Human Practices

Background:

As a very common disease in real life, scald is caused by non-flame high-temperature liquids (boiling water, hot oil, molten water), high-temperature solids (hot metal, etc.) or high-temperature vapor. After being scalded, people will feel very painful, and there will be a burning sensation and the skin will crack. From here, we got an inspiration.

Our group thought about whether we could make a product that is different from other products on the market. Spiders in the biological world can be used to make scald medicine.

Questionnaire

First of all, we made a questionnaire to find out how much the public knows about burns. Respondents need to answer on the questionnaire, such as whether they are familiar with burns and what they usually use to treat burns.

In addition, this questionnaire is used to popularize knowledge about scalds and how to treat scalds. In the questionnaire, people need to judge their understanding of burn treatment by answering the questions. In this way, the respondents can increase their awareness of burn treatment by combining questions and answers.

Analysis of results

The fourth question (Fig.1) of the questionnaire, " Do you know the treatment methods for burns?", we have set up two options, yes and no. Of the 205 people, 31.22% said they understood, and the remaining 68.78% said they did not. This shows that the public's understanding of the treatment of burns is very weak.

Among those who are familiar with treatment, the most commonly used external ointment is used to treat burns, reaching 96.88%. The second is skin transplantation, 84.38%. The third is surgery 54.69%. Later laser treatment, hyperbaric oxygen treatment, and pressure clothing were 34.38%, 26.56% and 15.63% respectively.

Fig.1 Do you know the treatment methods for burns?

Fig.2 Which kind of treatment methods have you heard of?

Fig.3 Have you ever heard of epidermal growth factor? I've heard of: 14.63%. Never heard of: 85.37%.

Fig.4 Have you ever heard of spider silk protein? I have heard of 37.07%, but I have never heard of 62.93%.

According to the survey results, we found that people know more about the traditional treatment of burns, and have no idea of any new treatment methods. Therefore，one of the purposes of this questionnaire is to popularize the knowledge that new spider silk protein can be used to treat burns. At the same time, it can also increase the popularity of synthic biology and our experiments to gain more support.

Fig.5 What do you think of our project?

In addition, 98.05% of people expressed support for our project, indicating that our project is very meaningful to them. 1.95% said it didn't matter. This may be because they don't know much about synthetic biology.

From the survey results, we have concluded that there is a great need for the popularization of knowledge about burn treatment. Because there are still people who lack knowledge about synthetic biology and new method for burn treatment. Our public knowledge popularization provides a good foundation. And it can also allow more people to participate in this project.

Interview with experts

During the summer vacation, we interviewed Professor Shan from the School of Animal Science at Zhejiang University and Professor Liu from institute of immunology. They both expressed that our project concept is very innovative and that it is a very interesting project. If we can achieve our goal, it‘s a better treatment plans for burn patients. Dr. Shan also stated that we can open up new ideas and our products can also be applied to the beauty field.

Fig.6 our team members were talking with Professor Shan（left）and Professor Liu（right）

In addition to experts in related fields, we also conducted interviews with doctors in dermatology and plastic surgery. Dr. Cai is from the Department of Orthopedic Surgery at Sir Run Run Shaw Hospital, Zhejiang University School of Medicine. We interviewed her on July 21st to learn about the usage the EGF materials and how can we test our results experimentally. Dr. Bu is the director of Hangzhou Xiaoshan Bu Dermatology Hospital. We interviewed him on line to learn what form should we make our products in and the environment for wound healing. They all expressed strong interest in our project and provided us with some product design ideas, expressing their hope of using our products one day.

Fig.7 our team members were talking with Dr. Cai（left）and Dr. Bu (online)（right）

Summary

Through the questionnaire, we learned that most of the public lack some common sense about burns and synthetic biology, so public education is in great need; most people, including a patient we interviewed, expressed their support for our research design. Through expert interviews and questionnaire data, we have learned that our project is necessary, and we are honored to receive the support of experts and people. We believe that after so many experiments, our project is the best.

Integtated Human Practices

Summary:

After brainstorming to determine the direction of our project, we divided into wet and dry teams to get started. Students in the wet team consulted literature to find ways to solve the problem, and students in the dry team contacted experts and doctors for guidance. Through literature research, we first targeted EGF protein. Bsaed on Professor Tan's interview and understanding of the extensibility and extensibility of spider silk, we decided to fuse EGF and spider silk protein together to achieve our purpose.

