Intergrated Human Practice

1. Overview

In our Human Practices journey, we have consistently adhered to the core principles emphasized by iGEM: reflection, responsiveness, and responsibility. Rather than treating Human Practices as a supplement to our experiments, we regarded it as a continuous process of dialogue and learning, where every round of research, theory and practice refined not only our project design but also our understanding of its broader impact.

Through engaging with patients, doctors, professors, enterprises, and international scholars, we did not simply collect opinions — we actively responded to their concerns, re-examined our assumptions, and adjusted our strategies accordingly. Each interaction became both a mirror and a compass, helping us to critically reflect on the feasibility, safety, and social value of our design, while guiding us toward more responsible choices.

This iterative process allowed us to build a project that is not only scientifically innovative but also socially grounded, aiming to bridge the gap between laboratory research and real-world needs. In this way, our Human Practices work fulfills iGEM's vision: to ensure that synthetic biology is developed responsibly and used for the good of the world.

Stakeholder Network Map

Our Human Practices journey involved extensive engagement with diverse stakeholders across multiple domains. Click on any stakeholder in the network below to learn about their contributions to our project development.

2. RTPNR Framework - Upgrade and Expansion of the TPNR Framework

2.1 Overview

In 2024, our OUC-Haide iGEM team proposed the TPNR framework—"Theory–Practice–Newtheory–Re-practice." This framework emphasized starting from theory, moving into practice, and then continuously refining and developing theory through reflection and verification. It helped us avoid being constrained by static assumptions, instead enabling dynamic progression in line with the vision of iGEM: "to make the world better with synthetic biology."

The inspiration for TPNR came from philosophical reflections on the dialectics of development. As Hegel pointed out, progress often unfolds in a spiral movement, where each stage can become a new beginning rather than a fixed end (Hegel, 1837/1956). From this perspective, theory and practice are not opposing forces, but mutually reinforcing elements in a continuous process of improvement.

Figure 1: The 2024 OUC-Haide TPNR cycle framework

However, by 2025, we gradually realized that the TPNR framework was still insufficient to encompass the full dimensions of Human Practices. If a project begins solely from "theory," it risks being disconnected from real-world needs and the voices of stakeholders. We recognized that the true starting point should be Research—listening, investigating, and collecting feedback from diverse groups. Research not only helps to identify the most pressing problems but also integrates expectations from people of different ages, backgrounds, and roles. In this way, the project remains grounded in its original mission: to use synthetic biology to respond to genuine human needs and ultimately make the world better.

This idea resonates strongly with John Dewey's philosophy of pragmatism. Dewey emphasized that knowledge does not emerge from abstract deduction, but from lived experience and the process of inquiry into real-world problems (Dewey, 1938/1991). He proposed a cycle of "inquiry–hypothesis–experiment–verification," in which truth is not a fixed entity but is gradually shaped through problem-solving and the refinement of experience. Dewey's pragmatism thus provides strong theoretical support for our principle of "research first."

Figure 2: The cycle of "exploration - hypothesis - experiment - verification"

Building on this understanding, in 2025 we proposed an upgraded framework: RTPNR (Research–Theory–Practice–New-theory–Re-practice/Re-search). It not only preserves the dynamic interaction between theory and practice but also emphasizes Research as the starting point, while highlighting the pivotal role of New-theory in driving the project upward.

Here, it is necessary to clarify the relationship between RTP and RTPNR. Every Human Practices activity we carried out essentially followed an RTP inner cycle (Research–Theory–Practice). This cycle is the basic unit of progress: through research we identify problems, we propose theory as the design framework, and we verify and refine it through practice. Each RTP cycle results in a New-theory. Yet, New-theory is not the end—it acts as a bridge, guiding us into a higher-level RTP cycle. In other words, RTP functions as the methodological "engine" of progress, while RTPNR is the overarching framework that links these engines together and ensures continuous upward advancement.

Figure 3: Basic RTP cycle

Figure 4: RTPNR spiral ascending framework

Over multiple iterations, the RTP cycles accumulate and connect. Each emergence of a new theory is a step upward in the spiral, allowing us to enter new research and practice with more mature ideas and deeper insights. After several rounds of RTP cycles, the project eventually arrives at the stage of Re-practice or Re-search, marking the entry into a higher-level cycle. In the context of iGEM, this does not simply mean repeating experiments—it represents the transition toward true clinical validation, patient adoption, and societal implementation. We refer to this as the "ultimate RTP cycle," in which Practice no longer refers solely to laboratory testing, but to the responsible application of synthetic biology within human society.

Figure 5： Spiral ascent

Figure 6: Ultimate RTP cycle

Therefore, RTPNR is not a replacement for RTP, but rather its extension and elevation. RTP provides the methodological units for each stage, while RTPNR ensures that these stages are not circular repetitions, but spiral ascents that progressively guide the project from initial research and theoretical conception toward higher levels of clinical and societal impact.

Throughout this process, our project remains reflective, responsive, and responsible. In each round, we generate new theories through reflection, use them to respond to real-world needs, and take on greater responsibility at higher levels. Ultimately, the RTPNR framework transforms feedback from Human Practices into sustained momentum, guiding the project step by step toward greater maturity and refinement. It reinforces our conviction that synthetic biology is not only a vehicle for scientific innovation but also a form of social responsibility. Only through the spiral ascent of RTPNR can we truly approach the world with greater responsibility and wisdom—and make it a better place.

Overview

The RTPNR framework provides us with a spiral pathway: starting from research, passing through theory and practice, and culminating in new theory, which then acts as a bridge to higher-level cycles. In this process, RTP serves as the fundamental unit of every Human Practices activity, while RTPNR functions as the overarching framework that connects and elevates these units. This mechanism ensures that our project does not remain confined to the laboratory but progresses step by step toward real-world application. Guided by this framework, we have consistently embedded reflection, response, and responsibility into every stage, enabling our project to advance not only in scientific innovation but also in social value—truly embodying the iGEM spirit of "making the world better with synthetic biology."

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3. From Research to Focus: Converging on Diabetic Wound Dressings Through Real Needs and Voices

3.1 R—From Personal Inspiration to Social Focus: The Prevalence and Urgency of Diabetic Chronic Wound Healing

Our Research began with a personal family story. A team member's grandmother, who suffers from diabetes and limited mobility, sustained a severe leg wound after a fall. Despite repeated attempts with common treatments—such as oral antibiotics and topical growth factor ointments—the wound showed no significant improvement. Instead, it lingered for months, complicated by recurring redness and inflammation. This experience made us deeply aware that chronic wounds in diabetic patients are not simple injuries, but rather persistent and often overlooked medical challenges.

Figure 7: Grandma was injured

Medical research further reveals the mechanisms behind this phenomenon. Chronic wounds in diabetic patients are often caused by vascular and nerve damage due to poor blood sugar control, combined with impaired circulation, hypoxia, and decreased immunity, which significantly delay healing (Wong et al., 2015). As clinical definitions highlight, such wounds are difficult to heal, highly susceptible to infection, and may even progress to gangrene or require amputation. Global epidemiological data underscores the severity of this issue: as of 2024, approximately 589 million adults worldwide live with diabetes, a number projected to rise to 853 million by 2050 (IDF, 2024). Among them, about 25% experience impaired wound healing, and 15% develop severe complications such as diabetic foot ulcers (Nunan et al., 2014). These chronic wounds not only diminish patients' quality of life but also place a heavy burden on families and healthcare systems.

Figure 8: Diabetic foot complications

Against this backdrop, we realized that for mid-to-late stage diabetic wounds—those extending into the dermis—traditional low-risk therapies are far from sufficient. While such methods may be suitable for minor injuries, they are often ineffective when facing severe chronic wounds. To better understand the prevalence of this issue in society, we conducted an online survey in the early stages of our project, focusing on diabetic patients and their wound healing experiences. A total of 251 valid responses were collected.

Figure 9: A total of 251 questionnaires were received

The results showed that nearly 80% of respondents were directly connected to diabetes, including patients themselves, family members, healthcare providers, and professionals in drug and device development. Family members made up the majority, accounting for over 60.16% of the sample. Meanwhile, 72.22% of respondents reported that they "frequently" or "occasionally" observed slow wound healing. These data confirmed once again the widespread and urgent nature of the problem.

Figure 10: Percentage distribution of respondents' identities

Figure 11: 72.22% respondents "frequently" or "occasionally" observed slow wound healing

To gain deeper first-hand insights, we also conducted field visits. In a diabetes specialty food store in Qingdao, we saw shelves filled with low-sugar, low-GI products, and patients enthusiastically purchasing them, reflecting the widespread awareness of blood glucose management. However, when we discussed chronic wounds with the store owner, he explained that most patients still relied on dietary supplements to boost immunity or improve metabolism, with little awareness that wounds require specialized care.

