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

Effects of periodontal ligament cells transplantation treated with human β defensin 3 gene transfection and gold nanoparticles on chronic periodontitis progress in SD rats

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。

Objective To investigate whether local injection of periodontal ligament cells which were transfected by hu- man β defensin 3 gene and then treated with gold nanoparticles can affect the chronic periodontifis 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. Ceils 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 immunohistochemi- cal staining were performed to observe the chronic periodontitis progress in SD rats. Results In ligation group, the alve- olar 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 injec- tion of periodontal ligament cells which were transfected by human β defensin 3 gene and then treated with gold nanopar-ticles 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 de- crease inflammatory cells and epithelial spikes hyperpla- sia in the gingival epithelium. Furthermore, situation of al- tered 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.