FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Vet Ophthalmol / Decorin-PEI nanoconstruct attenuates equine corneal fibrobla...
Veterinary ophthalmology2013; 17(3); 162-169; doi: 10.1111/vop.12060

Decorin-PEI nanoconstruct attenuates equine corneal fibroblast differentiation.

Abstract: To explore (i) the potential of polyethylenimine (PEI) nanoparticles as a vector for delivering genes into equine corneal fibroblasts (ECFs) using green fluorescent protein (GFP) marker gene, (ii) whether PEI nanoparticle-mediated decorin (DCN) gene therapy could be used to inhibit fibrosis in the equine cornea using an in vitro model. Methods: Polyethylenimine-DNA nanoparticles were prepared at nitrogen-to-phosphate (N-P) ratio of 15 by mixing 22 kDa linear PEI and a plasmid encoding either GFP or DCN. ECFs were generated from donor corneas as previously described. Initially, GFP was introduced into ECFs using PEI nanoparticles to confirm gene delivery, then DCN was introduced to evaluate for antifibrotic effects. GFP gene delivery was confirmed with real-time qPCR and ELISA. Changes in fibrosis after DCN therapy were quantified by measuring α-smooth muscle actin (αSMA) mRNA and protein levels with qPCR, immunostaining, and immunoblotting. Cytotoxicity was determined by evaluating cell morphology, cellular viability, and TUNEL assay. Results: Polyethylenimine-green fluorescent protein-treated cultures showed 2.2 × 10(4) GFP plasmid copies/μg of cellular DNA and 2.1 pg of GFP/100 μL of lysate. PEI-DCN delivery significantly attenuated TGFβ-induced transdifferentiation of fibroblasts to myofibroblasts (2-fold decrease of αSMA mRNA; P = 0.05) and significant inhibition of αSMA (49 ± 14.2%; P < 0.001) in immunocytochemical staining and immunoblotting were found. Furthermore, PEI-DNA nanoparticle delivery did not alter cellular phenotype at 24 h and cellular viability was maintained. Conclusions: Twenty-two kilo dalton Polyethylenimine nanoparticles are safe and effective for equine corneal gene therapy in vitro. PEI-mediated DCN gene delivery is effective at inhibiting TGFβ-mediated fibrosis in this model.
© 2013 American College of Veterinary Ophthalmologists.
Get Full Text
Publication Date: 2013-05-30 PubMed ID: 23718145DOI: 10.1111/vop.12060Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research suggests that polyethylenimine (PEI) nanoparticles could be effectively used for gene therapy in treating fibrosis in the equine cornea, essentially improving horse eye health.

Objective and Methods of the Research

  • The research aimed to assess the effectiveness of PEI nanoparticles as a tool for delivering genes into equine corneal fibroblasts (ECFs). Their desired outcome was to inhibit fibrosis in the equine cornea.
  • For this, they created PEI-DNA nanoparticles by combining 22 kDa linear PEI and a plasmid, which is a DNA molecule. It encoded either green fluorescent protein (GFP) or decorin (DCN).
  • The GFP was first introduced into ECFs using PEI nanoparticles to test if the gene delivery worked. Following this, DCN was introduced to look at its antifibrotic effects. The success was checked using real-time qPCR, ELISA, or immunostaining.

Results of the Study

  • The tests revealed successful inflow of the GFP gene via the PEI nanoparticles. The treated cultures exhibited 2.2 × 10(4) GFP plasmid copies/μg of cellular DNA and 2.1 pg of GFP/100 μL of lysate.
  • Further, the introduction of PEI-DCN successfully limited fibrosis. It decreased the transdifferentiation of fibroblasts to myofibroblasts, which were induced by TGFβ, by 2-folds.
  • Following the treatment, the cellular phenotype did appeared unaffected at 24 hours, and cellular viability was retained, marking the treatment safe.

Conclusions Drawn from the Research

  • From the study, it was concluded that 22 kilo Daltons PEI nanoparticles are safe and effective as a tool for gene therapy in treating equine corneas in vitro.
  • Evidently, the experimental PEI-mediated DCN gene delivery approach works in fighting TGFβ-mediated fibrosis in this model.

