Gene transfection achieved by utilizing antibacterial calcium phosphate nanoparticles for enhanced regenerative therapy
Gene transfection achieved by utilizing antibacterial calcium phosphate nanoparticles for enhanced regenerative therapy
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DOI：
10.1016/j.actbio.2020.11.003
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发表时间：
2021-01-01
影响因子：
9.7
通讯作者：
Sasaki, Keiichi
中科院分区：
文献类型：
--
作者：
Xiang, Chen;Tenkumo, Taichi;Sasaki, Keiichi
鱼精蛋白包被的多壳层磷酸钙（CaP）被开发为用于组织再生治疗的非病毒载体。将负载有不同量的编码骨形态发生蛋白2（BMP-2）和胰岛素样生长因子1（IGF-1）的质粒DNA的CaP纳米颗粒用于处理MC 3 T3 E1细胞，并在3天后使用ELISA测量释放的BMP-2或IGF-1的产量。将含有CaP纳米颗粒的胶原支架植入大鼠颅骨缺损中，并在基因转移后28天评估BMP-2和IGF-1产量、骨形成和骨矿物质密度增强。CaP纳米粒对变形链球菌和伴放线菌聚集杆菌的抗菌作用随鱼精蛋白剂量的增加而增加，而对金黄色葡萄球菌和牙龈卟啉单胞菌的抗菌作用则较低。在BMP-2和IGF-1的联合治疗中，BMP-2和IGF-1的浓度比是影响骨形成活性的重要因素。BMP-2：IGF-1浓度比为1：4的MC 3 T3 E1细胞的钙化活性和OCN mRNA在14天时较高。在基因转染处理过程中，BMP-2和IGF-1在基因转移后同时释放;编码IGF-1的质粒DNA的负载剂量不影响BMP-2或IGF-1产量或体外和体内新骨形成率。总之，两个生长因子释放系统开发使用抗菌基因转移载体，并在体外和体内测定加载的质粒DNA剂量和所得的生长因子产量之间的关系。(C)2020 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
Protamine-coated multi-shell calcium phosphate (CaP) was developed as a non-viral vector for tissue regeneration therapy. CaP nanoparticles loaded with different amounts of plasmid DNA encoding bone morphogenetic protein 2 (BMP-2) and insulin-like growth factor 1 (IGF-1) were used to treat MC3T3E1 cells, and the yield of the released BMP-2 or IGF-1 was measured using ELISA 3 days later. Collagen scaffolds containing CaP nanoparticles were implanted into rat cranial bone defects, and BMP-2 and IGF-1 yields, bone formation, and bone mineral density enhancement were evaluated 28 days after gene transfer. The antibacterial effects of CaP nanoparticles against Streptococcus mutans and Aggregatibacter actinomycetemcomitans increased with an increase in the protamine dose, while they were lower for Staphylococcus aureus and Porphyromonas gingivalis. In the combination treatment with BMP-2 and IGF-1, the concentration ratio of BMP-2 and IGF-1 is an important factor affecting bone formation activity. The calcification activity and OCN mRNA of MC3T3E1 cells subjected to a BMP-2:IGF-1 concentration ratio of 1:4 was higher at 14 days. During gene transfection treatment, BMP-2 and IGF-1 were released simultaneously after gene transfer; the loaded dose of the plasmid DNA encoding IGF-1 did not impact the BMP-2 or IGF-1 yield or new bone formation ratio in vitro and in vivo. In conclusion, two growth factor-releasing systems were developed using an antibacterial gene transfer vector, and the relationship between the loaded plasmid DNA dose and resultant growth factor yield was determined in vitro and in vivo. (C) 2020 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.