Composite artificial orifice / membrane tissue and preparation method thereof
By combining synthetic polymer materials with decellularized matrix scaffolds or collagen-based scaffolds to form an interpenetrating network structure and a smooth surface, the biocompatibility and healing problems of artificial cavity (membrane) materials in existing technologies are solved, achieving high-quality biological healing and biomechanical strength.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-31
- Publication Date
- 2026-03-10
AI Technical Summary
Existing artificial cavity (membrane) materials have problems such as poor biocompatibility, anastomotic leakage, artificial cavity stenosis, and difficulty in achieving biological fixation and permanent healing in vivo. In particular, because synthetic polymer materials are difficult to completely heal with the patient's cavity tissue, they lead to long-term inflammation of the anastomosis, easy anastomotic leakage, premature detachment, perforation, obstruction, thrombosis or eschar formation.
By combining synthetic polymer materials with decellularized matrix scaffolds or collagen-based scaffolds, an interpenetrating network structure is formed, which enhances biomechanical strength. Furthermore, a smooth surface is formed through secondary surface polymerization, which reduces immunogenicity and promotes the ingrowth of receptor cells and nerves.
It achieves high-quality physiological healing of artificial cavity (membrane) tissues, avoids thrombosis and blockage, reduces immune rejection, and improves biocompatibility and biomechanical strength.
Smart Images
Figure CN121623009A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of artificial cavity (membrane) tissue technology, specifically to a composite artificial cavity / membrane tissue and its preparation method. Background Technology
[0002] Clinically, after the removal of cavities such as the esophagus, blood vessels, and urethra, other tissues from the patient's own body, such as the intestines or blood vessels from other parts of the body, are often used as replacements. However, this method has the disadvantages of being highly invasive and causing serious complications. Current artificial replacement products suffer from poor biocompatibility, anastomotic leakage, artificial cavity stenosis, and difficulty in achieving biological fixation and permanent healing in vivo. The reason for this is that most current artificial cavities are made of biodegradable synthetic polymer materials. After implantation, although they can be surrounded by new epithelium and connective tissue, it is difficult for recipient tissues and cells to grow in, making it difficult to achieve complete healing with the patient's cavity tissue. This results in the artificial cavity itself having no blood supply, and insufficient blood supply to the surrounding area, leading to long-term inflammation of the anastomosis, anastomotic leakage, premature detachment, perforation, obstruction, thrombosis, or crusting, all of which affect the patency of the cavity and restrict the clinical application of artificial cavities.
[0003] Acellular matrix materials are natural biomaterials that can provide a natural biochemical environment for cells. Recipient cells and blood vessels can easily grow into them, ultimately completing the recellularization of the acellular matrix scaffold. It heals well with the patient's own tissues, and the structural reconstruction and functional restoration of artificial cavities (membranes) are expected to solve the bottleneck problems that restrict the clinical application of artificial cavities. Summary of the Invention
[0004] To overcome the problems of poor healing, low biomechanical strength, immune rejection and rough surface after transplantation of artificial cavity (membrane) tissue, this invention provides a composite artificial cavity / membrane tissue and its preparation method. This invention uses decellularized matrix scaffolds or collagen-based scaffolds combined with synthetic polymer materials to prepare composite artificial cavity (membrane) tissue.
[0005] The purpose of this invention is to provide a composite artificial cavity / membrane tissue, which is composed of synthetic polymer materials and natural polymer biomaterials. The synthetic polymer materials are selected from one or more of silicone, synthetic polymer hydrogels and acrylic polymers, and the natural polymer biomaterials are selected from one or more of decellularized matrix scaffold materials, non-decellularized matrix and non-decellularized collagen.
[0006] The composite artificial cavity / membrane tissue proposed in this invention involves the infiltration of synthetic polymer materials into natural polymer materials such as decellularized matrix. An interpenetrating network structure is formed between the two materials, enhancing the biomechanical strength of the composite cavity (membrane) and resulting in a firmly connected, integrated composite artificial cavity (membrane) tissue. The secondary surface polymerization of the synthetic polymer materials on the surface of natural polymer materials such as decellularized matrix improves smoothness, preventing thrombosis and secretion adhesion, thus avoiding blockage of the composite cavity (membrane). No synthetic polymer materials infiltrate the adjacent outer surface and surrounding areas of the decellularized matrix scaffold or pure natural collagen, preserving the maximum amount of natural components, which is beneficial for suture closure and the ingrowth of receptor cells and nerves, achieving high-quality physiological healing. The synthetic polymers encapsulate the antigens on the surface of the microfibers of the decellularized matrix scaffold material, further reducing the immunogenicity of the decellularized matrix scaffold.
