CN114681670A - Preparation method of bone repair scaffold with drug slow release and antibacterial effects - Google Patents
Preparation method of bone repair scaffold with drug slow release and antibacterial effects Download PDFInfo
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A—HUMAN NECESSITIES
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/58—Materials at least partially resorbable by the body
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/23—Carbohydrates
- A61L2300/236—Glycosaminoglycans, e.g. heparin, hyaluronic acid, chondroitin
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
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- A—HUMAN NECESSITIES
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
- A61L2300/414—Growth factors
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
- A61L2300/602—Type of release, e.g. controlled, sustained, slow
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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Abstract
The invention relates to the preparation of a bone repair bracket, in particular to a preparation method of a bone repair bracket with drug slow release and antibacterial effects, which comprises the steps of mixing MBG powder and PLLA powder, obtaining composite powder through ultrasonic dispersion, ball milling and drying, and obtaining PLLA/MBG bone bracket through selective laser sintering; immersing the PLLA/MBG bone scaffold into a hydrogel solution prepared by chitosan to prepare a PLLA/MBG/CS scaffold, and then immersing the PLLA/MBG/CS scaffold into a solution prepared by growth factors to obtain the PLLA/MBG/CS scaffold loaded with the growth factors, wherein a PLLA/MBG scaffold matrix provides mechanical properties and environments for cell migration and adhesion; the attached chitosan hydrogel on the scaffold provides a matrix for cell growth and has antibacterial and drug-loading capabilities; the Slit3 or PDGF-BB loaded on the scaffold can be connected with H-type blood vessels, so that the proliferation and differentiation of osteoprogenitor cells in bone marrow are promoted.
Description
Technical Field
The invention relates to preparation of a bone repair bracket, in particular to a preparation method of a bone repair bracket with drug slow release and antibacterial effects.
Background
Bone graft surgery is required for bone defects caused by trauma, tumors, infection, congenital diseases, spinal fusion, and the like. In the operation process, various problems exist, such as long operation time of autologous bone transplantation, limited supply, complication of donor parts, low compatibility between allogeneic bone transplantation and recipient tissues, easy occurrence of rejection reaction and the like. In order to solve these problems, artificial bone substitutes have appeared and developed rapidly, and bone grafting operations are gradually shifted to the use of synthetic bone substitutes. Polymer materials are considered a treatment for bone defects due to their good biological properties, providing conditions for cell migration and new tissue generation at the site of bone repair. Among them, poly-L-lactic acid (PLLA) is a biodegradable, biocompatible and non-toxic polymer material, and is considered as a promising bone scaffold material. However, PLLA bone scaffolds have delayed healing or no healing due to loss of cellular activity or death during bone repair after implantation into a bone defect site, and PLLA is brittle and hydrophobic, which is not conducive to adhesion and proliferation of cells on the scaffold, thus limiting its application to some extent.
Disclosure of Invention
Aiming at the technical problems, the invention provides a preparation method of a bone repair scaffold with drug slow release and antibacterial effects, and the PLLA/MBG/CS scaffold loaded with growth factors obtained by the method is beneficial to cell adhesion and proliferation.
The technical scheme adopted by the invention for solving the technical problems is as follows: a preparation method of a bone repair scaffold with drug slow release and antibacterial effects comprises the following steps:
(1) mixing MBG powder and PLLA powder, performing ultrasonic dispersion, ball milling and drying to obtain composite powder, and performing selective laser sintering on the composite powder to obtain a PLLA/MBG bone scaffold;
(2) immersing the PLLA/MBG bone scaffold into a hydrogel solution prepared by chitosan to prepare a PLLA/MBG/CS scaffold with chitosan attached on the PLLA/MBG bone scaffold;
(3) immersing the PLLA/MBG/CS scaffold in a solution configured by growth factors to obtain a growth factor-loaded PLLA/MBG/CS scaffold.
Preferably, the mass ratio of the MBG powder to the PLLA powder is 1 (5-10).
Preferably, the particle size of the composite powder is 50 to 100 micrometers.
Preferably, the hydrogel solution prepared by chitosan is a chitosan hydrogel solution with the concentration of 6g/L prepared by mixing chitosan and acetic acid.
Preferably, the PLLA/MBG bone scaffold is soaked in the chitosan hydrogel solution for 1 hour, then taken out, dried in an oven at 37 ℃, and washed.
Preferably, the dried PLLA/MBG/CS scaffolds are washed with ultrapure water, then with a 5% NaOH solution for 30min, and then repeatedly washed with ultrapure water.
Preferably, the growth factor is Slit3 or PDGF-BB, and the configured concentration is 2 g/L.
Preferably, the PLLA/MBG/CS scaffold is immersed in a solution prepared with growth factors for 1 hour at 37 ℃ and then taken out and frozen and dried at-20 ℃.
