Calcium phosphate nanoparticles(CaPNPs)have good biocompatibility as gene carriers;however,CaPNPs typically exhibit a low transfection efficiency.Cell penetrate peptide(TAT)can increase the uptake of nanoparticles but...Calcium phosphate nanoparticles(CaPNPs)have good biocompatibility as gene carriers;however,CaPNPs typically exhibit a low transfection efficiency.Cell penetrate peptide(TAT)can increase the uptake of nanoparticles but is limited by its non-specificity.Grafting adhesion peptide adhesion peptide on carriers can enhance their targeting.The Plekho1 gene encodes casein kinase-2 interacting protein-1(CKIP-1),which can negatively regulate osteogenic differentiation.Based on the above,we produced a Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA carrier system via hydrothermal synthesis,silanization and adsorption.The effects of this carrier system on cell endocytosis and biological effects were evaluated by cell culture in vitro.The results demonstrate that CaPNPs with 7%Mg(60 nm particle size,short rod shape and good dispersion)were suitable for use as gene carriers.The carrier system boosted the endocytosis of MG63 cells and was helpful for promoting the differentiation of osteoblasts,and the dual-ligand system possessed a synergistic effect.The findings of this study show the tremendous potential of the Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA carrier system for efficient delivery into cells and osteogenesis inducement.展开更多
P-15, a synthetic peptide of 15-amino acids, has been tested in clinical trials to enhance cell adhesion and promote osseointe-gration. This feature of P-15 has also inspired the development of designing new bone subs...P-15, a synthetic peptide of 15-amino acids, has been tested in clinical trials to enhance cell adhesion and promote osseointe-gration. This feature of P-15 has also inspired the development of designing new bone substitute materials. Despite the increasing applications of P-15 in bone graft alternatives, few studies focus on the mechanism of cell adhesion promoted by P-15 and the mechanical property changes of the cells interacting with P-15. In this article, we used atomic force microscope(AFM) based single cell indentation force spectroscopy to study the impact of P-15 on the stiffness and the adhesion ability of human jaw bone mesenchymal stem cells(HJMSCs) to hydroxyapatite(HA). We found that the stiffness of HJMSCs increases as the concentration of P-15 grows in short culture intervals and that the adhesion forces between HJMSCs and HA particles in both the presence and absence of P-15 are all around 30 p N. Moreover, by calculating the binding energy of HJMSCs to HA particles mixed with and without P-15, we proved that P-15 could increase the adhesion energy by nearly four times. Scanning electron microscope(SEM) was also exploited to study the morphology of HJMSCs cultured in the presence and absence of P-15 on HA disc surface for a short term. Apparent morphological differences were observed between the cells cultured with and without P-15. These results explain the probable underlying adhesion mechanism of HJMSC promoted by P-15 and can serve as the bases for the design of bone graft substitute materials.展开更多
基金Project(81571021)supported by the National Natural Science Foundation of ChinaProject(2018zzts944)supported by the Graduate Student Independent Exploration Innovation Fund of the Central South University,China+1 种基金Projects(2015WK3012,2018SK2017)supported by the Hunan Provincial Science and Technology Department,ChinaProject(20160301)supported by New Talent Project of the Third Xiangya Hospital of Central South University,China。
文摘Calcium phosphate nanoparticles(CaPNPs)have good biocompatibility as gene carriers;however,CaPNPs typically exhibit a low transfection efficiency.Cell penetrate peptide(TAT)can increase the uptake of nanoparticles but is limited by its non-specificity.Grafting adhesion peptide adhesion peptide on carriers can enhance their targeting.The Plekho1 gene encodes casein kinase-2 interacting protein-1(CKIP-1),which can negatively regulate osteogenic differentiation.Based on the above,we produced a Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA carrier system via hydrothermal synthesis,silanization and adsorption.The effects of this carrier system on cell endocytosis and biological effects were evaluated by cell culture in vitro.The results demonstrate that CaPNPs with 7%Mg(60 nm particle size,short rod shape and good dispersion)were suitable for use as gene carriers.The carrier system boosted the endocytosis of MG63 cells and was helpful for promoting the differentiation of osteoblasts,and the dual-ligand system possessed a synergistic effect.The findings of this study show the tremendous potential of the Mg-CaPNPs-RGD-TAT-CKIP-1 siRNA carrier system for efficient delivery into cells and osteogenesis inducement.
基金Nanjing Medical Transformation Center and the Project of the Nanjing Foundation for Development of Science and Technology(Grant No.201303024)
文摘P-15, a synthetic peptide of 15-amino acids, has been tested in clinical trials to enhance cell adhesion and promote osseointe-gration. This feature of P-15 has also inspired the development of designing new bone substitute materials. Despite the increasing applications of P-15 in bone graft alternatives, few studies focus on the mechanism of cell adhesion promoted by P-15 and the mechanical property changes of the cells interacting with P-15. In this article, we used atomic force microscope(AFM) based single cell indentation force spectroscopy to study the impact of P-15 on the stiffness and the adhesion ability of human jaw bone mesenchymal stem cells(HJMSCs) to hydroxyapatite(HA). We found that the stiffness of HJMSCs increases as the concentration of P-15 grows in short culture intervals and that the adhesion forces between HJMSCs and HA particles in both the presence and absence of P-15 are all around 30 p N. Moreover, by calculating the binding energy of HJMSCs to HA particles mixed with and without P-15, we proved that P-15 could increase the adhesion energy by nearly four times. Scanning electron microscope(SEM) was also exploited to study the morphology of HJMSCs cultured in the presence and absence of P-15 on HA disc surface for a short term. Apparent morphological differences were observed between the cells cultured with and without P-15. These results explain the probable underlying adhesion mechanism of HJMSC promoted by P-15 and can serve as the bases for the design of bone graft substitute materials.