Cite This Article

APA
Donnelly KS, Giuliano EA, Sharma A, Tandon A, Rodier JT, Mohan RR. (2013). Decorin-PEI nanoconstruct attenuates equine corneal fibroblast differentiation. Vet Ophthalmol, 17(3), 162-169. https://doi.org/10.1111/vop.12060

Publication

Veterinary ophthalmology
ISSN: 1463-5224
NlmUniqueID: 100887377
Country: England
Language: English
Volume: 17
Issue: 3
Pages: 162-169

Researcher Affiliations

Donnelly, Kevin S
  • Harry S. Truman Veterans Memorial Hospital, 800 Hospital Drive, Columbia, MO, 652012, USA; Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, 900 East Campus Drive, Columbia, MO, 65211, USA.
Giuliano, Elizabeth A
    Sharma, Ajay
      Tandon, Ashish
        Rodier, Jason T
          Mohan, Rajiv R

            MeSH Terms

            • Animals
            • Cells, Cultured
            • Cornea / cytology
            • Decorin / chemistry
            • Decorin / genetics
            • Decorin / metabolism
            • Fibroblasts / cytology
            • Fibroblasts / drug effects
            • Gene Transfer Techniques / veterinary
            • Genes, Reporter
            • Genetic Vectors
            • Green Fluorescent Proteins / genetics
            • Green Fluorescent Proteins / metabolism
            • Horses
            • Nanoparticles
            • Polyethyleneimine / chemistry
            • Polyethyleneimine / pharmacology

