中国实用口腔科杂志

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人β防御素3基因转染及金纳米颗粒处理后的牙周膜细胞对大鼠牙周炎形成影响研究

李凌俊1周靓1崔迪1谢晓婷1杨文荣2闫福华1   

  1. 1. 南京大学医学院附属口腔医院,南京市口腔医院,江苏  南京210008;2. School of Life and Environmental Science,Centre for Chemistry and Biotechnology,Deakin University,Geelong,Victoria 3216,Australia
  • 出版日期:2018-04-15 发布日期:2018-04-17
  • 基金资助:

    国家自然科学基金(81570982);江苏省科教强卫工程(CXTDB2017014);南京市科技发展计划国际合作项目(201605083)

  • Online:2018-04-15 Published:2018-04-17

摘要:

目的    探讨人β防御素3基因转染大鼠牙周膜细胞,并经金纳米颗粒处理后,局部注射对SD大鼠慢性牙周炎形成的影响。方法    丝线结扎法构建SD大鼠慢性牙周炎模型,酶消化法提取大鼠牙周膜细胞并鉴定,用携带人β防御素3基因的腺病毒转染细胞并经金纳米颗粒处理,在慢性牙周炎构建同期局部注射细胞悬液到腭侧牙龈,对照组分别注射生理盐水和空载体腺病毒转染后的细胞悬液。2周后处死大鼠取材。通过Micro-CT和免疫组化染色观察大鼠牙周炎形成情况。结果    单纯结扎组第二磨牙周围牙槽骨明显吸收,牙龈上皮钉突向内增生,棘层增厚,炎细胞浸润,胶原纤维变性、局部断裂。局部注射经转染和金纳米颗粒处理的牙周膜细胞后,结扎区域牙槽骨吸收水平降低,骨密度和骨体积分数增加;牙龈上皮炎症细胞浸润和上皮钉突增生减少,胶原纤维断裂和排列紊乱等病理变化得到改善。结论    人β防御素3基因转染及金纳米颗粒处理后的大鼠牙周膜细胞局部注射后,可减轻SD大鼠的牙周组织炎症,降低牙周组织破坏,从而在牙周炎的形成过程中起到保护作用。

关键词: 人&beta, 防御素3基因;金纳米颗粒;慢性牙周炎

Abstract:

Objective    To investigate whether local injection of periodontal ligament cells which were transfected by human β defensin 3 gene and then treated with gold nanoparticles can affect the chronic periodontitis progress in SD rats. Methods    Chronic periodontitis model was established by silk thread ligation. Periodontal ligament cells were cultured by enzyme digestion method and then identified. Cells after transfection and gold nanoparticles treatment were injected into the palatal gingival tissues after ligation. The control groups were injected with normal saline or cells transfected by empty vector adenovirus respectively. After 2 weeks,rats were executed and sampled. Micro-CT and immunohistochemical staining were performed to observe the chronic periodontitis progress in SD rats. Results    In ligation group,the alveolar bone around second molars was obviously absorbed,the gingival epithelial spikes hyperplasia and thickened stratum spinosum could also be observed,so did the inflammatory cell infiltration,altered and broken collagen fibers. Local injection of periodontal ligament cells which were transfected by human β defensin 3 gene and then treated with gold nanoparticles could significantly reduce the absorption of alveolar bone in the ligation area and increase the bone mineral density as well as bone volume fraction. It could also decrease inflammatory cells and epithelial spikes hyperplasia in the gingival epithelium. Furthermore,situation of altered and broken collagen fiber could be improved in those groups. Conclusion    Local injection of periodontal ligament cells transfected by human β defensin 3 gene and treated by gold nanoparticles can alleviate the inflammatory reaction and thus play a protective role in the progress of chronic periodontitis in SD rats.

Key words: human &beta, defensin 3 gene;gold nanoparticles;chronic periodontitis