Week 6:
Greetings Readers! This week our team has been tireless acquiring all of the materials that will be needed on our up coming test trials of our hydrogel design. So far we have obtained sodium alginate and calcium chloride from amazon, the fluorescent protein we will be using in the place of growth factors in our experiment, as well as a pH buffer solution and our dependent variable (cytidine monophosphate acid) curtsy of our assisting lab instructor Mohammad. We were even able to find a professor who had an instrument to measure the light given off by our fluorescent protein (Bovine Albumin Fluorescein) called a nanodrop, which functions very similar to a UV spectrophotometer and can accurately detect the light emitted when our hydrogel releases the protein. All that is left to acquire are sample containers to place our samples of fluorescent protein as they are released by our hydrogel device. So far we have reached out to a research lab to see if they have any spare test tubes or sterile containers they are not using, but if there is no other option we will purchase some from amazon for a small price. Once we have obtained all of these containers we plan to begin testing of our Alginate-cytidine hydrogel device by incorporating varying amounts of cytidine to different hydrogel trials. Cytidine is a natural monomer called a nucleotide, which is a component of RNA. it has a slight positive charge, which will allow it to bind to the negatively charged fluorescent protein and the negatively charged Alginate in the hydrogel. This binding will be further strengthened by cytidine's natural ability to form multiple hydrogen bonds, which will latch onto the oxygen, and nitrogen atoms on the fluorescent protein. This binding will allow the hydrogel to keep the protein trapped for a certain amount of time, until it is released at a very slow controlled rate (0.5 pg/ml over 16 days). By keeping the amounts of Alginate, calcium ions in solution, and fluorescent protein constant throughout our testing trials, while comparing the amount of fluorescent protein released based on the difference in how much cytidine was added, we will be able to obtain accurate data on the optimal amount of cytidine to add to our hydrogel for optimal release.
In our research we found that the optimal rate of release of TGF-beta, one of the most important growth factors in our original design, for wound healing was about 0.5 pg/ml over 16 days, so based on the importance of this specific growth factor in our design, we have chosen it as a suitable model for our goal release rate of our fluorescent protein. For obvious reasons we can not wait for 16 days to ensure that our protein release rate is at this value so we will account for this by measuring the intensity of the light emitted from our fluorescent protein in samples gathered every hour for a six hour period over four days. This consistent saving of samples for all trials will ensure that all samples will have been under the exact same conditions prior to measuring, leaving only the amount of cytidine added to the hydrogel mixture to be the altering factor. Having multiple release rate values will also ensure that any deviation between the release rates of a single sample are recorded and discussed upon. For more information on TGF or Bovine Albumin Fluorescein please click the links below.
Greetings Readers! This week our team has been tireless acquiring all of the materials that will be needed on our up coming test trials of our hydrogel design. So far we have obtained sodium alginate and calcium chloride from amazon, the fluorescent protein we will be using in the place of growth factors in our experiment, as well as a pH buffer solution and our dependent variable (cytidine monophosphate acid) curtsy of our assisting lab instructor Mohammad. We were even able to find a professor who had an instrument to measure the light given off by our fluorescent protein (Bovine Albumin Fluorescein) called a nanodrop, which functions very similar to a UV spectrophotometer and can accurately detect the light emitted when our hydrogel releases the protein. All that is left to acquire are sample containers to place our samples of fluorescent protein as they are released by our hydrogel device. So far we have reached out to a research lab to see if they have any spare test tubes or sterile containers they are not using, but if there is no other option we will purchase some from amazon for a small price. Once we have obtained all of these containers we plan to begin testing of our Alginate-cytidine hydrogel device by incorporating varying amounts of cytidine to different hydrogel trials. Cytidine is a natural monomer called a nucleotide, which is a component of RNA. it has a slight positive charge, which will allow it to bind to the negatively charged fluorescent protein and the negatively charged Alginate in the hydrogel. This binding will be further strengthened by cytidine's natural ability to form multiple hydrogen bonds, which will latch onto the oxygen, and nitrogen atoms on the fluorescent protein. This binding will allow the hydrogel to keep the protein trapped for a certain amount of time, until it is released at a very slow controlled rate (0.5 pg/ml over 16 days). By keeping the amounts of Alginate, calcium ions in solution, and fluorescent protein constant throughout our testing trials, while comparing the amount of fluorescent protein released based on the difference in how much cytidine was added, we will be able to obtain accurate data on the optimal amount of cytidine to add to our hydrogel for optimal release.
In our research we found that the optimal rate of release of TGF-beta, one of the most important growth factors in our original design, for wound healing was about 0.5 pg/ml over 16 days, so based on the importance of this specific growth factor in our design, we have chosen it as a suitable model for our goal release rate of our fluorescent protein. For obvious reasons we can not wait for 16 days to ensure that our protein release rate is at this value so we will account for this by measuring the intensity of the light emitted from our fluorescent protein in samples gathered every hour for a six hour period over four days. This consistent saving of samples for all trials will ensure that all samples will have been under the exact same conditions prior to measuring, leaving only the amount of cytidine added to the hydrogel mixture to be the altering factor. Having multiple release rate values will also ensure that any deviation between the release rates of a single sample are recorded and discussed upon. For more information on TGF or Bovine Albumin Fluorescein please click the links below.
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