According to the technical scheme, the PLLA/MBG/CS scaffold loaded with the growth factors can be prepared, wherein the PLLA/MBG scaffold matrix provides an environment for mechanical property, cell migration and adhesion; the attached chitosan hydrogel on the scaffold provides a matrix for cell growth and has antibacterial and drug-loading capabilities; the Slit3 or PDGF-BB loaded on the scaffold can be connected with H-type blood vessels so as to promote the proliferation and differentiation of osteoprogenitor cells in bone marrow; the combination of the three components greatly accelerates the bone repair.
Detailed Description
The present invention will be described in detail with reference to the following examples, which are illustrative of the present invention and are not to be construed as limiting the present invention.
The invention provides a preparation method of a bone repair scaffold with drug slow release and antibacterial effects, which comprises the following steps:
firstly, mixing MBG powder and PLLA powder, performing ultrasonic dispersion, ball milling and drying to obtain composite powder, and performing selective laser sintering on the composite powder to obtain a PLLA/MBG bone scaffold; the Mesoporous Bioactive Glass (MBG) is an inorganic material, and can obviously improve the hydrophobic property of a polymer and improve the biocompatibility after being compounded with a PLLA polymer, so that the Mesoporous Bioactive Glass (MBG) can provide good mechanical properties and environments for cell migration and adhesion as a bone scaffold material, and products generated after the MBG is degraded are nontoxic and harmless and can be metabolized by a human body. In the implementation process, the mass ratio of the MBG powder to the PLLA powder is 1 (5-10), and the particle size of the composite powder is 50-100 microns, so that the MBG has enough pores, and the scaffold has good mechanical properties.
Then, immersing the PLLA/MBG bone scaffold into a hydrogel solution prepared by chitosan to prepare the PLLA/MBG/CS scaffold with chitosan attached on the PLLA/MBG bone scaffold; chitosan (CS) not only has broad-spectrum antibacterial properties, but also is a slow-release drug delivery vehicle. Although CS itself has low mechanical strength, it is attached to the PLLA/MBG bone scaffold of the present invention to improve mechanical strength. Because the PLLA/MBG bone scaffold is immersed in the chitosan hydrogel, hydrogel molecules enter and are adsorbed in pores due to the existence of the pores in the MBG, and thus, the mechanical property is improved. In the implementation process, the chitosan hydrogel solution prepared by chitosan is prepared by mixing chitosan and acetic acid, and the concentration of the chitosan hydrogel solution is 6g/L, so that colloidal molecules can enter pores quickly. The PLLA/MBG bone scaffold is immersed in the chitosan hydrogel solution for 1 hour, then taken out and put into an oven for drying at 37 ℃, and then the dried PLLA/MBG/CS scaffold is rinsed with ultrapure water, then put into a 5% NaOH solution for 30min to neutralize the acidity of acetic acid, and then rinsed repeatedly with ultrapure water for at least 3 times, thereby obtaining the PLLA/MBG/CS scaffold.
Then, the PLLA/MBG/CS scaffold is immersed in a solution prepared by a growth factor, particularly at the temperature of 37 ℃, for 1 hour, and the PLLA/MBG/CS scaffold loaded with the growth factor can be obtained due to the existence of CS; then taking out, freezing at-20 deg.C, and drying for storage; in the implementation process, the growth factor is Slit3 or PDGF-BB, and the concentration of the growth factor is configured to be 2g/L, so that the Slit3 or PDGF-BB can accelerate the loading on CS. PDGF-BB is a platelet-derived growth factor, Slit3 is a pro-angiogenic growth factor with a remarkable effect, and the growth factor has high expression and secretion on endothelial cells and smooth muscle cells of blood vessels, and can promote the development of new blood vessels in vivo; in vitro, can promote the formation of endothelial cell vascular network and the sprouting of new blood vessels; and Slit3 and PDGF-BB have the capacity of promoting H-type blood vessels and bone formation, and the H-type blood vessels are capillary blood vessel subtypes which are related to osteogenesis and highly express CD31 and endogucin, are positioned near a metaphyseal growth plate, periosteum and intima, can promote the proliferation and differentiation of osteoprogenitor cells in bone marrow, and are important regulatory factors for bone regeneration. Therefore, when the PLLA/MBG/CS bracket loaded with Slit3 or PDGF-BB prepared by the invention is connected with an H-type blood vessel, the bone repair can be greatly accelerated. In the following, the Slit3 is used as an example, and the execution is referred to when PDGF-BB is used.
