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CN-117398515-B - High-efficiency load osteogenesis angiopoiesis multi-drug wood-based elastic artificial periosteum and preparation method and application thereof

CN117398515BCN 117398515 BCN117398515 BCN 117398515BCN-117398515-B

Abstract

The invention discloses a high-efficiency load osteogenesis angiopoiesis multi-drug wood-based elastic artificial periosteum, a preparation method and application thereof. The artificial periosteum takes wood-derived elastic wood as a basal layer, polydopamine as an adhesive layer and a layer-by-layer drug-loaded electrostatic self-assembly layer as a biological functional layer. The basal layer not only imitates the structural characteristics and mechanical anisotropy of a natural periosteum, but also is beneficial to clinical implantation of an artificial periosteum due to elasticity, the bonding layer effectively connects the basal layer and the biological function layer and simultaneously endows the artificial periosteum with certain adhesion performance, and the biological function layer can efficiently load multiple medicines to realize synergistic long-acting promotion of osteogenesis and vascularization biological functions in the bone repair process. The artificial periosteum prepared by the invention has the characteristics of anisotropy, high porosity, high elasticity and adhesion in terms of structure and mechanics, and the drug loading capacity and the mechanical property of the artificial periosteum can be regulated and controlled, so that the artificial periosteum is an ideal novel artificial periosteum material.

Inventors

  • LUO BINGHONG
  • Sun tianyi
  • CHEN CHUNHUA
  • WEN WEI
  • ZHOU CHANGREN

Assignees

  • 暨南大学

Dates

Publication Date
20260505
Application Date
20230914

Claims (10)

  1. 1. The high-efficiency load osteogenesis vascular multi-drug wood-based elastic artificial periosteum is characterized by comprising a basal layer, a bonding layer and a biological function layer which are sequentially connected, wherein the basal layer is elastic wood, the bonding layer is polydopamine, and the biological function layer is formed by self-assembling a positively charged layer loaded with drugs and a negatively charged layer loaded with drugs layer by layer; The elastic wood is obtained by removing lignin and hemicellulose from natural wood under alkaline conditions; the medicine in the positively charged layer loaded with the medicine is a negatively charged osteogenic medicine; the medicine in the negatively charged layer loaded with the medicine is positively charged angiogenic medicine.
  2. 2. The high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 1, wherein: the thickness of the elastic wood is 500-2000 mu m; the thickness of the bonding layer is 5-50 nm; the thickness of the biological functional layer is 10-1000 nm.
  3. 3. The high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 1, wherein: The biological functional layer is connected with the bonding layer through a positively charged layer loaded with a drug; The structure of the biological functional layer is shown as (a-b) n -a m , wherein a is a positively charged layer loaded with a drug, b is a negatively charged layer loaded with a drug, n is an integer more than 0, m is 0 or 1, and n cannot be 0.
  4. 4. The high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 1, wherein: the elastic wood is prepared by the following specific steps: (1) Preparing an alkaline solution; (2) Placing natural wood in an alkaline solution to remove bubbles in wood pore canals through vacuum impregnation, and releasing vacuum so as to fully immerse the mixed solution in the wood pore canals; (3) Heating the wood-containing solution obtained in the step (2), soaking and washing the wood obtained after the reaction with deionized water to remove residual chemical impurities, and finally freezing and drying to obtain elastic wood; the positively charged layer loaded with the medicine is obtained by encapsulating the medicine with a natural polymer with positive charges; The negative charge layer loaded with the medicine is obtained by coating the medicine with natural polymers with negative charges.
  5. 5. The high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 4, wherein: The alkaline solution in the step (1) is a mixed solution containing 3 mol/L NaOH and 0.5 mol/L Na 2 SO 3 ; The vacuum condition in the step (2) is negative pressure with the relative vacuum degree not more than-80 kPa; the soaking time in the step (2) is 10-25 min; The time for releasing the vacuum in the step (2) is 5-10 min; the heating reaction condition in the step (3) is that the reaction is carried out for 0.5-9 hours at 70-100 ℃; The soaking time in the step (3) is 24-48 hours, and water is replaced for multiple times for washing; The freezing condition in the step (3) is that the temperature is-20 ℃ and the freezing time is 6-24 hours; the drying mode in the step (3) is freeze drying; The natural wood comprises any one of modified or unmodified balsa wood, poplar, beech, basswood, walnut, elm and paulownia; the natural polymer with positive charges is chitosan; the natural polymer with negative charges is at least one of hyaluronate, alginate, gelatin and type I collagen.
  6. 6. The high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 1, wherein: the negative bone-forming medicament is pamidronate disodium; the positively charged angiopoiesis medicament is deferoxamine.
  7. 7. The method for preparing the high-efficiency load osteogenic vascular multi-drug wood-based elastic artificial periosteum according to any one of claims 1 to 6, which is characterized by comprising the following steps: 1) Soaking the elastic wood in a dopamine solution, washing after the dopamine forms a polydopamine layer on the surface of the elastic wood through an oxidation self-polymerization process to remove residual unreacted dopamine and impurities; 2) Preparing a natural polymer with positive charges into a solution A1 with a pH value of 1-6, adding a drug to obtain a solution A2, preparing a natural polymer with negative charges into a solution B1 with a pH value of 1-6, and then adding the drug to obtain a solution B2; 3) Soaking the elastic wood with the dopamine surface modified in a solution A2, and then washing with deionized water to obtain elastic wood with the polydopamine layer and the drug-loaded positively charged layer modified in sequence from inside to outside; 4) Soaking the elastic wood obtained in the step 3) in a solution B2, and then washing with deionized water to obtain the elastic wood with the polydopamine layer, the positively charged layer loaded with the medicine and the negatively charged layer loaded with the medicine, wherein according to actual requirements, the elastic wood finally obtained in the step 3) or the step 4) is the high-efficiency load osteogenesis multiple medicine wood-based elastic artificial periosteum, or the step 3) and the step 4) are repeatedly treated for a plurality of times to obtain the high-efficiency load osteogenesis multiple medicine wood-based elastic artificial periosteum.
  8. 8. The method for preparing the high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 7, further comprising the steps of: 5) And after the final layer is assembled, freezing and drying the obtained product to obtain the high-efficiency load osteogenesis vascular multiple drug wood-based elastic artificial periosteum.
  9. 9. The method for preparing the high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to claim 7 or 8, wherein the method comprises the following steps: the concentration of the dopamine solution in the step 1) is 0.2-3 g/L; The solvent in the dopamine solution in the step 1) is Tris-HCl buffer solution with the concentration of 8-12 mM and the pH value of 8.0-9.0; The soaking time in the step 1) is 1-30 hours; the concentration of the natural polymer with positive charges in the solution A1 in the step 2) is 1-10 mg/mL; the concentration of the drug in the solution A2 in the step 2) is 0.01-1 mg/mL; the Zeta potential of the solution A2 in the step 2) is +10 to +45 mV; The concentration of the natural polymer with negative charges in the solution B1 in the step 2) is 1-10 mg/mL; The concentration of the drug in the solution B2 in the step 2) is 20-200 mug/mL; the Zeta potential of the solution B2 in the step 2) is-5 to-45 mV; The soaking time in the step 3) and the step 4) is 15-60 min; The washing times in the step 3) and the step 4) are 2-3 times.
  10. 10. The use of the high-efficiency load osteogenic vascular multidrug wood-based elastic artificial periosteum according to any one of claims 1-6 in the preparation of bone repair materials.

