The importance of particle size and DNA condensation salt for calcium phosphate nanoparticle transfection.
Calcium phosphate has been used for over 30 years to deliver genetic material to mammalian cells. This vector has proven advantages over other transfection species such as viruses and dendrimers in terms of superior biocompatibility and reduced immune response. However, clinical application of calcium phosphate based transfection techniques is hampered by poor understanding of the key factors underlying its action. Despite widespread in vitro use, little attention has been given to the physico-chemical characteristics of the calcium phosphate particles mediating transfection. In this study parameters were optimised to produce calcium phosphate nanoparticles onto which plasmid DNA (pDNA) was adsorbed that were more effective than a commercial dendrimer vector in delivering pDNA to an osteoblastic cell line and compared favourably in a fibroblastic cell line without the need for special culture conditions such as cell cycle synchronization or glycerol shock treatment. Addition of the pDNA after nanoparticle synthesis allowed for characterisation of particle morphology, size, surface charge and composition. We found that the key parameters for effective calcium phosphate nanoparticle transfection were an optimal concentration of calcium and chloride ions and a nanosized non-agglomerated precipitate.
Pedraza
Claudio E
CE
Faculty of Dentistry, McGill University, Montréal, Québec, Canada.
Bassett
David C
DC
McKee
Marc D
MD
Nelea
Valentin
V
Gbureck
Uwe
U
Barralet
Jake E
JE
D016475:3T3 Cells; D000818:Animals; D002130:Calcium Phosphates; D049109:Cell Proliferation; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics / Q000302:isolation & purification; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D051379:Mice; D046529:Microscopy, Electron, Transmission; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D053758:Nanoparticles; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D010316:Particle Size; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection / Q000379:methods*
D002130:Calcium Phosphates; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D004247:DNA; C020243:calcium phosphate