Figure 12: Low-sugar and low-carbon foods in the store

Figure 13: The team member poses for a photo with the shop owner

In a nearby community hospital specializing in endocrinology, physicians echoed similar concerns: patients facing delayed wound healing typically relied on basic treatment methods and rarely used professional topical drugs or dressings. In fact, according to clinical definitions, such wounds are difficult to heal, prone to infection, and may further deteriorate into gangrene or require amputation. Low-risk conventional therapies can manage minor wounds, but once a wound progresses to deeper tissue damage, they are almost powerless to halt worsening conditions.

Figure 14: Qingdao Endocrine Diabetes Hospital

Figure 15: The team members had a conversation with the doctor from Qingdao Endocrine Diabetes Hospital

These findings gave us a clearer understanding of the gaps in both clinical and household care for diabetic chronic wounds and highlighted the broader societal shortcomings in this area. While many patients are well aware of the importance of blood sugar control, their approaches to wound care remain traditional and limited. As a result, we realized that developing a new wound dressing is not only a matter of scientific innovation but also a direct response to patients' genuine needs. In line with iGEM Human Practices principles, a responsible project must respectfully and openly listen to stakeholder voices and continuously adjust its direction through scientific research.

From a personal story to literature evidence, from an online survey to community fieldwork, we confirmed that diabetic wound healing challenges are not isolated cases but widespread medical and societal problems worldwide. This systematic Research stage allowed us to complete the "R" of the RTPNR framework, and also facilitated the transition from "Research → Theory": moving from identifying individual problems to summarizing collective pain points, and further elevating them into a researchable direction. This laid a solid foundation for our next step: the Theory stage, in which we propose to develop a novel hydrogel dressing to meet these pressing needs.

Figure 16: Research

3.2 T—From Needs to Solutions: The Conception of a Novel Bioactive Hydrogel Dressing

After completing our investigation of the problem, we moved into the "T: Theory" stage of the RTPNR framework. At this point, we needed to build upon the real-world needs we had identified and propose a feasible project concept, clarifying what kind of therapeutic dressing we would design to address the challenges of diabetic chronic wounds.

First, our questionnaire provided further insights into patient preferences regarding dressing forms. The results showed that 43.03% of respondents preferred gel/spray formats, while 34.26% favored smart monitoring patches. This indicates that patients value dressings with better cooling effects, absorbency, and adherence—products that not only cover wounds but also balance comfort and functionality.

Figure 17: The respondents' preference for the form of dressings

Building on this, we reviewed a large body of literature and gradually clarified that within the broad category of "gel" dressings, hydrogel dressings are the most suitable choice for diabetic chronic wounds. Unlike ordinary gels, hydrogels contain a high water content, allowing them to mimic natural tissue environments. They exhibit excellent moisturizing capacity, breathability, biocompatibility, and microenvironment-regulating functions. These properties not only relieve wound dryness and reduce external irritation but also help overcome healing stagnation caused by vascular damage, hypoxia, and persistent inflammation—thereby creating favorable conditions for cell migration and angiogenesis (Boateng, Matthews, Stevens, & Eccleston, 2008; Ahmed, 2015; Li, Mooney, & Xia, 2016). As a result, hydrogels have been widely used in the treatment of burns and chronic ulcers and are increasingly recognized as one of the most effective strategies for addressing impaired healing in diabetic wounds. This scientific conclusion, consistent with both survey data and fieldwork feedback, laid a solid foundation for our proposed solution.

Figure 18: Advantages of Hydrogel

At the same time, we conducted community-based fieldwork to further explore the habits and perceptions of elderly diabetic patients. Many participants admitted that severe wounds often required extremely long healing times. When asked about hydrogel dressings, most were aware of their effectiveness in treating burns and large wounds but still relied on traditional methods for daily injuries, without realizing that diabetic wounds required special care. They also noted that conventional commercial patches often had poor adhesion and caused irritation. These voices confirmed for us that diabetic chronic wounds are not only reflected in literature and statistics but also significantly impact the daily lives of the patients and family burdens.

Figure 19: The team members talked with the elderly people in the community

On this basis, we gradually realized that for mid-to-late stage diabetic wounds extending into the dermis, traditional low-risk therapies are far from sufficient. While such methods may work for minor injuries, they often fail to stop the progression of severe chronic wounds. Therefore, we proposed a new theoretical concept: to develop a bioactive hydrogel dressing capable of truly "filling" wound gaps and promoting tissue regeneration.

Unlike conventional spray-on or ointment products, our solution is a drop-in, filling-type hydrogel. This unique design not only allows firm adhesion to the wound surface but also penetrates deep into the wound microenvironment, improving conditions for tissue repair, enhancing cell proliferation and angiogenesis, accelerating healing, and reducing the risk of infection. Feedback from elderly community members about poor adhesion and irritation with traditional patches further validated the value of our proposed solution.

Figure 20: Injectable, filling-type hydrogel

Thus, moving from "Research" to "Theory," we completed the second step of the RTPNR framework: theoretical construction. Specifically, our theory is the proposal to develop a bioactive hydrogel dressing capable of truly "filling" wound gaps and promoting tissue regeneration. This concept is not only the natural outcome of scientific reasoning but also a direct response to the real needs of patients, families, and society. It also reflects the spiral-upward logic of the RTPNR framework: research drives the generation of theory, and theory in turn guides the next stage of practice and optimization.

Figure 21: Theory

3.3 P—From Solutions to Practice: Multi-Dimensional Exploration and Optimization of Therapeutic Yeast, Antimicrobial Peptides, and Hydrogel Materials

In the previous two stages, we had already completed the Research (R) and Theory (T) components of the RTPNR framework. First, through family experiences, questionnaire surveys, and on-site visits, we identified the prevalence and urgency of chronic wound healing disorders in diabetic patients, as well as the real challenges faced by patients and their families in wound care. Subsequently, we proposed a theoretical scheme to develop a bioactive, drip-injected hydrogel dressing capable of truly "filling" wound gaps and promoting tissue repair. Entering the Practice (P) stage, our task was to refine this theoretical concept into an implementable systemic design, and to explore and optimize it step by step at the levels of material, chassis, and functional molecules.

Figure 22: Practice

3.3.1 Understanding Pathological Complexity and the Necessity of Synthetic Biology

To address diabetic chronic wounds, it is first essential to understand their pathological complexity. Such wounds are often exposed to a high-glucose and high-ROS (reactive oxygen species) environment, which causes prolonged inflammation and prevents progression into the proliferative and remodeling phases. Wound healing is a dynamic process consisting of hemostasis, inflammation, proliferation, and remodeling, with each stage requiring specific cytokines: IL-6 and TNF-α clear pathogens in early inflammation; IL-10 suppresses inflammation and promotes repair in later stages; and VEGF drives angiogenesis during proliferation. Any disruption can stall healing. This realization underscored that single-factor drugs or external dressings cannot cover all phases simultaneously, highlighting the necessity of a living system capable of sensing the environment and dynamically adjusting outputs. Synthetic biology provides this unique advantage: through engineered chassis organisms, we can design closed-loop systems that integrate sensing and functional execution.

Figure 23: Pathological mechanism of diabetic chronic wounds

Against this background, we defined three essential tasks for our engineered yeast: antimicrobial, anti-inflammatory, and pro-healing functions. For antimicrobials, traditional antibiotics are limited by growing resistance, a concern we had already emphasized in 2024. Instead, we chose antimicrobial peptides (AMPs)—short peptides that are broad-spectrum, less toxic, and easier to express in chassis organisms. For anti-inflammatory function, we aimed for early secretion of IL-4 or IL-10 to induce macrophage polarization from M1 to M2 phenotypes, thereby resolving inflammation. Finally, for pro-healing, we targeted VEGF secretion after inflammation subsides to drive angiogenesis and tissue regeneration. To achieve such stage-specific responses, glucose-responsive promoters and heat-inducible promoters were introduced, ensuring dynamic molecular expression aligned with wound-healing rhythms.

3.3.2 Initial Exploration of Hydrogel Materials

Having defined functional requirements, we first explored candidate hydrogel materials. Systematic literature review led us to hydroxybutyl chitosan (HBC). Chitosan and its derivatives are widely applied in wound repair due to their unique structure and favorable properties (Jayakumar et al., 2011). As a modified chitosan-based gel, HBC offers multiple advantages: strong gelation and adherence, the ability to form a stable 3D network on wound surfaces, creation of a moist and protective microenvironment that reduces exudation and external irritation (Dutta et al., 2009), inherent biocompatibility and anti-inflammatory potential, and abundant chemical modification sites for functional expansion (Kong et al., 2010). These findings solidified our confidence in HBC as the foundational material for our hydrogel dressing design.

