Integrated Human Practices

Overview

The love of beauty is universal. Skin care has gradually become the daily life of many people, and the ways of skin care are almost the same. We hope to use the knowledge of synthetic biology to provide a new paradigm for skin care . Based on our project of artificial skin for burning treatment, we want to continue to give play to the moisturizing efficacy of bacterial cellulose(BC) on skin, and produce a live bacterial skincare product on this basis. We visited dermatologists, plastic surgeons, synthetic biologist and live bacterial enterprise, collected their views on our project, and improve our design of expereiments and product. We finally completed our project by focusing on the closed-loop mode of "Feedback - Improvement - Feedback".

Timeline

When we think about a problem, we usually fall into the "one-sided" mire, and systematic thinking can help people to see the problem more "comprehensively". In order to keep the team more efficient, we need to constantly make adjustments in the process of collecting information and feedbacks. To this end, we have organized a timeline circle of "Feedback - Improvement - Feedback".

Project Inspiration

Market data graph

Through network research, we concluded that people's demand for skin care is mainly reflected in the following aspects: pores-shirk, soothing redness, oil control, clearing acne blemishes, deep cleaning, anti-spot, repairing, brightening, anti-aging, moisturizing, etc. (Figure 3). In order to further grasp the current market demand, we conducted a questionnaire survey, and found that the skin care efficacy that people's mainly concerns are moisturizing, anti-aging, repairing, anti-spot, and brightening (Figure 4).

Interview with Watsons sales

Our team went to Watsons and interviewed Watsons sales(Figure 5). We learned that most of the products on the market have a single efficacy. To achieve multiple efficacy, you need to use different skincare products at the same time, such as Brightening Cream, Brightening Blemish Cream. Therefore, our team intends to create a multifunctional skincare product.

Interview with Winkey Pharmaceutical R&D Co., Ltd.

We need to find an ingredient with multiple skincare efficacy. Through investigation and research, we found that the three efficacy of brightening, anti-spot and anti-aging can be achieved through anti-oxidation. Therefore, we set the anti-oxidation of active ingredients as our goal, and fully considered the allergy problems that skincare products are prone to cause. Finally, we chose glutathione(GSH) as the ingredient of our product. Glutathione(GSH) is an important component of antioxidant in human body, and it can be naturally produced by human body. In order to have an deeper understanding of glutathione(GSH), our team came to Shenzhen Winkey Pharmaceutical Research and Development Co., Ltd. for an interview (Figure 7). The following is our conversation with Winkey:

Conclusion

After the interview with Winkey Pharmaceutical Research and Development Co., Ltd. , we brainstormed and finally decided to develop a new form of live bacterial skincare product. This new product will solve the practical problem that glutathione(GSH) is difficult to be directly applied to cosmetics. We allow users to release glutathione (GSH) through light control when using it, so as to ensure that the activity of GSH is not affected by production, transportation and storage.

Interview with Cosmetics Consumers

To better understand the market demand, our team conducted interviews with cosmetics consumers (Figure 8).We learned that cosmetics consumers (especially young consumers) are very concerned about "skin microbiome", in which they mentioned that the bacterial lysate has the efficacy of maintaining skin microbiome. In this case, we plan to develop a product , which produces bacterial lysate in the process of use. In addition, we found that consumers have a certain understanding of skin microbiome, but they do not fully understand synthetic biology. We hope to present synthetic biology in different forms to attract more people in learning synthetic biology. (See "Education" for more details)

Interview with Dermatologists and Plastic Surgeons

In order to improve our project, we consulted professionals about skin knowledge. Therefore, we interviewed and consulted with two dermatologists and two plastic surgeons offline to know more about the basics of skin (Figure 10,11). After the doctor's introduction, we found out that antioxidation is essential, and also learned that the antioxidant medical aesthetic methods on the market may cause secondary damage to the skin. We explained the preliminary idea of our project to the doctor. And then, they provided us with good suggestions for improvement. The following is the conversation process of our interview:

Conclusion

After listening to the preliminary idea of our project, the doctor raised two questions about our project:
1. Can glutathione(GSH) be fully absorbed by people's skin?
2. Can the product still be used on nonintact skin?

After certain research, we found that glutathione(GSH) small molecules can be absorbed by the skin, which again proves the feasibility of our project product. Also, considering the safety when users use our product on nonintact skin raised by the dermatologist, we decided to further improve the product. Later, we will indicate on our product package that "It is not recommended to use the product on nonintact skin". In addition, in order to avoid others' feeling that our project is too abstract, we have made different versions of brochures to better promote our project. (See "Communication" for more details)

Experimental Design

The lysis and safety module

Interview with Dr. Mingxing Tang

Based on our project from last year, we tried to optimize the lysis and safety modules. When we repeated the lysis experiment, we found that the constructed blue light reaction system did not work well in rapidly growing cells. Therefore, we contacted Dr. Mingxing Tang (Figure 12). He suggested removing the LVA tag from the repressor cI in pDawn to reduce the leaky expression of lysis genes, which might contribute to the stability of the blue light response system. (See "improvement" for more details)

Product Form

Interview with Yakult Factory

In order to find a better product form, we visited the Guangzhou, Yakult factory (Figure 14), a bottle of Yakult (65mL) contains 20 billion Lactobacillus casei Shirota strains. We communicated with a technician from the Yakult factory and she introduced the company and its product technology. And then, our team visited the factory's laboratory and the production workshop. The technician suggested that we should first determine the form of our product, whether it is bacterial liquid or emulsion. In the case of an emulsion, the emulsion formulation should be taken into account, which determines the toughness of the final film.

