This summary provides an overview of a document outlining homework assignments on a scientific research paper about protein adsorption and an experimental study on generating protein gradients:
Homework 6 is due on June 3, 2015 and involves summarizing a research paper on how oligo(ethylene oxide) resists protein adsorption and describing the motivation, experimental setup, and findings of an study on immobilizing protein gradients on hydrogel surfaces. The document provides questions to answer about the techniques, results, and conclusions of the two papers regarding protein-surface interactions and developing biomimetic systems to direct cell behavior through controlled microenvironments.
This summary provides an overview of a document outlining homework assignments on a scientific research paper about protein adsorption and an experimental study on generating protein gradients:
Homework 6 is due on June 3, 2015 and involves summarizing a research paper on how oligo(ethylene oxide) resists protein adsorption and describing the motivation, experimental setup, and findings of an study on immobilizing protein gradients on hydrogel surfaces. The document provides questions to answer about the techniques, results, and conclusions of the two papers regarding protein-surface interactions and developing biomimetic systems to direct cell behavior through controlled microenvironments.
This summary provides an overview of a document outlining homework assignments on a scientific research paper about protein adsorption and an experimental study on generating protein gradients:
Homework 6 is due on June 3, 2015 and involves summarizing a research paper on how oligo(ethylene oxide) resists protein adsorption and describing the motivation, experimental setup, and findings of an study on immobilizing protein gradients on hydrogel surfaces. The document provides questions to answer about the techniques, results, and conclusions of the two papers regarding protein-surface interactions and developing biomimetic systems to direct cell behavior through controlled microenvironments.
This summary provides an overview of a document outlining homework assignments on a scientific research paper about protein adsorption and an experimental study on generating protein gradients:
Homework 6 is due on June 3, 2015 and involves summarizing a research paper on how oligo(ethylene oxide) resists protein adsorption and describing the motivation, experimental setup, and findings of an study on immobilizing protein gradients on hydrogel surfaces. The document provides questions to answer about the techniques, results, and conclusions of the two papers regarding protein-surface interactions and developing biomimetic systems to direct cell behavior through controlled microenvironments.
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Homework 6. Due at the beginning of class June 3, 2015.
Prime et al. Adsorption of proteins onto surfaces containing end-attached oligo(ethylene
oxide): A model system using self-assembled monolayers 1. Please summarize, based on the discussion in the article, why adsorption of proteins occurs and how oligo(ethylene oxide) is thought to resist this process? Adsorption of proteins occur when the surfaces of the polymers and the proteins interact with each other like hydrophobic colloids do with surfaces. PEO's resist this process by compressing the polymer layers, making the chains have access to fewer configurations. Secondly, solvent is expelled from the surface layer. 2. Describe the motivation behind proposed experiments. Describe the experimental setup and the types of proteins and end-functional tested in this study? The motivation behind the proposed experiments were that they sought to define the length and number of EO chains needed to resist protein adsorption as well as a convenient way to test the effects of grafting density and polymer length upon the resistance properties of the surface. Gold films were immersed in ethanol solutions with different concentrations of proteins. Then XPS was used to determine the contact angle. The types of proteins used were fibrinogens, pyruvate kinese, lysozyme, and ribonuclease A. The thickness of the wafers were then measured. 3. How did authors characterize alkanethiol self-assembly and protein deposition? Please list the measurements they employed and briefly describe what those techniques are? They characterized alkanethiol self-assembly and protein deposition by finding the ellipsometric constants. They immersed the wafers in a solution of protein and then found the constants 4. Explain the meaning of Figure 4. What does refer to? Why is it when approaches 1 the dp is approaching zero? What conclusions about the effects of the length of ethylene glycol chain (number of mers) can you make from Figure 3 Figure 4 shows the relationship between wettability and x1. X refers to the mole fraction of the wafer. With the increasing X, the dp approaches 0 because if it is completely filled with the PEO then no proteins would adsorb. The adsorption of proteins decreases as the length of the chains increase.
5. What conclusions about ethylene glycol effects on protein adsorption did the authors reach? Was the length of polymer chain important? Was temperature important? Was MW of the protein important? They concluded that the ethylene glycol does indeed repulse or prevent protein adsorption. They found that long chains are optimal for making protein resistant surfaces. They also concluded that complete coverage of the surface longer chains are more effective. Temperatures between 4 and 25 degrees Celsius showed no difference while there was a significant difference between 25 and 37 degrees, where mixed SAMs adsorbed more protein at higher temperature. The MW of the protein was not important. Cosson et al. Capturing complex protein gradients on biomimetic hydrogels for cellbased assays 1) What is important of protein gradients in biology? Why are authors interested in recreating such gradients in vitro? Protein gradients are important in directing behavior of cells in numerous situations. They are established during tissue development to control cell behavior and induce specific tissue architectures. They are interested in recreating such gradients in vitro because they want to develop an approach of immobilizing protein gradients on more physiological cell culture substrates such as hydrogels. 2) What is Boyden chamber? Why are microfluidic gradient generators different from Boyden chamber? The Boyden chamber is used to study cell migration and invasion. It consists of a cylindrical cell culture where cells are seeded in the top and serum is placed in the well below. The cells then migrate through the pores toward the chemoattractant and their movement can be tracked. Microfluidic gradient generators provide a specific microenvironmental and spatiotemporal control which has been ignored in the Boyden chamber.
3) How do authors motivate their desire to work with hydrogel surfaces vs. glass or plastic? They believe that hydrogels have tissuelike diffusive and viscoelastic properties, while simultaneously providing an artificial microenvironment for cells. They can be decorated with bioactive ligands to make them smart. This closely mirrors that of the in vivo environment as contrasted with glass or plastic.
4) what are overlapping gradients, how were they generated and why were they generated? Overlapping gradients are a more complete composition of protein gradients because in cellular environments the gradients are more complex and proteins may interact with each other. They generated overlapping gradients with model proteins FITC-BSA-biotin and Alexa546-Fc fragments. They are generated in order to see how protein gradients would be like in vivo as compared to in vitro.
5) When discussing Figure 6 related to fibroblast migration on fibronectin gradients, the
authors state that cells residing on low or media concentrations of fibronectin where migrating towards high concentration but cells residing on high concentration where not migrating directionally. How do authors explain this behavior of cells? Authors explain this behavior of cells as the cells being fully saturated at that point. If they are at low or medium concentration they move towards the high concentration of proteins but are already saturated at the high concentration.
6) What are future uses of the technology being described in this paper? In the future, this technology can be used to discover the optimal concentration of factors that prevent stem cells from rapid and selective differentiation towards the desired specialization.
