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Charlotte Schramm, Martin S. Spitzer, Sigrid Henke-Fahle, Gabriele Steinmetz, Kai Januschowski, Peter Heiduschka, Jürgen Geis-Gerstorfer, Tilo Biedermann, Karl U. Bartz-Schmidt, Peter Szurman; The Cross-linked Biopolymer Hyaluronic Acid as an Artificial Vitreous Substitute. Invest. Ophthalmol. Vis. Sci. 2012;53(2):613-621. doi: 10.1167/iovs.11-7322.
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Biopolymers are promising substances in the development of a new vitreous substitute to overcome the drawbacks associated with current hydrophobic tamponade materials.
Different hydrogels were assembled by cross-linking hyaluronic acid either with adipic dihydrazide (ADH) by carboxylation with N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDCI) after hydrazation or by photocrosslinking with UV-light and N-vinyl-pyrrolidinone. The refractive index and rheologic properties of the obtained gels were investigated. To quantify the degradation of the hydrogels over time, free hyaluronic acid was measured photometrically by means of the degradation product uronic acid. For biocompatibility testing, the hydrogels were applied on top of cultured retinal pigment epithelial (RPE) cells and analyzed by the cell viability, MTT, and alamar blue viability cytotoxicity assays and flow cytometry, with Annexin V-FITC and propidium iodide co-staining. The in vivo biocompatibility of the hydrogels was tested in vitrectomized rabbit eyes for up to 6 weeks.
The synthesized hydrogels were all clear and transparent and had a refractive index similar to human vitreous. The rheologic measurements suggested sufficient viscosity and elasticity for intraocular use. Quantification of the degradation products revealed only a small decay of the gels over 1 month. However, the ADH cross-linked hydrogels induced mild cytotoxicity in the RPE cells. The UV cross-linked hydrogels showed no toxicity or induction of apoptosis. In vivo the UV cross-linked biogels remained in place for 6 weeks, and electrophysiology and histology showed excellent tissue biocompatibility.
Biopolymers based on UV cross-linked hyaluronic acid may be promising vitreous substitutes.
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