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Esteraseaktivität eines organotypischen humanen Kornea-Konstrukts (HCC) als In-vitro-Modell für Permeationsuntersuchungen

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Esterase activity of human organotypic cornea construct (HCC) as in vitro model for permeation studies

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Zusammenfassung

Organotypische Hornhautäquivalente werden als In-vitro-Modelle für Arzneistoffabsorptionsuntersuchungen verwendet. Viele ophthalmologische Wirkstoffe werden zur Erhöhung der Bioverfügbarkeit als Ester-Prodrugs eingesetzt. Es wurde die Esteraseaktivität von drei kornealen humanen Zelllinien (Epithel-, Stroma- und Endothelzellen) sowie von exzidierter porciner Kornea, humaner Spenderkornea und einem humanen Kornea-Konstrukt (HCC) untersucht und verglichen.

Die Esteraseaktivität von Zellen und Gewebe wurde unter Verwendung von p-Nitrophenylacetat und Hydrocortisonacetat (HCA) als Esterasesubstrate bestimmt. Vergleichende In-vitro-Permeationsuntersuchungen mit HCA mittels modifizierter Franz-Zellen wurden für porcine Korneae, humane Spenderhornhäute und HCC durchgeführt.

Die kornealen Epithelzellen zeigten die höchste Esteraseaktivität mit geringen Unterschieden zu Keratozyten und Endothelzellen. Die kornealen Gewebe wiesen sehr ähnliche Permeationseigenschaften für HCA auf mit leichten Unterschieden in der Metabolisierungsrate (porcine Kornea > HCC > humane Spenderkornea).

Permeations- und Metabolisierungsuntersuchungen zeigen, dass das In-vitro-Permeationsmodell HCC in der Lage ist, das Ester-Prodrug Hydrocortisonacetat adäquat in Hydrocortison umzusetzen.

Abstract

Organotypic cornea equivalents are used as in vitro models for permeation studies. Many ophthalmic drugs are applied as ester prodrugs to achieve a higher bioavailability. The esterase activity of three corneal human cell lines (epithelial, stromal, endothelial cells) as well as of excised porcine cornea, human donor cornea and human cornea construct (HCC) was investigated and compared.

Esterase activity was determined using p-nitrophenyl acetate and hydrocortisone acetate (HCA) as esterase substrates. Hydrocortisone acetate permeation across porcine cornea, human donor cornea and HCC was studied in vitro using Franz-diffusion cells.

Corneal epithelial cells showed the highest esterase activity and only small differences to keratocytes and endothelial cells were detectable. The permeation barrier properties of the different corneal tissues were very similar in the case of HCA permeation whereas HCA metabolism rates were in the ranking order of porcine cornea > HCC > human donor cornea.

Permeation and metabolism studies indicate that the in vitro permeation model HCC is able to adequately convert hydrocortisone acetate to hydrocortisone.

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Danksagung

Wir danken der Bundesforschungsanstalt für Landwirtschaft, D-Braunschweig, für die Unterstützung mit porcinem Korneamaterial sowie Fa. Nestec (Nestec Ltd., Nestlé Research Center Lausanne) für die Überlassung der CEPI-Zelllinie.

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Der korrespondierende Autor versichert, dass keine Verbindungen mit einer Firma, deren Produkt in dem Artikel genannt ist, oder einer Firma, die ein Konkurrenzprodukt vertreibt, bestehen.

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Authors and Affiliations

  1. Institut für Pharmazeutische Technologie der Technischen Universität Braunschweig

    L. Meyer, C. C. Müller-Goymann &  S. Reichl

  2. Klinik und Poliklinik für Augenheilkunde des Universitätsklinikums Hamburg Eppendorf

    J. Bednarz

  3. Institut für Pharmazeutische Technologie, TU Braunschweig, Mendelssohnstr. 1, 38106 , Braunschweig

    S. Reichl

Authors
  1. L. Meyer
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  2. J. Bednarz
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  3. C. C. Müller-Goymann
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  4. S. Reichl
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Corresponding author

Correspondence to S. Reichl.

Additional information

Teile des Artikels wurden auf der Jahrestagung der Deutschen Pharmazeutischen Gesellschaft DPhG 2004 in Regensburg präsentiert.

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Meyer, L., Bednarz, J., Müller-Goymann, C.C. et al. Esteraseaktivität eines organotypischen humanen Kornea-Konstrukts (HCC) als In-vitro-Modell für Permeationsuntersuchungen. Ophthalmologe 102, 971–980 (2005). https://doi.org/10.1007/s00347-005-1200-z

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  • DOI: https://doi.org/10.1007/s00347-005-1200-z

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