Figure 24: Characteristics of HBC hydrogel dressings

3.3.3 Chassis Selection and the Establishment of Yarrowia lipolytica

With the goal of developing a living material-based therapy, we recognized that choosing the right chassis organism was the cornerstone of the design. Only after selecting a chassis could subsequent literature exploration and functional module design proceed effectively. We required a eukaryotic expression platform capable of producing and secreting complex proteins—such as interleukins and VEGF—that are only functional in humans. Compared with bacteria, yeasts as eukaryotes offer robust post-translational modification, including correct disulfide bond formation and glycosylation, as well as rapid growth and established culture conditions.

We initially considered three candidates: the widely used Saccharomyces cerevisiae, the protein production powerhouse Pichia pastoris, and Schizosaccharomyces pombe, which more closely resembles mammalian cells in expression. To evaluate feasibility, we consulted Professor Guanglei Liu, an expert in marine yeasts. He noted that S. cerevisiae has limited protein secretion and differences in processing that may reduce activity; P. pastoris, though advanced in vector systems, cannot efficiently utilize glucose—the main carbon source in diabetic wounds—and is costly to modify. Instead, he recommended Yarrowia lipolytica, which features a well-developed secretion system (Madzak, 2015), versatile carbon metabolism including glucose utilization (Ledesma-Amaro & Nicaud, 2016), and a mature molecular toolbox of standardized plasmids and pathways. Following this advice, we conducted further review and confirmed Y. lipolytica as the optimal chassis, reflecting both practical and conceptual refinement under the RTPNR framework.

Figure 25: The team members exchanged ideas with Professor Liu

Figure 26: Yarrowia lipolytica

3.3.4 AMP Screening and the Establishment of Pexiganan

With material and chassis selected, we focused on antimicrobial factors. Diabetic wounds are especially vulnerable to infection, most notably from Staphylococcus aureus. We sought an AMP both effective against such pathogens and compatible with yeast expression.

Initially, we shortlisted Pediocin PA-1, Turgencin A, and Pexiganan based on prior yeast expression reports. However, manual literature screening proved time-consuming, and our wet-lab team needed rapid candidates. The dry-lab team proposed using a large language model (LLM) to accelerate AMP discovery. This greatly improved efficiency.

Figure 27: Team meeting

To validate our choices, we consulted Associate Professor Pengfei Cui, an AMP expert. He emphasized two key selection criteria: cytotoxicity level and antimicrobial spectrum. While our candidates were reasonable, he warned that narrow-spectrum AMPs may be inadequate for complex wounds. He also highlighted the importance of LLM-assisted AMP libraries as a future strategy to combat antibiotic resistance.

Figure 28: The team members exchanged ideas with Professor Choi

Incorporating his advice, combined with LLM output and literature validation, we finalized Pexiganan. This AMP has demonstrated broad efficacy in vitro (Ge et al., 1999) and comparable therapeutic outcomes to antibiotics in clinical trials for diabetic foot ulcers (Lipsky et al., 2008). Moreover, studies suggest AMPs may also modulate inflammation and enhance cell migration, accelerating healing (Mangoni et al., 2016). Thus, Pexiganan not only met our requirements for low-toxicity and broad activity but also validated the clinical potential of AMPs in diabetic wound care. At this stage, our engineered yeast was designed to secrete IL-4/IL-10 early, release Pexiganan to clear pathogens, and later express VEGF to promote regeneration.

3.3.5 Functional Limitations of HBC and the L-DOPA Modification Strategy

In subsequent experiments, we evaluated HBC under heat-induced conditions. Results showed that while HBC had good adhesion and gelation, it heated slowly, taking ~30 minutes to reach 37 °C—too slow for timely activation of heat-inducible promoter s. Since our hydrogel relied on thermal triggers for gene expression, this limitation had to be overcome.

We consulted Professor Ya Liu, an expert in hydrogels, who advised targeted modification rather than redesign. She proposed leveraging DOPA polymerization chemistry DOPA polymerization chemistry to enhance photothermal conversion and adhesion.

Figure 29: Have a conversation with Professor Liu

Based on literature and expert advice, we adopted L-DOPA modified HBC (L-HBC). L-DOPA introduces extra amino and carboxyl groups for amide reactions with HBC and, under ROS in wounds, converts to DOPA and undergoes polymerization. This significantly enhances photothermal conversion and tissue adhesion (Lee et al., 2014; Liu, Ai, & Lu, 2014; Zhao et al., 2020). This strategy not only resolved the performance bottleneck of HBC but also embodied responsible Human Practices: avoiding unnecessary risks while ensuring clinical feasibility.

Figure 30: The L-DOPA-modified hydroxybutyl chitosan structure

Figure 31: L-DOPA modified hydroxybutyl chitosan hydrogel

3.3.6 Summary of Practice Stage

With the chassis yeast selection gradually clarified, the final determination of antimicrobial peptides, and the targeted optimization of hydrogel materials, our project design began to take shape: centered on Yarrowia lipolytica as the chassis, engineered to secrete antimicrobial peptides and pro-healing factors, and combined with a modified hydrogel carrier, we constructed an integrated therapeutic system with antibacterial, anti-inflammatory, and regenerative capabilities.

Figure 32: Yeast Medics - Hydrogel dressings infused with engineered yeast, aiding wound regeneration in diabetic patients

Looking back, this process was not only a multidimensional exploration and optimization at the experimental level but also a complete RTP inner cycle : starting from research and theoretical assumptions, then repeatedly tested and refined through experiments and expert consultations, we gradually formed new insights and refined strategies. This spiral-upward cycle transformed our initial concept into a more feasible and clinically promising solution, laying a solid foundation for subsequent experimental validation and further optimization.

Figure 33: RTP cycle

3.4 Overview

In this section From Research to Focus: Converging on Diabetic Wound Dressings Through Real Needs and Voices, we accomplished three interlinked stages:

R — From Personal Inspiration to Social Focus: The Prevalence and Urgency of Diabetic Chronic Wound Healing: Through personal stories, surveys, and community visits, we confirmed the prevalence and urgency of diabetic chronic wound healing challenges.

T — From Needs to Solutions: The Conception of a Novel Bioactive Hydrogel Dressing: Based on real needs, we proposed the theoretical design of a drop-in, filling-type bioactive hydrogel dressing.

P — From Solutions to Practice: Multidimensional Exploration and Optimization of Therapeutic Yeast, Antimicrobial Peptides, and Hydrogel Materials: In practice, through iterative testing and refinement of materials, chassis, and antimicrobial peptides, we advanced our project toward a viable prototype.

Together, these three parts formed a complete Research–Theory–Practice (RTP) inner cycle , which ultimately generated our New-Theory:

By constructing an integrated therapeutic system based on engineered Yarrowia lipolytica and L-DOPA–modified hydrogel, it is possible to achieve multi-level regulation of antibacterial, anti-inflammatory, and pro-healing functions within the complex microenvironment of diabetic chronic wounds.

This New-Theory not only represents the natural elevation of our earlier research and practice but also acts as a bridge for spiral advancement—marking the transition to a higher-level RTP cycle. It provides the theoretical foundation for the next major section, "Theory-Guided Iteration: Continuous Testing and Reflection in Practice", ensuring that the RTPNR framework continues to spiral upward.

Figure 34: Research

4. Theory-Guided Iteration: Continuous Testing and Reflection in Practice

4.1 R — Academic Guidance: Consultation with Professors and Project Refinement

In the previous stage, we proposed the new theory of "a comprehensive therapeutic system based on engineered Yarrowia lipolytica and L-DOPA–modified hydroxybutyl chitosan hydrogel ." This marked the culmination and elevation of the earlier RTP cycle. However, the new theory was not an endpoint; rather, it served as a bridge propelling us into a higher-level cycle. With this new theoretical foundation, we entered the Research phase, conducting a more rigorous and data-driven re-investigation and reassessment of our design.

Figure 35: Research

During this process, we consulted Associate Professor Zhan Yuanchao, an expert in both microbiology and pharmacy. She fully affirmed the overall feasibility of our design in addressing clinical needs, but also sharply highlighted several critical issues that demanded further attention: Was the drug production rate of yeast sufficient? Were the diffusion and release rates of active factors within the hydrogel appropriate? How should the replacement frequency of the dressing be scientifically determined in clinical settings? She reminded us that without rigorous characterization of these key indicators, the therapeutic efficacy of our hydrogel dressing could easily be called into question.

Figure 36: Have a conversation with Associate Professor Zhan

Figure 37: Member and Associate Professor Zhan take a group photo together

This feedback compelled us to engage in deeper reflection and reactivated the Research → Theory link within the new RTP cycle. We realized that our initial theoretical design must be substantiated and refined through a dual strategy combining dry experiments and computational modeling. Accordingly, we introduced a two-layer validation framework.

At the macro level, we integrated results from dry-lab experiments and modeling to compare the dynamics of multiple biochemical factors in untreated chronic wounds versus treated wounds (see figure below). The analysis revealed that after intervention, the accumulation rate of AGEs and ROS was significantly reduced, NF-κB activity was effectively suppressed, fibroblast and keratinocyte proliferation was notably enhanced, and bacterial load was greatly decreased due to the introduction of antimicrobial peptides. These findings suggest that our system design holds promising potential for stabilizing the wound microenvironment at the macro scale.

