Conventional surgical sutures are frequently associated with significant postoperative challenges, including risks of infection, inflammatory responses, and the painful necessity of suture removal. These issues pose particular difficulties for patients undergoing procedures with high recovery demands, such as cesarean sections. To address these long-standing clinical problems, we have engineered ReGenStitch, a novel, multifunctional bioactive suture.

Our project is founded on a synthetic biology approach, featuring the construction of three core systems: a Bacterial Cellulose (BC) synthesis system, a Chitosan/Chitosan Oligosaccharide (COS) bioconversion system, and a Curcumin biosynthesis system. ReGenStitch organically integrates these three bioactive materials to achieve a full-cycle, synergistic effect encompassing mechanical support, antimicrobial and hemostatic action, and anti-inflammatory and tissue-regenerative properties. Our experimental results demonstrate that our design not only effectively mitigates the risk of postoperative infection but also promotes high-quality tissue regeneration, while sparing patients the ordeal of secondary suture removal. ReGenStitch presents an efficient, safe, and environmentally sustainable solution to the persistent challenges in clinical suturing.

Figure 1: The components of ReGenStitch and their origins.

Our project was inspired by a deeply personal family experience. Last year, a team member's sister underwent a cesarean section. As we gathered to celebrate the new arrival, she shared the arduous details of her postoperative recovery: the persistent pain and pruritus at the suture site, and her profound anxiety over potential infection and hypertrophic scarring. Her evident distress during dressing changes and the final, painful suture removal resonated with us, prompting a fundamental question: Must the process of surgical recovery be so arduous?

Figure 2: Common complications following a cesarean section.

During our visit, her sister graciously served us homemade Kombucha. As she poured the sweet and sour beverage from a large glass jar, our attention was drawn to the translucent, uniquely textured, gel-like biofilm floating within.

"What is this?" we asked, intrigued.

"That's the 'SCOBY'," she explained with a smile, "the 'mother' used to brew Kombucha. It acts like a living guardian, protecting the tea below from contamination by airborne microbes (Chin et al., 2015)."

This concept of a "biological protective barrier" immediately captured our attention. We drew an instant parallel to one of the greatest challenges in postoperative care—wound infection. If this natural, microbially-created barrier could protect a jar of tea, could it also protect a human wound? Could its tough yet flexible texture hold the potential to become an ideal surgical suture?

Figure 3: Kombucha and its mother culture (SCOBY).

（https://en.wikipedia.org/wiki/File:SCOBY_mushroom.jpg）

To explore this possibility, we conducted a preliminary assessment of the biofilm's physical properties, which revealed unexpected mechanical strength and flexibility. This discovery prompted an immediate turn to scientific literature to define its chemical composition and biological characteristics.

Our subsequent literature review confirmed our initial hypothesis: the primary structural component of this biofilm is Bacterial Cellulose (BC). Further investigation revealed the multifaceted advantages of BC as a biomaterial:

1. High Purity & Unique Nanostructure: Unlike plant cellulose, BC is free of lignin and hemicellulose. Its three-dimensional nanofibrillar network endows it with exceptional physical properties.
2. Excellent Biocompatibility: BC exhibits minimal immunogenicity, reducing the risk of foreign body reactions or inflammation when in contact with human tissue.
3. Promotion of Cellular Adhesion: Its biomimetic structure provides an ideal scaffold for cellular migration and proliferation.

These properties converged to a clear conclusion: Bacterial Cellulose is an ideal candidate material for engineering a new generation of high-performance surgical sutures. It could provide not only reliable physical closure but also has the potential to act as an "active biological scaffold," compatible with and supportive of the tissue healing process.

This became the starting point for our project, ReGenStitch. Our goal is to harness the power of synthetic biology to transform this remarkable biomaterial, born from a natural fermentation phenomenon, into a high-performance surgical suture that addresses real-world clinical needs.

