Jefferson Institute of Molecular Medicine, Thomas Jefferson University, 233 S. 10th Street, Philadelphia, PA 19107-5541, USA.
Rheumatology (Oxford). 2010 Jun;49(6):1069-75. doi: 10.1093/rheumatology/keq024. Epub 2010 Mar 4.
To obtain an objective, unbiased assessment of skin fibrosis in patients with SSc for use in clinical trials of SSc disease-modifying therapeutics.
Skin biopsies from the dorsal forearm of six patients with diffuse SSc and six healthy controls, and skin biopsies from the forearm of one patient with diffuse SSc before and following 1 year treatment with mycophenolate mofetil were analysed by confocal laser scanning microscopy (CLSM) with specific antibodies against collagen types I and III or fibronectin. The integrated density of fluorescence (IDF) was calculated employing National Institutes of Health-ImageJ software in at least four different fields per biopsy spanning the full dermal thickness.
The intensities of collagen types I and III and fibronectin IDF were 174, 147 and 139% higher in SSc skin than in normal skin, respectively. All differences were statistically significant. The sum of the IDF values obtained for the three proteins yielded a comprehensive fibrosis score. The average fibrosis score for the six SSc samples was 28.3 x 10(6) compared with 18.6 x 10(6) for the six normal skin samples (P < 0.0001). Comparison of skin biopsies obtained from the same SSc patient before treatment and after 12 months of treatment with mycophenolate mofetil showed a reduction of 39% in total fibrosis score after treatment.
CLSM followed by quantitative image analysis provides an objective and unbiased assessment of skin fibrosis in SSc and could be a useful end-point for clinical trials with disease-modifying agents to monitor the response or progression of the disease.
获得一种客观、无偏倚的评估系统性硬化症（SSc）患者皮肤纤维化的方法，用于 SSc 疾病修饰治疗的临床试验。
采用共聚焦激光扫描显微镜（CLSM），使用针对 I 型和 III 型胶原或纤维连接蛋白的特异性抗体，对 6 例弥漫性 SSc 患者和 6 例健康对照者的前臂背侧皮肤活检标本以及 1 例弥漫性 SSc 患者治疗前和治疗 1 年后前臂皮肤活检标本进行分析。采用 NIH-ImageJ 软件，在每个活检标本的至少 4 个不同视野中，对跨越整个真皮厚度的区域进行荧光积分密度（IDF）计算。
SSc 皮肤中 I 型和 III 型胶原以及纤维连接蛋白的 IDF 强度分别比正常皮肤高 174%、147%和 139%，差异均有统计学意义。三种蛋白的 IDF 值之和得出综合纤维化评分，6 例 SSc 样本的平均纤维化评分为 28.3×10^6，6 例正常皮肤样本的平均纤维化评分为 18.6×10^6（P＜0.0001）。对同一 SSc 患者治疗前和治疗 12 个月时的皮肤活检标本进行比较，发现治疗后总纤维化评分降低了 39%。
CLSM 结合定量图像分析为 SSc 皮肤纤维化提供了一种客观、无偏倚的评估方法，可能成为监测疾病反应或进展的疾病修饰药物临床试验的有用终点。