Figure 38: Macro comparison of factor concentrations before vs. after intervention

At the micro level, we conducted molecular dynamics–based simulations to examine the diffusion and activity of VEGF, IL-4, IL-10, and antimicrobial peptides in heterogeneous wound environments (see figure below). The results showed distinct spatial patterns: VEGF and IL-4 maintained high concentrations in granulation tissue, promoting angiogenesis and resolution of inflammation; IL-10 diffused more effectively at necrotic margins, alleviating excessive inflammation; while antimicrobial peptides formed a protective layer on the wound surface, effectively preventing bacterial colonization. These micro-level findings provided quantitative insights into the dynamic behavior of therapeutic molecules in complex wound environments.

Figure 39: diffusion modeling of VEGF, IL-4, IL-10, and antimicrobial peptides

This dual-track approach of dry-lab validation and modeling projection not only strengthened the scientific robustness and feasibility of our design but also gradually facilitated the generation of new theoretical hypotheses. Looking ahead, the wet-lab group will design more phenotypic experiments to verify therapeutic performance in realistic wound models, while the dry-lab group will continue refining modeling parameters and approaches. Through this "experiment–modeling" synergy, we aim to build a more persuasive body of evidence to support the feasibility of our therapeutic hydrogel dressing.

From a Human Practices perspective, this process was not only academic guidance but also a profound reminder of responsibility. It emphasized that our project cannot rely solely on conceptual designs or isolated data points; rather, it must integrate laboratory design, clinical requirements, and societal responsibility. Through this renewed round of research and reflection, we established a solid foundation for the next Theory stage. In this higher-level cycle, the professor's insights not only filled critical gaps in our validation strategy but also guided us toward more advanced theoretical thinking and iterative refinement.

4.2 T— Cross-Cultural Exchange: Discussions with Professors at the University of Adelaide

After the previous stage of Research, we entered the Theory phase. To further examine and refine our design, on April 10 we held an offline seminar with professors from the University of Adelaide who specialize in metabolic biochemistry, immunology, and virology. With extensive expertise in the pathogenesis of diabetes, immune regulation, and drug development, their feedback provided us with a unique cross-disciplinary and cross-cultural perspective.

Figure 40: Theory

During the discussion, we systematically presented our design of an integrated dressing based on engineered yeast, antimicrobial peptides (AMPs), and hydrogel, along with our modeling and hardware exploration pathways. The professors recognized the potential of our project in addressing clinical needs, but also raised three critical questions:

(1) Why choose surface expression of AMPs on yeast rather than directly immobilizing them in the hydrogel?

(2) How will the non-toxicity of AMPs be validated?

(3) Could the secretion of inflammatory factors IL-4 and IL-10 disrupt immune balance?

Figure 41: Team members delivered a presentation on the project

Figure 42: The professor posed questions and offered suggestions to the team members

To address the first question, we reviewed the literature and found that strategies for immobilizing AMPs in hydrogels have been reported (Mao et al., 2017). However, our objective is not only to provide antimicrobial activity but also to ensure that engineered yeast can remain stably embedded within the hydrogel matrix. This requires careful consideration of the interaction between yeast and hydrogel. Against this background, we adopted the strategy of surface display of AMPs on yeast , which enhances its affinity with the hydrogel carrier while maintaining antimicrobial activity and system stability.

For the second question, we found reports indicating that AMPs such as Pexiganan exhibit relatively low cytotoxicity at the mammalian cell level (Ge et al., 1999), supporting their potential clinical safety. For the third question, we decided to employ a heat-inducible promoter to achieve conditional expression, thereby reducing the risk of immune imbalance caused by IL-4 and IL-10 overexpression (Chen et al., 2018).

Figure 43: Comparison of the expression patterns of antimicrobial peptides

Based on this feedback, our Theory can be clearly articulated as follows: By engineering yeast to dynamically secrete AMPs and inflammatory factors on its surface, combined with an L-DOPA–modified hydrogel carrier that enables conditional regulation, we aim to achieve simultaneous antimicrobial, anti-inflammatory, and pro-healing effects in the complex microenvironment of diabetic chronic wounds.

This stage not only allowed us to respond to critical questions raised by international scholars but also further strengthened the academic rigor and clinical feasibility of our theoretical design. Moving forward, we will enter the Practice stage, working with enterprises and application scenarios to continue testing and refining our approach.

Figure 44: Group photo

4.3 P — Industry Engagement: Collaborations with Medical Device and Chitosan Companies

In the previous Theory stage, we further clarified the feasibility of our project through cross-cultural academic exchanges and proposed the theoretical framework of a "composite therapeutic system based on engineered yeast and L-DOPA-modified hydroxybutyl chitosan hydrogels." However, theory alone is not sufficient; only through continuous testing and refinement in practice can it move closer to real-world application. With this in mind, we entered the Practice stage, bringing our laboratory concepts into real industrial environments by engaging with medical device and biomaterial companies, in order to understand how scientific innovations can be translated into clinical reality under the frameworks of compliance, safety, and resource utilization.

Figure 45: Practice

4.3.1 Visiting a Medical Device Company: Rethinking Standardization and Safety

On July 4, our team visited the modern factory of Qingdao Prisen Pharmaceutical Technology Co., Ltd., a subsidiary of Qidu Pharmaceuticals located in Linzi, to gain first-hand insights into the production process of medical consumables, quality control systems, and standardized workshop regulations. For the first time, members closely observed the entire process—from raw material preparation, equipment filling, sterilization, to packaging and inspection—which offered a vivid understanding of the rigorous details and technical requirements behind "standardized production."

Figure 46: Factory visit

During the exchange, company researchers emphasized that the development of medical products means more than just "innovation"; it requires countless rounds of biochemical and safety testing. Only after repeated verification can products finally enter clinical use. This realization prompted us to reflect: our therapeutic yeast hydrogel dressing cannot remain at the level of conceptual functionality; it must also overcome the critical barrier of safety. As the project matures, we recognize the necessity of conducting multidimensional validation, including material cytotoxicity tests, engineered yeast stability assessments, sterility checks, and long-term stability studies. More importantly, a mindset of compliance and safety must be embedded at the earliest stages of research. This is not only a regulatory requirement but also a fundamental responsibility toward patients.

Figure 47: Factory staff introduced the products and standards

Inside the sample production and factory workshops, we also experienced the high standards of medical product operations. Staff explained how the company follows the ISO 13485 medical device quality management system and CE international certification to implement full-process quality assurance, ensuring products meet market-entry regulations. These observations reinforced our understanding that for synthetic biology innovations to truly serve society, compliance and safety must be integrated into the research mindset from the outset, rather than treated as an afterthought.

Figure 48: ISO and CE Marking

4.3.2 Visiting a Chitosan Company: Rethinking Material Selection and Resource Utilization

If the visit to the medical device company highlighted the importance of "compliance and safety," then the field investigation of a chitosan company deepened our understanding of "material selection and resource responsibility." On July 11, our team visited Shandong Hongtai Bioengineering Co., Ltd. to study raw material processing and supply chains. The hydrogel material we employ—L-DOPA-modified hydroxybutyl chitosan (HBC)—is derived from the recycling of marine crustaceans, embodying not only the principle of "ocean-enabled health" but also the commitment to sustainable resource utilization.

Figure 49: SHANDONG HONGTAI BIOENGINEERING CO·, LTD

Company experts introduced the characteristics of different raw materials: mainstream production currently relies on animal-derived chitosan, with sources such as Indonesian krill tails, Korean red crab shells, and Alaskan snow crab shells. These materials are stable and suitable for large-scale production. In contrast, shellfish-based sources have been phased out due to issues of low viscosity and poor degradability. This revealed to us that materials appearing feasible in the lab may struggle to reach industrialization if they lack stable raw material supply chains and adaptable processing methods.

Figure 50: Team members visited the factory to see the raw materials 1

Figure 51: Team members visited the factory to see the raw materials 2

We also learned about the classification and application differences of chitosan: industrial grade (low cost, high yield, used in low-value fields like pesticides), food grade (used in health supplements but inadequate for medical use), and medical grade (requiring extremely high standards of purity, performance, and verification). This reality reinforced the notion that for our modified hydrogel to enter clinical applications, it must target the medical-grade standard, rather than compromise for lower thresholds. Such a requirement entails higher costs and stricter validation but, more importantly, underscores the principle that "patient safety must always come first."

Figure 52: Application of Chitosan

Additionally, we discovered how different chemical modifications of chitosan affect its antibacterial properties and potential applications. For instance, chitosan lactate exhibits significant antibacterial effects within 24 hours, making it suitable for military trauma use, while carboxymethyl chitosan tends to allow bacterial regrowth after 24 hours, necessitating cautious application. These differences made us realize that materials cannot follow a "one-size-fits-all" approach but must instead be tailored to the unique pathological features of diabetic chronic wounds.

