L. De Franceschi, Ph. D. , L. Roseti, Ph. D. , G. Desando, Ph. D. , A

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A molecular and histological characterization of cartilage from patients with Morquio syndrome L. De Franceschi, Ph.D., L. Roseti, Ph.D., G. Desando, Ph.D., A. Facchini, M.D., B. Grigolo, Ph.D. Osteoarthritis and Cartilage Volume 15, Issue 11, Pages 1311-1317 (November 2007) DOI: 10.1016/j.joca.2007.04.008 Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Real-Time PCR analysis of collagens I and II and aggrecan mRNAs' expression in chondrocytes isolated from patients with Morquio syndrome and from controls. Data were normalized to GAPDH and expressed as mRNA copies/%GAPDH. Osteoarthritis and Cartilage 2007 15, 1311-1317DOI: (10.1016/j.joca.2007.04.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 Representative patient light microscopy evaluation after Safranin-O staining showing a poorly organized morphological structure with larger and not homogeneously distributed cells both in the superficial layer and in the deep zone (A) (bar=200μm) compared to normal hyaline cartilage (B) (bar=320μm). Osteoarthritis and Cartilage 2007 15, 1311-1317DOI: (10.1016/j.joca.2007.04.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Immunostaining for collagen type II in cartilage from a patient with Morquio disease shows a weak positivity which is confined to the extracellular matrix of both the superficial layer (A) (bar=200μm) and the deep zone while the cells are negative (B) (bar=500×). The healthy tissue shows a positivity which is evident both for the extracellular matrix (C) (bar=160μm) and the cells (D) (bar=500×). Collagen type II was developed using new fuchsine (red is positive stain). Osteoarthritis and Cartilage 2007 15, 1311-1317DOI: (10.1016/j.joca.2007.04.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Immunostaining for collagen type I shows a great positivity at cellular level both in the superficial and in the deep zones (A) (bar=200μm). The control tissue is negative (B) (bar=320μm). Collagen type I was developed using new fuchsine (red is positive stain). Osteoarthritis and Cartilage 2007 15, 1311-1317DOI: (10.1016/j.joca.2007.04.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Immunostaining for total proteoglycans is highly positive inside the cells and in the extracellular matrix also at the superficial layer (A) (bar=200μm). The control shows a normal staining and the superficial layer is almost negative (B) (bar=200μm). Proteoglycans were developed using new fuchsine (red is positive stain). Osteoarthritis and Cartilage 2007 15, 1311-1317DOI: (10.1016/j.joca.2007.04.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions

Fig. 6 Immunostaining for keratan sulfate showed a diffuse positivity (A) (bar=200μm), while the control is negative (B) (bar=200μm). The image was acquired under green fluorescence and shows only those nuclei that are positive. Osteoarthritis and Cartilage 2007 15, 1311-1317DOI: (10.1016/j.joca.2007.04.008) Copyright © 2007 Osteoarthritis Research Society International Terms and Conditions