Title: A Microfluidic Cell Culture Based Platform for Simulating Periodontal Microenvironment
Sam Elcik (Presenter)
Roseman University of Health Sciences
Rachana Hegde, Roseman University of Health Sciences
Venkata Yellepeddi, Roseman University of Health Sciences
Objectives: Currently, there are no in-vitro models available that can effectively simulate the periodontal microenvironment and provide an adequate setting to develop new strategies for diagnosis and treatment of periodontal disease. Based on a recent report issued by the Centers for Disease Control and Prevention, one out of every two American adults age 30 and over has periodontal disease. Moreover, 64.7 million (47.2%) American adults have been diagnosed with mild to severe periodontitis. Additionally, the intrinsic complexity of the periodontal microenvironment that includes high GCF turnover (~50 times/hour), low volume of GCF (~0.4µL), the presence of host-derived enzymes, inflammatory mediators, and tissue breakdown products, confounds the development of new approaches. Therefore, we fabricated a microfluidic chip-based device with chambers containing living cells that are continuously perfused with media to simulate tissue- and organ level-physiology.
Methods: To validate the applicability of a prototype microfluidic chip-based device for simulating the periodontal microenvironment, we cultured primary human gingival keratinocytes (HGK) and seeded them onto the polycarbonate membrane at 1 X 105/cm2 density inside the microfluidic chip. To evaluate viability, cells were stained using a LIVE/DEADTM cell imaging kit and analyzed under a fluorescence microscope.
Results: The results showed that the HGK cells were successfully injected through microchannels without any obstruction and were seeded on the membrane. The fluorescence microscopy images showed that more than 95% of the HGK cells were viable for up to a period 4 hours after seeding on chip membranes and that the cells did not adhere to the walls of the microchannels.
Conclusions: In conclusion, a novel microfluidic-chip based device simulating the periodontal microenvironment was successfully validated for the feasibility of growing HGK cells. The microfluidic-chip device reported in this research can thus be utilized for various applications related to periodontal disease diagnosis, periodontal drug development, and individualization of periodontal therapy.