In light of these insights, we gained a deeper appreciation of both the value and the challenges of L-DOPA-modified hydroxybutyl chitosan. On one hand, it enhances adhesion and mechanical properties through biomimetic chemistry, making it well-suited for filling chronic diabetic wounds that extend into the dermis. On the other hand, its raw material origin from marine crustaceans ensures biological activity and reflects sustainable resource recycling. Yet, we must also acknowledge the reality: even if such a material performs well in laboratory settings, clinical application still requires compliance with all medical-grade standards and rigorous regulatory approval.

Figure 53: Communicated with experts

This experience reminded us that in advancing experimental designs, industrial feasibility and clinical safety must be given equal priority alongside scientific innovation , ensuring that our project truly has the potential for real-world translation.

4.4 Overview

N: New-Theory Through surface-engineered Yarrowia lipolytica dynamically secreting antimicrobial peptides and inflammatory regulators, combined with an L-DOPA–modified hydroxybutyl chitosan hydrogel, we can achieve conditional and dynamic regulation of antibacterial, anti-inflammatory, and pro-healing functions in the diabetic chronic wound microenvironment. This new theory represents the project's spiral advancement, elevating it into a higher-level RTPNR cycle.

In this major section, "Theory-Guided Iteration: Continuous Testing and Reflection in Practice," we completed a new RTPNR inner cycle.

Figure 54: RTP inner cycle

In the Academic Guidance (Research) section, we conducted further investigation based on the previously established new theory. Associate Professor Zhan Yuanchao's feedback highlighted the need to verify key issues such as drug production rate, diffusion of active factors, and the replacement cycle of dressings through both experimental validation and modeling, thereby advancing our initial assumptions into a more rigorous scientific framework.

In the Cross-Cultural Dialogue (Theory) section, discussions with professors from the University of Adelaide further refined our theoretical design. In response to questions on antimicrobial peptide immobilization, toxicity validation, and immune regulation, we clarified a more precise theoretical framework.

In the Enterprise Engagement (Practice) section, we entered medical device and chitosan companies, where we gained insights into compliance, safety, and sustainable resource utilization. These experiences reminded us that even the most refined theoretical designs must ultimately withstand industrial standards and clinical regulations.

Therefore, the new theory generated in this stage is not only a validation and optimization of previous work but also signifies the project's transition from localized conceptual design to a more systematic framework. It marks the beginning of a higher-level cycle, serving as the bridge to the next RTPNR round and laying the foundation for the upcoming Practice stage.

Figure 55: Theory

5. Practical Innovation: Optimization Ideas under Project Problems and Limitations

5.1 R — Challenges Unveiled: Market Entry, Clinical Acceptance, and Application Barriers

In the previous stage, we proposed "integrated therapeutic system based on engineered Yarrowia lipolytica and L-DOPA–modified hydroxybutyl chitosan hydrogel" as our new theory, and we continuously refined the design through academic dialogue and industry engagement. Yet, the emergence of this new theory did not mark the end of our journey; instead, it pushed us into a higher-level cycle. As we move toward application and translation, we must directly confront the new challenges posed by market demands, clinical acceptance, and regulatory oversigh t. In this phase of research , our task was to place the theory into a more realistic context, focusing on the barriers and risks that could hinder the pathway from concept to practice.

Figure 56: Research

During our discussions with medical device companies, we also consulted an expert in hydrogel dressing development. She introduced us to commonly used hydrogel materials on the market, such as collagen, alginate, chitosan, and hyaluronic acid. After hearing about our innovative design that employs a thermosensitive hydroxybutyl chitosan (HBC) hydrogel as a carrier for therapeutic yeast, she recognized the novelty and confirmed that chitosan is indeed a strong material candidate due to its intrinsic anti-inflammatory properties. She also noted that our design of L-DOPA–modified HBC further enhanced the adhesive and conformal properties of the material.

Figure 57: The expert introduced the hydrogel market to the team members

At the same time, she pointed out the key challenges that must be addressed for clinical translation. Currently, most chitosan-based hydrogels on the Chinese market are classified as Class II medical devices—single-material products without pharmacological activity. In contrast, our novel design, which integrates engineered yeast and antimicrobial peptides, would be classified as a Class III medical device because of its pharmacological functions. This classification is extremely rare in the market and is subject to strict regulation not only in China, but also under CE standards in the EU and FDA guidelines in the United States. She advised us to engage early with relevant standards and guidelines to prepare for potential clinical translation.

The expert further explained that the circulation model for Class III devices differs significantly from our initial assumptions: such products generally require physician prescriptions and cannot be freely purchased in pharmacies or community clinics. This means that, even if a wound reaches the dermis, the dressing could only be used after a doctor's recommendation. The actual adoption of a new product therefore depends not only on scientific merit but also on whether physicians are willing to prescribe it and patients are willing to accept it. We came to realize that, beyond meeting scientific and regulatory requirements, clinical acceptance must be cultivated through preclinical validation, evidence-based studies, and academic dissemination. In other words, the ultimate success of translation is deeply tied to social trust, physician attitudes, and patient willingness.

Figure 58: Class III medical devices:Prescription-only flow

The expert also reminded us that market acceptance depends not only on material performance but also on factors such as conformability, dosage, and production design. If these dimensions are overlooked, even high-performing materials may struggle to compete in real markets. From this, we began to recognize that our next step is not only to refine the experimental plan but also to strengthen our understanding of clinical application scenarios through direct dialogue with physicians and patients, while complementing these insights with modeling and hardware exploration to optimize product design for practical use.

Figure 59: Factors influencing the market acceptance of hydrogel products

At this stage, our research revealed that clinical acceptance and regulatory compliance are the true bottlenecks of translation — that is, the process of moving scientific innovations from laboratory design into real clinical practice. These challenges themselves constitute the starting point of our next theoretical refinement.

By identifying these barriers, we became aware that theoretical predictions alone cannot capture the complexity of real-world application. Thus, we shifted our focus toward frontline contexts, entering communities and hospitals to consult directly with doctors and patients, so as to validate these challenges in real usage environments and gather the most authentic feedback. In this way, research not only grounded our design in practice but also laid the foundation for the next stage of Theory within the RTPNR framework, driving the project upward into a higher spiral of development.

5.2 T— Stakeholder Co-Creation: Multi-Dimensional Dialogue with Doctors and Patients

In the previous stage of Research, we recognized that regulatory, market, and clinical acceptance are the key barriers preventing scientific outcomes from reaching real applications. To further address these challenges, we turned back to our core stakeholders—doctors and patients—hoping that their voices could help us refine our understanding of clinical adoption and societal acceptance. This marked our entry into the Theory stage, where multi-dimensional feedback was used to improve and elevate our design.

Figure 60: Theory

During conversations in community hospitals, doctors reported that they had indeed encountered elderly diabetic patients whose large wounds, caused by falls or collisions, were difficult to heal. Yet they also admitted several practical challenges: on the one hand, many elderly patients are highly cautious about medication use and reluctant to try new therapies—an attitude that directly weakens the link between public awareness and clinical adoption; on the other hand, community doctors themselves often lack experience in handling complex chronic wounds and usually refer patients to larger hospitals. Meanwhile, many patients prefer home care, but the precise filling required for novel hydrogel dressings poses real difficulties for older adults. These exchanges made us realize that if a product cannot adapt to the habits and abilities of elderly users, its clinical value will be limited regardless of functional performance. This pushed us to consider elderly-friendly designs, and even hardware assistance (e.g., robotic arms for dressing application), to bridge the gap between laboratory solutions and daily wound care.

Figure 61: The team member communicated with the community doctor

Figure 62: Group photo

In interviews with doctors at large hospitals, we received more clinically grounded feedback. Surgeons noted that chronic diabetic wounds often receive little attention until they progress to severe complications such as diabetic foot, where treatment still relies on traditional antibiotics. They emphasized that if our hydrogel dressing can undergo rigorous clinical trials and demonstrate proven efficacy, they would be willing to recommend it to patients. This highlighted for us that doctors are not only gatekeepers of clinical adoption but also key bridges of patient trust.

Figure 63: Team member communicated with the doctor

Survey data further reinforced these insights. Results showed that 90.84% of respondents regarded safety as their primary concern, far outweighing cost or convenience. Meanwhile, 45.02% expressed stronger trust in hospital and physician prescription channels than in e-commerce platforms. This aligns closely with the doctors' feedback, indicating that both clinically and from the patient side, safety and physician endorsement are decisive factors for whether a new dressing can be adopted.

Figure 64: 90.84% of respondents regarded safety as their primary concern

Figure 65: 45.02% expressed stronger trust in hospital and physician prescription channels

In addition, we summarized the patients' detailed open-ended feedback into a visual chart:

Figure 66: Feedback

These responses clustered into five core expectations: treatment course and cost control, elderly-friendly usability, clinical trial assurance, convenient access channels, and smart functionality. This showed that patients' expectations for a new hydrogel dressing go beyond healing efficacy to include practical usability and reliability in daily life.

