Effects of Mesenchymal Stromal Cells and Human Recombinant Nerve Growth Factor Delivered by Bioengineered Human Corneal Lenticule on an Innovative Model of Diabetic Retinopathy

Overview

Journal Front Endocrinol (Lausanne)

Specialty Endocrinology

Date 2024 Oct 30

PMID 39473506

Authors

Letizia Pelusi

Jose Hurst

Nicola Detta

Caterina Pipino

Alessia Lamolinara

Gemma Conte

Rodolfo Mastropasqua

Marcello Allegretti

Nadia Di Pietrantonio

Tiziana Romeo

Mona El Zarif

Mario Nubile

Laura Guerricchio

Sveva Bollini

Assunta Pandolfi

Sven Schnichels

Domitilla Mandatori

Affiliations

Soon will be listed here.

Abstract

Introduction: Diabetic retinopathy (DR) is a microvascular complication of diabetes in which neurodegeneration has been recently identified as a driving force. In the last years, mesenchymal stromal cells (MSCs) and neurotrophins like Nerve Growth Factor (NGF), have garnered significant attention as innovative therapeutic approaches targeting DR-associated neurodegeneration. However, delivering neurotrophic factors directly in the eye remains a challenge. Hence, this study evaluated the effects of MSCs from human amniotic fluids (hAFSCs) and recombinant human NGF (rhNGF) delivered by human corneal lenticule (hCL) on a high glucose (HG) induced model simulating the molecular mechanisms driving DR.

Methods: Porcine neuroretinal explants exposed to HG (25 mM for four days) were used to mimic DR . hCLs collected from donors undergoing refractive surgery were decellularized using 0.1% sodium dodecyl sulfate and then bioengineered with hAFSCs, microparticles loaded with rhNGF (rhNGF-PLGA-MPs), or both simultaneously. Immunofluorescence (IF) and scanning electron microscopy (SEM) analyses were performed to confirm the hCLs bioengineering process. To assess the effects of hAFSCs and rhNGF, bioengineered hCLs were co-cultured with HG-treated neuroretinal explants and following four days RT-PCR and cytokine array experiments for inflammatory, oxidative, apoptotic, angiogenic and retinal cells markers were performed.

Results: Data revealed that HG-treated neuroretinal explants exhibit a characteristic DR-phenotype, including increased level of NF-kB, NOS2, NRF2 GFAP, VEGFA, Bax/Bcl2 ratio and decreased expression of TUBB3 and Rho. Then, the feasibility to bioengineer decellularized hCLs with hAFSCs and rhNGF was demonstrated. Interestingly, co-culturing hAFSCs- and rhNGF- bioengineered hCLs with HG-treated neuroretinal explants for four days significantly reduced the expression of inflammatory, oxidative, apoptotic, angiogenic and increased retinal markers.

Conclusion: Overall, we found for the first time that hAFSCs and rhNGF were able to modulate the molecular mechanisms involved in DR and that bioengineered hCLs represents a promising ocular drug delivery system of hAFSCs and rhNGF for eye diseases treatment. In addition, results demonstrated that porcine neuroretinal explants treated with HG is a useful model to reproduce the DR pathophysiology.

Citing Articles

Future Directions in Diabetic Retinopathy Treatment: Stem Cell Therapy, Nanotechnology, and PPARα Modulation.

Kapa M, Koryciarz I, Kustosik N, Jurowski P, Pniakowska Z J Clin Med. 2025; 14(3).

PMID: 39941353 PMC: 11818668. DOI: 10.3390/jcm14030683.

References

Yerra V, Negi G, Sharma S, Kumar A . Potential therapeutic effects of the simultaneous targeting of the Nrf2 and NF-κB pathways in diabetic neuropathy. Redox Biol. 2013; 1:394-7. PMC: 3757712. DOI: 10.1016/j.redox.2013.07.005. View

Ipp E . Diabetic Retinopathy and Insulin Insufficiency: Beta Cell Replacement as a Strategy to Prevent Blindness. Front Endocrinol (Lausanne). 2021; 12:734360. PMC: 8667804. DOI: 10.3389/fendo.2021.734360. View

Zhong Q, Mishra M, Kowluru R . Transcription factor Nrf2-mediated antioxidant defense system in the development of diabetic retinopathy. Invest Ophthalmol Vis Sci. 2013; 54(6):3941-8. PMC: 3676188. DOI: 10.1167/iovs.13-11598. View

Schnichels S, Paquet-Durand F, Loscher M, Tsai T, Hurst J, Joachim S . Retina in a dish: Cell cultures, retinal explants and animal models for common diseases of the retina. Prog Retin Eye Res. 2020; 81:100880. DOI: 10.1016/j.preteyeres.2020.100880. View

