Classical osteoblastoma, atypical osteoblastoma, and osteosarcoma: a comparative study based on clinical, histological, and biological parameters

OBJECTIVE: To investigate the biological behavior of classical and atypical osteoblastomas in comparison to osteosarcomas. METHODS: Based on histological parameters, 30 osteoblastomas were subclassified as classical osteoblastomas (23/30) or atypical osteoblastoma (high cellularity, prominent blue osteoid, and epithelioid osteoblasts-7/30). Comparative immunohistochemical and clinical analysis was performed in 17 cases of patients with high-grade osteosarcoma. Formalin-fixed, paraffin-embedded archival tissue was immunostained for p53 and proliferation cell nuclear antigen. Tumors with positive p53 stain underwent molecular analyses for fragments of exon 10. RESULTS: The mean proliferating cell nuclear antigen indexes for classical osteoblastoma, atypical osteoblastoma, and osteosarcoma were 33 percent, 61 percent, and 79 percent, respectively. The atypical subgroup showed similar results to those of the osteosarcoma group (P < 0.001). p53 protein was detected in 4 (13 percent) osteoblastomas (3 of these were atypical osteoblastoma), and 4 osteosarcomas (23 percent) also showed p53 positivity. DNA mutation performed in p53-positive cases was confirmed in exon 10 in 2 atypical osteoblastomas (2/3), 1 classical osteoblastoma (1/1), and 1 osteosarcoma (1/4). Disease recurrence was correlated with p53 expression (P = 0.009), atypical subtype (P = 0.031), spiculated blue bone on histology (P = 0.018), and proliferatingcell nuclear antigen labeling index > 40 (P = 0.015). CONCLUSION: These results validate atypical osteoblastoma as an entity, with p53 and proliferation cell nuclear antigen immunoexpression closer to that of osteosarcoma than of classical osteoblastoma. Proliferating cell nuclear antigen labeling index and p53 may be useful predictors of recurrence.

OBJETIVOS: Investigar o comportamento biológico de osteoblastomas clássicos e atípicos comparados com osteossarcomas. MÉTODOS: Com base em parâmetros histológicos classificamos um grupo de 30 osteoblastomas nos subgrupos de osteoblastomas clássicos (23/30) e de osteoblastomas atípicos (que apresentam como característica grande celularidade, osteóide azul proeminente e osteoblastos epitelióide-7/30). Como efeito de comparação dos resultados imunohistoquímicos e análise clínica, avaliamos 17 pacientes com osteosarcoma de grau avançado. Os cortes histológicos com bloco de parafina fixado em formalina foram imunocorados para p53 e antígeno nuclear de célula em proliferação. Tumores com coloração positiva para p53 tiveram análise molecular para fragmentos do exon 10. RESULTADOS: O índice médio de antígeno nuclear de célula em proliferação para osteoblastoma clássico, osteoblastoma atípico e osteosarcoma foram de 33 por cento, 61 por cento e 79 por cento, respectivamente. O subgrupo atípico demonstrou resultados similares aos dos osteosarcomas (p<0,001). Foram detectadas proteína p53 em 4 (13 por cento) osteoblastomas; 3 desses foram osteoblastomas atípicos, sendo que 4 osteosarcomas (23 por cento) também demonstraram p53 positivo. A mutação do DNA nos casos positivos de p53 foi confirmada no exon 10 em dois osteoblastomas atípicos (2/3), um osteoblastoma clássico (1/1) e um osteosarcoma (1/4). A recorrência da doença foi correlacionada com a expressão do p53 (p=0,009), subtipo atípico (p=0,031), osso azul espiculado no resultado da histologia (p=0,018), e índice de marcação pelo antígeno nuclear de célula em proliferação > 40 (p=0,015). CONCLUSÃO: Esses resultados validam os osteoblastomas atípicos como entidade real, com imunoexpressão das proteínas p53 e antígeno nuclear de célula em proliferação mais perto do osteosarcoma do que do osteoblastoma clássico. O índice de marcação pelo antígeno nuclear de célula em proliferação e o p53 podem...

Non-radiometric immunoassays [fluoroimmunoassay (FIA) and fluorometric enzyme immunoassay (FEIA) with radioimmunoassay (RIA) for evaluation of adrenal function in normal and hypercortisolemic dogs

Non-radiometric immunoassays offer many advantages over radiometric assays, such as higher stability of kit compounds and absence of potential hazardous effects for users and environment. The comparison of cortisol measurements by fluoroimmunoassay (FIA) and fluorometric enzyme immunoassay (FEIA) with radioimmunoassay (RIA) in adrenal function evaluation of normal (n=50) and hypercortisolemic dogs (n=12) was proposed. Serum concentrations of cortisol were measured in basal conditions and 8 hours after dexamethasone (DEX) suppression (0.01mg/kg/IV). All our reference values were based on the 5th and 95th percentile. The values for basal cortisol of healthy dogs were 0.20 to 2.35mg/d for FIA, 0.30 to 5.39mg/d for FEIA, and 0.65 to 4.64mg/d for RIA. After DEX suppression the values were <0.87mg/d, <0.30mg/d and < 0.80mg/d for FIA, FEIA and RIA, respectively. In hypercortisolemic dogs, the values of cortisol (mean ± SD) in basal and post-DEX conditions were 2.71 + 0.41mg/d and 1.73 + 1.15mg/d for FIA, 7.05 + 2.85mg/d and 4.93 + 2.26mg/d for FEIA, and 4.80 + 1.43mg/d and 3.52 + 1.08mg/d for RIA. Statistically significant differences (p<0.05) between the normal and the hypercortisolemic groups (Kruskal-Wallis test) were observed in the three methods, and between basal and post-DEX values (Wilcoxon test) using RIA and FEIA methods but not with FIA. Cortisol determinations by FEIA and RIA methods at DEX suppression test showed 100% of sensitivity and specificity for the diagnosis of hyperadrenocorticism in dogs. The results demonstrate that serum cortisol concentrations measurements by FEIA is a suitable alternative to the traditional RIA method for adrenal function evaluation in dogs.