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CN-121971708-A - Keratinized gingival repair material and preparation method and application thereof

CN121971708ACN 121971708 ACN121971708 ACN 121971708ACN-121971708-A

Abstract

The invention discloses a keratinized gingival repair material, and a preparation method and application thereof, and belongs to the technical field of periodontal and biomedical materials. The keratinized gingival repair material body is composed of an inner material layer and an outer material layer, wherein the inner material layer and the outer material layer are nanofiber membranes formed by polycaprolactone, gelatin and sodium hyaluronate, and interface components of the inner material layer and the outer material layer are matched, the inner material layer nanofiber membrane is of a non-oriented structure and is used for simulating an inherent layer of the keratinized gingiva and is close to a tissue surface, the outer material layer nanofiber membrane is of an oriented structure and is used for simulating an epithelial layer of the keratinized gingiva and is close to an oral cavity surface, and the integrated keratinized gingival repair material body is formed. The invention establishes a bionic integrated material structure, realizes rapid epithelialization, effectively seals wound surfaces, simultaneously facilitates cell spreading, migration and growth on the wound surfaces, can guide host cells to construct mature tissues similar to natural keratinized gingiva in structure and function, shortens repair time, and is convenient for batch production and clinical transformation application.

Inventors

  • TANG XIAOLIN
  • ZHAO YUAN
  • YANG JINGXIN
  • YANG MAN

Assignees

  • 中国医科大学附属口腔医院

Dates

Publication Date
20260505
Application Date
20260126

Claims (10)

  1. 1. The keratinized gingival repair material is characterized in that the keratinized gingival repair material body is composed of an inner material layer and an outer material layer, wherein the inner material layer and the outer material layer are nanofiber membranes formed by polycaprolactone, gelatin and sodium hyaluronate, and interface components of the inner material layer and the outer material layer are matched, the inner material layer nanofiber membrane is of a non-oriented structure and simulates an inherent layer of the keratinized gingiva to be close to a tissue surface, the outer material layer nanofiber membrane is of an oriented structure and simulates an epithelial layer of the keratinized gingiva to be close to an oral cavity surface, and the integrated keratinized gingival repair material body is formed.
  2. 2. The keratinized gingival repair material of claim 1 wherein the sodium hyaluronate has a weight average molecular weight of 10-20 kilodaltons, a gelatin freezing force of 240Bloom g, and a polycaprolactone number average molecular weight of 8-10 kilodaltons.
  3. 3. A keratinized gingival repair material as set forth in claim 1 wherein the thickness ratio of the inner layer of material to the outer layer of material nanofiber membrane, i.e., the ratio of non-oriented structure to oriented structure, is (5-8): 2-5; The nanofiber membrane is an electrostatic spinning fiber membrane, the diameter of the oriented structure fiber is 200nm-500nm, and the diameter of the unoriented structure fiber is 100nm-800nm.
  4. 4. The method for preparing the keratinized gingival repair material according to claim 1, wherein A C-B-A reverse order liquid preparation method is adopted to prepare A complex homogeneous electrostatic spinning solution, wherein C represents sodium hyaluronate, B represents gelatin, A represents polycaprolactone, and then an inner layer non-oriented structure nanofiber membrane and an outer layer oriented structure nanofiber membrane are respectively prepared through an electrostatic spinning method, and electrostatic spinning technological parameters and thicknesses of the inner layer electrostatic spinning fiber membrane and the outer layer electrostatic spinning fiber membrane are regulated and controlled.
  5. 5. The method for preparing the keratinized gingival repair material according to claim 4, comprising the following steps: preparing sodium hyaluronate solution, namely dissolving sodium hyaluronate in a binary/compound solvent system, and stirring until the sodium hyaluronate is completely dissolved; Preparing sodium hyaluronate-gelatin solution, namely adding gelatin into the sodium hyaluronate solution, and stirring until the gelatin is completely dissolved; preparing a sodium hyaluronate-gelatin-polycaprolactone electrostatic spinning solution, namely adding polycaprolactone into the sodium hyaluronate-gelatin solution, and stirring until the polycaprolactone is completely dissolved; Injecting a sodium hyaluronate-gelatin-polycaprolactone electrostatic spinning solution into an electrostatic spinning equipment injector pump, setting positive voltage, negative voltage and propulsion rate parameters, adjusting the distance and the rotating speed from an electrostatic spinning nozzle to the edge of a roller collector, and spinning to obtain a material inner layer nanofiber membrane with a non-oriented structure; The method comprises the steps of preparing a three-component material outer layer directional structure by electrostatic spinning, namely adjusting the rotating speed of a roller collector, keeping the rest electrostatic spinning parameters unchanged, continuing spinning, and generating a material outer layer nanofiber membrane of the directional structure on a material inner layer nanofiber membrane of a non-directional structure to obtain a keratinized gingival repair material body; and (3) drying the electrostatic spinning film, namely vacuum drying the keratinized gingival repair material body at room temperature for later use.
  6. 6. The method for preparing the keratinized gingival repair material according to claim 5, wherein the binary/composite solvent system is hexafluoroisopropanol/formic acid mixed solvent in sodium hyaluronate solution, wherein the mass volume ratio of sodium hyaluronate to solvent is (0.5-1) to (100); Preparing a sodium hyaluronate-gelatin solution, wherein the mass volume ratio of gelatin to solvent is mg (3.3-3.9) to mL (100); Preparing a sodium hyaluronate-gelatin-polycaprolactone electrostatic spinning solution, wherein the mass volume ratio of the polycaprolactone to the solvent is mg (7.7-9.1) to 100; the environment temperature is 25 ℃ plus or minus 1 ℃ and the humidity is 50% plusor minus 5% in the electrostatic spinning process; in the drying of the electrostatic spinning film, the vacuum drying time is more than 72 hours.
  7. 7. The method of preparing a keratinized gingival repair material according to claim 6, wherein the volume ratio of hexafluoroisopropanol to formic acid is (7-8): 2-3.
  8. 8. The method for preparing the keratinized gingival repair material according to claim 5, wherein in the electrostatic spinning preparation of the three-component material inner layer non-directional structure, parameters of positive voltage 18kV-20kV, negative voltage 2kV-3kV, propulsion rate 1.08mL/h-1.32mL/h are set, the distance from an electrostatic spinning nozzle to the edge of a roller collector is adjusted to be 13cm-15cm, the rotating speed is 300rpm-500rpm, and the spinning time is 5h-8h.
  9. 9. The method for preparing the gingival repair material according to claim 5, wherein in the preparation of the three-component material outer layer directional structure by electrostatic spinning, the rotation speed is adjusted to 3000rpm-4000rpm, and the spinning time is 2h-5h.
  10. 10. Use of a keratinized gingival repair material according to claim 1 for repairing local gingiva and surrounding keratinized tissue widening and thickening and vestibular sulcus deepening of a natural tooth or implant.

