CN114558176A - Chitosan-chondroitin sulfate nanoparticle and drug-loaded joint lubricant - Google Patents
Chitosan-chondroitin sulfate nanoparticle and drug-loaded joint lubricant Download PDFInfo
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- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/042—Polysaccharides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
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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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—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
- 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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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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
- A61L2300/232—Monosaccharides, disaccharides, polysaccharides, lipopolysaccharides
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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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- 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
- A61L2300/40—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
- 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus
Abstract
The invention provides chitosan-chondroitin sulfate nanoparticles and a drug-loaded joint lubricant, and belongs to the technical field of joint lubricants. In the invention, the chitosan has the advantages of good biocompatibility, blood compatibility, biodegradation, human body metabolism and the like, can be used as a safe and reliable drug carrier for treating arthritis, and has good bactericidal performance. The chondroitin sulfate contains a large amount of hydrophilic sulfonic groups, so that the chitosan nanoparticle can be endowed with excellent water lubrication performance, and the chitosan-chondroitin sulfate nanoparticle has a good antifriction effect; the chondroitin sulfate has a cartilage repair function, can promote the growth of cartilage, and lays a good foundation for the drug-loaded treatment and repair functions of the joint lubricant. Because the chitosan nano-particles have a soft network structure microscopically, when the chitosan nano-particles are used as a drug carrier, drugs wrapped in the chitosan nano-particles can be slowly released at a damaged joint interface, and an excellent stress-induced slow release effect is presented.
Description
Technical Field
The invention relates to the technical field of joint lubricants, in particular to chitosan-chondroitin sulfate nanoparticles and a drug-loaded joint lubricant.
Background
Osteoarthritis is a disease that causes malfunction of the knee joint, and is mainly associated with degeneration and damage of the articular cartilage. Hundreds of millions of people suffer from arthritis in China, and more than half of the patients are old people. Patients with severe arthritis require artificial joint replacement surgery, which is a great pain and economic burden to the patients (see Calcif. tissue int.2014,95, 495- & 505.). However, for patients with less early symptoms, injecting an artificial joint lubricant into the joint cavity to reduce wear of the damaged cartilage interface may be a better option.
In the course of the injectable treatment of arthritis, joint lubricants are required to have good friction-reducing lubricating properties and biocompatibility. Most of the traditional lubricating drug carriers are silicon dioxide (SiO)2) And inert hard nanoparticles such as gold (Au) have poor biocompatibility, so that the application of the inert hard nanoparticles in joint lubricants is limited.
Disclosure of Invention
In view of this, the invention aims to provide a chitosan-chondroitin sulfate nanoparticle and a drug-loaded joint lubricant, and the chitosan-chondroitin sulfate nanoparticle provided by the invention has good antifriction lubricity and biocompatibility.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a chitosan-chondroitin sulfate nanoparticle, which comprises a chitosan nanoparticle and chondroitin sulfate loaded on the surface of the chitosan nanoparticle.
Preferably, the particle size of the chitosan nanoparticle is 50-300 nm; the deacetylation degree of the chitosan nanoparticles is 75-85%, and the viscosity is 5-200 mpa.s.
Preferably, the mass ratio of the chitosan nanoparticles to the chondroitin sulfate is 1.5-3: 1.
The invention provides a preparation method of the chitosan-chondroitin sulfate nano-particles, which comprises the following steps:
mixing chondroitin sulfate, a carboxyl activating agent and a buffer solution, and performing carboxyl activation to obtain carboxyl-activated chondroitin sulfate;
and mixing the carboxyl-activated chondroitin sulfate with the chitosan nanoparticles, and performing chemical grafting reaction to obtain the chitosan-chondroitin sulfate nanoparticles.
Preferably, the carboxyl activating agent comprises 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and succinimide;
the mass ratio of the chondroitin sulfate to the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 1: 1-1.2;
the mass ratio of the chondroitin sulfate to the succinimide is 1: 1.2-1.5.
