Modulating acrylic acid content of nanogels for drug delivery & biocompatibility studies.

Dual stimuli-responsive nanogels (NGs) have gained popularity in the field of bio medicine due to their versatile nature of applicability. Poly(N-isopropylacrylamide)-co-poly(acrylic acid) (pNIPAm-pAAc)-based NGs provide such dual stimuli-response with pNIPAm and pAAc providing thermal and pH-based responses, respectively. Studying the growth of these NGs, as well as, understanding the effect of the incorporation of pAAc in the NG matrix, is important in determining the physico-chemical properties of the NG. Studies have been conducted investigating the effect of increasing pAAc content in the NGs, however, these are not detailed in understanding its effects on the physico-chemical properties of the pNIPAm-pAAc-based NGs. Also, the biocompatibility of the NGs have not been previously reported using human whole blood model. Herein, we report the effect of different reaction parameters, such as surfactant amount and reaction atmosphere, on the growth of pNIPAm-pAAc-based NGs. It is shown that the size of the NGs can be precisely controlled from ~130 nm to ~400 nm, by varying the amount of surfactant and the reaction atmosphere. The effect of increasing incorporation of pAAc in the NG matrix on its physico-chemical properties has been investigated. The potential of these NGs as drug delivery vehicles is investigated by conducting loading and release studies of a model protein drug, cytochrome C (Cyt C) from the NGs at temperature above the volume phase transition temperature (VPTT) and acidic pH. An ex vivo human whole blood model was used to investigate biocompatibility of the NGs by quantifying inflammatory responses during NG exposure. The NGs did not induce any significant production of chemokine IL-8 or pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α), and the cell viability in human whole blood was maintained during 4 h exposure. The NGs did neither activate the complement system, as determined by low Terminal Complement Complex (TCC) activation and Complement Receptor 3 (CR3) activation assays, thereby overall suggesting that the NGs could be potential candidates for biomedical applications.

The effects of oil-in-water nanoemulsion polyethylene glycol surface density on intracellular stability, pharmacokinetics, and biodistribution in tumor bearing mice.

PURPOSE: Lipid-based nanoparticles are extensively studied for drug delivery. These nanoparticles are often surface-coated with polyethylene glycol (PEG) to improve their biodistribution. Until now, the effects of varying PEG surface density have been studied in a narrow and low range. Here, the effects of high and a broad range of PEG surface densities on the in vivo performance of lipid-based nanoparticles were studied. METHODS: Oil-in-water nanoemulsions were prepared with PEG surface densities of 5-50 mol%. Confocal microscopy was used to assess intracellular disintegration in vitro. In vivo pharmacokinetics and biodistribution in tumor bearing mice were studied using a small animal optical imager. RESULTS: PEG surface density did not affect intracellular nanoemulsion stability. Surprisingly, circulation half-lives decreased with increasing PEG surface density. A plausible explanation was that nanoemulsion with high (50 mol%) PEG surface density activated the complement in a whole blood assay, whereas nanoemulsion with low (5 mol%) PEG density did not. In vivo, nanoemulsion with low PEG surface density was mostly confined to the tumor and organs of the mononuclear phagocyte system, whereas nanoemulsion with high PEG density accumulated throughout the mouse. CONCLUSIONS: Optimal PEG surface density of lipid-based nanoparticles for tumor targeting was found to be below 10 mol%.

Combined cell wall polysaccharide, mycotoxin and bacterial lipopolysaccharide exposure and inflammatory cytokine responses.

Human exposure to environmental microbes occurs regularly. Microbial compounds may interact with each other to affect cellular responses. We hypothesized that interactions between microbial compounds could modulate inflammatory cytokine responses in vitro. We investigated monocyte production of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF-alpha) and the regulatory cytokine interleukin-10 (IL-10) after combined exposure to the fungal cell wall polysaccharide mannan and to the beta-glucan laminarin, the mycotoxin citrinin and bacterial lipopolysaccharide (LPS). Interactions between the cell wall microbial compounds were estimated statistically in a general linear mixed model. We found that LPS (100 ng/ml) and the used beta-glucan (up to 1000 microg/ml) significantly interacted with each other to reduce TNF-alpha production. Mannan (up to 100 microg/ml) did not interact with the beta-glucan, but interacted with LPS. IL-10 production was induced by LPS only. The mycotoxin citrinin did not induce cytokine production, but was toxic to the cells in a dose- and time-dependent manner. However, non-toxic doses of citrinin reduced LPS-induced IL-10 production while LPS-induced TNF-alpha production was not similarly reduced by citrinin. In conclusion, interactions between microbial compounds can modulate cellular inflammatory cytokine production and experimental investigations of one compound at a time could give misleading conclusions about these combined effects.