April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Nanotechnology for improvement of stem cell therapy of laser damaged retina in an animal model
Author Affiliations & Notes
  • Simona Nicoara
    Ophthalmology, "Iuliu Hatieganu" University of Medicine and Pharmacy, Cluj-Napoca, Romania
  • Cristina Cristian
    Ophthalmology, "Iuliu Hatieganu" University of Medicine and Pharmacy, Cluj-Napoca, Romania
  • Cristian Berce
    Center of Experimental Medicine, "Iuliu Hatieganu" University of Medicine and Pharmacy, Cluj-Napoca, Romania
  • Flaviu Tabaran
    Pathology, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania
  • Oana Tudoran
    Radiobiology and Tumor Biology, "Ion Chiricuta" Institute of Oncology, Cluj-Napoca, Romania
  • Sanda Boca-Farcau
    Radiobiology and Tumor Biology, "Ion Chiricuta" Institute of Oncology, Cluj-Napoca, Romania
    Nanobiophotonics and Microspectroscopy Center, Babes-Bolyai University, Cluj-Napoca, Romania
  • Simion Astilean
    Nanobiophotonics and Microspectroscopy Center, Babes-Bolyai University, Cluj-Napoca, Romania
  • Olga Soritau
    Radiobiology and Tumor Biology, "Ion Chiricuta" Institute of Oncology, Cluj-Napoca, Romania
  • Footnotes
    Commercial Relationships Simona Nicoara, None; Cristina Cristian, None; Cristian Berce, None; Flaviu Tabaran, None; Oana Tudoran, None; Sanda Boca-Farcau, None; Simion Astilean, None; Olga Soritau, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 4624. doi:
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      Simona Nicoara, Cristina Cristian, Cristian Berce, Flaviu Tabaran, Oana Tudoran, Sanda Boca-Farcau, Simion Astilean, Olga Soritau; Nanotechnology for improvement of stem cell therapy of laser damaged retina in an animal model. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4624.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract
 
Purpose
 

We aimed to develop an animal model of retinal injury and repair by the intravitreal administration of adult mesenchymal stem cells (MSCs) using gold nanoparticles (GNPs) as delivery system for neuronal specific growth factors (NGFs).

 
Methods
 

Isolated MSCs from the bone marrow of CD1 mice were characterized by immuno-fenotyping and RT-PCR for stem cell markers. We produced GNPs and conjugated them with NGF-beta in order to attain a specific functionality. 20 nm GNPs were chemically synthesized by citrate reduction of gold salt and functionalized with NGF-beta through nonspecific electrostatic interaction between the nanoparticles and the protein. GNPs were characterized by several techniques: transmission electron microscopy, UV-VIS absorption spectroscopy, dynamic light scattering and zeta potential. In vitro differentiation ability of MSCs into retinal progenitors was investigated using a multistep protocol with exploration of the most suitable combination of growth factors and GNPs conjugated with NGF-beta, as well as the most suitable cell substrate. The inducing of the retinal lesions by indirect diode laser photocoagulation in CD1 mice was followed by the intravitreal administration of fluorescent MSCs and GNPs conjugated with NGF. The evaluation of MSC grafting and retinal morphology was performed at 1 and 3 weeks after the procedure, by confocal microscopy and hystopathology.

 
Results
 

Isolation and cultivation of MSCs from bone marrow of CD1 mice was possible even without substrate. The substrate supports better the cell proliferation and reduces the percentage of cell death during differentiation. Isolated cells were adult type MSCs. The multistep protocol requires further investigation, mainly in stage 4 where cells might require a richer medium. Growing these cells on substrates that mimic the extracellular matrix (laminin and collagen) supported the viability and cell proliferation. MSCs in CD1 mice replaced the degenerated retinal cells and the association with GNPs as delivery system for NGFs improved the differentiation potential of MSCs.

 
Conclusions
 

The administration of cells in the vitreous cavity has an acceptable invasiveness with low risk of infection. GNPs conjugated with NGFs offer a proper microenvironment for MSCs differentiation into retinal cells and is an alternative delivery system to viral vectors.

 
 
Differentiated MSCs
 
Differentiated MSCs
 
 
Quantitative analysis of MSCs in the retina
 
Quantitative analysis of MSCs in the retina
 
Keywords: 688 retina • 607 nanotechnology • 721 stem cells  
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