Preferably, the temperature for activating the carboxyl is 25-75 ℃, and the time is 1-2 h;
the temperature of the chemical grafting reaction is 25-37 ℃, and the time is 12-36 h.
The invention provides a drug-loaded joint lubricant, which comprises the chitosan-chondroitin sulfate nanoparticle and a drug loaded on the surface of the chitosan-chondroitin sulfate nanoparticle.
Preferably, the medicament is one or more of diclofenac sodium, isomonoside, mangiferin, fenoprofen, aspirin, loxoprofen, ibuprofen and nimesulide.
Preferably, the drug-loaded joint lubricant has a drug loading amount of 10-40 wt%.
The invention provides a preparation method of the drug-loaded joint lubricant, which comprises the following steps:
mixing chitosan-chondroitin sulfate nanoparticles, a medicament and a buffer solution, carrying the medicament to obtain the medicament-carrying joint lubricant.
The invention provides a chitosan-chondroitin sulfate nanoparticle, which comprises a chitosan nanoparticle and chondroitin sulfate loaded on the surface of the chitosan nanoparticle. In the invention, the Chitosan (CS) has the advantages of good biocompatibility, blood compatibility, biodegradation, human body metabolism and the like, can be used as a safe and reliable drug carrier for treating arthritis, and has good bactericidal performance. The chondroitin sulfate contains a large amount of hydrophilic sulfonic groups, so that the chitosan nanoparticle can be endowed with excellent water lubrication performance, and the chitosan-chondroitin sulfate nanoparticle has a good antifriction effect; chondroitin sulfate (CHI) has cartilage repair function, can promote the growth of cartilage, and lays a good foundation for drug-loaded treatment and repair function of joint lubricants. Because the chitosan nano-particles have soft network structures microscopically, when the chitosan nano-particles are used as drug carriers, the drugs wrapped in the chitosan nano-particles can be slowly released at damaged interfaces of joints in the dynamic loading and shearing processes, and an excellent stress-induced slow release effect is presented.
The invention provides a preparation method of the chitosan-chondroitin sulfate nanoparticle, which comprises the steps of firstly carrying out carboxyl activation on chondroitin sulfate, and then adding chitosan nanoparticles for chemical grafting to obtain the chitosan-chondroitin sulfate nanoparticle. The preparation method provided by the invention is simple to operate and is easy for industrial batch production. Meanwhile, the chitosan nanoparticle is chemically connected with the chondroitin sulfate, so that the chitosan-chondroitin sulfate nanoparticle has good stability.
The invention provides a drug-loaded joint lubricant, which comprises the chitosan-chondroitin sulfate nanoparticle and a drug loaded on the surface of the chitosan-chondroitin sulfate nanoparticle. The drug-loaded joint lubricant provided by the invention integrates water-collecting lubrication, drug-loaded treatment and cartilage repair performance, has good biocompatibility and drug slow-release effect, and has good bone joint treatment application prospect.
Drawings
FIG. 1 is a transmission electron micrograph of CS and CS-CHI nanoparticles;
FIG. 2 shows the dissolution and dispersion of the CS-CHI nanoparticle lubricant in different physiological media;
FIG. 3 is a biocompatibility assessment of CS-CHI nanoparticle lubricants;
FIG. 4 is a friction test curve of CHI biomacromolecule and CS-CHI nanoparticle lubricant on the surface of artificial joint material Ti6Al 4V;
FIG. 5 is a graph of the coefficient of friction of saline, CS nanoparticles, and CS-CHI nanoparticle lubricants on the surface of natural bovine articular cartilage;
fig. 6 is a graph of drug release profiles of diclofenac sodium loaded CS nanoparticles and CS-CHI nanoparticle lubricant in buffer at pH 7.4.
Detailed Description
The invention provides a chitosan-chondroitin sulfate nanoparticle, which comprises a chitosan nanoparticle and chondroitin sulfate loaded on the surface of the chitosan nanoparticle.