[0007] Preferably, the decellularized matrix scaffold material, non-decellularized matrix, and non-decellularized collagen are allogeneic or xenogeneic decellularized matrix scaffold tissues or organs.
[0008] Preferably, the acrylic polymer is poly(hydroxyethyl methacrylate).
[0009] Preferably, the composite artificial cavity / membrane tissue includes a central surface area, a peripheral area, and an outer surface area connected in sequence. The central surface area is a synthetic polymer material, the peripheral area is a composite of synthetic polymer material and natural polymer biomaterial, and the outer surface area is a natural polymer biomaterial.
[0010] This invention also protects the method for preparing the composite artificial cavity / membrane tissue, comprising the following steps: using a natural polymer material as a scaffold, synthesizing a polymer material on the scaffold, and obtaining a composite artificial cavity / membrane tissue in which the synthetic polymer material and the natural polymer material interpenetrate through two polymerization processes.
[0011] Preferably, the natural polymeric biomaterial is a decellularized matrix scaffold, and the synthetic polymeric material is poly(hydroxyethyl methacrylate).
[0012] Further preferred, the preparation method specifically includes the following steps: using hydroxyethyl methacrylate (HEMA) as raw material, ammonium persulfate / sodium metabisulfite (APS / SMBS) as initiator, and triethylene glycol dimethacrylate (TEGDMA) as crosslinking agent, and stirring with water until uniform, the mixture is coated onto the inner surface / membrane surface of the decellularized matrix scaffold, allowing the hydroxyethyl methacrylate monomer to penetrate into the decellularized cavity or membrane tissue. The reaction is carried out in a constant temperature water bath at 40℃~50℃ for 30~42 h, inducing polymerization within the decellularized matrix scaffold to form a composite tissue in which poly(hydroxyethyl methacrylate) hydrogel and the decellularized matrix scaffold interpenetrate. The above materials are added again, and through secondary polymerization, a smooth surface layer containing only poly(hydroxyethyl methacrylate) is formed on the surface of the composite tissue. The tissue is then immersed in deionized water to remove unreacted small molecules, thus obtaining a composite artificial cavity / membrane tissue in which poly(hydroxyethyl methacrylate) and the decellularized matrix scaffold interpenetrate.
[0013] Further preferably, the molar ratio of hydroxyethyl methacrylate to the initiator is 10. 4 ~10 5 1. The molar ratio of hydroxyethyl methacrylate to crosslinking agent is 10:1. 4 ~10 5 :1.
[0014] The specific steps for the composite artificial cavity / membrane tissue are as follows: (1) purify hydroxyethyl methacrylate by vacuum distillation; (2) prepare an initiator solution with a concentration of 0.4~1.5wt% by dissolving ammonium persulfate and sodium metabisulfite in deionized water. Add HEMA / H2O (75 / 25 (w / w)), crosslinking agent triethylene glycol dimethacrylate (TEGDMA) (1.0 wt%), initiator APS / SM BS (0.5 wt%) and water in sequence to obtain the PHEMA prepolymerization solution. (3) Fix the decellularized matrix scaffold on the mold and use the mold to pressurize and fix the peripheral area of the decellularized matrix scaffold to reduce or even avoid the PHEMA prepolymerization solution from seeping into the peripheral area of the decellularized matrix scaffold; (4) Apply the PHEMA prepolymerization solution to the inner surface of the lumen of the decellularized matrix tubular scaffold or the epithelial surface of the decellularized matrix membrane scaffold (in special cases, the non-epithelial surface, such as composite artificial cornea). The PHEMA prepolymerization solution seeps into the decellularized matrix scaffold. Under constant temperature water bath conditions of 40℃~50℃, after 30~42 h, hydroxyethyl methacrylate gradually polymerizes into polyhydroxyethyl methacrylate hydrogel, forming an interpenetrating network structure with the decellularized matrix scaffold material. (5) Apply PHEMA prepolymer solution to the inner surface of the decellularized matrix scaffold lumen or the surface of the decellularized matrix membrane scaffold again, and form a smooth surface containing only PHEMA components on the surface of the new composite tissue through secondary HEMA polymerization; (6) Soak in deionized water for 7 days, changing the water every 24 hours to remove unreacted small molecules in the composite material, and obtain a composite material that interpenetrates with the decellularized matrix scaffold and the PHEMA hydrogel network.
