Subject(s)
Equipment Safety , Filtering Surgery/instrumentation , Glaucoma/surgery , Minimally Invasive Surgical Procedures , Ophthalmology/organization & administration , Therapies, Investigational , United States Food and Drug Administration , Congresses as Topic , Endpoint Determination , Humans , Intraocular Pressure , Societies, Medical , United States , Visual FieldsSubject(s)
Endpoint Determination , Lenses, Intraocular , Ophthalmology/standards , Societies, Medical/standards , United States Food and Drug Administration/standards , Academies and Institutes , Cataract Extraction , Device Approval/legislation & jurisprudence , Diffusion of Innovation , Equipment Design/economics , Humans , Medical Device Legislation , United StatesABSTRACT
Contact lenses and lens care regimens are an important part of eyecare practices and vital to lens-wearing patients. New contact lens materials and cleaning options continue to come to market and affect how patients wear and care for their lenses. In this section we look at how the contact lens and lens solution revolution started, how it has evolved over the last 40 years, and how standards have evolved and impacted these new offerings.
Subject(s)
Contact Lens Solutions/standards , Contact Lenses/standards , Contact Lenses/trends , Equipment Contamination/prevention & control , Practice Guidelines as Topic , Contact Lens Solutions/history , Contact Lenses/history , History, 20th Century , History, 21st CenturyABSTRACT
Chitosans with various degrees of deacetylation were synthesized by acetylation with acetic anhydride. These chitosans were evaluated for efficacy of nanoparticle formation, DNA binding efficiency, morphology, and in vitro and in vivo gene transfection efficiency. DNA binding efficacy was reduced as degree of deacetylation was decreased, therefore requiring an increased +/-ratio to effect complete DNA complexation. For chitosan with a molecular weight of 390 kDa, the +/-ratio to achieve complete DNA complexation for degrees of deacetylation of 90%, 70% and 62% was 3.3:1, 5.0:1, and 9.0:1, respectively. The size and morphology of these nanoparticle formulations were not significantly different. The decreased degree of deacetylation results in a decrease in overall luciferase expression levels in HEK293, HeLa, and SW756 cells due to particle destabilization in the presence of serum proteins. However, intramuscular luciferase expression levels increased with decreasing deacetylation over the time points tested. Degree of chitosan deacetylation is an important factor in chitosan-DNA nanoparticle formulation as it affects DNA binding, release and gene transfection efficiency in vitro and in vivo.
Subject(s)
Chitosan/chemistry , Gene Expression Regulation , Gene Transfer Techniques , Cell Line , DNA/chemistry , HeLa Cells , Humans , Luciferases/chemistry , Luciferases/metabolism , Microscopy, Electron, Transmission , Models, Chemical , Muscles/metabolism , Nanotechnology , Spectrophotometry , Time Factors , Transfection , Ultraviolet RaysABSTRACT
Poly(propyl acrylic acid) (PPAA) is a polymer specifically designed to disrupt lipid bilayer membranes within a sharply defined pH range. The pH sensitivity can be used to enhance the release of endocytosed drugs into the cytoplasmic compartment of the cell. By incorporating this polymer in a polymeric gene carrier, chitosan, the release of plasmid DNA from the endosomal compartment was enhanced. In vitro transfection studies confirmed that the incorporation of PPAA into the chitosan-DNA nanoparticles enhanced gene expression in both HEK293 and HeLa cells compared to chitosan nanoparticles alone. The dose and time at which PPAA was incorporated during the complex formation affected the release of DNA and transfection efficiency. The optimal dose of PPAA incorporated into the chitosan nanoparticles was determined to be 10 microg, corresponding to a PPAA/DNA weight ratio of 1:1. At this dose, the ternary complexes are approx. 400 nm in size with a net negative surface charge of -17.4 mV. Intracellular trafficking studies confirmed the association of PPAA, DNA and chitosan at 24 h post-transfection and the subsequent release of DNA and PPAA from the chitosan at 48 h. The diffuse appearance of the majority of the DNA and the PPAA at later time points suggests that the PPAA triggered membrane disruption resulting in the release of DNA from the endosomal compartment. Finally, the lack of colocalization between PPAA and Lysotracker indicated that the PPAA-loaded nanoparticles were not trafficked through a lysosomal pathway. This study suggests the promising strategy of including PPAA in the formulation of polymer-DNA complexes for non-viral gene delivery.