Examples
Mixing MBG powder and PLLA powder according to the mass ratio of 1:10, performing ultrasonic dispersion, performing ball milling to obtain composite powder with the average particle size of about 100 microns, drying for later use, and performing selective laser sintering to obtain a cylindrical PLLA/MBG bone scaffold with the diameter of about 5mm and the height of 10 mm; then, immersing the PLLA/MBG bone scaffold into 6g/L chitosan hydrogel solution for 1 hour, then putting the bone scaffold into an oven for drying at 37 ℃, taking out the bone scaffold and washing the bone scaffold with ultrapure water, then putting the bone scaffold into 5% NaOH solution for 30min, and then repeatedly washing the bone scaffold with ultrapure water for 3 times to obtain the PLLA/MBG/CS scaffold; then, the PLLA/MBG/CS bracket is immersed in a 2g/L Slit3 solution for 1 hour at the temperature of 37 ℃, then the PLLA/MBG/CS bracket is taken out and frozen and dried at the temperature of-20 ℃ to obtain the PLLA/MBG/CS bracket loaded with Slit3, the PLLA/MBG/CS bracket is cut into a small section to expose the cross section, the pores of the MBG are basically filled with CS through electron microscope detection, and the Slit3 loaded on the CS is distributed more uniformly; the rest section has the advantages of compressive strength of about 6.3MPa, compressive modulus of about 78.3MPa and excellent mechanical property through mechanical detection; and then soaking the rest section in a buffer solution at 37 ℃, measuring the content of Slit3 in the buffer solution at days 1, 5, 10, 20 and 40, and obtaining the cumulative release amounts of 5.6%, 28.3%, 67.2%, 89.7% and 98.2% respectively by calculation, thereby showing that the slow release effect is obvious.
Claims (8)
1. A preparation method of a bone repair scaffold with drug slow release and antibacterial effects is characterized by comprising the following steps:
(1) mixing MBG powder and PLLA powder, performing ultrasonic dispersion, ball milling and drying to obtain composite powder, and performing selective laser sintering on the composite powder to obtain a PLLA/MBG bone scaffold;
(2) immersing the PLLA/MBG bone scaffold into a hydrogel solution prepared by chitosan to prepare a PLLA/MBG/CS scaffold with chitosan attached on the PLLA/MBG bone scaffold;
(3) immersing the PLLA/MBG/CS scaffold in a solution configured by growth factors to obtain a growth factor-loaded PLLA/MBG/CS scaffold.
2. The method for preparing the bone repair scaffold with the drug slow release and antibacterial effects according to claim 1 is characterized in that: the mass ratio of the MBG powder to the PLLA powder is 1 (5-10).
3. The method for preparing the bone repair scaffold with the drug slow release and antibacterial effects according to claim 1 is characterized in that: the particle size of the composite powder is 50-100 microns.
4. The method for preparing the bone repair scaffold with the drug slow release and antibacterial effects according to claim 1 is characterized in that: the hydrogel solution prepared from chitosan is a chitosan hydrogel solution with the concentration of 6g/L prepared by mixing chitosan and acetic acid.
5. The method for preparing the bone repair scaffold with the drug slow-release and antibacterial effects according to claim 4 is characterized in that: and soaking the PLLA/MBG bone scaffold into the chitosan hydrogel solution for 1 hour, taking out the bone scaffold, drying the bone scaffold in an oven at 37 ℃, and cleaning the bone scaffold.
6. The method for preparing the bone repair scaffold with the drug slow-release and antibacterial effects according to claim 5 is characterized in that: when cleaning, the dried PLLA/MBG/CS stent is washed by ultrapure water, then is put into a 5% NaOH solution for 30min, and then is washed by ultrapure water repeatedly.
7. The method for preparing the bone repair scaffold with the drug slow release and antibacterial effects according to claim 1 is characterized in that: the growth factor is Slit3 or PDGF-BB, and the configured concentration is 2 g/L.
8. The method for preparing a bone repair scaffold with drug sustained release and antibacterial effects according to claim 7, wherein the method comprises the following steps: the PLLA/MBG/CS scaffold was immersed in a solution of growth factor for 1 hour at 37 deg.C, removed, frozen at-20 deg.C and dried.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210142120.XA CN114681670A (en) | 2022-02-16 | 2022-02-16 | Preparation method of bone repair scaffold with drug slow release and antibacterial effects |
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| CN202210142120.XA CN114681670A (en) | 2022-02-16 | 2022-02-16 | Preparation method of bone repair scaffold with drug slow release and antibacterial effects |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105311676A (en) * | 2015-11-10 | 2016-02-10 | 北京大清生物技术有限公司 | Hard tissue engineering scaffold material having bioactivity and preparation method of hard tissue engineering scaffold material |
| US20190134262A1 (en) * | 2017-11-06 | 2019-05-09 | Purdue Research Foundation | Bioactive Glass-Polymer Composite Bone Scaffolds |
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2022
- 2022-02-16 CN CN202210142120.XA patent/CN114681670A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105311676A (en) * | 2015-11-10 | 2016-02-10 | 北京大清生物技术有限公司 | Hard tissue engineering scaffold material having bioactivity and preparation method of hard tissue engineering scaffold material |
| US20190134262A1 (en) * | 2017-11-06 | 2019-05-09 | Purdue Research Foundation | Bioactive Glass-Polymer Composite Bone Scaffolds |
Non-Patent Citations (1)
| Title |
|---|
| GUOWEN QIAN等: "《Silver-doped bioglass modified scaffolds: A sustained antibacterial efficacy》", 《MATERIALS SCIENCE AND ENGINEERING： C》 * |
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