Through these exchanges, we gradually recognized that the essence of R&D lies not only in technical breakthroughs, but also in ensuring that products can be accepted by clinical systems and truly integrated into patients' daily routines. Stakeholders' voices not only validated our research pathway but also urged us to respond to their needs in a more responsible way. These insights will continue to shape our optimization efforts, providing a solid foundation for New-Theory generation and Re-practice within the RTPNR framework.

At this stage, our theory is that a feasible hydrogel dressing must not only integrate antibacterial, anti-inflammatory, and pro-healing functions, but also meet patient usability requirements (especially for elderly groups) and clinical acceptability (with safety and evidence-based validation as prerequisites). Survey data revealed that over 90% of users prioritize safety, while nearly half trust hospital prescription channels, reinforcing this theory through both patient and clinical perspectives. This shift transforms our work from pure scientific innovation into a systematic, application-oriented theory, paving the way for the next stage: Practice (practical validation).

5.3 P— Shifts in Perspective and Emerging Solutions: Expert Opinions and Survey Feedback

In the previous Theory stage, through feedback from doctors, patients, and surveys, we gradually clarified a new theoretical understanding: for hydrogel dressings to truly enter clinical practice, they must be built upon safety and clinical acceptability, while also incorporating elderly-friendly design and intelligent monitoring. However, theory can only be consolidated through the test of practice. With this awareness, we entered the Practice stage, bringing these theoretical insights into broader investigations and academic exchanges, where we reflected on and optimized the positioning of our project.

Figure 67: Practice

At the 2025 CCiC Conference in Beijing, we had the privilege of meeting China iGEM Ambassador Zhang Xiaohan. After listening to our presentation on project progress and real-world challenges, he offered highly insightful feedback. He emphasized that the issues encountered during our research and practice were not obstacles but rather opportunities that could drive the project toward maturity. It was precisely because we faced regulatory barriers and uncertainties in clinical acceptance that we realized more clearly that the project's future could not remain at the level of laboratory concepts or competition demonstrations, but must instead focus on long-term clinical application and societal implementation. He further suggested that we should firmly position our hydrogel dressing as a higher-standard medical product and Class III medical device, rather than as a simple Class II product or competition concept. This advice helped us redefine our project positioning and reminded us that the very existence of challenges serves as a catalyst for reflection and improvement.

Figure 68: The team members talked with the ambassador

Figure 69: The team members talked with the ambassador

Carrying this reflection forward, we revisited the three sections of this subsection—Challenges Identified, Stakeholder Co-creation with Doctors and Patients, and Expert Opinions with Survey Feedback. Together, they form an RTPNR mini-cycle. In the initial investigation, we identified regulatory barriers, clinical trust, and market application obstacles (Research → Theory); through frontline interviews with doctors and patient surveys, we distilled a new theoretical understanding, namely that safety is the primary threshold for clinical acceptance, physician endorsement is the key to patient trust, and elderly-friendly as well as intelligent functions are the directions for future optimization (Theory); supported by expert insights and quantitative validation, this theory was carried into reflective practice and optimization, gradually transforming into a more systematic positioning and development strategy.

Figure 70: RTP cycle

Throughout this process, data provided solid support for theory. Survey results revealed that 90.84% of respondents prioritized safety, far above price (less than 20%) or convenience (around 30%). At the same time, 45.02% of users preferred obtaining new dressings through hospitals/physician prescriptions, significantly higher than those who would choose e-commerce platforms (29%). These findings closely aligned with doctors' interviews: clinicians stressed that only after rigorous clinical trials and evidence-based research could innovative products be recommended to patients. Both data and clinical perspectives demonstrated that safety and physician endorsement are not only prerequisites for acceptance but also the core conditions for real clinical adoption.

Figure 71: Elderly-Friendly intelligent

Through this stage of practical exploration, we completed a clear mini-cycle: from identifying regulatory and clinical barriers (Research → Theory), to distilling the theoretical recognition of safety priority, physician endorsement, and elderly-friendly/intelligent directions (Theory), and finally to incorporating expert feedback and quantitative data into reflective practice and positioning upgrades (Practice → New theory). This process made us realize that project development cannot remain limited to technical breakthroughs but must also address the complex realities of clinical demand and regulatory environments. The voices of experts, doctors, and patients not only validated our research pathway but also guided us toward new directions for optimization.

5.4 Overview

Under this major section "Practical Innovation: Optimization Ideas under Project Problems and Limitations," we have completed a full RTP cycle.

N: New-Theory Our new theoretical insight is that a hydrogel dressing with true clinical potential must not only achieve antibacterial, anti-inflammatory, and pro-healing functions, but also be built on the foundation of safety, gain clinical endorsement from physicians, ensure operability for patients—especially the elderly, and progressively incorporate intelligent features to meet future development needs.

This new theory represents yet another spiral ascent within the RTPNR framework, pushing our project to a higher-level starting point. It bridges the tension between scientific research and real-world application, moving us from what is merely "feasible" to what is "trustworthy and usable."

At the same time, this section occupies the Practice (practical validation) stage within the overall Human Practices framework. It not only consolidates and refines the theories developed earlier in real-world contexts, but also lays the groundwork for subsequent re-practice. Building on this foundation, we now move to the next section, "Practical Innovation: Optimization Ideas under Project Problems and Limitations," where we will focus on specific optimization strategies, such as elderly-friendly design, intelligent monitoring, and mechanical-arm-assisted application systems. Through this continuous cycle of reflection and re-practice, we aim to strike a true balance between scientific innovation and societal needs, steadily advancing toward the goal of being "good for the world."

Figure 72: Practice

6. Practical Innovation: Optimization Ideas under Project Problems and Limitations

6.1 R— Patients' Needs and Application Prospects

In the earlier stages of investigation, experimentation, and optimization, we continuously refined our design through the RTPNR cycles. Each cycle resembled an upward spiral: beginning with personal stories and broad social surveys, we confirmed the prevalence and urgency of diabetic chronic wounds; we then proposed an integrated hydrogel dressing platform based on Yarrowia lipolytica and functionalized hydrogels, which was iteratively improved through expert advice, modeling, and industry consultations. This ultimately led us to a new theoretical insight: safety must be the foundation, physician endorsement the key to patient trust, and elderly-friendly as well as intelligent functions the future directions of development. This new theory acted as a "bridge" in the previous cycle, allowing the project to ascend to a higher level.

However, as iGEM Ambassador Xiaohan Zhang reminded us during the 2025 CCiC Conference in Beijing, the true value of a project cannot remain confined to competition-level presentations. Its long-term significance lies in how innovations can leave the laboratory, enter patients' lives, and become medical products that are not only usable but also trustworthy and sustainable. Inspired by this, we realized that after completing several cycles, the project had entered a new critical stage—Re-research. At this stage, the focus was no longer limited to laboratory verification or theoretical deduction, but to once again return to patients and society to seek new insights from their voices.

Figure 73: Re-search

Therefore, in the mid-to-late phase of our iGEM project, we brought our optimized design back to society and launched a more targeted re-research. Unlike our earlier surveys that asked broad questions such as "Does this need exist?" , this round focused on future user experience and directions for improvement. Our goal was to identify specific insights that could directly influence future experimental pathways and product design, ensuring that the project truly moves from "scientific conception" toward "real-world application."

Figure 74: The exploration and practice of new theories are initiated based on patient feedback

6.1.1 Survey Feedback: Mapping Social Expectations and Clinical Needs

In this round of research, we conducted online surveys and included open-ended questions that invited respondents to share their expectations for a new diabetic wound hydrogel dressing. Compared with our earlier surveys, the feedback here was more concrete and directly revealed the issues patients care about most in daily use and future applications.

Figure 75: Practice

Although diverse in detail, these responses revealed a crucial fact: the very shortcomings of current hydrogel dressings define the directions for future optimization. For example, insufficient adhesiveness and poor breathability are widely reported problems, and patient feedback once again confirmed their significance. Meanwhile, the demand for enhanced antimicrobial performance made us realize that beyond material improvement, the environment during dressing replacement and application is equally important. This insight became a key motivation behind our later concept of a "mechanical arm–assisted dressing system" to ensure more sterile operating conditions.

Figure 76: Word cloud map

6.1.2 Home Visits: Insights from Real-Life Scenarios

In addition to the online survey, we also conducted home visits to better understand patient experiences in real-world settings.

During one such visit, we met an elderly diabetic patient who shared how her chronic wounds not only caused physical pain but also createdM psychological burdens and anxiety. Living alone, she often worried whether she could properly use new medical products. While she expressed strong anticipation for our hydrogel dressing, she also admitted: if the operation is too complicated, she might find it overwhelming.

