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lauren M. James, Hari Jayaram, Phillippa Cottrill, Karen Eastlake, Peng T. Khaw, Robert A. Brown, Karl Matter, G Astrid Limb; Development Of Multifunctional Collagen Constructs For Retinal Cell Transplantation Of müLler Stem Cell Derived Rgc. Invest. Ophthalmol. Vis. Sci. 2012;53(14):309.
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We aimed to design multifunctional collagen constructs containing therapeutic agents to facilitate the migration, integration and survival of transplanted retinal ganglion cells (RGC) derived from human Müller stem cells (hMSC).
Triamcinolone and Chondroitinase ABC (ChABC) were incorporated into type I collagen constructs; following polymerisation and subsequent plastic compressed to form cellular scaffolds. Following pH neutralisation of collagen solutions, a suspension of triamcinolone containing 4mg of the drug was added to the collagen, as well as the addition of alginate microparticles containing ChABC. Inclusion of ChABC into alginate particles was achieved using an encapsulator, at a voltage of 9KV and an air gap distance of 7cm. Following polymerisation at 37°C, collagen was then compressed under 150g for 5 minutes to form ultrathin scaffolds to which RGC derived from hMSC could adhere.Electron microscopy was used to examine the microstructure of the collagen matrices containing the agents to assess any morphological alterations to the fibrillar architecture of the scaffolds. Cellular viability of hMSC grown on the collagen scaffolds was examined using a hexosaminidase assay. Optical density of the various scaffolds was also determined over a range of 350nm-800nm using a spectrophotometer. Cellular with RGC derived from hMSC were transplanted onto explants of cadaveric human retina.
Scanning electron microscopy showed that cells seeded onto collagen scaffolds were able to develop firm attachments to the collagen matrices. Cellular viability was maintained throughout 7 days in culture; where cells were unaffected under the varying culturing microenvironments when compared to MatrigelTM alone. Scaffolds containing therapeutic doses of both triamcinolone and ChABC did not exclude any light within the visual spectrum. Transplantation of cellular scaffolds onto the inner surface of human retinal explants ex vivo showed that cells were capable of migrating into the RGC layer, as determined by immunostaining of retinal sections prepared from the in vitro transplanted retina.
The results suggest that plastic compressed collagen constructs provide suitable vehicles for drug and cell delivery into the eye. These composite multifunctional constructs can be produced consistently under sterile conditions, providing a viable method of supplying transplanted cells with pro-survival factors when transplanted into a host environment. These therefore constitute a viable strategy for transplantation of Müller stem cell derived RGC into the retina.
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