About the Project
Neutrophils are the most abundant white blood cell in circulation and are the first line for innate defence. This means that these cells locate to a site of injury and/or infection very quickly and initiate the body’s response. This migration is mediated by cell movement and follows a chemoattractant gradient to the area where they are needed. These chemoattractants are either of host or microbial origin and can molecules either released from injured or activated cells, in the case of the host, or are cellular components or virulence factors in the case of microbes. Once neutrophils arrive at the site of injury they degranulate proteins and produce reactive species, such as reactive oxygen species (ROS), for the disarmament of microbes. These proteins form a secretome which can change with time and type of stimulation.
Periodontal disease is a highly prevalent disease of the oral cavity which is initiated by plaque bacteria and which is exacerbated by the immune response. We have found that the migration of neutrophils in patients with periodontitis, a severe form of gum disease, is slower  and also that these cells also produce more ROS  and greater quantities of cytokines  in the secretome. This means that there is a greater quantity of tissue destruction during the migration of these cells through the tissue to the site of plaque accumulation. Currently all of these analyses need to be performed in separate assays and the real time production of the secretome with analysis of migration is not possible.
This project will develop a leaky waveguide (LW)  based assay for the following purposes:
Study migration of cells: LWs comprise of a few micron thick hydrogel film deposited on a glass substrate. We will use the evanescent field of light propagating in these hydrogel films to illuminate cells present on top of hydrogel films. As cells present on top of the hydrogel films will scatter light, they will appear as bright spots in the images captured by a camera placed underneath a trapezoidal prism . The images will be captured in real-time to study migration of cells. Suitable proteins such as serum albumin may be adsorbed/chemically attached to hydrogel films to allow cells to move freely. A microfluidic gradient generator flow cell will be mounted on top of the hydrogel film to study the migration of cells in response to different types of signalling molecules and their concentrations .
Quantify secretome components of cells: We will immobilise antibodies towards secretome components of cells in hydrogel films. The binding of secretome components to antibodies will cause the resonance angles of LWs to change. The resonance angles will be monitored to determine the concentration of the secretome components of cells . Cells may be cultured in-situ in the microfluidic flow cell mounted on top of LWs so that spatial and temporal profiles of secretome components produced by cells can be studied.
For informal enquiries please contact Dr Ruchi Gupta at firstname.lastname@example.org. Please include a CV and transcripts in your email.
2. Matthews et al, Neutrophil Hyper-responsiveness in Periodontitis, Journal of Dental Research, (2007), 718.
3. M.R. Ling et al, Peripheral Blood Neutrophil Cytokine Hyper-Reactivity in Chronic Periodontitis, Innate Immunity, (2015), 714.
4. R. Gupta et al, Leaky Waveguides (LWs) for Chemical and Biological Sensing − A Review and Future Perspective, Sensors and Actuators B, (2020), 128628.
5. M. Zourob et al, Bacteria detection Using Disposable Optical Leaky Waveguide Sensors, Biosensors and Bioelectronics, (2005), 293.
6. T. Ishida et al, Investigation of the Influence of Glucose Concentration on Cancer Cells by Using a Microfluidic Gradient Generator without the Induction of Large Shear Stress, Micromachines, (2016), 155.
7. B.R. Andrew et al, Label-free Leaky Waveguide for VEGF Detection, 22nd International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS, (2018), 940.
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