Möglichkeiten und Grenzen der Diagnose Kupferspeicherkrankheit aus Biopsiematerial der Leber

Abstract

IntroductionThe morphological diagnosis of Wilson's disease cannot be made from the histological section alone and is aided by the quantitative analysis of copper in liver tissues. While, according to Sternlieb et al. (1968) , for this determination the liver tissue must be removed and processed under certain given conditions, in this investigation we examined the question as to whether it is possible to obtain and adequately reliable diagnosis using the tissue removed and fixed in formalin in accordance with conventional techniques, and whether this diagnosis is also possible in residual liver embedded in paraffin for histological examination. Material and methodsLiver tissue, obtained per needle biopsy or with the scalpel was examined with respect to its content of copper using the technique of atomic absorption spectrophotometry. Copper determinations in 107 cadaver livers were evaluated. In each of 10 autopsy cases with morphologically intact livers, a single specimen of liver tissue, removed with the scalpel, and also 2–10 needle biopsies from each right liver lobe were processed a) after simple drying b) after fixation in formalin and drying c) after fixation in formalin and embedding in Autotechnicon as for histological processing, followed by de-paraffining in xylol and acetone and drying. The tissue specimens were subjected to acid hydrolysis and then measured against and aqueous standard solution. In the same manner, the hepatic copper was determined in residual material taken out of paraplast block in 90 morphologically intact liver cylinders of all age groups. This material had been sent to the laboratory for histological examination. The values obtained were compared with cases of Wilson's disease that had been confirmed either clinically or at autopsy. Resultats and discussionThe average copper content in 97 biopsy material samples removed with the scalpel and subjected to multiple determinations was 3.2 mg of copper per 100 g dry weight. The tissue of the biopsy specimens obtained with untreated needles was examined using the same method at a considerably stronger dilution of the measured samples and was found to give 10 mg per 100 g of dry liver tissue. The figures for dry liver tissue fixed in formalin are only slightly higher than those for unfixed tissue. Thus it is established that the demonstration of copper with adequate diagnostic information content is possible even in tissue fixed in formalin. The demonstration of copper in residual material from liver cylinders embedded in paraffin provides no diagnostically useful results. The reason for this is largely to be found in the embedding technique commonly employed at our institute. In the case of biopsy material removed with the scalpel and embedded in paraffin, which has had no opportunity of being contaminated exogenously with copper, the diagnosis of disturbed copper metabolism can be made unequivocally.