International journal of oral biology : official journal of the Korean Academy of Oral Biology and the UCLA Dental Research Institute最新文献

英文中文

Human β-defensin-2 gene transduction of dental pulp cells: A model for pulp antimicrobial gene therapy. 牙髓细胞的人类 β防御素-2 基因转导：牙髓抗菌基因治疗模型。

International journal of oral biology : official journal of the Korean Academy of Oral Biology and the UCLA Dental Research Institute

Pub Date : 2004-01-01

George T-J Huang, Hai-Bo Zhang, Chunyi Yin, Sang Hyuk Park

The objective of this study was to determine whether cells from human pulp can be transduced to express the antimicrobial peptide--human β-defensin-2 (HBD-2). Primary human pulp cells and gingival fibroblasts from normal tissue, as well as two mouse cell lines (NIH 3T3 and AT-84) and a human cell line SCC-9 were transduced with a retroviral vector carrying HBD-2 cDNA. ELISA and Northern blot analyses were performed to detect HBD-2 expression by these transduced cells. Antimicrobail assays using recombinant HBD-2 were performed on two caries-associated bacteria Streptococcus mutnas and Lactobacillus acidophilus. The results showed that transduced pulp cells secreted 62.4 ± 27.2 ng/3 days of HBD-2, which was comparable to that by NIH 3T3 (78.0 ± 14.1 ng/4 days), and higher than those by gingival fibroblasts (17.9 ± 7.9 ng/3 days), AT-84 (2.6 ± 1.0 ng/3 days), and SCC-9 (47.6 ± 9.9 ng/3 days). Northern blot analysis showed that the levels of HBD-2 mRNA expression correlated with their protein secretion levels. There was approximately 50% reduction of growth when S. mutans and L. acidophilus were exposed to HBD-2 at 1 µM. Pulp cells appear to be suitable for HBD-2 transduction using retroviral vectors, suggesting a potential for use in controlling pulpal infections.

本研究的目的是确定人类牙髓细胞能否转导表达抗菌肽--人β防御素-2（HBD-2）。研究人员用携带 HBD-2 cDNA 的逆转录病毒载体转导了原代人类牙髓细胞、正常组织的牙龈成纤维细胞、两种小鼠细胞系（NIH 3T3 和 AT-84）以及人类细胞系 SCC-9。为检测这些转导细胞的 HBD-2 表达，进行了 ELISA 和 Northern 印迹分析。使用重组 HBD-2 对两种龋齿相关细菌--突变链球菌和嗜酸乳杆菌进行了抗菌试验。结果显示，转导的牙髓细胞在 62.4 ± 27.2 纳克/3 天内分泌出 HBD-2，与 NIH 3T3 的分泌量（78.0 ± 14.1 纳克/4 天）相当，高于牙龈成纤维细胞（17.9 ± 7.9 纳克/3 天）、AT-84（2.6 ± 1.0 纳克/3 天）和 SCC-9（47.6 ± 9.9 纳克/3 天）。Northern 印迹分析表明，HBD-2 mRNA 的表达水平与其蛋白质分泌水平相关。当突变酵母菌和嗜酸乳杆菌暴露于 1 µM 的 HBD-2 时，其生长速度降低了约 50%。牙髓细胞似乎适合使用逆转录病毒载体进行 HBD-2 转导，这表明它有可能用于控制牙髓感染。

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