TGFβ Treated Placenta Derived Mesenchymal Stem Cells Selectively Promote Anti-Adipogenesis in Thyroid-Associated Ophthalmopathy

Abstract: Background Thyroid-associated ophthalmopathy (TAO) includes several ocular manifestations of the systemic autoimmune process, Graves’ disease. The key pathogenic cells are orbital fibroblasts (OFs). The OFs of most TAO patients are differentiated from pre-adipocytes to mature adipocytes, increased lipid and fat expansion are characteristic of ophthalmic disease. Human placental mesenchymal stem cells (hPMSCs) immunomodulate the pathogenesis. We previous showed that hPMSCs were therapeutic. Here, we prepared transforming growth factor β (TGFβ) treated hPMSCs (TGFβ-hPMSCs) to enhance adipogenesis inhibition in vitro and TAO mice. Methods To investigate anti-adipogenic effects in vitro, primary OFs were grown in differentiation medium for 2, 4, and 10 days. After co-cultur with hPMSCs and TGFβ-hPMSCs, the OFs were analyzed via quantitative real-time polymerase chain reaction, Oil red O staining and western blotting. Human PMSCs and TGFβ-hPMSCs were injected into the left orbits of TAO mice. We explored the anti-adipogenic pathway in play using SB431542, a TGFβ receptor kinase inhibitor. Results TGFβ-hPMSCs suppressed trrancription of the adipogenic, lipogenic, and fibrotic genes PPARγ, C/EBPα, LEPTIN, AP2, SREBP2, HMGCR, α-SMA, FIBRONECTIN and IL-17 more so than did hPMSCs when adipogenesis-induced primary TAO OFs. Moreover, in TAO mice injected with TGFβ-hPMSCs, the adipose area was reduced compared to that after injection, of hPMSCs or a steroid, and the expression levels of the Pparγ and C/ebpα were more effectively suppressed. These anti-adipogenic effects were mediated by the SMAD 2 and SMAD3 pathways both in vitro and in vivo. TGFβ-hPMSCs inhibited inflammation as effectively as did a steroid both in vitro and in vivo, and reduced in vitro fibrosis. Conclusion TGFβ-hPMSCs reduced adipogenesis and lipogenesis in in vitro and TAO mice with experimental indicating that TGFβ-hPMSCs could serve as cellular therapy for TAO patients. Furthermore, TGFβ-hPMSCs exhibited anti-inflammatory and anti-fibrotic functions and thus, could be used to treat muscle fibrosis in TAO patients.