Conclusion

After talking, we got the preliminary design of the product form, and we will consider the effect of formulation on the quality of bacterial cellulose film.

Interview with Shannxi Lifegen Co., Ltd.

During the communication and cooperation with the NWU-China-A, we helped each other to find the company we needed ( Dr. Penggao Dai, Technical Director Shannxi Lifegen Co., Ltd.). Due to the severe of epidemic, we interviewed Dr. Penggao Dai of Shannxi Lifegen Co., Ltd. online (Figure 15, 16). He provided us with useful information from the perspective of working in the field for almost 20 years. Here's the conversation we had:

Conclusion

Considering that the bacterial emulsion is easier to use, we finally decided not to use freeze-dried powder as our product form.

Interview with Bode Chuangyan (Guangzhou) Biotechnology Co., Ltd.

Is there any better methods can we use to evaluate the efficacy of the product? How to adjust the bacterial emulsion formulations? Therefore, we interviewed and consulted the technician of Bode Chuangyan company offline(Figure 17, 18). Our team visited Bode's lab and presented our project to the technician of Bode Chuangyan. She suggested that we may improve the toughness of the bacterial cellulose film by adjusting the ratio of the thickening agent.

Conclusion

The product form originally envisaged was the bacterial emulsion, but after many formulation and performance experiments, our product is presented in the form of live bacterial emulsion/gel. This is more conducive to later use and product efficacy. (See "Proof of Concept" for more details) In addition, we learned that zebrafish eggs can be used for antioxidant efficacy assays. However, Considering the welfare of animals, we finally decided to abandon the method of using zebrafish eggs for the detection of antioxidant efficacy, but used a free radical kit (in vitro). (See "Experiments" for more details) This interview pushed our project forward.

Equipment Design

Since our product require red and blue light illumination for use, our team discussed and designed a device with dual-light control. We got the inspiration for the device design from an interview with Ms. Lee (Figure 19), a skin health specialist at Naturade. Considering the user usage scenarios, we designed a large-scale device for the beauty salon (Figure 20), which can be used with live bacterial products. Finally, a portable device is designed for other usage scenarios (Figure 21).

Questionnaire survey

At the reminder of the skin health specialist at Naturade, we thought the users' acceptance of the live bacterial skincare product should be taken into account. According to statistics, stakeholders' knowledge of live bacteria mainly comes from food, and they are willing to try live bacterial skincare product. The main suggestion was that the product should be easy and effective to use.

Summary

When we started to implement human practices, the question we always thought about was "How can our project contribute to the world?" The significance of our project is not only to prove how excellent we are, but also to prove the significance of the project to the world. To this end, we identified the stakeholders of our project, shaped and improved the design of our experiments and product through in-depth communication with them. Thanks again to our stakeholders who have given us helps and cares during the project exploration!

References

[1] GAO Yuhao, WU Yongjie, ZHU Yaxin, et al. Construction and energy regulation of engineered glutathione-producing Pichia pastoris[J]. Food and FermentationIndustries, 2021,47(7):21-26.
[2] Wang Xiaowei, Zhang Hongyan, Liu Rui, Wang Dongkai. Research Progress of Glutathione [ J ]. Chinese Journal of Pharmaceutics ( Web Edition ), 2019, 17 ( 04 ) : 141-148.
[3] WANG Wei-wei, Tang Liang, ZHOU Wen-long, YANG Yan, GAO Bo, ZHAO Yun-Feng, WANG Wei. Progress in the Biosynthesis and Metabolism of Glutathione. China Biotechnology, 2014, 34(7): 89-95.
[4] LIU Xiaodong. Directed Evolution of Glutathione Bifunctional Synthase and Enzymatic Synthesis of Glutathione [ D ]. Zhejiang University, 2018.
[5] Li. Preparation and Characterization of Glutathione Liposome [ D ]. Shenzhen University, 2018.
[6] Zhu Yifu. Study on the stability of reduced glutathione [ J ]. Modern Food Technology, 2011, 27 ( 08 ) : 919-923.
[7] HU Jing.Study on the purification and stability of reduced glutathione [ D ].Hubei University of Technology, 2007.
[8] Zhang Yi, Ye Sheng. Physiological function of reduced glutathione and its clinical application [ J ]. Chemistry of life, 2020, 40 ( 12 ) : 2226-2235.
[9] Wu Xujun.Preparation and functional product development of biological cellulose mask [ D ]. Donghua University, 2014.
[10] Yang Yuxin. China 's skin care industry status and consumer demand analysis [ J ]. Value Engineering, 2019, 38 ( 17 ) : 258-260.
[11] Li Zhaofeng.Optimization of bacterial cellulose production and study on the structure and mechanical properties of bacterial cellulose-hemicellulose complex [ D ].Guangdong University of Technology, 2021.
[12] Jia Qinghui, Lu Hongmei, Chen Li, Zhang Liping. Effects of synergistic factors on the changes of substances in bacterial cellulose fermentation broth produced by Gluconacetobacter hansenii [ J ].Chinese Brewing, 2016,35 ( 01 ) : 14-18.
[13] Gong Shuhui. Analysis of the current situation and prospects of microecological skin care products in China [ J ].Science of Daily Chemicals, 2020,43 ( 09 ) : 1-4.
[14] https://marketing.hd.weibo.com/article/view/5525