Description

Keratinized gingival repair material and preparation method and application thereof Technical Field The invention belongs to the technical field of periodontal and biomedical materials, and particularly relates to a keratinized gingival repair material, and a preparation method and application thereof. Background The loss of keratinized gingiva is a difficult problem to be solved urgently in periodontal and implant clinical treatment. The main stream substitute materials in the prior art for repairing the keratinized gingiva comprise Acellular Dermal Matrix (ADM) and heterologous collagen matrix (XCM), but have ethical problems and the like, and the clinical common use is autologous soft tissue free gingiva transplantation, epithelial connective tissue transplantation and the like, wherein a second operation area needs to be opened up, and the pain degree of patients is increased along with the bleeding and swelling risks of the supply area. At present, the prior keratinized gingival repair membrane material still has the technical bottlenecks that in structure, a single-layer membrane structure cannot simulate the epithelial layer and the inherent layer structure of the keratinized gingiva, and the design of the multi-layer membrane has the problem of interlayer interface separation and the cytotoxicity risk caused by using a cross-linking agent. In terms of components, the traditional single or composite component membrane material cannot achieve both mechanical strength and biological activity, and cannot form synergy between physical and chemical signals. In the process, the combination of multiple components and multiple structures often needs to adopt complex technology, the preparation process is complex, and the production efficiency is low. Therefore, how to obtain the bionic repair material of natural keratinized gingival components and structures is a technical field yet to be developed. Clinical applications of polycaprolactone (Polycaprolactone, PCL) are mainly focused on the fields of bone tissue engineering, soft tissue regeneration and the like, and an electrostatic spinning membrane prepared by the polycaprolactone has good biocompatibility, degradability and mechanical strength, is similar to an extracellular matrix of natural connective tissue in morphology, but the hydrophobicity of the polycaprolactone electrostatic spinning membrane can limit cell adhesion and proliferation. Gelatin (Gel) is a product of collagen partial hydrolysis, has hydrophilicity, is the most important natural polymer in connective tissue extracellular matrix, has chemical sequence groups which are easily identified and adhered by cells, and can promote cell adhesion and proliferation, but Gelatin is difficult to form continuous fibers under a high electric field of electrospinning, and is easy to cause phenomena of broken filaments and uneven fiber morphology. Hyaluronic Acid (HA) acts on various cell types of keratinocytes, vascular endothelial cells, etc., stimulates proliferation of cells under various mechanisms by activating cell surface receptors and regulating the structure and function of extracellular matrix to support tissue repair, but its mechanical strength is insufficient. In addition, in the electrostatic spinning production process, the hydrophilic-hydrophobic difference between polycaprolactone, gelatin and hyaluronic acid causes difficulty in uniform blending of spinning solutions, continuous fibers cannot be formed in the electrospinning process of hyaluronic acid due to the excessively high surface tension and low viscosity of hyaluronic acid, the cell biocompatibility can be influenced by using an emulsifying agent and a cross-linking agent, and the problems of high cost, high complexity and the like exist when two sets of solutions or two sets of equipment are used in coaxial or conjugated electrospinning. Thus, from a clinical point of view, an ideal keratinized gingival repair material should simultaneously meet the following conditions: (1) Structurally, the material mimics the epithelium and lamina propria of the keratinized gums. The epithelial layer is mainly formed by keratinocytes to form a compact brick wall structure, the main cell component of the natural layer is fibroblasts, collagen fibers are mainly synthesized, and the fibers are interwoven to form a reticular structure, so that the structure gradient is provided. (2) Compositionally, the material is capable of mimicking the composition of keratinized gingival tissue. Wherein collagen is the main part of gingival connective tissue, and extracellular matrix is proteoglycan, wherein hyaluronic acid is the main component. (3) The material can efficiently guide the growth of keratinized gingiva, and has good biocompatibility, no toxicity and no sensitization. (4) The material has certain mechanical property and proper degradation rate. (5) The material production and preparation process should be simplified, and batch production