Modelling Whole Smoke exposure: a superior tool for smoke research.
Two models are currently commonly used to study the effect of cigarette smoke on Human Bronchial epithelial cells: Cigarette Smoke Extract (CSE) and Whole Smoke model. In the CSE model, cells are directly submersed in medium containing dissolved CSE. With regards to Human Bronchial epithelial cells, CSE impairs development of cilia, epithelial damage repair and gene expression. In the Whole Smoke model, cells are placed in an air/liquid interface (ALI). The ALI method results in culture medium on the basolateral side and air/smoke dilution on the apical side. The Whole Smoke model allows for the induction of cilia formation and exposure to both soluble and insoluble particles (in CSE model, only soluble particles are retained by the liquid). Thus the Whole Smoke model allows for a more accurate ex vivo model.
The Whole Smoke Chamber
The device is calibrated to software, which controls the distance that the pneumatic cylinder moves, and hence volumes of cigarette smoke drawn back. This quantified volume is then exhaled over A549 cells on ALI insert, which are fixed inside a desiccator unit.
Smoke Exposure increases IL-6 concentration in A549 cells.
At 6 hours, both control and smoked A549 cells had a similar concentration of IL-6 produced, with the smoked plate marginally more. By 48 hours, the smoked plate had a clear increase in IL-6 concentration.
Smoking with RSV infection
Current work has included the infection of Respiratory Syncytial virus (RSV) post smoke exposure to A549 cells.
Further Work
Further work is required to calibrate the machine and software to optimise volume and dilution of smoke environment. Once this has been done, work to develop Human Bronchial epithelial cells on ALI will begin on 24th November 2014.
Bio:
Oliver Reed
Year: BM4 Y2
Interests in medicine: Respiratory medicine
Interests outside of medicine: Cycling
About Oliver's project: Undertaken over the summer (2014) for 3 months with the Tom Wilkinson Research group in Pulmonary Immunology. I was given the responsibility to design a smoke machine to model the effect of cigarette smoke in vitro.
Personal experience of research so far: I thoroughly enjoyed the summer internship and it has given me an insight into the exciting area of academic medicine. You need to be patient and organised, but the rewards are worth it.
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