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The Utility of Alizarin Red S Staining in Calcium Pyrophosphate Dihydrate Crystal Deposition Disease
KOJI YAMAKAWA, HIROSHI IWASAKI, IKUKO MASUDA, YUKO OHJIMI, ITSUO HONDA, KAZUHIKO SAEKI, JINGFAN ZHANG, EISUKE SHONO, MASATOSHI NAITO, and MASAHIRO KIKUCHI
ABSTRACT. Methods. Paraffin sections of CPPD crystal-bearing tissues of 31 patients were stained with hematoxylin and eosin (H&E) and Alizarin red S (ARS). For H&E, the sections were treated with Mayer's hematoxylin (pH 2.3) for 5 min and with eosin alcohol (pH 4.1) for 1 min. For ARS, 1% ARS dissolved in distilled water was adjusted to pH 6.4 by adding 0.1% ammonia solution drop by drop while stirring. As controls, unstained sections were soaked in 1% citric acid monohydrate solution (CAMS, pH 2.3) for 5 or 10 min. The histological preparations were examined under a compensated polarized light using a first-order red compensator. We counted the number of weakly positive birefringent CPPD crystals in 3 high power fields (HPF, 0.272 mm2). Results. CPPD crystals were seen clearly in most specimens stained with ARS, but were markedly reduced in tissue sections stained with H&E or CAMS. The number of CPPD crystals detected in sections stained by ARS (1723 ± 683 per 3 HPF, mean ± standard deviation) was significantly higher compared with H&E, CAMS (5 min), and CAMS (10 min) (401 ± 374, 1022 ± 616, and 494 ± 636 per 3 HPF, respectively; p < 0.001, each). Conclusion. Standard H&E staining reduces the number of visible CPPD crystals, probably due to the strong acidity of both hematoxylin and eosin solutions, whereas the ARS stain seems to preserve a large number of CPPD crystals. The utility of ARS staining may improve the identification of CPPD crystals and contribute to a correct diagnosis of CPPD crystal deposition. (J Rheumatol 2003;30:1032-5) Key Indexing Terms:
CALCIUM PYROPHOSPHATE DIHYDRATE CRYSTAL DEPOSITION DISEASE
From the Departments of Pathology and Orthopedic Surgery, Fukuoka University School of Medicine, Fukuoka, Japan; the Department of Medicine, Division of Rheumatology, Medical College of Wisconsin, Milwaukee, Wisconsin, USA; and the Department of Pathology, Kumamoto Orthopedic Hospital, Kumamoto, Japan. Dr. Masuda's work was supported by NIH grant AR44862. K. Yamakawa, MD, PhD; Y. Ohjimi, MD, PhD, Instructor; H. Iwasaki, MD, PhD, Chief Professor; M. Kikuchi, MD, PhD, Chief Professor, Department of Pathology, Fukuoka University School of Medicine; K. Saeki, MD, PhD, Instructor; J. Zhang, MD, PhD, Instructor; E. Shono, MD, PhD, Instructor; M. Naito, MD, PhD, Chief Professor, Department of Orthopedic Surgery, Fukuoka University School of Medicine; I. Masuda, MD, PhD, Assistant Professor, Department of Medicine, Division of Rheumatology, Medical College of Wisconsin; I. Honda, MD, PhD, Chief, Department of Pathology, Kumamoto Orthopedic Hospital. Address reprint requests to Dr. K. Yamakawa, Department of Pathology, Fukuoka University School of Medicine, Nanakuma 7-45-1, Jonan-ku, Fukuoka 814-0180, Japan. E-mail: 6789@d3.dion.ne.jp Submitted June 27, 2002; revision accepted October 4, 2002. |