silicone IOL materials has the potential to decrease the incidence of PCO.
