Evaluation of hbeta;D-1 and hbeta; D-2 levels in saliva of patients with oral mucosal diseases / Evaluación de los niveles de hbeta; D-1 y hbeta;D-2 en la Saliva de pacientes con enfermedades de la mucosa oral

OBJECTIVE: This study aimed to determine a possible correlation between oral mucosal disease and salivary concentrations of the antimicrobial peptides human beta-defensin-1 (hβD-1) and human betadefensin- 2 (hβD-2). METHOD: The present work focussed on the establishment of a reversed phase-high performance liquid chromatography (RP-HPLC) procedure to quantify human beta-defensins (hβD-1 and hβD-2) in saliva samples of patients with oral diseases such as lichen planus (n = 10), Behçet (n = 10) and recurrent apthous stomatitis (n = 10). RESULTS: Linear calibration range for hβD-1 and hβD-2 defensins was 1.67−200 µg mL-1 and 3.13− 100 µg mL-1 with R2 values of 0.9998 and 0.996, correspondingly. The concentration of beta-defensins in saliva was determined by comparing the peak areas of eluted hβD-1 and hβD-2 with that of their standards. The variation of the amount of beta-defensins was evaluated by comparisons of the results obtained from the patients with oral mucosal diseases before and after treatments and the control subjects. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 1.62 µg mL- 1 and 5.39 µg mL-1 for hβD-1 and 0.94 µg mL-1 and 3.13 µg mL-1 for hβD-2, respectively. CONCLUSION: The salivary beta-defensin concentration was significantly higher in patients with oral mucosal diseases than in healthy volunteers; furthermore, in patients with oral mucosal diseases, the concentration was significantly higher before treatment than after treatment.

OBJETIVO: Este estudio tuvo por objeto determinar una posible correlación entre la enfermedad de la mucosa oral y las concentraciones salivales de la beta-defensina humana 1 (hβD-1) y la beta-defensina humana 2 (hβD-2) de los péptidos antimicrobianos. MÉTODO: El presente trabajo estuvo encaminado al establecimiento de un procedimiento de cromatografía líquida de alta eficacia de fase reversa (RP-HPLC) para cuantificar las beta-defensinas humanas (hβD-1 y hβD-2) en muestras de saliva de pacientes con enfermedades orales como el liquen plano (n = 10), Behçet (n = 10), y la estomatitis aftosa recurrente (n = 10). RESULTADOS: El rango de calibración lineal de las defensinas hβD-1 y hβD-2 fue 1.67-200 µg mL-1 y 3.13-100 µg mL-1 con valores R2 de 0.9998 y 996, respectivamente. La concentración de beta-defensinas en la saliva se determinó utilizando el área de sus estándares. La variación de la cantidad de beta defensinas fue evaluada por comparaciones de los resultados obtenidos de los pacientes con enfermedades de la mucosa oral, antes y después de los tratamientos y los sujetos de control. Se halló que el límite de detección (LDD) y el límite de cuantificación (LDC) fueron 1.62 µg mL-1 y 5.39 µg mL- 1 para hβD-1 y 0.94 µg mL-1 y 3.13 µg mL-1 hβD-2, respectivamente. CONCLUSIÓN: La concentración de beta-defensina salival fue significativamente mayor en los pacientes con enfermedades de la mucosa oral que en los voluntarios sanos. Además, en pacientes con enfermedades de la mucosa oral, la concentración fue significativamente mayor antes del tratamiento que después del tratamiento.

Direct Immunofluorescence in Behcet's Disease: A Controlled Study with 108 Cases

PURPOSE: Behcet's disease (BD) is a disease of unknown etiology, which has multisystemic involvement. This multisystemic involvement might be the clue for an autoimmune pathogenesis. In order to evaluate an autoimmune pathogenesis, we examined immunoreactans depositions in the skin of BD patients. MATERIALS AND METHODS: The skin samples of 108 BD patients (28 perilesional skin, 44 positive pathergy test site, 22 negative pathergy test site, 14 normal skin) were examined for the depositions of immunoglobulin (Ig)M, IgG, IgA, complement 3 (C3), and fibrinogen (F) using direct immunofluorescence (DIF). The data were statistically compared to the DIF of 36 systemic lupus erythematosus (SLE) patients and 20 healthy controls using chi-square Fisher exact test. RESULTS: Highly significant immunoreactans depositions were obtained in BD (deposition rates: IgM 70.3%, IgG 0%, IgA 20.3%, C3 62.9%, F 83.3%). The comparison with SLE revealed no differences in IgM, IgA, and C3. However, IgG deposition was higher in SLE while F deposition was higher in BD. In both BD and SLE, the Ig depositions were highly significant when the data were compared with the healthy controls. CONCLUSION: The significant deposition of immunoreactans in BD, especially in the negative pathergy and the normal skin sites, were observed. This study is the first controlled study revealing positive Ig depositions in BD, and it is expected to help us to reconsider the autoimmune pathogenesis in BD.

Direct Immunofluorescence in Behcet's Disease: A Controlled Study with 108 Cases

PURPOSE: Behcet's disease (BD) is a disease of unknown etiology, which has multisystemic involvement. This multisystemic involvement might be the clue for an autoimmune pathogenesis. In order to evaluate an autoimmune pathogenesis, we examined immunoreactans depositions in the skin of BD patients. MATERIALS AND METHODS: The skin samples of 108 BD patients (28 perilesional skin, 44 positive pathergy test site, 22 negative pathergy test site, 14 normal skin) were examined for the depositions of immunoglobulin (Ig)M, IgG, IgA, complement 3 (C3), and fibrinogen (F) using direct immunofluorescence (DIF). The data were statistically compared to the DIF of 36 systemic lupus erythematosus (SLE) patients and 20 healthy controls using chi-square Fisher exact test. RESULTS: Highly significant immunoreactans depositions were obtained in BD (deposition rates: IgM 70.3%, IgG 0%, IgA 20.3%, C3 62.9%, F 83.3%). The comparison with SLE revealed no differences in IgM, IgA, and C3. However, IgG deposition was higher in SLE while F deposition was higher in BD. In both BD and SLE, the Ig depositions were highly significant when the data were compared with the healthy controls. CONCLUSION: The significant deposition of immunoreactans in BD, especially in the negative pathergy and the normal skin sites, were observed. This study is the first controlled study revealing positive Ig depositions in BD, and it is expected to help us to reconsider the autoimmune pathogenesis in BD.