Nano-Sized Calcium Phosphate Particles for Periodontal Gene Therapy
Nano-Sized Calcium Phosphate Particles for Periodontal Gene Therapy
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DOI：
10.1902/jop.2012.120012
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发表时间：
2013-01-01
影响因子：
4.3
通讯作者：
Amiji, Mansoor
中科院分区：
文献类型：
--
作者：
Elangovan, Satheesh;Jain, Shardool;Amiji, Mansoor
背景资料：生长因子如血小板衍生生长因子（PDGF）具有显著增强的牙周治疗结果，具有高度的可变性，主要是由于缺乏持续供应所需的时间段。克服这一障碍的一种方法是基因治疗。本体外研究的目的是评估PDGF-B基因在成纤维细胞中使用纳米尺寸的磷酸钙颗粒（NCaPP）作为vectors.Methods：NCaPP纳入绿色荧光蛋白（NCaPP-GFP）和PDGF-B（NCaPP-PDGF-B）质粒的交付使用建立的沉淀系统和其特征在于使用透射电子显微镜和1.2%琼脂糖凝胶电泳。分别使用细胞毒性测定和荧光显微镜评价纳米复合物在成纤维细胞中的生物相容性和转染。用聚合酶链反应和酶联免疫吸附试验检测不同时间点处理后PDGF-B的转染情况，用细胞增殖试验检测PDGF-B的转染功能。结果：合成的NCaPP纳米复合物为球形，直径约30 ~ 50 nm。凝胶电泳证实了纳米复合物中的DNA掺入和稳定性，3-（4，5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑鎓试剂测定证明了它们在成纤维细胞中的生物相容性。体外转染研究揭示了NCaPP-PDGF-B处理后更高和更长的持续转染，其持续长达96小时。显着增强成纤维细胞增殖观察NCaPP-PDGF-B处理的细胞证实了这些nanoplexes.Conclusion的功能：NCaPP表现出更高水平的生物相容性和有效地转染PDGF质粒成纤维细胞在体外条件下描述。J Periodontol 2013;84：117-125.
Background: Growth factors such as platelet-derived growth factor (PDGF) have significantly enhanced periodontal therapy outcomes with a high degree of variability, mostly due to the lack of continual supply for a required period of time. One method to overcome this barrier is gene therapy. The aim of this in vitro study is to evaluate PDGF-B gene delivery in fibroblasts using nano-sized calcium phosphate particles (NCaPP) as vectors.Methods: NCaPP incorporating green fluorescent protein (NCaPP-GFP) and PDGF-B (NCaPP-PDGF-B) plasmids were synthesized using an established precipitation system and characterized using transmission electron microscopy and 1.2% agarose gel electrophoresis. Biocompatibility and transfection of the nanoplexes in fibroblasts were evaluated using cytotoxicity assay and florescence microscopy, respectively. Polymerase chain reaction and enzyme-linked immunosorbent assay were performed to evaluate PDGF-B transfection after different time points of treatments, and the functionality of PDGF-B transfection was evaluated using the cell proliferation assay.Results: Synthesized NCaPP nanoplexes incorporating the genes of GFP and PDGF-B were spherical in shape and measured about 30 to 50 nm in diameter. Gel electrophoresis confirmed DNA incorporation and stability within the nanoplexes, and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium reagent assay demonstrated their biocompatibility in fibroblasts. In vitro transfection studies revealed a higher and longer lasting transfection after NCaPP-PDGF-B treatment, which lasted up to 96 hours. Significantly enhanced fibroblast proliferation observed in NCaPP-PDGF-B-treated cells confirmed the functionality of these nanoplexes.Conclusion: NCaPP demonstrated higher levels of biocompatibility and efficiently transfected PDGF plasmids into fibroblasts under described in vitro conditions. J Periodontol 2013;84:117-125.