RESUMO
The immunogenicity and toxicity of N-2-Hydroxypropyl trimethyl ammonium chloride chitosan/N, O-carboxymethyl chitosan nanoparticles (N-2-HACC/CMCS NPs) as a universal vaccine adjuvant/delivery system remains unclear. The present study indicated that the positively charged N-2-HACC/CMCS NPs showed a regular spherical morphology, with a particle size of 219 ± 13.72 nm, zeta potential of 37.28 ± 4.58 mV, had hemocompatibility and biodegradation. Acute toxicity, repeated dose toxicity, abnormal toxicity, muscle stimulation, whole body allergic reaction evaluation in vitro, and cytotoxicity in vivo confirmed N-2-HACC/CMCS NPs is safe and non-toxic. N-2-HACC/OVA/CMCS NPs were prepared to evaluate the immunogenicity, which showed a particle size of 248.1 ± 15.53 nm, zeta potential of 17.24 ± 1.28 mV, encapsulation efficiency of 92.43 ± 0.96 %, and loading capacity of 42.97 ± 0.07 %. Oral or intramuscular route with the N-2-HACC/OVA/CMCS NPs in mice not only induced higher IgG, IgG1, IgG2a, and sIgA antibody titers, but also significantly produced higher levels of IL-6, IL-4, IFN-γ, and TNF-α, demonstrating that the N-2-HACC/OVA/CMCS NPs enhance humoral, cellular, and mucosal immune responses. Our results not only support the N-2-HACC/CMCS NPs to be a safe and potential universal nano adjuvant/delivery system in vaccine development, especially mucosal vaccines, but also rich the database knowledge of adjuvant/delivery systems, and provide new direction to introduce more licensed adjuvants.
Assuntos
Quitosana , Nanopartículas , Vacinas Virais , Adjuvantes Imunológicos/farmacologia , Cloreto de Amônio , Animais , Galinhas , Quitosana/farmacologia , Derivados da Hipromelose , Imunoglobulina A Secretora , Imunoglobulina G , Interleucina-4 , Interleucina-6 , Camundongos , Fator de Necrose Tumoral alfaRESUMO
Kenaf stalk was pretreated by the white-rot fungus Pleurotus sajor-caju incubated in solid-state kenaf stalk cultivation medium. Delignification and subsequent enzymatic saccharification and fermentation of kenaf stalk were investigated in order to evaluate effects of microbial pretreatment on bioconversion of kenaf lignocellulose to fuel ethanol production. The highest delignification rate of 50.20% was obtained after 25-35 days cultivation by P. sajor-caju, which could improve subsequent enzymatic hydrolysis efficiency of kenaf cellulose. And the saccharification rate of pretreated kenaf stalk reached 69.33 to 78.64%, 4.5-5.1 times higher than the control. Simultaneous saccharification and fermentation (SSF) with microbial-pretreatment kenaf stalk as substrate was performed. The highest overall ethanol yield of 68.31% with 18.35 to 18.90 mg/mL was achieved after 72 h of SSF.