Nebulizer-spray chamber apparatus and method for production of multi-component particle types and their use in affecting cellular responses

ABSTRACT

A particle generation apparatus and methodology is described for its application to prepare heterogeneous particles of defined chemical composition and their subsequent delivery to human lung cell cultures. A concentric type nebulizer was used together with a single pass spray chamber that was operated at two different temperature settings, either room temperature or to produce intact particles, at nonuniform temperature with the central axis measuring 54°C. Silica (SiO₂) particles in the micrometer size range were produced from the aggregation of nanoparticles. Tumor necrosis factor (TNF)-α, a well-characterized cytokine, was incorporated as a secondary soluble component within host silica particles for application as a proof-of-concept heterogeneous particle type. The overall particle deposition and wetting efficiency into media was 2.0 ± 0.4%, and the average size of particles that impacted and sank in the growth medium was 3.4 µm. The particle dose, reported as particle-to-cell ratio, spanned 0.1 to ∼2.5. Co-cultures of A549 and differentiated human monocytic cells (THP-1*) exposed to silica particles caused IL-6 upregulation, but not IL-8. For the silica plus TNF-α particle type, ICAM-1 signal increased in A549 mono-cultures as the particle-to-cell ratio increased, yet an estimated 90% of the TNF-α was denatured or possibly bound to the silica particle host, and was therefore bio-unavailable. Method quantitation using particles having two different chemical compositions has been described, illustrating capability for subsequent systematic investigation of the role of particles having different chemical composition in the human health aspects of the particulate air pollution issue.

Copyright © 2016 American Association for Aerosol Research

EDITOR:

• Tiina Reponen

Acknowledgments

We are thankful to Dr. Geoff Coleman for discussion and nebulizer donation, Meinhard Inc. CO, USA, I.-H. Sherry Lu and Haley Mitchell for preliminary work, and 4D labs for electron microscopy.