With the rising volume of surgeries globally, the surgical suture market is projected to reach an impressive $9.83 billion by 2032, reflecting a continuous and expanding demand. However, behind this massive market lies a persistent dilemma: existing suture technologies fail to meet the clinical need for high-quality postoperative recovery. Both absorbable and non-absorbable sutures, the two main categories, present significant drawbacks:

1. Absorbable Sutures: Natural materials (e.g., catgut) are known to provoke strong inflammatory responses and have unpredictable degradation rates. While synthetic alternatives (e.g., PGA) offer improvements, their degradation byproducts can still create a localized acidic microenvironment, interfering with the healing process.
2. Non-absorbable Sutures: Although they provide high tensile strength, most require a secondary surgical procedure for removal, subjecting patients to additional trauma, time, and financial costs. If left in the body permanently (e.g., silk), they can become a nidus for bacterial colonization, leading to chronic infections.
3. Monofunctional Nature: Over 90% of sutures available on the market are designed solely for passive mechanical closure. They lack active biological properties, such as antimicrobial, anti-inflammatory, or pro-regenerative functions, leaving them ill-equipped to manage the complex postoperative microenvironment.

To more clearly illustrate these limitations, we have summarized them in the table below:

Table 1: Characteristics of Different Types of Surgical Sutures

Category	Main Material	Advantages	Disadvantages	Applicable Tissues
Absorbable Sutures	Natural (Catgut, Collagen)	Absorbed by the body, no removal needed.	1. Strong tissue/inflammatory reaction: Leads to wound redness, pain, and delayed healing. 2. Unpredictable absorption time: Premature absorption can cause wound dehiscence; delayed absorption increases inflammation risk. 3. Rapid loss of tensile strength: Unsuitable for tissues requiring long-term support.	Subcutaneous tissue, Mucosa
	Synthetic (PGA, PLA)	1. Relatively predictable absorption time. 2. Longer maintenance of tensile strength.	1. Inflammation from degradation byproducts: Accumulation of substances like lactic acid can trigger sterile inflammation. 2. Higher cost: Increases the economic burden on patients. 3. Suboptimal flexibility and knot security.	Fascia, Internal organs
Non-absorbable Sutures	Natural (Silk, Cotton)	Good handling properties, low cost.	1. Prone to bacterial colonization: The multifilament structure provides a haven for bacteria, increasing infection risk. 2. Foreign body reaction: As a permanent foreign object, it can cause chronic inflammation or granuloma formation.	Skin closure, Ligation
	Synthetic (Polypropylene, Nylon)	1. Extremely high strength, minimal tissue reaction. 2. Chemically stable.	1. Requires a second surgery for removal: Causes secondary trauma, pain, and costs. 2. Tissue cutting effect: The stiff texture may cut through fragile tissues. 3. No bioactive properties: Cannot actively combat infection or promote repair.	Cardiovascular, Tendon, Nerve repair

To overcome the limitations of traditional sutures, we have designed and engineered ReGenStitch—a next-generation surgical suture that integrates high-strength support, broad-spectrum antimicrobial activity, intelligent anti-inflammatory control, and active tissue repair capabilities. Through three synergistic biological systems, ReGenStitch upgrades surgical suturing from a passive mechanical linkage to an active tissue healing management system.

Figure 4: The three synergistic biological systems of ReGenStitch.

• System 1: Bacterial Cellulose Production - Serves as the "structural backbone" of the suture, providing mechanical strength and biocompatibility.
• System 2: Chitosan/COS Bioconversion - Acts as the "antimicrobial shield" and "healing initiator," enabling immediate hemostasis, long-lasting antimicrobial effects, and accelerated tissue regeneration.
• System 3: Curcumin Biosynthesis - Functions as the "inflammation regulator," precisely modulating the postoperative inflammatory response to prevent excessive inflammation and inhibit scar formation.

System 1: Bacterial Cellulose - The High-Strength Biocompatible Scaffold

Core Material & Advantages: Bacterial Cellulose (BC) is a natural polymer synthesized by bacteria such as Komagataeibacter xylinus. Its primary component is a straight-chain cellulose composed of β-1,4-glucan units. Compared to plant cellulose, BC possesses higher purity, as it is free from impurities like lignin and hemicellulose. It features a unique three-dimensional nanofibrillar network structure, high crystallinity, a large specific surface area, and strong water absorption capacity. Its physical properties are outstanding, exhibiting excellent mechanical strength, biocompatibility, and biodegradability, while being easily processable into various forms. Owing to these characteristics, BC has found wide-ranging applications, particularly in biomedicine as wound dressings, tissue engineering scaffolds, and drug delivery vehicles. Due to its green and sustainable production methods and exceptional performance, bacterial cellulose is increasingly becoming a focal point of research and application in biomaterials (Prilepskii et al., 2023).

