What is the Invention? (Background & Purpose)

• This invention is a regenerative sleeve designed to enclose the wound site of an amputated or injured appendage.
• It creates a sealed, controlled, and moist environment that promotes tissue regeneration and reduces scar formation.
• The device combines pharmaceutical treatment (via a treatment fluid) with biophysical stimulation (electrical stimulation) to mimic natural regenerative processes seen in certain animals.
• It is intended to help trigger cells to re-enter a mitotically active state, ultimately leading to the regrowth of tissues such as bone, muscle, and skin.

Components of the Regenerative Sleeve

• Tubular Sleeve (Outer Body):
  • A hollow, cylindrical structure that encloses the end of the appendage and wound site.
  • Constructed from a transparent, rigid material to allow monitoring of the wound and maintain a consistent internal volume.
• Cuff:
  • A hollow cylindrical element that is positioned at one end of the sleeve.
  • Designed to conform to the shape and size of the appendage, ensuring a tight, sealed fit to prevent leakage.
• Access Port:
  • Located on the outer body, it enables the administration and drainage of treatment fluids.
  • Made of a self-sealing material so that when punctured (e.g., by a syringe), it automatically closes to maintain the sealed environment.
• Electrical Stimulation Device:
  • Includes a pair of electrodes: an anode and a cathode.
  • The electrodes are connected to a power source that delivers a low-level current (up to around 10 µA) to mimic natural bioelectric signals.
  • This stimulation aids in triggering cellular dedifferentiation and proliferation.
• Additional Features:
  • An annular seal supports the cuff and helps maintain the sealed closure.
  • The sleeve may incorporate telescoping portions to adjust the wound space volume as regeneration progresses.
  • An optional rigid outer cover can be added to protect the device from external damage or animal tampering.
  • A treatment fluid is housed within the sealed space to chemically stimulate tissue regeneration by altering the ionic properties of the cells.

How the Regenerative Sleeve Works (Step-by-Step)

• Preparation:
  • The device is pre-selected in the correct size and configuration for the targeted appendage.
  • All components (sleeve, cuff, access port, and electrical device) are assembled and checked for integrity.
• Application:
  • The sleeve is applied directly to the wound site immediately after amputation or injury.
  • The cuff is adjusted to fit snugly around the appendage, forming a sealed wound space.
• Fluid Administration:
  • A predetermined treatment fluid is introduced into the wound space through the access port using a syringe.
  • This fluid maintains constant moisture and contains regenerative agents that help control the ionic environment of the cells.
  • The treatment fluid may include substances like porcine urinary bladder matrix (UBM) pepsin digest or specially formulated depolarization/hyperpolarization agents.
• Electrical Stimulation:
  • An electrical stimulation device is connected so that a low-level current flows from the anode to the cathode.
  • The current helps drive an internal wound stump current and provides guidance cues for cell migration and innervation.
  • This stimulation is applied periodically (for example, on days 0, 1, and 3) or continuously, depending on the treatment protocol.
• Environmental Control & Adjustments:
  • The sealed design maintains an optimal, hydrated environment for the wound.
  • If needed, a fluid pump may be connected to continuously replenish the treatment fluid.
  • The telescoping design of the sleeve allows for dynamic adjustments in volume as tissue regeneration progresses.
  • An optional protective shroud can be added to prevent tampering, especially in small animal models.

Treatment Fluid and Its Role

• The treatment fluid is formulated to modulate the ionic properties of the cells at the wound site.
• It stimulates the cells to become mitotically active (i.e., to start dividing and regenerating tissue).
• Examples of treatment fluids include:
  • Depolarization compositions (with elevated sodium and potassium levels) that push cells into a regenerative state.
  • Hyperpolarization agents that open ATP-sensitive potassium channels, making the cell interior more negative.
  • Extracellular matrix digests (such as UBM pepsin digest) that provide a physical scaffold and biochemical cues.
• The fluid may be replaced or used in sequential treatments to match different stages of the regeneration process.
• A continuous moist environment is critical to support cell proliferation and migration.