[0015] This interpenetrating network structure tightly binds the two materials, enhancing their biomechanical strength. Furthermore, the PHEMA encapsulates the antigens in the decellularized matrix, further reducing immunogenicity. Secondary HEMA polymerization forms a smooth surface containing only PHEMA on the surface of the composite artificial cavity (membrane) tissue, preventing secretion adhesion and thrombus formation. The surrounding decellularized matrix, with little or no PHEMA infiltration, facilitates the ingrowth of recipient cells and nerves, enabling excellent post-transplantation healing.
[0016] The novel composite artificial cavity (membrane) tissue proposed in this invention has the advantages of good post-transplant healing, low immunogenicity, and smooth surface. It can effectively solve the problems of insufficient biomechanical strength, poor postoperative healing, and easy formation of thrombosis or scab due to the rough surface of the material in collagen-based artificial materials.
[0017] Preferably, the reaction is carried out in a constant temperature water bath at 45°C for 36 hours.
[0018] Compared with existing technologies, this invention has the following advantages: The composite artificial cavity / membrane tissue and its preparation method, which integrate natural and synthetic polymeric biomaterials, better reflect the advantages of both materials. The interpenetrating network structure between the synthetic polymeric material and the decellularized matrix material further enhances the biomechanical strength of the product, improving the biomechanical strength of the composite artificial cavity / membrane tissue and providing a smooth surface. The synthetic polymeric material encapsulates the surface antigens of the microfibers in the decellularized matrix scaffold material, further reducing the immunogenicity of the decellularized matrix material. The natural polymeric biomaterial facilitates the ingrowth of receptor cells and nerve fibers, maximizing biological healing. Attached Figure Description
[0019] Figure 1 This is a schematic diagram of the composite artificial membrane tissue structure proposed in this invention;
[0020] Figure 2 This is a schematic diagram of the composite artificial cavity structure proposed in this invention;
[0021] Explanation of reference numerals in the attached diagram: 1. Central surface area; 2. Peripheral area; 3. Outer surface area; 4. Lumen. Detailed Implementation
[0022] The following embodiments are further illustrations of the present invention, but not limitations thereof.
[0023] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention. Unless otherwise specified, the experimental materials and reagents used herein are commercially available products conventional in this technical field. Welding processes, welding materials, and welding equipment are all acceptable as long as they can realize the multi-layer, multi-pass model proposed in this invention.
[0024] like Figure 1 As shown, the composite artificial membrane tissue proposed in this invention includes a central surface region 1, a peripheral region 2, and an outer surface region 3 connected in sequence. The central surface region is a synthetic polymer material, the peripheral region is a composite of synthetic polymer material and natural polymer biomaterial (i.e., the polymer material penetrates into the shallow surface layer of the biomaterial), and the outer surface region is a natural polymer biomaterial.
[0025] like Figure 2 As shown, the composite artificial cavity tissue proposed in this invention includes a central surface area 1, a peripheral area 2, and an outer surface area 3 connected in sequence. A lumen 4 is provided in the central surface area. The central surface area is made of synthetic polymer material, the peripheral area is made of a composite of synthetic polymer material and natural polymer biomaterial (i.e., the polymer material penetrates into the shallow surface layer of the biomaterial), and the outer surface area is made of natural polymer biomaterial.
[0026] The synthetic polymer material is selected from one or more of silicone, synthetic polymer hydrogels and acrylic polymers, and the natural polymer biomaterial is selected from one or more of decellularized matrix scaffold materials, non-decellularized matrix and non-decellularized collagen.
[0027] The following examples or comparative examples of vascular decellularization treatment include the following steps: cleaning animal lumens or tissue slices, scraping off the surface cell layer, drying in an oven, and then immersing in pure water to prepare pretreated biological tissue slices (lumens) for later use; immersing the pretreated biological tissue slices (lumens) in sodium periodate buffer solution, shaking repeatedly, and after 30 minutes, transferring them to a colloidal polymer osmotic equilibrium solution for 10 minutes to terminate the reaction; removing the biological tissue, placing it in sodium deoxycholate carbonate buffer solution, and treating it with shaking in a water bath at room temperature for 2 hours; then treating it with carbonate buffer solution, shaking it in a water bath at 37°C for 12 hours; finally, treating it with carbonate buffer solution for repeated extraction and washing to remove the above decellularization agents, and then dehydrating and storing it.