Description

High-efficiency load osteogenesis angiopoiesis multi-drug wood-based elastic artificial periosteum and preparation method and application thereof Technical Field The invention belongs to the field of biomedical materials, and in particular relates to a high-efficiency load osteogenesis vascular multiple drug wood-based elastic artificial periosteum, a preparation method and application thereof. Background Critical-size bone defects caused by trauma, tumor resection surgery remain a major challenge to orthopedics, and bone defects themselves have been considered excessively and the importance of periosteum has been ignored. Periosteum is a layer of highly vascularized connective tissue membrane that covers the surface of bone, delivering oxygen and nutrients to the bone and adjacent tissues by providing a blood supply. The ideal artificial periosteum material has the bionic characteristics of both structure and function, and is structurally divided into an outer fibrous layer and an inner layer, wherein the outer fibrous layer mainly consists of collagen and high-density elastin which are axially arranged, so that the periosteum has anisotropy and elasticity in mechanics, and the inner layer (also called as an osteogenesis layer) is tightly adhered to bone, and the osteogenesis process is coordinated by releasing cytokines. The artificial periosteum disclosed at present mainly comprises a decellularized periosteum bracket, an electrostatic spinning membrane, a micro/nano fiber membrane carrying a growth factor and a hydrogel membrane. Structurally, the existing artificial periosteum often neglects the structural and mechanical anisotropy of the natural periosteum, has certain flexibility and elasticity and is limited in adhesion. The national patent application publication of application number 202210193993.3 discloses a preparation method and application of a double-layer bionic artificial periosteum, which has a double-layer structure of a natural periosteum and a certain biological functionality, but neglects the anisotropism of the natural periosteum in structure and mechanics and the requirement of the natural periosteum on the adhesion of the artificial periosteum in practical application. Natural wood is abundant in source, is mainly composed of aligned fibers and has high porosity and many tubular channels, which is essential for rapid transmission of nutrients and moisture. It is particularly important that the naturally occurring anisotropic structure makes the wood a "natural template" for the anisotropic collagen fiber layer in the simulated natural periosteum. However, wood is too strong and lacks flexibility, and would cause mechanical damage as an implant material. According to researches, the wood-based cellulose fiber material prepared by completely or partially removing lignin and hemicellulose components of the wood after alkaline treatment can not only retain the anisotropic mechanical and structural characteristics of the wood, but also improve the flexibility of the wood. In the prior art, we have disclosed a high-efficiency antibacterial agent combination, a natural wood-based hydrogel periosteum material and an application (application number 202210550879.1), wherein an artificial periosteum prepared by immersing a hydrogel loaded with a drug into a means of completely removing lignin and hemicellulose white wood channels has certain antibacterial and osteogenic effects, but the completely removed lignin and hemicellulose white wood does not have morphological stability in a wet environment, and the hydrogel needs to be introduced to endow the artificial periosteum with stable morphology and good flexibility. However, the introduction of hydrogel occupies the unique channel structure of wood, and the periosteum design ignores vascularization and adhesion characteristics, and the sufficient blood supply and adhesion characteristics are the basic precondition and guarantee for ensuring the effective repair of damaged bone tissues when the artificial periosteum is clinically applied. Therefore, the anisotropic fiber structure of the bionic periosteum by using wood is an effective method, however, how to design a wood-based artificial periosteum, which can not only efficiently load the osteogenic and angiogenic drugs, but also retain the unique channel structure of the wood, and has certain elasticity and adhesion characteristics required by clinical practical use, becomes a problem to be solved urgently. Disclosure of Invention The invention aims to overcome the defects and shortcomings of the prior art and provide a high-efficiency load osteogenesis vascular multi-drug wood-based elastic artificial periosteum. The invention also aims to provide a preparation method of the high-efficiency load osteogenesis vessel multi-drug wood-based elastic artificial periosteum. Still another object of the present invention is to provide the use of the above-mentioned high-efficiency load osteogenic v