Experimental Method and Findings (Animal Study)

• Study Overview:
  • The device was tested in a murine (mouse) digit amputation model.
  • Mice were divided into two treatment groups: one received a control treatment (neutral pepsin buffer) and the other received a UBM digest treatment.
• Surgical Procedure:
  • Mice were anesthetized and prepared with sterile techniques under a microscope.
  • Digit amputation was performed at the distal phalange, and the regenerative sleeve was immediately applied to enclose the wound site.
• Treatment Protocol:
  • Electrical stimulation was administered on specified days (e.g., days 0, 1, and 3) at a current of approximately 6.4 µA for 15 minutes per session.
  • The treatment fluid was injected through the access port; care was taken to avoid air bubbles and ensure a full, moist environment.
• Outcome Measures:
  • Histological analysis evaluated wound hydration, cell proliferation, new gland formation, and evidence of bone remodeling.
  • Regenerative sleeves using UBM digest with electrical stimulation showed enhanced organization of cells and collagen deposition compared to control treatments.
  • The study demonstrated that a well-hydrated, electrically stimulated environment significantly improves tissue regeneration.
• Study Conclusion:
  • The regenerative sleeve effectively creates a protective, moist microenvironment that supports tissue regrowth.
  • Electrical stimulation further enhances the regenerative process by mimicking natural bioelectric signals.
  • This method shows promise for application to other types of wounds beyond murine digits.

Key Advantages and Future Applications

• Provides a closed, controlled, and hydrated environment optimal for tissue regeneration.
• Integrates pharmaceutical treatment with electrical stimulation for a synergistic regenerative effect.
• Design is flexible and can be adapted or scaled for various wound sizes, including limbs and organs.
• Simplifies the surgical process by reducing the number of components and assembly time.
• Potential applications include limb regeneration, treatment of congenital defects, and repair of traumatic injuries.
• Supports both temporary and permanent electrical stimulation setups, offering versatility in clinical use.

Summary of Key Points

• The regenerative sleeve is a novel device that stimulates tissue regeneration by combining a sealed, moist environment with treatment fluid delivery and electrical stimulation.
• Its components include a tubular sleeve, a conforming cuff, a self-sealing access port, and an electrical stimulation device.
• The application process involves preparing the device, applying it to the wound immediately post-injury, administering a regenerative treatment fluid, and providing electrical stimulation to activate cell proliferation.
• Animal studies have demonstrated that this method enhances tissue regeneration, showing improved cell organization, new gland formation, and early signs of bone remodeling.
• The device’s flexible design and multifunctional approach offer significant potential for future medical applications in various regenerative therapies.

发明简介（背景和目的）

• 该发明是一种再生袖套，专为包裹截肢或受伤部位的伤口设计。
• 它创造了一个密封、受控且湿润的环境，有助于组织再生并减少瘢痕形成。
• 该设备结合了药物处理液和生物电刺激，模拟某些动物自然再生过程，促使细胞恢复分裂活性。
• 目的在于激活细胞再生，最终实现骨骼、肌肉及皮肤等组织的重建。

再生袖套的组成部分

• 管状袖套（外壳）：
  • 空心圆柱形结构，用于包裹伤口部位。
  • 采用透明且坚硬的材料制成，便于观察伤口且能维持稳定的内腔体积。
• 袖口：
  • 位于袖套的一端，为空心圆筒形，设计上贴合截肢部位的形状与尺寸。
  • 帮助形成紧密的密封，防止处理液泄漏。
• 进出液口：
  • 设置在外壳上，允许注入和排出处理液，维持封闭的伤口环境。
  • 采用自封材料，当针头穿刺后能自动闭合，确保密封性。
• 电刺激装置：
  • 包括阳极和阴极，通过低电流（最高约10微安）来模拟生物电信号。
  • 这种电流有助于激活细胞，使其进入再生状态，并为细胞迁移提供指导信号。
• 其他功能：
  • 环形密封件支撑袖口，确保密封闭合。
  • 袖套可能采用伸缩设计，以便在再生过程中调整伤口内腔体积。
  • 可选的硬质外盖用于保护设备免受外界损伤或动物干扰。
  • 密封腔内储存的处理液含有促进细胞再生的活性成分，通过调节细胞离子环境发挥作用。