Figure 77: Communicated with the elderly

Her words deeply moved us. For elderly patients, questions like "Can I use it?" and "Do I know how to use it?" are just as important as "Is it effective?" . She further suggested that future dressing design should incorporate a senior-friendly scheme—one that is gentle enough for fragile skin while also simple and intuitive to operate, thereby reducing both psychological and practical burdens.

This reinforced an important realization: elderly-friendly design is not an optional feature but a core requirement. Without it, even the most advanced scientific breakthroughs may fail to integrate into patients' everyday lives.

6.1.3 From Re-research to Future Practice

Through this re-research, we gathered a wealth of patient and social feedback. These insights not only revealed the limitations of current hydrogel dressings but also illuminated clear directions for improvement. They reminded us that in order to truly "translate" scientific innovations, hydrogel dressings must integrate safety, antimicrobial performance, adhesiveness, and elderly-friendly usability to meet real patient needs.

More importantly, this process taught us that Human Practices is not an external supplement but a true driver of scientific iteration. It was the patients' voices that helped us identify previously overlooked problems in the later stages of the project; it was the community's feedback that showed us pathways for future optimization. Through this re-research, we not only collected essential inputs for improvement but also completed another mini-cycle within the RTPNR framework.

This stage has effectively laid the groundwork for the upcoming New-theory generation. A consensus has emerged within the team: the future direction of our project must incorporate elderly-friendly design, intelligent monitoring, and mechanical arm–assisted application systems to ensure that our hydrogel dressing can truly advance toward clinical translation and societal adoption.

Figure 78: Future hydrogel dressing

Thus, this Re-research phase serves not only as a response to our earlier work but also as a new starting point for the future. Within the spiral of upward progression, it has once again elevated us to a higher stage, preparing us to embrace the next phase of New-theory and Re-practice.

6.2 R— Doctors' Expectations and Clinical Vision

In the previous stage of re-research, through questionnaires, home visits, and social feedback, we realized that patients' voices are the key driving force behind the project's upward progression. Their expectations regarding safety, antimicrobial performance, elderly-friendly usability, and intelligent features reshaped the direction of our design. Yet, patients' perspectives are only one part of the application landscape. For our product to truly enter the clinic, it was equally essential to listen to another crucial group of stakeholders—doctors. With our refined plan and the concept of a "robotic arm-assisted drug delivery system," we once again stepped into hospitals, engaging in in-depth discussions with clinicians. This time, the focus was not merely on verifying feasibility but rather on understanding doctors' visions for future clinical applications and their genuine expectations for intelligent dressings and robotic arm systems.

Figure 79: Communication between team members and doctors

Figure 80: Intelligent monitoring

The doctors first acknowledged the potential of our integrated design, but emphasized that to make it truly viable, we must further address ease of operation and functional accuracy. They reminded us that hydrogel dressings could not only monitor wound healing via pH and temperature but should also integrate biomarkers capable of more precisely reflecting inflammatory status. In current clinical practice, physicians lack a reliable indicator to clearly determine when the inflammatory phase has ended. As a result, treatment decisions regarding whether to increase or reduce medication often rely heavily on clinical experience. If our intelligent dressing could incorporate such monitoring, it would provide doctors with a more scientific basis for adjusting drug dosage. This feedback highlighted that an intelligent dressing cannot stop at measuring basic physical parameters; it must evolve into an interdisciplinary tool capable of precise inflammation recognition and dynamic therapeutic regulation.

Figure 81: The team member communicated with the doctor

Figure 82: Group photo of team member and doctor

At the same time, doctors expressed strong enthusiasm for the robotic arm-assisted drug delivery system, stressing that its potential applications extend far beyond our initial expectations.

On the patient side, they pointed out that the system would not only benefit elderly diabetics living alone by ensuring sterile drug delivery but could also serve patients with limited mobility or disabilities, as well as those required to manage complex wounds at home, who risk secondary infections due to improper handling. In this sense, the robotic arm's value lies not only in convenience but also in its ability to create a relatively sterile environment, thereby reducing complications.

Figure 83: A robotic arm helps an elderly person living alone apply medicine

On the doctor side, they observed that the system could prove useful in diverse healthcare settings. In rural areas or aging communities, physicians often avoid handling complex wounds due to limited resources. With robotic arm assistance, they could safely perform interventions that would otherwise be too risky. Meanwhile, in large hospitals where surgeons and nurses face heavy workloads, the system could help alleviate the burden of wound care and dressing changes, enabling more rational allocation of limited medical resources.

Figure 84: The robotic arm helps doctors treat wounds

These insights expanded our imagination of the product's possible applications and reminded us to rethink the responsibility dimension of project design. Previously, our considerations were mainly patient-centered. The doctors' advice made it clear that true clinical translation must simultaneously satisfy patient usability and physician trust. Within the RTPNR framework, this signifies a shift from earlier cycles of research and testing (Research → Theory → Practice) toward new theoretical generation (New theory). In other words, intelligent dressings and robotic arm systems represent not only scientific innovations but also direct responses to real-world challenges in healthcare systems.

Through this stage of clinical dialogue, our understanding once again ascended. In the spiral of Human Practices, this process integrated previously scattered demands into a more systematic vision: at the patient level, products must be safe, easy to use, and elderly-friendly; at the doctor level, they must provide clinical reliability, evidence-based validation, and real support for reducing workload. From this synthesis, we gradually reached a new theoretical consensus: future hydrogel dressings should evolve into an integrated platform that combines intelligence, elderly-friendly design, and robotic arm assistance—not only meeting individual patients' needs but also serving broader healthcare contexts such as aging societies, rural clinics, and overburdened hospitals.

Doctors' feedback thus marked a transformation within the RTPNR cycle, moving us from New-Theory → Re- Practice. It not only broadened our perspective on application scenarios but also clarified how to make the project more clinically credible. This theoretical consolidation now guides our next steps toward re-practice (Re-practice), and it lays the groundwork for the upcoming section, "Technological Expansion: Introducing Robotic Arms and Other New Approaches into Experiments."

Figure 85: New-Theory

6.3 P— Technological Expansion: Introducing Robotic Arms and Other New Approaches

In the previous stage of our discussions and theoretical refinement, we gradually reached a consensus: for a hydrogel dressing to truly demonstrate clinical potential, it must be developed on the foundation of safety and physician endorsement, while also incorporating elderly-friendly usability and intelligent monitoring functions. This new theory, derived from both patient and physician feedback, represented a spiral advancement in our Human Practices framework. Yet the true value of theory lies only in its practice-based testing. With this realization, we formally entered the stage of new practice exploration (Re-practice), attempting to transform our earlier concepts into concrete technological solutions that directly address patient difficulties and physician expectations.

Figure 86: Re-practice

We came to understand that having a high-performance hydrogel dressing alone was not sufficient to solve the real challenges of diabetic chronic wound care. Social research and physician interviews consistently reminded us that in rural and community hospitals, the scarcity of medical resources often prevents patients from receiving timely and standardized treatment. For elderly patients living alone, the need to frequently visit hospitals for debridement and dressing changes not only imposes significant economic and time costs but also brings about physical inconvenience and psychological anxiety. For disabled or mobility-impaired patients, dressing changes often rely on relatives or caregivers, and the exposure of wounds during this process can generate shame and loss of dignity, hindering psychological recovery. These realities made us realize: without a solution that allows patients to achieve "safe, sterile, and autonomous" care at home or in resource-limited communities, our innovation would not truly address their needs. It was through such reflection that we proposed the concept of a low-cost, open-source robotic arm for automatic drug application combined with an intelligent wound monitoring system.

Figure 87: The team members were connecting the mechanical arm circuit 1

Figure 88: The team members were connecting the mechanical arm circuit 2

The robotic arm was first introduced to respond to the "dressing application challenge." Chronic diabetic wounds require extremely strict sterility, and traditional manual dressing changes not only involve heavy labor but also carry a risk of contamination and procedural error. Based on this, we designed an automated system centered on a six-degree-of-freedom robotic arm, capable of recognizing wound boundaries, applying hydrogel automatically, and maintaining full contact-free operation. In early simulations, we built a digital twin environment with ROS and Gazebo, validating wound recognition, path planning, and sterile application. The robotic arm used camera and sensor inputs to reconstruct wound geometry in 3D and generate a precise "approach–apply–withdraw" trajectory, ensuring even and safe drug coverage. This process significantly reduced errors such as under-application or secondary injury caused by human handling, laying a strong foundation for future real-world testing.

As the project advanced, we also broadened the scope of potential applications. Physician feedback highlighted that the system could benefit not only elderly individuals living alone but also a wider range of patients—such as those with disabilities who cannot manage their own wound care, or patients who need to handle severe wounds at home. For doctors, this system could also provide support. In remote clinics, where resources and training are limited, robotic assistance could enable safe treatment of complex wounds. In large hospitals, where surgeons and nurses often carry heavy workloads, robotic arms could help reduce their burden during peak hours, ensuring more efficient allocation of medical resources. Thus, the robotic system evolved from being just a "care tool" into a bridge connecting patient usability and physician trust, embodying our Human Practices commitment to humanistic care and social responsibility.

