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Evaluation of antioxiodant drugs for the treatment of ochronotic alkaptonuria in an in vitro human cell model
Author(s) -
Tinti Laura,
Spreafico Adriano,
Braconi Daniela,
Millucci Lia,
Bernardini Giulia,
Chellini Federico,
Cavallo Giovanni,
Selvi Enrico,
Galeazzi Mauro,
Marcolongo Roberto,
Gallagher James A.,
Santucci Annalisa
Publication year - 2010
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.22199
Subject(s) - alkaptonuria , homogentisic acid , in vitro , ochronosis , viability assay , chemistry , ascorbic acid , chondrocyte , pharmacology , apoptosis , in vivo , biochemistry , medicine , biology , pathology , microbiology and biotechnology , food science
Alkaptonuria (AKU) is a rare autosomal recessive disease, associated with deficiency of homogentisate 1,2‐dioxygenase activity in the liver. This leads to an accumulation of homogentisic acid (HGA) and its oxidized derivatives in polymerized form in connective tissues especially in joints. Currently, AKU lacks an appropriate therapy. Hence, we propose a new treatment for AKU using the antioxidant N ‐acetylcysteine (NAC) administered in combinations with ascorbic acid (ASC) since it has been proven that NAC counteracts the side‐effects of ASC. We established an in vitro cell model using human articular primary chondrocytes challenged with an excess of HGA (0.33 mM). We used this experimental model to undertake pre‐clinical testing of potential antioxidative therapies for AKU, evaluating apoptosis, viability, proliferation, and metabolism of chondrocytes exposed to HGA and treated with NAC and ASC administered alone or in combination addition of both. NAC decreased apoptosis induced in chondrocytes by HGA, increased chondrocyte growth reduced by HGA, and partially restored proteoglycan release inhibited by HGA. A significantly improvement in efficacy was found with combined addition of the two antioxidants in comparison with NAC and ASC alone. Our novel in vitro AKU model allowed us to demonstrate the efficacy of the co‐administration of NAC and ASC to counteract the negative effects of HGA for the treatment of ochronotic arthropathy. J. Cell. Physiol. 225: 84–91, 2010. © 2010 Wiley‐Liss, Inc.