再生袖套的工作原理（步骤解析）

• 准备阶段：
  • 选择合适尺寸和配置的设备，并检查各部件的完整性。
• 应用步骤：
  • 在截肢或受伤后立即将袖套应用于伤口处，形成一个密封的伤口空间。
  • 调整袖口，使其紧密贴合受伤部位，确保环境封闭。
• 处理液注入：
  • 通过进出液口使用注射器将预定的处理液注入伤口空间。
  • 处理液保持伤口持续湿润，并含有调控细胞离子环境及激活细胞分裂的成分。
  • 处理液可能包括尿膀胱基质（UBM）消化液或特定的去极化/超极化调控剂。
• 电刺激：
  • 连接电刺激装置，使低电流从阳极流向阴极，激活伤口区域细胞。
  • 电刺激可以周期性（如第0、1、3天，每次15分钟）或连续应用，以增强再生效果。
• 环境控制与调整：
  • 密封设计保证伤口环境始终湿润，预防感染。
  • 必要时可连接液体泵，实现处理液的持续补充。
  • 伸缩设计允许根据再生进程动态调整伤口空间体积。
  • 可加装防护罩以防止动物干扰，特别适用于小动物模型。

处理液及其作用

• 处理液的主要作用是调控伤口细胞的离子特性，从而激活细胞分裂和再生。
• 可使用高浓度钠、钾离子的去极化液或ATP敏感钾通道开放剂使细胞内环境改变，促进再生。
• 例如，尿膀胱基质（UBM）消化液可作为物理支架和生化信号源促进细胞增殖。
• 处理液可根据再生阶段进行顺序或周期性替换，保持伤口持续湿润，支持细胞增殖与迁移。

实验方法及研究发现（动物研究）

• 研究概述：
  • 采用鼠足截肢模型测试再生袖套的效果。
  • 研究分为两组：对照组（使用中性缓冲液）和实验组（使用UBM消化液）。
• 手术过程：
  • 对小鼠进行麻醉和严格的无菌操作，在显微镜下完成数字截肢。
  • 截肢后立即应用再生袖套，将伤口区域密封起来。
• 治疗方案：
  • 在预定的时间（例如第0、1、3天）对伤口施加电刺激（约6.4微安，15分钟/次）。
  • 通过进出液口注入处理液，确保无气泡形成，维持伤口湿润。
• 结果评估：
  • 通过组织学分析观察伤口湿润程度、细胞增殖、新腺体形成及骨重塑情况。
  • 实验组（UBM消化液加电刺激）显示出较对照组更好的细胞组织排列、胶原沉积和再生迹象。
  • 研究证明，密封且电刺激的伤口环境显著促进了组织再生。
• 研究结论：
  • 再生袖套能有效创建一个保护、湿润的微环境，促进伤口组织再生。
  • 电刺激进一步增强了这一过程，通过模拟生物电信号激活细胞。
  • 该方法不仅适用于鼠足，也具备推广到其他伤口再生治疗的潜力。

主要优势与未来应用前景

• 提供一个封闭、受控且持续湿润的伤口环境，有利于细胞再生。
• 将药物处理和电刺激相结合，形成协同再生效应。
• 设计灵活，可根据不同伤口尺寸和类型进行调整和扩展。
• 简化了手术操作，减少组件组装时间和复杂性。
• 未来可应用于肢体再生、器官修复以及先天性缺陷的治疗。
• 适用于临时或长期的电刺激配置，提供多样化的临床应用可能。

关键要点总结

• 再生袖套是一种新颖设备，通过封闭、湿润的环境及药物与电刺激的综合作用，促进伤口处组织再生。
• 主要部件包括管状袖套、紧密贴合的袖口、自封进出液口以及电刺激装置。
• 使用流程包括设备准备、伤口应用、处理液注入和电刺激治疗，步骤清晰如同“烹饪食谱”。
• 动物实验显示，该方法能显著改善细胞组织排列、新腺体形成及骨重塑，验证了其再生效果。
• 整体设计多功能且灵活，未来在多种医疗再生领域具有广阔应用前景。