Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy

Natasha Josifovska, Dóra Júlia Szabó, Richárd Nagymihály, Zoltán Veréb, Andrea Facskó, Ketil Eriksen, Morten C. Moe, G. Petrovski

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Purpose Development of ex vivo model to study pathogenesis, inflammation and treatment modalities for pterygium. Methods Pterygium obtained from surgery was cultivated (3 months). Gravitational attachment method using viscoelastic facilitated adherence of graft and outgrowing cells. Medium contained serum as the only growth supplement with no use of scaffolds. Surface profiling of the multi-layered cells for hematopoietic- and mesenchymal stem cell markers was performed. Examination of cells by immunohistochemistry using pluripotency, oxidative stress, stemness, migration and proliferation, epithelial and secretory markers was performed. The effect of anti-proliferative agent Mitomycin C upon secretion of pro-inflammatory cytokines IL-6 and IL-8 was assessed. Results Cells showed high expression of migration- (CXCR4), secretory- (MUC1, MUC4) and oxidative damage- (8-OHdG) markers, and low expression of hypoxia- (HIF-1α) and proliferation- (Ki-67) markers. Moderate and low expression of the pluripotency markers (Vimentin and ΔNp63) was present, respectively, while the putative markers of stemness (Sox2, Oct4, ABCG-2) and epithelial cell markers- (CK19, CK8-18) were weak. The surface marker profile of the outgrowing cells revealed high expression of the hematopoietic marker CD47, mesenchymal markers CD90 and CD73, minor or less positivity for the hematopoietic marker CD34, mesenchymal marker CD105, progenitor marker CD117 and attachment protein markers while low levels of IL-6 and IL-8 secretion ex vivo, were inhibited upon Mitomycin C treatment. Conclusion Ex vivo tissue engineered pterygium consists of a mixture of cells of different lineage origin, suitable for use as a disease model for studying pathogenesis ex vivo, while opening possibilities for new treatment and prevention modalities.

Original languageEnglish
Pages (from-to)283-292
Number of pages10
JournalContact Lens and Anterior Eye
Volume40
Issue number5
DOIs
Publication statusPublished - Oct 1 2017

Fingerprint

Pterygium
Mitomycin
Interleukin-8
Interleukin-6
Therapeutics
Vimentin
Cell Lineage
Hematopoietic Stem Cells
Mesenchymal Stromal Cells
Oxidative Stress
Epithelial Cells
Immunohistochemistry
Cytokines
Inflammation
Transplants
Growth
Serum
Proteins

Keywords

  • IL-6
  • IL-8
  • Long-term cultures
  • Mitomycin C
  • Pterygium
  • Tissue engineering

ASJC Scopus subject areas

  • Ophthalmology
  • Optometry

Cite this

Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy. / Josifovska, Natasha; Szabó, Dóra Júlia; Nagymihály, Richárd; Veréb, Zoltán; Facskó, Andrea; Eriksen, Ketil; Moe, Morten C.; Petrovski, G.

In: Contact Lens and Anterior Eye, Vol. 40, No. 5, 01.10.2017, p. 283-292.

Research output: Contribution to journalArticle

Josifovska, N, Szabó, DJ, Nagymihály, R, Veréb, Z, Facskó, A, Eriksen, K, Moe, MC & Petrovski, G 2017, 'Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy', Contact Lens and Anterior Eye, vol. 40, no. 5, pp. 283-292. https://doi.org/10.1016/j.clae.2017.04.002
Josifovska, Natasha ; Szabó, Dóra Júlia ; Nagymihály, Richárd ; Veréb, Zoltán ; Facskó, Andrea ; Eriksen, Ketil ; Moe, Morten C. ; Petrovski, G. / Cultivation and characterization of pterygium as an ex vivo study model for disease and therapy. In: Contact Lens and Anterior Eye. 2017 ; Vol. 40, No. 5. pp. 283-292.
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AU - Szabó, Dóra Júlia

AU - Nagymihály, Richárd

AU - Veréb, Zoltán

AU - Facskó, Andrea

AU - Eriksen, Ketil

AU - Moe, Morten C.

AU - Petrovski, G.

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N2 - Purpose Development of ex vivo model to study pathogenesis, inflammation and treatment modalities for pterygium. Methods Pterygium obtained from surgery was cultivated (3 months). Gravitational attachment method using viscoelastic facilitated adherence of graft and outgrowing cells. Medium contained serum as the only growth supplement with no use of scaffolds. Surface profiling of the multi-layered cells for hematopoietic- and mesenchymal stem cell markers was performed. Examination of cells by immunohistochemistry using pluripotency, oxidative stress, stemness, migration and proliferation, epithelial and secretory markers was performed. The effect of anti-proliferative agent Mitomycin C upon secretion of pro-inflammatory cytokines IL-6 and IL-8 was assessed. Results Cells showed high expression of migration- (CXCR4), secretory- (MUC1, MUC4) and oxidative damage- (8-OHdG) markers, and low expression of hypoxia- (HIF-1α) and proliferation- (Ki-67) markers. Moderate and low expression of the pluripotency markers (Vimentin and ΔNp63) was present, respectively, while the putative markers of stemness (Sox2, Oct4, ABCG-2) and epithelial cell markers- (CK19, CK8-18) were weak. The surface marker profile of the outgrowing cells revealed high expression of the hematopoietic marker CD47, mesenchymal markers CD90 and CD73, minor or less positivity for the hematopoietic marker CD34, mesenchymal marker CD105, progenitor marker CD117 and attachment protein markers while low levels of IL-6 and IL-8 secretion ex vivo, were inhibited upon Mitomycin C treatment. Conclusion Ex vivo tissue engineered pterygium consists of a mixture of cells of different lineage origin, suitable for use as a disease model for studying pathogenesis ex vivo, while opening possibilities for new treatment and prevention modalities.

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