[0028] Example 1
[0029] This embodiment discloses a method for preparing composite amniotic tissue for use as a biological dressing, including the following steps:
[0030] The surface three layers of amnion tissue (epithelial cell layer, basement membrane layer, and compact layer) were harvested and fixed with the epithelial side facing down on a fixation frame. The HEMA prepolymerization solution (hydroxyethyl methacrylate (HEMA), ammonium persulfate / sodium metabisulfite (APS / SMBS), and triethylene dimethacrylate (TEGDMA)) was mixed with deionized water and stirred until homogeneous. The molar ratio of HEMA:APS / SMBS:TEGDMA:water was 10. 5 :1:1:10 5 A 1:1 molar ratio of ammonium persulfate and sodium metabisulfite was used to coat the dense layer of the amnion. After 36 hours in a constant-temperature water bath at 45°C, HEMA was induced to polymerize into poly(hydroxyethyl methacrylate) hydrogel (PHEMA) within the amnion, completing the interpenetrating polymerization between PHEMA and the amnion matrix. The pre-polymerized HEMA solution was then coated again onto the surface of the dense amnion layer. Through secondary HEMA polymerization, a smooth surface containing only PHEMA was formed on the tissue surface. The surface was then soaked in deionized water for 7 days, with the water changed every 24 hours to remove unreacted small molecules, resulting in a novel composite PHEMA amnion. This was fixed and stored for later use. In application, the composite amnion was fixed with the epithelial side facing down (the side in contact with human tissue) as a biological dressing to cover ulcers, promoting wound healing and reducing scarring.
[0031] Comparative Example 1
[0032] Currently, there are two main types of amniotic membrane products, both collected from human placental amniotic membranes delivered via cesarean section, and preserved using either dry or wet methods, respectively. In clinical use, they are fixed to the wound or ulcer surface with sutures. Because the amniotic membrane used clinically only comprises the outer three layers of the fetal membranes, with a thickness of only 0.02–0.1 mm, the procedure during transplantation and the cutting force of the sutures can easily cause the amniotic membrane to tear or break, especially when the suture force is uneven.
[0033] The composite amniotic membrane obtained in Example 1 improves the thickness and toughness of the amniotic membrane, without affecting the physiological function of the amniotic membrane, and can also avoid the breakage and tearing of the new amniotic membrane. If used in conjunction with a biomembrane fixation device, it can be fixed without sutures.
[0034] Example 2
[0035] This embodiment discloses a method for preparing composite amniotic membrane tissue for use as a transplant material. The steps are roughly similar to those in Embodiment 1, except for the composite position of the polymer material and the amniotic membrane. In this embodiment, the epithelial surface of the amniotic membrane tissue is fixed upwards on a fixation frame, and the remaining operations are the same as in Embodiment 1. In use, the composite amniotic membrane is fixed with the epithelial surface upwards (not in contact with human tissue) to cover the ulcer surface as a transplant material. The purpose is to provide a basement membrane for epithelial cell growth, thereby healing the wound and ulcer.
[0036] Comparative Example 2
[0037] The current amniotic membrane used in comparison is insufficient compared to control group 1.
[0038] Example 3
[0039] A method for preparing a composite artificial blood vessel includes the following steps:
[0040] Using hydroxyethyl methacrylate (HEMA) as a raw material, ammonium persulfate / sodium metabisulfite (APS / SMBS) as an initiator, and triethylene glycol dimethacrylate (TEGDMA) as a crosslinking agent, a prepolymerization solution of HEMA was prepared by mixing with water. The molar ratio of HEMA:APS / SMBS:TEGDMA:water was 10. 5 :1:1:10 5The molar ratio of ammonium persulfate to sodium metabisulfite was 1:1. Using the same type of blood vessel, after decellularization, the pre-polymerization solution of HEMA was applied to the inner surface / membrane surface, allowing hydroxyethyl methacrylate monomer to penetrate into the superficial tissue of the decellularized blood vessel lumen. Under constant temperature water bath conditions of 45℃, after 36 hours, polymerization was induced within the decellularized vascular matrix scaffold to form a novel composite tissue with interpenetrating poly(hydroxyethyl methacrylate) hydrogel (PHEMA) and the decellularized vascular matrix scaffold. The above material was added again, and through secondary surface polymerization, a smooth surface layer containing only PHEMA was formed on the inner surface of the novel composite artificial blood vessel. The vessel was then immersed in deionized water for 7 days, with the water changed every 24 hours to remove unreacted small molecules, thus obtaining the novel composite artificial blood vessel with interpenetrating PHEMA and the decellularized matrix scaffold.
[0041] This composite artificial blood vessel has a tensile strength of 20-31 MPa, a biomechanical strength destructive test result of 128 mmHg, and a sphere pressure resistance test. It can withstand pressure above 80 mmHg for 48 hours without pressure drop or liquid leakage, maintaining structural integrity. The maximum pressure range is 90.4-187.6 mmHg, with an average of 128.0±34.2 mmHg. It also has pressure resistance with 10-0 nylon sutures.