            Grant Funding

            • R01EY017294 / NEI NIH HHS

            Citations

            This article has been cited 19 times.
            1. Mahaling B, Low SWY, Ch S, Addi UR, Ahmad B, Connor TB, Mohan RR, Biswas S, Chaurasia SS. Next-Generation Nanomedicine Approaches for the Management of Retinal Diseases. Pharmaceutics 2023 Jul 22;15(7).
              doi: 10.3390/pharmaceutics15072005pubmed: 37514191google scholar: lookup
            2. Sarkar S, Panikker P, D'Souza S, Shetty R, Mohan RR, Ghosh A. Corneal Regeneration Using Gene Therapy Approaches. Cells 2023 Apr 28;12(9).
              doi: 10.3390/cells12091280pubmed: 37174680google scholar: lookup
            3. Fuchs AA, Balne PK, Giuliano EA, Sinha NR, Mohan RR. Evaluation of a novel combination of TRAM-34 and ascorbic acid for the treatment of corneal fibrosis in vivo. PLoS One 2022;17(1):e0262046.
              doi: 10.1371/journal.pone.0262046pubmed: 35007294google scholar: lookup
            4. Gupta S, Sinha NR, Martin LM, Keele LM, Sinha PR, Rodier JT, Landreneau JR, Hesemann NP, Mohan RR. Long-Term Safety and Tolerability of BMP7 and HGF Gene Overexpression in Rabbit Cornea. Transl Vis Sci Technol 2021 Aug 12;10(10):6.
              doi: 10.1167/tvst.10.10.6pubmed: 34383876google scholar: lookup
            5. Amador C, Shah R, Ghiam S, Kramerov AA, Ljubimov AV. Gene Therapy in the Anterior Eye Segment. Curr Gene Ther 2022;22(2):104-131.
              doi: 10.2174/1566523221666210423084233pubmed: 33902406google scholar: lookup
            6. Mohan RR, Martin LM, Sinha NR. Novel insights into gene therapy in the cornea. Exp Eye Res 2021 Jan;202:108361.
              doi: 10.1016/j.exer.2020.108361pubmed: 33212142google scholar: lookup
            7. Mohan RR, Tripathi R, Sharma A, Sinha PR, Giuliano EA, Hesemann NP, Chaurasia SS. Decorin antagonizes corneal fibroblast migration via caveolae-mediated endocytosis of epidermal growth factor receptor. Exp Eye Res 2019 Mar;180:200-207.
              doi: 10.1016/j.exer.2019.01.001pubmed: 30611736google scholar: lookup
            8. Zahir-Jouzdani F, Soleimani M, Mahbod M, Mottaghitalab F, Vakhshite F, Arefian E, Shahhoseini S, Dinarvand R, Atyabi F. Corneal chemical burn treatment through a delivery system consisting of TGF-β(1) siRNA: in vitro and in vivo. Drug Deliv Transl Res 2018 Oct;8(5):1127-1138.
              doi: 10.1007/s13346-018-0546-0pubmed: 29869292google scholar: lookup
            9. Gupta S, Fink MK, Ghosh A, Tripathi R, Sinha PR, Sharma A, Hesemann NP, Chaurasia SS, Giuliano EA, Mohan RR. Novel Combination BMP7 and HGF Gene Therapy Instigates Selective Myofibroblast Apoptosis and Reduces Corneal Haze In Vivo. Invest Ophthalmol Vis Sci 2018 Feb 1;59(2):1045-1057.
              doi: 10.1167/iovs.17-23308pubmed: 29490341google scholar: lookup
            10. Marlo TL, Giuliano EA, Tripathi R, Sharma A, Mohan RR. Altering equine corneal fibroblast differentiation through Smad gene transfer. Vet Ophthalmol 2018 Mar;21(2):132-139.
              doi: 10.1111/vop.12485pubmed: 28685927google scholar: lookup
            11. Marlo TL, Giuliano EA, Sharma A, Mohan RR. Development of a novel ex vivo equine corneal model. Vet Ophthalmol 2017 Jul;20(4):288-293.
              doi: 10.1111/vop.12415pubmed: 27471196google scholar: lookup
            12. Fink MK, Giuliano EA, Tandon A, Mohan RR. Therapeutic potential of Pirfenidone for treating equine corneal scarring. Vet Ophthalmol 2015 May;18(3):242-50.
              doi: 10.1111/vop.12194pubmed: 25041235google scholar: lookup
            13. Young KAS, Schnabel LV, Gilger BC. Cell and Gene Therapy in Equine Ocular Disease. Vet Ophthalmol 2026 Mar;29(2):e70151.
              doi: 10.1111/vop.70151pubmed: 41623202google scholar: lookup
            14. Routh BL, Tripathi R, Giuliano E, Lujin P, Sinha PR, Mohan RR. Anti-fibrotic effects of lisinopril (ACE inhibitor) and fasudil (ROCK inhibitor) in combination for canine corneal fibrosis in vitro. Vet Ophthalmol 2026 Jan;29(1):e13304.
              doi: 10.1111/vop.13304pubmed: 39592228google scholar: lookup
            15. Sorrentino FS, Gardini L, Culiersi C, Fontana L, Musa M, D'Esposito F, Surico PL, Gagliano C, Zeppieri M. Nano-Based Drug Approaches to Proliferative Vitreoretinopathy Instead of Standard Vitreoretinal Surgery. Int J Mol Sci 2024 Aug 9;25(16).
              doi: 10.3390/ijms25168720pubmed: 39201407google scholar: lookup
            16. Posarelli M, Romano D, Tucci D, Giannaccare G, Scorcia V, Taloni A, Pagano L, Borgia A. Ocular-Surface Regeneration Therapies for Eye Disorders: The State of the Art. BioTech (Basel) 2023 Jun 15;12(2).
              doi: 10.3390/biotech12020048pubmed: 37366796google scholar: lookup
            17. Gesteira TF, Verma S, Coulson-Thomas VJ. Small leucine rich proteoglycans: Biology, function and their therapeutic potential in the ocular surface. Ocul Surf 2023 Jul;29:521-536.
              doi: 10.1016/j.jtos.2023.06.013pubmed: 37355022google scholar: lookup
            18. Kumar R, Sinha NR, Mohan RR. Corneal gene therapy: Structural and mechanistic understanding. Ocul Surf 2023 Jul;29:279-297.
              doi: 10.1016/j.jtos.2023.05.007pubmed: 37244594google scholar: lookup
            19. Mohan RR, Kempuraj D, D'Souza S, Ghosh A. Corneal stromal repair and regeneration. Prog Retin Eye Res 2022 Nov;91:101090.
              doi: 10.1016/j.preteyeres.2022.101090pubmed: 35649962google scholar: lookup
            Subscribe to our newsletter
            Join 99,034 horse owners like you who receive our email updates on horse health and nutrition.