Figure 89: Future application scenarios of robotic arms

Alongside automation, we integrated an intelligent monitoring module to address the demand for real-time feedback. Survey data indicated that patients and doctors expected the dressing not only to cover and treat wounds but also to monitor the healing process. Based on this, our monitoring module was designed to continuously detect pH and temperature, later extending to ROS (reactive oxygen species) levels. pH and temperature trends help determine whether wounds are healing or at risk of infection, while ROS levels provide more direct insights into inflammatory status. By combining these parameters, both patients and doctors can make informed decisions about dressing changes and treatment adjustments, avoiding both "delays" and "over-frequent changes." Furthermore, we plan to connect the system to a mobile application, enabling remote sharing of wound data and facilitating telemedicine follow-up and personalized care.

Figure 90: PCB diagram of PH and temperature

Figure 91: Design schematic diagram

This entire exploration was not a stroke of inspiration but rather another complete cycle within the RTPNR framework. Starting from social research (Research), we identified real challenges in wound care. Through interdisciplinary dialogue and theoretical modeling (Theory), we conceptualized the robotic arm and monitoring system. Simulation and validation (Practice) refined the design. Feedback from doctors and patients expanded the application scenarios and value (New theory), which then led us into new practice (Re-practice). At present, we are preparing to introduce this system into more realistic testing environments to evaluate its feasibility and adaptability across diverse patient groups and medical settings.

Figure 92: Ultimate PCR cycle

The introduction of robotic arms and intelligent monitoring marks the transition of our project from material-based research to a holistic solution integrating technology and humanistic care. It not only enables elderly and mobility-impaired patients to receive dignified and safe care, but also alleviates the workload of healthcare providers while promoting fairer allocation of medical resources. Within the RTPNR framework, this exploration sits at the New theory → Re-practice stage, symbolizing an upward spiral from patient needs to technological expansion.

Figure 93: New theory → Re-search/Re-practice

More importantly, this stage reinforced our understanding that iGEM is not the end of the project. The low-cost, open-source robotic arm and intelligent monitoring system are not just designed for diabetic patients—they also represent a forward-looking response to global challenges such as unequal distribution of medical resources, population aging, and healthcare workforce well-being. By continually cycling through reflection and optimization, we aim for our project to evolve from a competition deliverable into a responsible, future-oriented solution. Only by striking a balance between scientific innovation and social demand can synthetic biology truly achieve "good for the world."

7. Summary and Reflection: Spiral Ascent under the RTPNR Framework

Looking back at the entire Human Practices process, we gradually realized that any iGEM project aiming to achieve good for the world cannot remain confined to functional design within the laboratory. It must instead find a balance between scientific innovation and social responsibility. For us, the RTPNR framework has proven to be not only a methodological tool but also a responsibility logic: it enabled us to continuously engage in Research–Theory–Practice cycles, and through the emergence of New-theory, to ascend into higher-level explorations (Re-search/Re-practice), thus forming a spiral upward trajectory.

Figure 94: RTPNR spiral upward graph

In the section "Starting from Research: Focusing on Diabetic Wound Dressings," we completed our first RTP cycle. Beginning with family experiences and literature reviews (Research), we gradually formulated a hydrogel dressing concept (Theory), and then validated demand through surveys and field visits (Practice). This cycle not only revealed the real pain points of diabetic chronic wounds but also laid the social value foundation of the project. The New-theory generated here was that diabetic chronic wounds urgently require a bioactive hydrogel dressing that combines patient usability with genuine medical necessity.

In the section "Project Exploration and Optimization," we moved further. After selecting Yarrowia lipolytica as the chassis organism (Research), we proposed a systematic plan to integrate antimicrobial peptides and engineered yeast (Theory), and refined this design through wet-lab/dry-lab iterations and expert consultations (Practice). This round advanced us from fragmented ideas to a holistic strategy, generating a New-theory: the feasibility of therapeutic hydrogels depends on the coordinated optimization of materials, chassis, and functional modules.

In the section "Theory-Led Iteration," the cycle advanced once again. Internal professors reminded us of the gaps in drug release rate and diffusion profiling (Research), cross-cultural dialogues allowed us to understand the credibility required in an international context (Theory), while enterprise consultations made us reflect on compliance and safety barriers (Practice). From this, we formed another New-theory: a clinically viable hydrogel must simultaneously pass scientific validation, achieve international credibility, and meet regulatory compliance standards.

In the section "Problems and Limitations: Toward Optimization," expert feedback made us face the realities of regulatory hurdles, physician acceptance, and market cost structures (Research). We then generated a refined theoretical understanding (Theory): clinical translation is not only a matter of science but is profoundly shaped by institutions, trust, and social environments. Combined with physician and patient interviews (Practice), this stage generated a New-theory: clinical acceptance and societal trust are decisive conditions for the hydrogel's successful adoption.

In the section "New-Theory Exploration and Future Practice," we moved into Re-research. Through surveys and home visits, we identified new expectations: elderly-friendly design, simpler operation, longer replacement intervals, intelligent monitoring, and even mechanical-arm-assisted application. Doctors emphasized the urgent need for biomarkers to clearly signal the end of inflammation. This stage of Re-research generated a New-theory: hydrogel dressings must integrate safety and efficacy with elderly usability, intelligent functions, and assisted application systems.

In the section "Technological Expansion," we translated this New-theory into Re-practice. We designed an open-source, low-cost robotic arm system for sterile drug delivery and an intelligent monitoring module capable of tracking pH, temperature, and ROS levels. Dry-lab modeling verified diffusion behavior and delivery accuracy, while future wet-lab experiments were planned for phenotypic validation. Doctors and patients further expanded the application scenarios: from helping elderly or disabled individuals at home, to empowering rural physicians, to reducing the burden on nurses in busy hospitals. This Re-practice applied our New-theory in concrete systems and marked the project's step toward true social application.

Thus, every section of Human Practices contained its own RTP cycle—Research → Theory → Practice. Each cycle generated a New-theory, which served as a bridge to a higher-level cycle. The accumulation of multiple such cycles eventually converged into the ultimate cycle of Re-research/Re-practice → New-theory → Practice, which constitutes the true essence of the RTPNR framework. The entire Human Practices content therefore unfolds as a spiral upward path of RTPNR.

Throughout Human Practices, we consistently upheld the following values:

(1) Social Value: Responding to patient needs and reducing the burden on families and physicians. For instance, community doctors' feedback on elderly patients' difficulties inspired us to prioritize elderly-friendly design.

(2) Scientific Value: Using synthetic biology to construct a system that integrates antibacterial, anti-inflammatory, and pro-healing functions. Examples include the selection of Pexiganan and Y. lipolytica.

(3) Ethical Responsibility: Emphasizing safety as a baseline, validated through plans for biocompatibility testing, yeast stability studies, and compliance frameworks.

(4) Sustainability: Leveraging marine shell-based resources to create L-DOPA-modified chitosan, enhancing both performance and resource reutilization.

(5) Humanistic Care: Designing for elderly, disabled, and solitary patients by proposing intelligent monitoring and robotic-assisted application systems.

Figure 95: Project values compass

To ensure these values were appropriate, we consulted patients, family members, doctors, professors, enterprise experts, and cross-cultural scholars, collecting a wealth of survey and interview data. These resources not only confirmed the project's direction but also provided evidence that our work is responsible.

We believe the project's impacts will manifest at multiple levels:

(1) End-users: Diabetic patients, especially the elderly and those with non-healing wounds.

(2) Usage Pathways: Clinical adoption through physician prescriptions, complemented by intelligent hardware for home and community scenarios.

(3) Social Dissemination: Open-source platforms and educational outreach to extend benefits to communities and remote regions.

At the same time, Human Practices made us acutely aware of our limitations. It revealed unresolved challenges and urged us to seek solutions through continuous feedback and cross-disciplinary collaboration. We recognized key next steps: moving into clinical applications, improving hydrogel adhesion and breathability, integrating robotic arms with mobile applications, and expanding into complex wound care scenarios. These remind us that the end of iGEM is not the endpoint of the project, but the beginning of Yeast Medics' journey into reality.

Figure 96: Stakeholder engagement map

In conclusion, the entire Human Practices process itself embodies a spiral ascent of RTPNR. Each inner RTP cycle pushed us upward; each New-theory became a stepping stone; and the final convergence into Re-research/Re-practice marked the higher-level cycle. From personal stories, to cross-cultural dialogues, to enterprise and clinical challenges, to technological expansion and future implementation, we have answered iGEM's fundamental Human Practices requirement: our work is responsible, responsive, future-oriented, and committed to making synthetic biology truly good for the world.

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