Difference between revisions of "Team:SSTi-SZGD/Human Practices"

Our team mainly consists of female team members (7 out of 12) who have interest in natural cosmetic and skin care products. In addition, our department not only has instructor in the cosmetics industry with rich teaching experience and professional knowledge, but also has a club to develop cosmetics, which has professional equipment and space for research. At the beginning of the brainstorming process, we wanted to take advantages of the existing resources and facilities, combining with our own interest, to focus the project on cosmetics.

By doing background research and visiting numerous cosmetics stores, supermarkets, drug store, we learned that the current cosmetic market is dominated by functional cosmetics, especially anti-wrinkle, whitening and moisturizing products are most popular. During the visits, we also learned a lot about the ingredients of the products, we found out that most anti-wrinkle and moisturizing products contain hyaluronic acid (HA). HA, a polysaccharide that naturally exists in human bodies, has exceptional water retention ability that keeps the skin moist, weakening wrinkles and restoring skin physiological function. As far as we have observed, HA has become a selling point in recent years due to the increasing demand of moisturizing products, especially in Asian countries. In addition, we also found that HA is a biochemical drug of high clinical value, which is widely used in various ophthalmic operations as well as in treating arthritis and accelerating wound healing.

In order to understand how efficient HA is in medicine and wrinkle removal. We visited the plastic surgery department of Shenzhen People's Hospital. We had informative conversation with Dr. Xiaojun Li. From the conversation we learned that the current methods used for wrinkle removal mainly include surgery, filler injection and laser treatment, however, each of them has certain limitations: Surgical procedures can lead to facial infections, subcutaneous bleeding, scarring and numbness caused by nerve damage; filler injection primarily using HA and botulinum toxin, while botulinum toxin can lead to local functional inhibition, facial paralysis, drug dispersion and stiffness; HA is part of body's own substance, it is relatively safe but has limited effect when injected. Laser treatment is mainly used for the treatment of skin defects and has requirements on the quality of the treatment equipment. Poor equipment quality can result to skin burns or scars. Also, these methods are expensive, painful and require considerable amount of time to complete. Plastic surgeries are the last resources for people who could not get satisfactory results from topical application of cosmetic products. There is no middle ground available.

In order to take words from more than one person, and continue seeking alternative methods to achieve better skin, we went to the PoK Oi Hospital, a specialized beauty clinic that has many franchisers in Guangdong province, China. By consulting with the chief physician at Pok Oi, we learned extensive information about HA and its applications. For example, most patients came here for wrinkle removal procedures were in their late 20s to 50s; generally middle and low molecule weight HA were selected for this procedure, and these HA products are mainly manufactured in Korea or Switzerland and great in purity.

In addition to filler injection, physicians in Pok Oi sometimes used golden microneedle devices for wrinkle removal. These devices are made of metal, and usually used to facilitate the delivery of nutrient ingredients into the skin. Compare to ordinary needles, these devices are with tiny needles, infiltrate the skin in a non-invasive and painless way, but are now rarely used because the susceptibility to infection. It was the first time we came across the concept of micro-needle, which opened up a new direction for our project development. After a few more rounds of brainstorm, we decided to develop a novel device using HA to prepare microneedle for wrinkle removal.

By doing another round of background research, and consulting with our advisers who had expertise in cosmetic development, we understood that most of the HA product in the market is produced by the fermentation of Streptococcus species. Although Streptococcus has natural HA secreting pathway, these species have high risk of pathogenicity, which may influence the safety of the end product. We decided to use genetically engineered food-grade B.subtilis to produce HA, taken the advantage of existing precursor pathway and genetic methods. We thought that our idea was of great economical value, not only optimize the microbial production of HA, but also develop a novel approach for wrinkle removal. Therefore by completing the business proposal, we participated in the “Inspiration Cup” innovation and entrepreneurship competition organized by the local government in Shenzhen.
At the first round of the competition, our project was praised by the judges for its innovative application, but they needed further information to be ensured that the project generates sufficient economical value. We were troubled by these problems and eager to find a solution.

Through the introduction of our instructors, we visited Bontac biological engineering company, which is manufacturing company specialized in growth factor production using synthetic biology technology. The chief technical officer, Dr. Zhao, received us and provided solutions to some of the problems that troubled us:

1. enzyme (hyaluronidase) purification can be achieved by a series steps of filtration and chromatography.
2. Yield can be improved through promoter optimization and metabolic engineering.
3. Some third party agents can help with HA molecular weight determination.

Based on his advice, we designed a 6His-tag for enzyme purification via affinity chromatography, decided to up-regulate a number of native genes of B.subtilis in order to increase HA production rate, and optimized HA separation method to remove all impurities. We are very grateful to Bontac and Dr. Zhao for their construction advice that helped us improving our project.

Genetic modification is the key to our project, however, currently in China, GMO is still a concern to a lot of people. As working on a manufacturing project, it is very unlikely that our genetic modified organism is directly exposed to human or the environment. In our proposal, we drafted a plan that the key ingredient, HA and hyaluronidase, were to be produced in a strictly regulated manufacturing plant and undergo process of separation, purification, QA testing, etc, before commencing the HA microneedle production. In order to further elaborate on this issue, we visited Shenzhen Watsin Genetech Co. Ltd, who is specializing in microbial fermentation of a natural polymer that is similar to our HA.

Ms Lin, the head of technology department, gave us advice on biosafety issues. Watsin has a well-equipped manufacturing plant coupled with a whole set of production procedures with national standards to be complied. General speaking, in production, after the fermentation stage, initial inactivation of microorganisms via cell breakage or acid and alkali treatment is carried out. Followed by secondary ultra-high temperature sterilization, and the residues will be treated at the sewage treatment station. In addition, each individual product has specially designated fermentor that is regularly checked for leakage. Taking every step together, there is little chance for bio-waste leakage or contamination to human or the environment to take place in their production plant. We were convinced by the stringent control in their plant, and also came up with the idea of collaborating with Watsin in the future for production outsourcing opportunity.

Followed to the previous competition, our project further got invited to participate in the “Entrepreneur Star” competition in Nanshan District, Shenzhen. This round of competition was slight different from the last ones because there were judges from different fields for project review. We received feedback from judges with respect to business planning, financial analysis and so on.

Participating in the competitions helping us identifying pitfalls in the project. After the competition, we went over again the business plan, and tried to reformulate it in a way that fits the industrial production box. This time we needed more professional guidance from the field experts. Therefore we visited Shenzhen Sinobioway Xinpeng Biomedicine Co. Ltd, who is the leading supplier to a series of enzyme products with customers all around the world.

By talking with their financial analysts and the head of production, we presented our problems with regards to transition from laboratory R&D to industrial manufacturing. They advised us on how to analyze management expenses, sales expenses, manufacturing expenses, direct materials and interest expenses, and eventually make a complete set of financial planning (ref BP).