Histopathology 2008, 52, 877–884. DOI: 10.1111/j.1365-2559.2008.03051.x
Detection of spirochaetal microorganisms by focus floating microscopy in necrobiosis lipoidica in patients from central Europe K Eisendle, M Baltaci, H Kutzner1 & B Zelger Department of Dermatology and Venerology, Innsbruck Medical University, Innsbruck, Austria, and 1Dermatohistological Private Laboratory, Friedrichshafen, Germany Date of submission 6 September 2007 Accepted for publication 11 January 2008
Eisendle K, Baltaci M, Kutzner H & Zelger B (2008) Histopathology 52, 877–884
Detection of spirochaetal microorganisms by focus floating microscopy in necrobiosis lipoidica in patients from central Europe Aims: Necrobiosis lipoidica (NL) is a chronic inflammatory skin disease with unknown aetiology. The aim was to determine the presence of spirochaetal microorganisms in NL. Methods and results: Focus-floating microscopy (FFM) is a modified immunohistochemical technique that was developed to detect borrelial spirochaetes within tissue sections. It has proven to be more sensitive for the detection of spirochaetes than polymerase chain reaction (PCR). Fifty-six cases of NL as well as 44 negative and 33 positive controls were investigated for the presence of Borrelia within tissue specimens. Using FFM, Borrelia could be detected in 42 cases
(75.0%) and were seen significantly more often in histologically active inflammatory-rich (38 ⁄ 41, 92.7%) than in inflammatory-poor (4 ⁄ 15, 26.7%) cases of NL (P < 0.001). Seven cases investigated with a Borreliaspecific PCR (23s-RNA) remained negative. In contrast, FFM was positive in 30 of 33 (90.9%) positive controls of acrodermatitis chronica atrophicans and 15 of the positive controls (45.5%) were also positive with PCR, whereas no negative controls revealed any microorganisms. Conclusions: Detection of spirochaetes in NL points to a specific involvement of B. burgdorferi or other similar strains in the development of or trigger for this disease.
Keywords: Borrelia burgdorferi, focus-floating microscopy, immunhistochemistry, Lyme disease, necrobiosis lipoidica, PCR Abbreviations: ACA, acrodermatitis chronica atrophicans; FFM, focus-floating microscopy; NL, necrobiosis lipoidica; Osp, outer surface protein; PCR, polymerase chain reaction
Introduction Necrobiosis lipoidica (NL) is an inflammatory skin disorder of unknown cause, occurring three times more often in women than in men. The indurated, sometimes ulcerated, yellow–red–brown plaques and nodules mainly involve the lower extremities (Figure 1A), and much less frequently other localizations such as the Address for correspondence: Klaus Eisendle, Clinical Department of Dermatology and Venerology, Innsbruck Medical University, Anichstr 35, A-6020 Innsbruck, Austria. e-mail:
[email protected];
[email protected] 2008 The Authors. Journal compilation 2008 Blackwell Publishing Limited.
trunk or upper extremities. The name necrobiosis refers to the type of inflammation that occurs when lesions are examined histologically, namely a diffuse dermal to subcutaneous infiltrate of variably xanthomatized macrophages, fibroblasts and occasional giant cells surrounding a zone of rather amorphous disordered, degenerate collagen plus fibrin and mucin (including necrosis of cellular components) seen besides lymphocytes and plasma cells (Figure 1B,C). Lipoidica reflects the yellow appearance of lesions due to lipid deposits occurring secondary to inflammation and due to atrophy, allowing the subcutaneous fat to be better seen.1,2
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Figure 1. Necrobiosis lipoidica. A, Necrobiosis lipoidica on both shins reveals well-demarcated, irregularly shaped, indurated plaques with central atrophy, erosion and yellow pigmentation. B–C, Histopathology of necrobiosis lipoidica at different magnifications shows infiltrate of lymphocytes, plasma cells and macrophages as well as fibrosclerotic (‘necrobiotic’) degeneration of collagen with increased numbers of fibrocytes involving the full thickness of the dermis with extension into the subcutis (H&E).
NL can be associated with diabetes mellitus, although
