凝血酶对培养的人牙周韧带成纤维细胞生长、蛋白质合成、附着、聚集及碱性磷酸酶活性的影响。

C P Chan, C P Lin, M C Chang, C C Hsieh, C C Hsu, C L Lin, J H Jeng
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摘要

先前的研究表明凝血酶可以激活髓细胞,包括成纤维细胞。由于牙髓细胞和牙周韧带(PDL)成纤维细胞可以表达凝血酶受体mRNA,因此本研究的具体目的是确定凝血酶是否可以激活培养的人PDL成纤维细胞的生长、附着、蛋白质合成、碱性磷酸酶活性和细胞聚集。凝血酶可以刺激PDL成纤维细胞的生长,并呈剂量依赖性(MTT法分析)。凝血酶浓度为5和10 U/ml时,培养5 d后,凝血酶使细胞数量分别比对照增加141%和153%。凝血酶(5-20 U/ml)也使蛋白质合成率(以[3H]脯氨酸掺入测定)达到对照的1.88-2.13倍。然而,用凝血酶(1 ~ 20 U/ml)预处理PDL成纤维细胞不能促进PDL成纤维细胞对I型胶原和纤维连接蛋白的附着。凝血酶在5 ~ 20 U/ml浓度范围内可诱导成纤维细胞聚集。而凝血酶(1 ~ 20 U/ml)对细胞碱性磷酸酶活性既无刺激作用,也无抑制作用。总之,凝血酶的存在似乎在细胞生长、蛋白质合成和细胞聚集方面对PDL成纤维细胞有影响。这提示凝血酶可能在牙周手术后牙周组织的早期愈合过程中起重要作用。
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Effects of thrombin on the growth, protein synthesis, attachment, clustering and alkaline phosphatase activity of cultured human periodontal ligament fibroblasts.

Previous studies have indicated that thrombin can activate pulp cells, including fibroblasts. Because pulp cells and periodontal ligament (PDL) fibroblasts can express thrombin receptor mRNA, the specific aim of this study was to determine whether thrombin can activate the growth, attachment, protein synthesis, alkaline phosphatase activities and cellular clustering of cultured human PDL fibroblasts. Thrombin can stimulate the growth of PDL fibroblasts in a dose dependent manner (as analyzed by MTT assay). At concentrations of 5 and 10 U/ml, thrombin increased the cell numbers to 141% and 153% greater than that of the control after 5 days of incubation, respectively. Thrombin (5-20 U/ml) also stimulated the protein synthesis rate (assayed by [3H]proline incorporation) to 1.88-2.13 fold that of the control. However, pretreatment of PDL fibroblasts with thrombin (1-20 U/ml) could not promote the attachment of PDL fibroblasts to type I collagen and fibronectin. Moreover, thrombin could induce clustering of PDL fibroblasts within a concentration range of 5-20 U/ml. However, thrombin (1-20 U/ml) exerted neither stimulatory nor inhibitory effect on cellular alkaline phosphatase activities. In conclusion, it appears that the presence of thrombin seems to have effects on PDL fibroblasts in terms of cell growth, protein synthesis and cell clustering. This suggests that thrombin might be important in the early healing process of periodontium following periodontal surgery.

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