Decellularized human corneal lenticule as a delivery system of recombinant human nerve growth factor and human amniotic fluid stem cells factors for posterior ocular disease treatment.

Mandatori, Domitilla; Pelusi, Letizia; Ghelardoni, Maddalena; Agnifili, Luca; Di Pietrantonio, Nadia; Hurst, Jose; Allegretti, Marcello; Schnichels, Sven; Mastropasqua, Leonardo; Bollini, Sveva; Nubile, Mario; Pandolfi, Assunta

Aims/Purpose: Posterior ocular diseases, such as glaucoma and diabetic retinopathy (DR), are the main causes of blindness worldwide. Among the possible therapeutic approaches particular interest was turned to neurotrophic factors, such as Nerve Growth Factor (NGF), and to mesenchymal stem cells (MSCs)‐derived factors. Nevertheless, the local drugs and factors delivery in the eye remains a challenge. Therefore, using an in vitro model of glaucoma and an ex vivo model of DR, this study aimed to test the feasibility to use decellularized human corneal lenticule (hCL) as natural delivery system able to vehicle neurotrophic and MSCs‐regenerative factors. Methods: hCLs (thickness 100–120 μm) collected from donors undergoing refractive surgery, were decellularized (0.1% SDS) and engineered with MSCs from human amniotic fluid (hAFSCs) and/or with microparticles containing rhNGF (rhNGF‐MPs). Human Trabecular Meshwork cells (HTMCs), damaged with Transforming Growth Factor‐β2 (TGF‐β2; 5 ng/mL for 4 days), were employed to mimic glaucoma condition in vitro. DR disease was reproduced using an ex vivo model of porcine retinal explants exposed to high glucose (HG; 25 mM for 4 days). Results: Results deriving from glaucoma model showed that the deposition of extracellular matrix and the specific protein expression of COL4a1, COL1a1 and ACTA1 were reduced following the HTMCs incubation with hCL engineered with hAFSCs or rhNGF‐MPs. Likewise, in the DR model, the HG‐increased Tumour Necrosis Factor α (TNF‐α) and Nuclear factor erythroid 2–Related Factor 2 (NRF2) levels were reduced following the incubation with hCL engineered with hAFSCs and rhNGF‐MPs. Of note, as demonstrated by the increase of structural (TUBB3 and Rho) and apoptotic (BAX/Bcl2 and p21) markers, a neuroprotective effect of engineered‐hCLs was observed in the DR model. Conclusions: Overall, our results demonstrated that this innovative bio‐engineered scaffold may represent a potential therapeutic approach for posterior ocular diseases treatment.

AMNIOTIC liquid; NERVE growth factor; NUCLEAR factor E2 related factor; STEM cell factor; CORNEAL transplantation; THERAPEUTICS; STEM cell transplantation; HUMAN growth

Acta Ophthalmologica (1755375X), 2024, Vol 102, pN.PAG

1755-375X

Academic Journal

10.1111/aos.15999