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2019
OMIG Abstract Methods: Corneas of 6-8 week-old male wildtype (WT) C57BL/6 mice underwent suture placement to induce corneal NV. Splenic GFP+ pDCs from DPE-GFP×RAG1-/- mice and WT CD11b+ myeloid cells were isolated. After trephination, 104 pDCs, CD11b+ cells, or PBS control were locally applied onto the corneas using Tisseel fibrin sealant. On day 7, corneas were stained for CD31 (vascular marker) and underwent confocal microscopy. Length of NV and the density of adoptively-transferred GFP+ pDCs were measured by ImageJ. Relative mRNA level of anti-angiogenic molecule endostatin was quantified in the corneas using qRT-PCR. ANOVA with LSD post-hoc test was used to assess statistical significance. p<0.05 was considered significant.. Results: Confocal microscopy confirmed successful transfer of GFP+ pDCs to both central (452.8±39.1 cells/mm2) and peripheral corneas (435.1±52.6) on day 2 following local application of pDCs. qRT-PCR showed that local adoptive transfer of pDCs was accompanied by 4.7-fold increase in the mRNA level of anti-angiogenic molecule endostatin compared with fibrin sealant-only control (p=0.009) and 2.3-fold increase compared with adoptive transfer of CD11b+ cells (p=0.03). One-time adoptive transfer of pDCs significantly reduced NV length on day 7 following suture placement (350.1±43.4 µm), compared with transfer of CD11b+ cells (477.0±33.9 µm; p=0.004) as well as fibrin sealant-only controls (454.1±36.5 µm; p=0.01).
Conclusion: Local adoptive transfer of pDCs can limit corneal NV following suture placement and may serve as a novel cell-based therapeutic approach to treat corneal NV.
Disclosure: S: NIH-R01- EY022695 (PH), NIH-R21- EY025393 (PH), NIH-R01- EY026963 (PH), NIH-R01- EY029602 (PH), Tufts Medical Center Institutional Support (PH).
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