During the summer vacation, students from the wet team conducted experiments in the laboratory. Students from the dry team interviewed seven experts, including Lawyer Hu from the intellectual property field, Dr. Cai from the Department of Orthopedic Surgery, and Ms. Miao from a pharmaceutical company. They provided guidance and suggestions on our patent application, product production, and target groups. So, we started thinking about what our product would look like. Professor Liu also suggested that egf, spider silk protein, and egf fusion spider silk protein should be compared to obtain better results. Our results later showed that egf fusion with spider silk had a greater effect on skin damage. Through the communication with Professor Shan and Doctor Bu, as well as the suggestions of Ms. Miao from the pharmaceutical company, we finally decided on the form of our product - hydrogel.

After determining the product form, we contacted Professor Sun from Zhejiang Academy of Agriculture Science, who explained the production method of hydrogel in detail. We have conducted experiments on our products and preliminarily formed the hydrogel shape. The experiment is very successful. We will continue to optimize in the future.

1. Interview with Lawyer Hu

Why can't human intellectual achievements be shared? Because through the protection of patents, it can promote the development of this intellectual achievement, give these developers the corresponding privileges, and give them the corresponding rewards, which can promote invention and creation. When in a specific situation, human beings face specific difficulties and need to be solved with this patent to open up, then the state has the right to ask you to open up, and of course, it will be paid accordingly.

Lawyer Hu proposed that the patent is to give the inventor and applicant of the patent an exclusive protection. It is not the product itself that applies for a patent, but the technology. In fact, the more important thing about the patent is the technical solution, that is, the invention patent, the utility model patent and the design patent, which are the three patents in China. Therefore, products and patents are controlled by different laws. Of course, our patents should be novelty, practicality and creativity. Moreover, whether this patent application can be solved by the Patent Law, and your application will be approved according to the National Patent Office to see if it can grant you a patent.

The so-called novelty, practicality and creativity of patents are, first of all, novelty that has not appeared in any public media public literature; second, it is called creativity that is updated compared with existing technologies and existing schemes; and finally, it is called practicality that can serve the society.

For security, in addition to applying for a patent, there is another way to keep it confidential and not make it public casually, just like Coca-Cola.

This gives us an inspiration for patent applications. For us, what we need to apply for a patent is the method of making fusion protein. This is creative for the current market, because it is an application update; it is also practical, which can be put into production and use. This action may further develop our project, expand the scope of influence, and make it possible for us to profit from it.

2. Interview questions Dr. Shan

Dr. Shan is the professor and Doctoral Supervisor, from School of Animal Science, Zhejiang University and we interviewed him on July 19th to learned about usage of materials from organisms.

He reminded us of some of the key points in terms of papers, patents, products, etc. Also, he raised some questions which we should think about:whether the protein generated is active; whether it can be produced; how it can be used; whether it is innovative; how to optimize the feasibility; and whether there are alternatives.

In addition, he told us that there are now studies on the use of animal molecules to treat wounds or for cosmetic purposes, such as the traditional "ejiao" and silk masks. We were really inspired and benefited from the discussion.

3. Interview with Professor Liu

Professor Liu is is a lecturer and master tutor at the institute of immunology, Zhejiang University. On July 19th, we interviewed professor Liu to mainly understand the knowledge about the rejection reaction of alloprotein and the human immune system at Zhejiang University’s Zijingang campus.

Professor Liu mentioned that many existing dressings use collagen as matrix, and the recombinant protein of spider silk protein and EGF should consider its solubility, and if another material is used as a matrix, the value of spider silk protein cannot be reflected. He suggested that we should do several sets of control experiments: spider silk recombinant protein, EGF recombinant protein, spider silk protein plus EGF fusion protein three sets of experiments to test the superiority of our fusion protein, and whether it can better promote wound healing to carry out further experiments to verify.

He said that in the shallow part of the wound, the rejection reaction is not strong, and the treatment is simple, just remove the reaction causing the substance and clean the wound, and treat it with hormonal ointment. The deep wound rejection response is stronger and needs to be tested experimentally. He proposed that rejection is inevitable, depending on the time and location, no product will be perfect, all have side effects, for example, the treatment of tumor therapy has large side effects, but still needs to be used because its side effects are negligible compared to the disease.

