Week 7:
This week the research group conducted its first set of experimental trails using 0.2 mg of the Albumin Fluorescein protein and approximately 0.2 mg of cytidine residue. First we synthesized our hydrogels for the two experimental trials we conducted, one control without any cytidine residue for data comparison, and one with 20 mg cytidine to measure the effect that a positively charged component will have on the release rate of a negatively charge protein based drug (Fluorescein). We did this by creating a 1% Alginate solution with 1 g of Alginate placed in 100 ml of water, adding half of this solution to one designated container and the other half to the adjacent container. One of these containers were added with 20 mg of cytidine for comparison of it's effects on the normal rate of release of an Alginate hydrogel, and both were given the Albumin Fluorescein protein diluted in a 0.1 mg/ml concentration solution with water to ensure that both the control and 20 mg experimental trials could have their release rates measured. We then used the the eyedropper to take beads-sized droplets of these Alginate solutions and added them each to a 2% calcium chloride solution made from 1 gram of calcium and 50 ml of distilled water. The calcium chloride served as the primary activator of cross-linking between the monomers of the Alginate in the solutions which then caused the two hydrogels to form. Once this happened the hydrogel beads containing the different concentrations of cytidine were extracted from the calcium chloride solution and placed in test tubes containing a pH buffer, which served to maintain a general pH of about 7.0, the optimal level for the Fluorescein protein to stay viable. Finally samples were taken in different increments of time (5 min, 15 min, 30 min, 2 hr, 4, hr, 6 hr, 8 hr, etc.) over the course of two days (48 hr) and placed in test tubes that labeled the hydrogel and the time which they were taken. This was done for both hydrogels in order to obtain viable data of the release rate of the protein Fluorescein for the different conditions of the two hydrogels, which we hope to gain by analyzing the concentration of protein (mg/ml) in each of the samples and noting how it changes as time goes on through the use of a nanodrop device generously lent to this team by a researching professor at the Drexel College of Medicine. This team expects the protein concentration to increase as time does because as the protein will be released it will begin to accumulate in the pH buffer solution where the samples were taken, which should gain more and more protein as it is released.
This week the research group conducted its first set of experimental trails using 0.2 mg of the Albumin Fluorescein protein and approximately 0.2 mg of cytidine residue. First we synthesized our hydrogels for the two experimental trials we conducted, one control without any cytidine residue for data comparison, and one with 20 mg cytidine to measure the effect that a positively charged component will have on the release rate of a negatively charge protein based drug (Fluorescein). We did this by creating a 1% Alginate solution with 1 g of Alginate placed in 100 ml of water, adding half of this solution to one designated container and the other half to the adjacent container. One of these containers were added with 20 mg of cytidine for comparison of it's effects on the normal rate of release of an Alginate hydrogel, and both were given the Albumin Fluorescein protein diluted in a 0.1 mg/ml concentration solution with water to ensure that both the control and 20 mg experimental trials could have their release rates measured. We then used the the eyedropper to take beads-sized droplets of these Alginate solutions and added them each to a 2% calcium chloride solution made from 1 gram of calcium and 50 ml of distilled water. The calcium chloride served as the primary activator of cross-linking between the monomers of the Alginate in the solutions which then caused the two hydrogels to form. Once this happened the hydrogel beads containing the different concentrations of cytidine were extracted from the calcium chloride solution and placed in test tubes containing a pH buffer, which served to maintain a general pH of about 7.0, the optimal level for the Fluorescein protein to stay viable. Finally samples were taken in different increments of time (5 min, 15 min, 30 min, 2 hr, 4, hr, 6 hr, 8 hr, etc.) over the course of two days (48 hr) and placed in test tubes that labeled the hydrogel and the time which they were taken. This was done for both hydrogels in order to obtain viable data of the release rate of the protein Fluorescein for the different conditions of the two hydrogels, which we hope to gain by analyzing the concentration of protein (mg/ml) in each of the samples and noting how it changes as time goes on through the use of a nanodrop device generously lent to this team by a researching professor at the Drexel College of Medicine. This team expects the protein concentration to increase as time does because as the protein will be released it will begin to accumulate in the pH buffer solution where the samples were taken, which should gain more and more protein as it is released.
The team using a milliliter scale to measure the appropriate amounts of reagents (calcium chloride) to create the solutions used to synthesize our hydrogels
These masses of regents then had to be diluted into distilled water to create a solution with a predetermined concentration of the reagent. Above you can see the mixing of the calcium chloride solution
Here you can see the Albumin Fluorescein protein (yellow) in solution being added to the Alginate solutions prior to hydrogel synthesis in order to incorporate this protein into its cross-linked structure
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