Optimization of DNA Origami Sensors to Assess the Microenvironment of the Intervertebral Disc
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Date
2023-05
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The Ohio State University
Abstract
Throughout the day, tissue osmolarity varies in the intervertebral discs (IVD) as water enters and leaves the system due to applied loads on the spine. This variation in osmotic pressure occurs on a consistent cycle throughout the day and is essential in regulating cellular metabolism. Due to these consistent changes, the exact osmotic environment in-vivo is still unknown. However, advances in nanotechnology have shown promise in measuring changes in ion concentration using biosensors created through the process of DNA origami. DNA origami is a method that utilizes a viral single-stranded DNA molecule known as a scaffold to self-assemble with multiple smaller single-stranded DNA molecules known as staples. A specific biosensor known as the NanoDyn is created through the process of DNA origami and has been shown to assess physiologically relevant ion concentrations that are present in the IVD. This study aims to optimize the spatial resolution of several variations of the NanoDyn biosensor with the ultimate goal of using the biosensors to assess the osmotic environment of the IVD in-situ. Variations of the device were created by changing the number of base pair interactions constraining a linker and varying the number of linkers that were constrained. Devices with 3 to 7 base pair interactions constraining each linker were tested. The results show that the NanoDyn device prefers the closed stable state as the number of base pair interactions increases. Devices with 3, 4, and 5 base pair interactions were chosen for further testing. The results show that an increase in the number of constrained linkers and an increase in salt concentration of the base buffer resulted in the device having a preference towards the closed stable state. The physiologically relevant range for the IVD is estimated to be between 400-550 mOsm. Future testing will evaluate the resolution for the device with 3, 4, and 5 base pair interactions using a fluorometer to access the device's sensitivity in the desired range. The device that fits the given range will be tested on a coverslip with an IVD tissue sample to measure the osmotic environment in-situ.
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Keywords
DNA Origami, Biomechanics, Intervertebral Disc, Nanotechnology