Isolation of alkaline phosphatase-positive gingival fibroblasts from patients with chronic inflammatory periodontal disease
Isolation of alkaline phosphatase-positive gingival fibroblasts from patients with chronic inflammatory periodontal disease
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DOI：
10.1111/j.1600-0765.1996.tb00495.x
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发表时间：
1996-05-01
影响因子：
3.5
通讯作者：
Maeda, K
中科院分区：
文献类型：
--
作者：
Abe, T;Hara, Y;Maeda, K
我们最近报道，膜碱性磷酸酶（ALP）活性表达增加是位于慢性炎症性牙周病变中牙龈成纤维细胞的表型特征。为了了解这些细胞的细胞特性，我们从成人牙周炎患者中分离出 ALP 阳性牙龈成纤维细胞，并评估其增殖潜力。使用酶消化程序，我们制备了含有 ALP 阳性成纤维细胞的牙龈细胞悬液，而不影响其 ALP 活性。然后将这些细胞悬液进行 1 g 沉降，然后让细胞粘附到基质上。使用此程序，71.9% 的分离细胞呈 ALP 阳性。通过细胞化学和免疫细胞化学检查 ALP 阳性成纤维细胞的解离和非成纤维细胞污染。通过监测重复传代后 ALP 阳性细胞的比例以及用溴脱氧尿苷 (BrdU) 标记 DNA 合成细胞来评估培养物中 ALP 阳性成纤维细胞的增殖能力。在细胞培养传代过程中，ALP 阳性成纤维细胞的比例下降，而 ALP 阳性表型没有明显变化。 ALP 阳性成纤维细胞中 BrdU 阳性细胞的百分比显着低于 ALP 阴性成纤维细胞。这些结果表明慢性炎症性牙周病变中ALP阳性成纤维细胞的生长潜力较低。我们认为它们体外生长能力的降低反映了体内炎症条件下诱导的更加分化的状态。
We have reported recently that increased expression of membrane alkaline phosphatase (ALP) activity is a phenotypical characteristic of gingival fibroblasts located in chronic inflammatory periodontal lesions. To understand the cellular properties of these cells, we isolated ALP-positive gingival fibroblasts from patients with adult periodontitis and evaluated their proliferative potential. Using an enzymatic digestion procedure, we prepared gingival cell suspensions containing ALP-positive fibroblasts without affecting their ALP activities. These cell suspensions were then subjected to 1 g sedimentation, followed by allowing cells to adhere to substrata, Using this procedure, 71.9% of isolated cells were ALP-positive. Dissociation of ALP-positive fibroblasts and contamination by non-fibroblastic cells were examined by cytochemical and immunocytochemical analyses, The proliferative capacity of ALP-positive fibroblasts in culture was assessed by monitoring the proportion of ALP-positive cells after repeated subculture passages and by labelling DNA-synthesizing cells with bromodeoxyuridine (BrdU). The proportion of ALP-positive fibroblasts decreased during cell culture passages without an apparent change in the ALP-positive phenotype. The percentage of BrdU-positive cells was significantly lower among ALP-positive than among ALP-negative fibroblasts. These results indicate that ALP-positive fibroblasts in chronic inflammatory periodontal lesions have low growth potential. We suggest that their reduced capacity to grow in vitro reflects a more differentiated state induced under inflammatory conditions in vivo.