According to studies, the immunogenicity of spider silk protein is similar to the one from collagen, which is within the acceptable range, so it can be preliminarily considered feasible, but it is still necessary to test to make a conclusion. He suggested that we could test the feasibility of our products using animal models such as mice and miniature pigs.

He told us that the drugs used clinically to relieve rejection are immunosuppressants, but its principle is to reduce rejection effects by making people's immune system being weakened, so it cannot be used for prevention before surgery.

For our project, he believes that fusion proteins will change the properties of proteins, then EGF would be unable to exert its original effect, whether it is necessary to do fusion and whether the spider silk protein structure can be maintained still questions worth thinking about . He told us that in the recent journal called Science, it was mentioned that artificial recombinant proteins form a network in the liquid phase, and whether EGF can be added to this.

He mentioned that if the spider silk protein is a polyform, large structure, so that it can be better used, but whether it can become a multimer, and how his water solubility is, also needs to consider.

Finally, he reminded us that in the project, we should highlight the key of our design concept, and our products should be able to make up for the shortcomings of other products on the market, and so if there are advantages, there is value.

4. Interview with Dr. Cai

Dr. Cai is from the Department of Orthopedic Surgery at Sir Run Run Shaw Hospital, Zhejiang University School of Medicine. We interviewed her on July 21st to learn about the usage the EGF materials and how can we test our results experimentally.

Dr. Cai told us that growth factors, antibacterial materials, etc. are used in plastic surgery to promote the growth of wound granulation, and now more humanoid protein, porcine protein and bovine protein are used, but like pig protein and bovine protein will be allergenic and rejective.

For the so-called EGF flowing into the blood will cause human cells to become cancerous, Dr. Cai said that "leaving the concentration to talk about the negative effects are hooliganism", EGF to promote the healing of the wound, that must promote cell regeneration, that after overexpression after the regeneration is not controlled there will be tumors, these are two-sided.

Dr. Cai believes that what we call "immobilization of EGF" is physical fixation, and the fusion protein we are doing is a biochemical gene loading rather than a simple package. We also need to talk to the instructor about this issue.

EGF is generally used in plastic surgery for moles, abrasions and some refractory wounds, and can be used in large and small areas. These drugs are already widely available, available online and affordable.

EGF can enhance its proliferation ability in cells and can be verified by cloning experiments. The expression of EGF in different cells is also different, for example, the expression in tumor cells will be higher than in normal cells, so it can grow wildly.

Animal experiments generally use mice and rabbits, and now diabetic wounds are mostly used in plastic surgery topics, because diabetes is a difficult wound (which is related to microvascular damage caused by diabetes), but due to our limited funds, we can only raise it in the defense.

5. Interview with Dr. Bu

Dr. Bu is the director of Hangzhou Xiaoshan Bu Dermatology Hospital. We interviewed him on July 17th to learn what form should we make our products in and the environment for wound healing.

Dr. Bu said that because the factory protects employees better, there are fewer accidental burns at work, but there are more allergies in daily life. Burns belong to the category of surgery. Dermatology mainly treats some patients with chronic ulcers and some deep wounds. He proposed that the dressing should prevent infection, preferably in the form of a spray, wet compress to promote issue growth, and to be breathable and humid to speed up wound healing. EGF spray (refrigerated) and ointment (at room temperature) have no side effects. Chronic burns are difficult to heal. Those with large wounds and poor physique should be assisted by EGF. He mentioned that he has not studied spider silk protein now, and silk has studied masks. The function of Band-Aid is to stop the bleeding and prevent the skin from being exposed in the air. For chronic patients, EGF can be used for adjuvant treatment. Dr. Bu suggested that the human body may have allergies and rejection reactions of spider silk protein, so two kinds of dressings can be made. One has egf and the other is spider silk excipients (similar to Band-Aids and Bandings) for chronically added EGF.

For the popularity, Dr. Bu said that it can be popularised if it is made into a band-aid similar product (no egf and have egf) combined with anti-inflammatory and anti-inflammatory and anti-infection materials,This can make our projects more productization , and team member in our design team can also design logos for Band-Aids.Although the cost of spider silk protein is relatively high, the strength of the product, breathability and the effect of rapid haemostasis for acute trauma are good publicity points. Generally speaking, EGF is not added for acute, and EGF is added for chronic use(patients with large wounds).Silk is more skin-friendly, less exclusive, and more comfortable to use. For EGF, due to patent issues, there are fewer domestic manufacturers and more expensive ones. Now they are basically made in China.