[0042] Comparative Example 3
[0043] Current artificial blood vessels made of polymer materials lack biological activity, making it difficult to heal with the patient's blood vessels at the implantation site, leading to leakage and even rupture. The composite artificial blood vessel proposed in Example 3 demonstrates the advantages of artificial blood vessels on its inner surface. Furthermore, due to the inclusion of decellularized blood vessels, it possesses excellent biological properties, allowing for effective healing with the implantation site of the patient's blood vessel, thus avoiding the aforementioned shortcomings.
[0044] Comparative Example 4
[0045] Current decellularized blood vessels can heal well with the patient's blood vessels, but because their lumen lacks epithelial cells, they are prone to thrombosis or dissolution. The composite artificial blood vessel proposed in Example 3, with its decellularized tissue wall, heals well with the patient's blood vessels. Its advantage lies in the presence of a PHEMA layer on its inner surface, which can avoid or reduce adverse events such as thrombosis or dissolution after transplantation.
[0046] Example 4
[0047] Same as Example 1, except that the reaction was carried out in a constant temperature water bath at 40°C for 42 h, and the molar ratio of hydroxyethyl methacrylate to initiator was 10:1. 4 1. The molar ratio of hydroxyethyl methacrylate to crosslinking agent is 10:1. 4 :1.
[0048] Example 5
[0049] Same as Example 1, except that the reaction was carried out in a constant temperature water bath at 50°C for 30 h, and the molar ratio of hydroxyethyl methacrylate to initiator was 10:1. 5 1. The molar ratio of hydroxyethyl methacrylate to crosslinking agent is 10:1. 5 :1.
[0050] The above description of the embodiments is only for the purpose of helping to understand the technical solution and core idea of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made to the present invention without departing from the principle of the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.
Claims
1. A composite artificial lumen / membrane tissue, characterized by, The synthetic polymer material is selected from one or more of silicone, synthetic polymer hydrogel and acrylic polymer, and the natural polymer biomaterial is selected from one or more of decellularized matrix scaffold material, non-decellularized matrix and non-decellularized collagen.
2. The composite artificial body orifice / membrane tissue of claim 1, wherein, The decellularized matrix scaffold tissue or organ is allogeneic or xenogeneic.
3. The composite artificial body orifice / membrane tissue according to claim 1 or 2, wherein, The acrylic polymer is polymethyl hydroxyethyl methacrylate.
4. The composite artificial body orifice / membrane tissue according to claim 1 or 2, wherein, The composite artificial lumen / membrane tissue comprises a central surface region, a peripheral region and an outer surface region connected in sequence, wherein the central surface region is a synthetic polymer material, the peripheral region is a composite of synthetic polymer material and natural polymer biomaterial, and the outer surface region is a natural polymer biomaterial.
5. The preparation method of the composite artificial body orifice / membrane tissue according to claim 1, characterized in that, The method comprises the following steps: using a natural polymer material as a scaffold, synthesizing a synthetic polymer material on the scaffold, and obtaining a composite artificial lumen / membrane tissue with the synthetic polymer material and the natural polymer material interpenetrating through twice polymerization.
6. The production method according to claim 5, wherein The natural polymer biomaterial is a decellularized matrix scaffold, and the synthetic polymer material is polymethyl hydroxyethyl methacrylate.
7. The production method according to claim 6, wherein The method comprises the following steps: using polymethyl hydroxyethyl methacrylate as a raw material, ammonium persulfate / sodium metabisulfite as an initiator, and triethylene glycol dimethacrylate as a crosslinking agent, stirring the materials with water to be uniform, coating the mixture on the inner surface of the decellularized matrix scaffold / membrane surface, allowing the polymethyl hydroxyethyl methacrylate monomer to penetrate into the decellularized lumen or membrane tissue, and reacting for 30-42 h in a constant-temperature water bath at 40-50℃, so as to induce polymerization into a composite tissue with polymethyl hydroxyethyl methacrylate hydrogel and decellularized matrix scaffold interpenetrating, adding the above materials again, forming a smooth surface layer containing only polymethyl hydroxyethyl methacrylate on the surface of the composite tissue through twice polymerization, and soaking the composite tissue in deionized water to remove unreacted small molecules, thereby obtaining a composite artificial lumen / membrane tissue with polymethyl hydroxyethyl methacrylate and decellularized matrix scaffold interpenetrating.
8. The production method according to claim 7, characterized by, Molar ratio of hydroxyethyl methacrylate to initiator is 10 4 ~10 5 :
1. Molar ratio of hydroxyethyl methacrylate to crosslinker is 10 4 ~10 5 :
1.
9. The preparation method according to claim 7, characterized in that, The reaction is performed in a constant-temperature water bath at 45℃ for 36 h.