Figure 5: Application of a microbial cellulose dressing to an injured hand.

（https://doi.org/10.1016/j.biomaterials.2005.07.035）

Figure 6: Schematic diagram of bacterial cellulose synthesis.

（https://pubs.acs.org/doi/10.1021/acssynbio.4c00615）

Biosynthesis System: In this project, we selected the clinically proven probiotic, Escherichia coli Nissle 1917 (EcN), as our chassis cell. As a probiotic with decades of clinical validation, EcN has an outstanding safety record, being non-pathogenic and non-endotoxic. Choosing it as our production chassis is intended to ensure, from the source, that our final product—bacterial cellulose—meets the stringent safety requirements for medical-grade materials. Although its protein expression efficiency may be slightly lower than that of the commonly used laboratory strain BL21 under certain conditions, its clinical-grade safety profile effectively prevents potential contamination with harmful impurities, making it a superior choice for constructing medical-grade biomaterials (Malc et al., 2024).

The core engineering challenge we then faced was how to efficiently activate bacterial cellulose synthesis in a non-native host. The complete biosynthetic pathway for BC is quite complex, regulated by a multi-gene bcs operon. Building this entire operon from scratch and ensuring its functionality would be an engineering challenge of immense difficulty and uncertainty. However, through in-depth research, we discovered that many Enterobacteriaceae, including E. coli, possess an endogenous, complete bacterial cellulose synthesis gene cluster within their genomes. While this system is typically "silent" or expressed at low levels under standard laboratory conditions, it provides an excellent starting point for our engineering modifications.

Therefore, we adopted a more strategic "metabolic bottleneck-breaking" engineering approach rather than a complex "de novo construction." Our logic was that since the basic "production line" (the full set of genes) already exists, the most efficient modification is to identify and amplify the core "engine" and "throttle" of this line—the key rate-limiting steps. Extensive research confirms that bcsA and bcsB play precisely these central roles (Sajadi et al., 2019). We activated the endogenous cellulose synthesis pathway in EcN by introducing these two key genes from a high-yield strain, Gluconacetobacter xylinus:

1. The bcsA gene: Encodes the catalytic core subunit of cellulose synthase. This protein is the "molecular machine" of cellulose synthesis. It directly uses intracellular UDP-glucose (UDPG) as a substrate to catalyze the formation of β-1,4-glycosidic bonds, polymerizing glucose monomers into long chains that form the basic structure of cellulose. It is the direct executor of production.
2. The bcsB gene: Encodes the cyclic di-guanylate (c-di-GMP) binding protein, which acts as the master regulatory switch for the entire synthesis system. Intracellularly, c-di-GMP is a crucial second messenger that transmits environmental signals. When c-di-GMP levels rise, it binds to the BcsB protein, inducing a conformational change that, in turn, allosterically activates the coupled BcsA subunit. Therefore, overexpressing BcsB enhances the system's sensitivity to the c-di-GMP signal, thereby more effectively "switching on" cellulose synthesis.

Working Mechanism: Upon receiving appropriate physiological signals (leading to c-di-GMP production), the overexpressed BcsB protein in our engineered strain efficiently captures these signals and potently activates the catalytic activity of BcsA. The activated BcsA "engine" then operates at high speed, utilizing the abundant intracellular UDP-glucose substrate to continuously synthesize and secrete long cellulose chains.

Through this precise and efficient engineering strategy, we achieved maximal activation of the host's endogenous metabolic pathway with minimal genetic burden, thereby constructing a safe and highly efficient bacterial cellulose production system.

Figure 7: Schematic diagram of the bacterial cellulose production system.

This system enables the efficient, controllable, and toxin-free production of bacterial cellulose, suitable for the development of biomedical materials such as wound dressings and tissue engineering scaffolds.

System 2: Chitosan & COS - The Antimicrobial and Healing Shield

Core Material & Advantages: We transform seafood waste—shrimp and crab shells—into two powerful bioactive substances. Chitosan, a large molecule, possesses excellent film-forming properties and a positive charge, allowing it to rapidly form a protective barrier on the wound surface. It disrupts bacterial cell membranes through electrostatic interactions and promotes platelet aggregation, making it particularly suitable for scenarios requiring rapid hemostasis, such as cesarean sections. Its degradation product, Chitosan Oligosaccharide (COS), is a small, water-soluble molecule that can penetrate deep into tissues. It reinforces antimicrobial effects by inhibiting bacterial DNA replication and actively accelerates wound healing by stimulating the proliferation of fibroblasts and vascular endothelial cells.

