Abstract
Diseases and injuries that compromise the ocular surface cause considerable patient distress and have long term consequences for their quality of life. Treatment modalities that can address the delicate balance of tissue regeneration, inflammation and maintenance of corneal transparency are therefore needed. We have recently formulated two novel eye drops from placental tissues: cord blood platelet lysate (CBED) and amniotic membrane extract eye drops (AMED), which can be used to treat severe ocular disorders. Here we characterise these two preparations by measuring: (a) growth factors (GF) and cytokines composition, (b) promotion of human corneal epithelial cell (HCEC) growth and (c) effects on immune cells in a lymphocyte culture assay. Finally, their bioavailability was assayed in an ex vivo porcine corneal model. We show that both preparations contain GF and cytokines that were able to promote the in vitro growth of HCEC and support repair in an in vitro scratch test. When assessed in a lymphocyte culture, both favoured immune suppression reducing the cellular expression of NKG2D and CD107a as well as the production of interferon gamma (IFN-γ) in natural killer, NKT and T cells. Regarding bioavailability, CBED active molecules were found mainly in the pre-corneal fraction with some penetration into the corneal fraction, in an ex vivo model. In summary, both placental-derived allogeneic preparations, CBED and AMED, display regenerative and immunomodulatory capabilities. These results will help define mechanisms of action and the best indications and doses of each product for use in a particular patient and support the development of off-the-shelf therapies for ocular surface pathologies in which wound healing defects and inflammatory events are contributing factors.
Highlights
Diseases and injuries that compromise the ocular surface cause considerable patient distress and have long term consequences for their quality of life
We describe the immunomodulatory and regenerative properties of CBED and amniotic membrane extract eye drops (AMED), two eye-drop preparations that have been recently used for the treatment of severe ocular surface diseases
The benefits of this protein profile is widely described in the literature; growth factors (GF) such as PDGF, transforming growth factor beta (TGF-β), bFGF and vascular endothelial growth factor (VEGF) have been shown to enhance regeneration in other tissues[24]
Summary
Diseases and injuries that compromise the ocular surface cause considerable patient distress and have long term consequences for their quality of life. We have recently formulated two novel eye drops from placental tissues: cord blood platelet lysate (CBED) and amniotic membrane extract eye drops (AMED), which can be used to treat severe ocular disorders We characterise these two preparations by measuring: (a) growth factors (GF) and cytokines composition, (b) promotion of human corneal epithelial cell (HCEC) growth and (c) effects on immune cells in a lymphocyte culture assay. Less well characterised is CB platelet rich plasma, CBED’s starting material, which appears to be a more potent immune regulator than adult p lasma[15], in part due to presence of the soluble Natural Killer group 2D ligands (sNKG2DLs) such as MHC class I polypeptide-related sequence (MICA/B) and unique long 16 binding protein (ULBP1), which we have been previously shown to have an immunosuppressive effect on immune cells[16] These sNKG2DLs are barely detectable in healthy individuals, but cell surface expression is induced when cells have been infected by viruses or become malignantly transformed. We first measured their GF and cytokine profile and tested their regenerative potential and bioavailability in ex vivo model and we further tested their immune modulatory effects using a lymphocyte assay
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