An effective functionalization method was investigated to take full advantage of the exceptional performance of both carbon nanotubes and epoxy polymer for composite application. Epoxy polymer curing agent, EPI-W, was grafted to the single-walled carbon nanotubes through diazotization. Fourier transformed infrared spectroscopy, Raman spectroscopy, differential scanning calorimetry, dynamical mechanical analysis and thermo-gravimetric analysis were performed to characterize the functionalization effect. The degree of functionalization was estimated to be 1 in 50 carbons in the nanotube framework. The elastic modulus of the nanocomposite was enhanced 24.6% with only 0.5 wt% loading of functionalized carbon nanotubes, in contrast to the 3.2% increase of un-functionalized carbon nanotube reinforced composite. This significant improvement suggested an effective way to realize an industrial application of nanotubes reinforcing epoxy composite.

Nucleic acids are used in many therapeutic modalities, including gene therapy, but their ability to trigger host immune responses in vivo can lead to decreased safety and efficacy. In the case of adeno-associated viral (AAV) vectors, studies have shown that the genome of the vector activates Toll-like receptor 9 (TLR9), a pattern recognition receptor that senses foreign DNA. Here, we engineered AAV vectors to be intrinsically less immunogenic by incorporating short DNA oligonucleotides that antagonize TLR9 activation directly into the vector genome. The engineered vectors elicited markedly reduced innate immune and T cell responses and enhanced gene expression in clinically relevant mouse and pig models across different tissues, including liver, muscle, and retina. Subretinal administration of higher-dose AAV in pigs resulted in photoreceptor pathology with microglia and T cell infiltration. These adverse findings were avoided in the contralateral eyes of the same animals that were injected with the engineered vectors. However, intravitreal injection of higher-dose AAV in macaques, a more immunogenic route of administration, showed that the engineered vector delayed but did not prevent clinical uveitis, suggesting that other immune factors in addition to TLR9 may contribute to intraocular inflammation in this model. Our results demonstrate that linking specific immunomodulatory noncoding sequences to much longer therapeutic nucleic acids can “cloak” the vector from inducing unwanted immune responses in multiple, but not all, models. This “coupled immunomodulation” strategy may widen the therapeutic window for AAV therapies as well as other DNA-based gene transfer methods.