Figure 8: The defensive role of shrimp shells against environmental pathogens.

Biosynthesis System: Traditionally, the production of COS using engineered bacteria faces a critical bottleneck: chitosanase is typically expressed intracellularly. This means that before catalysis can occur, a series of cumbersome and expensive steps are required, including cell lysis, enzyme extraction, and purification. This not only increases costs but also leads to a loss of enzyme activity during extraction, and the enzyme cannot be reused.

To overcome this limitation, we designed an innovative system based on Surface Display technology. We fused the high-efficiency chitosanase, encoded by the CHI-1 gene, to a specific anchor protein, stably displaying it on the outer membrane of E. coli BL21. This transforms the process from a "single-use extraction" to "continuous catalysis."

The revolutionary aspect of this design is that our engineered bacterium itself becomes a living, self-regenerating, and reusable biocatalyst. During production, we do not need to kill or lyse the cells. We simply mix the live engineered bacteria with the chitosan substrate under specific conditions to continuously and efficiently produce the target product—bioactive COS of 2-6 units—under mild conditions. This "mix-and-use" model completely eliminates the steps of cell lysis and enzyme purification, dramatically simplifying the production process, reducing costs, and enabling easy catalyst recovery and continuous production, thereby significantly enhancing the economic viability and sustainability of the entire system (Chen et al., 2022).

Figure 9: The advantages of the surface display system.

Working Mechanism:

1. Raw Material Pretreatment: Waste shrimp and crab shells are pulverized using a blender, sieved, and dried for later use.
2. Chitosan Preparation: In a culture flask containing the shell powder and other nutrients, an environmentally friendly deproteinization and demineralization process is carried out using microbial fermentation (e.g., with Bacillus subtilis and Acetobacter sp.) to obtain high-purity chitin, which is then chemically deacetylated to yield chitosan.
3. Enzymatic Hydrolysis: The chitosan solution is mixed with the engineered bacteria expressing CHI-1. The chitosanase displayed on the cell surface precisely hydrolyzes the long chitosan chains into COS with optimal biological activity (2-6 sugar units).

Figure 10: The process from shrimp shells to chitosan oligosaccharides.

Table 2: Information on microbial strains used in the Chitosan & COS production process.

Stage	Microorganism	Selection Logic
Deproteinization (DP)	Bacillus subtilis	Naturally secretes protease; metabolic traits suit protein degradation in shrimp shells, no genetic modification required, simplifying the process.
Demineralization (DM)	Acetobacter sp.	Naturally produces organic acids, efficiently dissolves calcium carbonate, uses natural metabolic pathways to reduce cost.
Enzymatic Hydrolysis (COS)	Engineered E. coli BL21 strain	High expression of CHI-1 gene; surface display avoids enzyme loss and improves reuse.

This system enables the valorization of shrimp and crab shell waste, overcomes the monofunctional limitations of traditional sutures, and offers both environmental and medical value.

System 3: Curcumin - The Anti-inflammatory and Scar-Regulating Agent

Core Material & Advantages: Curcumin is a natural polyphenolic compound renowned for its potent anti-inflammatory, antioxidant, and scar-inhibiting properties. In the later stages of wound healing, an excessive inflammatory response is a primary cause of scar formation. By incorporating curcumin into the suture, we can actively downregulate pro-inflammatory factors and upregulate anti-inflammatory factors, creating a low-inflammation, "pro-regenerative" microenvironment for the wound (Hasanzadeh et al., 2020).

Biosynthesis System: This curcumin biosynthetic pathway is constructed using a synthetic biology strategy, achieving targeted synthesis of curcuminoids from simple precursors through three sequential, catalytically-driven core gene modules.

Working Mechanism:

• Input Substrates: Feruloyl-CoA (produced by the adenylation and thioesterification of ferulic acid) and Malonyl-CoA (a primary metabolite from the same pathway).