Kong J, Zheng D, Chen S, Duan H, Wang Y, Dong M . A comparative study on the transplantation of different concentrations of human umbilical mesenchymal cells into diabetic rats. Int J Ophthalmol. 2015; 8(2):257-62. PMC: 4413587. DOI: 10.3980/j.issn.2222-3959.2015.02.08. View

Di Fulvio P, Pandolfi A, Formoso G, Di Silvestre S, Di Tomo P, Giardinelli A . Features of endothelial dysfunction in umbilical cord vessels of women with gestational diabetes. Nutr Metab Cardiovasc Dis. 2014; 24(12):1337-45. DOI: 10.1016/j.numecd.2014.06.005. View

Rocco M, Calza L, Aloe L . NGF and Retinitis Pigmentosa: Structural and Molecular Studies. Adv Exp Med Biol. 2021; 1331:255-263. DOI: 10.1007/978-3-030-74046-7_17. View

Sun F, Sun Y, Zhu J, Wang X, Ji C, Zhang J . Mesenchymal stem cells-derived small extracellular vesicles alleviate diabetic retinopathy by delivering NEDD4. Stem Cell Res Ther. 2022; 13(1):293. PMC: 9284871. DOI: 10.1186/s13287-022-02983-0. View

Costa A, Balbi C, Garbati P, Palama M, Reverberi D, de Palma A . Investigating the Paracrine Role of Perinatal Derivatives: Human Amniotic Fluid Stem Cell-Extracellular Vesicles Show Promising Transient Potential for Cardiomyocyte Renewal. Front Bioeng Biotechnol. 2022; 10:902038. PMC: 9214211. DOI: 10.3389/fbioe.2022.902038. View

10.

Kensler T, Wakabayashi N, Biswal S . Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. Annu Rev Pharmacol Toxicol. 2006; 47:89-116. DOI: 10.1146/annurev.pharmtox.46.120604.141046. View

11.

Yam G, Bandeira F, Liu Y, Devarajan K, Binte M Yusoff N, Htoon H . Effect of corneal stromal lenticule customization on neurite distribution and excitatory property. J Adv Res. 2022; 38:275-284. PMC: 9091752. DOI: 10.1016/j.jare.2021.09.004. View

12.

Costa A, Ceresa D, de Palma A, Rossi R, Turturo S, Santamaria S . Comprehensive Profiling of Secretome Formulations from Fetal- and Perinatal Human Amniotic Fluid Stem Cells. Int J Mol Sci. 2021; 22(7). PMC: 8038201. DOI: 10.3390/ijms22073713. View

13.

Stitt A, Curtis T, Chen M, Medina R, Mckay G, Jenkins A . The progress in understanding and treatment of diabetic retinopathy. Prog Retin Eye Res. 2015; 51:156-86. DOI: 10.1016/j.preteyeres.2015.08.001. View

14.

Colafrancesco V, Coassin M, Rossi S, Aloe L . Effect of eye NGF administration on two animal models of retinal ganglion cells degeneration. Ann Ist Super Sanita. 2011; 47(3):284-9. DOI: 10.4415/ANN_11_03_08. View

15.

Lambiase A, Aloe L, Centofanti M, Parisi V, Bao S, Mantelli F . Experimental and clinical evidence of neuroprotection by nerve growth factor eye drops: Implications for glaucoma. Proc Natl Acad Sci U S A. 2009; 106(32):13469-74. PMC: 2726400. DOI: 10.1073/pnas.0906678106. View

16.

Yam G, Binte M Yusoff N, Goh T, Setiawan M, Lee X, Liu Y . Decellularization of human stromal refractive lenticules for corneal tissue engineering. Sci Rep. 2016; 6:26339. PMC: 4876320. DOI: 10.1038/srep26339. View

17.

Soni D, Sagar P, Takkar B . Diabetic retinal neurodegeneration as a form of diabetic retinopathy. Int Ophthalmol. 2021; 41(9):3223-3248. DOI: 10.1007/s10792-021-01864-4. View

18.

Toda N, Nakanishi-Toda M . Nitric oxide: ocular blood flow, glaucoma, and diabetic retinopathy. Prog Retin Eye Res. 2007; 26(3):205-38. DOI: 10.1016/j.preteyeres.2007.01.004. View

19.

Wang Q, Sheng W, Yi C, Lv H, Cheng B . Retrobulbarly injecting nerve growth factor attenuates visual impairment in streptozotocin-induced diabetes rats. Int Ophthalmol. 2020; 40(12):3501-3511. DOI: 10.1007/s10792-020-01537-8. View

20.

Alio Del Barrio J, Chiesa M, Garagorri N, Garcia-Urquia N, Fernandez-Delgado J, Bataille L . Acellular human corneal matrix sheets seeded with human adipose-derived mesenchymal stem cells integrate functionally in an experimental animal model. Exp Eye Res. 2015; 132:91-100. DOI: 10.1016/j.exer.2015.01.020. View