6. Interview with Ms. Miao

Miss Miao is a marketing manager of pharmaceutical company. We interviewed her on July 31st to understand how to create a complete market plan and increase the brand awareness of our products.

Ms. Miao said that creating a complete market plan requires executing a summary, market analysis, market strategy, and action plan. At the same time, she also suggested that if conditions permit, we can conduct a policy update to help us focus on patient flow, pay attention to patient treatment paths, target markets, and identify target population. To find leverage points (based on policy dynamics). The most important point is to conduct SWOT analysis to identify product strengths and weaknesses, such as spidroin, which has the advantage of high toughness but the disadvantage of high cost. So at this point, we need to look for a doctor to introduce the benefits of spidroin to them, and then use the doctor to persuade consumers to give up the antiscald drugs they pass on and choose higher priced spidroin. At the same time, it is necessary to establish a good market strategy to determine the corresponding departments and patients (based on SWOT analysis to determine the future development and sales strategy of the product).

In order to increase the brand awareness of our product, we need to decide on the implementation plan and conduct clinical trials to demonstrate how our spidroin is better than the leading product. This requires doctors and patients to participate and integrate into this activity. Determine the target audience to increase sales. If you want to sell products, you still need to find a suitable distributor. Find academic leaders and authoritative experts in this field to endorse the product. You can introduce spidroin to experts at the academic annual meeting, which will allow more experts to come to the product platform. At the same time, training should also be provided to the sales team to increase their confidence in the product and enable them to better introduce and sell the product. If you have the strength, you can publish articles to let more people see spidroin, in order to increase product awareness.

7. Interview with Professor Sun

Professor Sun is from Zhejiang Academy of Agriculture Science and we interviewed him on August 3rd in the collage. We mainly learned that there are also many types of hydrogels and learned how to make them, and the combination of 3d printing and bio-ink that we mentioned. The professor also contacted us to add some pieces of spider silk.

Professor Sun mentioned that the gel is mainly a polymer with high water content. There are also many types of hydrogels. The specific method is very simple. First, use a spoon to add the initiator (transparent catalyst, potassium permanganate), then add the main spider silk fusion protein (add whatever you do), then add deionized water, shake it until it dissolves, and illuminate the lamp gel. The main principle of this step is that under the stimulation of light, the key will open and become a long chain, from small points. The son becomes a macromolecule. Finally, add the gel and make a net of wires. At the same time, it is suggested that the finished product should be physically close to human skin, flexible and elastic, and not toxic in itself. The structure should also be relatively close to the human body, which can induce the formation of new human skin. Because the drugs used on the human body are very strict and cannot involve ethics, Professor Sun also explained to us how to test the implementability of the finished product after it comes out. First, in vitro experiments, starting with human dermal cells in vitro, to see if the cells can survive and proliferate. If there is no problem, you can proceed to the next step, animal experiments, cut off a piece of the animal's skin, and then paste the dressing to observe the situation. If there is no problem, you can go to the clinic and find a volunteer. At the same time, he proposed that he recently saw someone combined with 3D printing, added some biological ink, and finally printed it into a three-dimensional biological material. In response to the question we asked, he replied that our finished product will not have much rejection. We are not a transplant, but a tissue project. We build a tissue framework in vitro and then transplant it to people. This process can also be called regenerative medicine. For allergic reactions, experiments can be carried out in vitro to see if there is rejection and make statistics at the same time. Do it according to the number of days to see if the growth is reasonable (it needs to be on the instrument). Finally, we asked how to preserve the hydrogel. The professor said that it should be placed in a sealed bag and put the air in a constant temperature or 3-5 degrees. Generally speaking, the temperature should not be too high. Some gels are relatively absorbent, but they are generally not so strong in biology. They can be put in liquids, but it is best to put them in safer solvents such as buffer and glycerin. Remember to put dry ice to prevent them from mutating. The professor finally raised a question. The output of spider silk is so small, not as much as silk, and the supply may be insufficient. He suggested that we add some silk fragments.