1. Carbon Chain Extension (DCS): The polyketide chain extension module is mediated by the DCS gene from turmeric (encoding a type III polyketide synthase, EC 2.3.1.). This enzyme uses Feruloyl-CoA as a starter unit and Malonyl-CoA as an extender unit, driving decarboxylative carbon-carbon bond formation via a Claisen condensation reaction to construct an intermediate with a C6-C7-C6 basic skeleton, feruloyl-diketide-CoA. This builds the "core skeleton" for curcumin.
2. Assembly (CURS1): The product assembly module is mediated by the CURS1 gene from turmeric (encoding Curcumin Synthase 1). This enzyme catalyzes the synthesis of curcumin by catalyzing the condensation of two feruloyl-diketide-CoA intermediates.

This pathway achieves efficient, targeted biosynthesis of the high-value secondary metabolite curcumin from primary metabolic precursors through the rational combination of genes across species (Chen et al., 2024).

Figure 11: The synthetic pathway of curcumin

How These Three Systems Work?

·System 1 (Bacterial Cellulose, BC) provides continuous support throughout the healing process. By offering a high-strength, flexible scaffold, it promotes directed cell migration and growth while minimizing foreign body reactions. It serves as a stable mechanical platform and biological interface for all functional modules.

·System 2 (Chitosan/COS Layer) activates the body's self-repair mechanisms at the wound site, delivering broad-spectrum antimicrobial and rapid hemostatic effects.

·System 3 (Curcumin) is activated in the presence of an excessive inflammatory response (e.g., elevated pro-inflammatory cytokines). It effectively curbs the "cytokine storm" and remodels the healing microenvironment by downregulating pro-inflammatory factors and upregulating anti-inflammatory ones.

Core Synergy:

·System 1 (Support/Biocompatibility) provides the ideal scaffold for cell growth.

·System 2 (Antimicrobial/Healing Initiation) activates the body's repair functions, reduces bacterial infection, and provides a significant hemostatic effect.

·System 3 (Anti-inflammatory) protects the body from inflammatory damage.

Together, these three systems achieve a full-cycle, high-quality healing process encompassing "Mechanical Support - Antimicrobial & Hemostasis - Inflammation Regulation."

Proposed Implementation

1. Target Clinical Applications:

ReGenStitch is primarily aimed at patient populations and surgical types with high demands for postoperative recovery quality, including but not limited to:

1. Cesarean Sections: For new mothers who have weaker postoperative recovery capabilities, experience frequent movement that pulls on the abdominal wound, and have high aesthetic requirements for scarring, the anti-infection, anti-inflammatory, and scar-inhibiting functions of ReGenStitch can significantly enhance the recovery experience.
2. Plastic and Dermatologic Surgery: In fields with extremely high aesthetic demands, its potential to inhibit hypertrophic scarring is of immense value.

2. Production and Quality Control Pipeline:

We envision our novel suture technology not just remaining in the laboratory but being implemented in actual clinical applications. To this end, we have conceptualized a complete industrialization pathway from lab to production:

1. Upstream Fermentation: In a GMP-compliant facility, engineered microbial strains are cultured on a large scale to produce bacterial cellulose, a mixture of chitosan and COS, and curcumin, respectively.
2. Midstream Purification and Compounding: The three core biomaterials are efficiently and aseptically extracted and purified. Subsequently, the three raw materials are mixed with other components in specific ratios, and the chitosan/COS and curcumin are uniformly compounded onto the bacterial cellulose scaffold to create a composite solution.
3. Downstream Forming and Sterilization: The composite material, in a moist state, is formed into standard-sized surgical sutures through spinning or cutting. It then undergoes final sterilization and is packaged in medical-grade sterile packaging, labeled with its functional parameters.

Figure 12: Conceptual diagram of the industrial production of surgical sutures.

3. Safety Assurance:

Safety is the cornerstone of any medical material. Our design incorporates rigorous risk control from the source to the final product:

• Source Material Safety: The "backbone" of the suture, bacterial cellulose, is produced using the clinically validated, non-pathogenic E. coli Nissle 1917. The introduced genes are solely responsible for cellulose synthesis and do not produce harmful byproducts. The chitosan/COS, which provides antimicrobial and hemostatic functions, is derived from natural shrimp shell waste, and the microorganisms used in its production, such as Bacillus subtilis and Acetobacter sp., are also safe strains, with no harmful substances involved in the entire process.
• Controllable Production Process: The entire production process is sterile. Fermentation and purification parameters are strictly controlled to ensure batch-to-batch consistency and high purity, eliminating harmful byproducts. Packaging utilizes medical-grade materials to ensure the suture remains stable and safe during storage and use.

This project successfully utilizes synthetic biology to construct an innovative, multifunctional surgical suture named "ReGenStitch." By organically integrating three bioactive systems, it achieves a synergy of structural support, antimicrobial and hemostatic action, and anti-inflammatory repair. Compared to existing surgical sutures on the market (such as nylon, silk, or polylactic acid), ReGenStitch demonstrates breakthrough advantages in multiple aspects.

1. Superior Mechanical Properties and Biocompatibility

First, the suture is built on a core backbone of bacterial cellulose. Its unique three-dimensional nanofibrillar network structure endows the material with mechanical strength and flexibility far exceeding that of plant cellulose. At the same time, its ultra-high purity ensures excellent biocompatibility, preventing immune rejection. This design effectively addresses the risks of tissue cutting caused by the excessive rigidity of traditional materials or the chronic inflammation triggered by unpredictable degradation rates, thereby significantly reducing postoperative complications and enhancing long-term patient comfort.

2. Dual Antimicrobial and Active Pro-healing Capabilities

Second, we have integrated a system of chitosan and chitosan oligosaccharides derived from shrimp and crab shell waste. These two substances work synergistically to form a powerful bioactive layer:

• The macromolecular nature of chitosan provides excellent film-forming and adhesive properties, allowing it to rapidly form a physical barrier on the wound surface for immediate hemostasis and broad-spectrum antimicrobial action.
• Chitosan oligosaccharide (COS), its degradation product, uses its small molecular size to penetrate deep into the tissue, reinforcing the antimicrobial effect and actively promoting wound healing by stimulating fibroblast proliferation.

This dual mechanism not only effectively prevents infection, avoiding the risk of antibiotic resistance associated with additional antibiotic use, but also transforms the suture from a passive closure tool into an active healing promoter.

3. Precise Anti-inflammatory Regulation and Scar Inhibition

Furthermore, we have introduced curcumin, produced via biosynthesis, as the suture's "inflammation regulator." It can precisely modulate the wound microenvironment post-surgery, effectively alleviating excessive inflammatory responses and inhibiting fibroblast over-proliferation, thereby reducing the likelihood of scar formation and chronic pain. This addresses the limitation of traditional sutures, which generally lack active anti-inflammatory functions, and is key to achieving high-quality, "scarless" healing.

In conclusion, ReGenStitch is not merely a product innovation but a holistic solution based on synthetic biology. Through the valorization of waste and green microbial manufacturing, it achieves environmentally friendly, sustainable production. Through multifunctional integration, it provides a forward-thinking approach to the design of medical materials. This project holds significant scientific value and vast clinical application potential, poised to bring about a revolution in the field of modern surgery by significantly improving patient outcomes and enhancing overall medical efficiency.

Moreover, ReGenStitch is a complete synthetic biology engineering practice. We followed the DBTL (Design-Build-Test-Learn) engineering cycle: In the Design phase, we identified bacterial cellulose, chitosan/COS, and curcumin as the core functional modules through literature review and clinical needs analysis. In the Build phase, we selected the safe probiotic E. coli Nissle 1917 and the common engineered strain BL21 as chassis, and introduced and combined the key gene modules. In the Test phase, through in vitro product detection and functional validation, we demonstrated the effectiveness of the three systems in mechanical support, antimicrobial/hemostasis, and anti-inflammatory repair. In the Learn phase, we analyzed the differences in yield and safety between EcN and BL21, the impact of different TDC sources on metabolite production, and the advantages of the surface display system in continuous catalysis. These lessons will directly guide subsequent iterative optimizations.

This series of engineering explorations demonstrates that ReGenStitch achieves environmentally friendly, sustainable production through waste valorization and green microbial manufacturing, and provides a forward-thinking approach to medical suture design through multifunctional integration. We believe that ReGenStitch has significant scientific value and vast clinical application potential. Through continuous iteration and clinical validation, it is poised to ultimately transform into an innovative solution that improves patient outcomes and enhances medical efficiency.

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