In the invention, the particle size of the chitosan nanoparticle is preferably 50-300 nm, and more preferably 100-200 nm. In the invention, the deacetylation degree of the chitosan nanoparticle is preferably 75-85%, and more preferably 80%; the viscosity is preferably 5 to 200mPa.s, more preferably 20 to 150mPa.s, and further preferably 50 to 100 mPa.s.
The present invention has no special requirements for the preparation method of the chitosan nanoparticle, and can use the chitosan nanoparticle which is conventionally sold in the market in the field or can be prepared by itself, and when prepared by itself, the preparation method preferably comprises the following steps:
mixing chitosan, acetic acid solution, sodium tripolyphosphate and a cross-linking agent, and carrying out cross-linking reaction to obtain a cross-linked product;
and mixing the cross-linked product with a reducing agent, and carrying out reduction reaction to obtain the chitosan nano-particles.
The invention mixes chitosan, acetic acid solution, sodium tripolyphosphate and cross-linking agent to carry out cross-linking reaction, and obtains cross-linked product. In the invention, the pH value of the acetic acid solution is preferably 3-5, and more preferably 4; the volume ratio of the mass of the chitosan to the volume of the acetic acid solution is preferably 100mg: 50 mL.
In the invention, the mass ratio of the chitosan to the sodium tripolyphosphate is preferably 1: 0.3-0.5, and more preferably 0.4. In the present invention, the crosslinking agent is preferably glutaraldehyde. In the invention, the glutaraldehyde is preferably added in the form of an aqueous solution, the volume concentration of the glutaraldehyde aqueous solution is preferably 50%, and the ratio of the mass of the chitosan to the volume of the glutaraldehyde aqueous solution is preferably 100mg: 1-5 mL, and more preferably 100mg: 2-3 mL.
In the invention, the temperature of the crosslinking reaction is preferably 0-10 ℃, more preferably 4-6 ℃, and the time is preferably 6-24 hours, more preferably 10-20 hours. In the present invention, the crosslinking reaction is preferably carried out under mechanical stirring conditions, and the stirring speed is preferably 300 rpm.
After the cross-linked product is obtained, the cross-linked product is mixed with a reducing agent for reduction reaction to obtain the chitosan nano-particle. In the present invention, the reducing agent is preferably sodium borohydride. In the present invention, the mass ratio of the chitosan to the reducing agent is preferably 100mg:55 mg; in the invention, the temperature of the reduction reaction is preferably 20-10 ℃, and the time is preferably 2-6 h, and more preferably 2-4 h.
After the reduction reaction, the present invention preferably performs a post-treatment on the obtained reduction reaction solution, and the post-treatment preferably includes:
and (3) sequentially centrifuging, dialyzing and freeze-drying the reduction reaction solution to obtain the chitosan nanoparticle solid.
In the present invention, the rate of the centrifugation is preferably 10000 rpm. In the present invention, the dialysis is preferably performed in a PBS buffer solution having a pH of 7.4, and the time period of the dialysis is preferably 2 days. The present invention does not require any particular manner of freeze-drying, and any manner of freeze-drying known to those skilled in the art may be used.
In the invention, the mass ratio of the chitosan nanoparticle to the chondroitin sulfate is preferably 1.5-3: 1, and more preferably 2-2.5: 1.
Different from traditional medicine carriers such as hard silicon dioxide particles and the like, the soft swelling type nano particles prepared by taking natural macromolecular chitosan as a raw material are taken as the medicine carriers, so that the biocompatibility, the cartilage surface abrasion resistance, the medicine controllable release property and the blood compatibility are more excellent; the chitosan nano-particles can be degraded in a human body and cannot damage the human body.
In the invention, chondroitin sulfate macromolecules grafted on the surfaces of the chitosan nanoparticles contain a large number of hydrophilic sulfonic groups, so that the chitosan nanoparticles have excellent water lubrication performance; the chitosan-chondroitin sulfate nano-particles prepared by combining the cartilage repair function of chondroitin sulfate macromolecules integrate the water lubrication, drug-loaded treatment and cartilage repair performances, and are suitable to be used as a carrier component of a joint lubricant.
The invention provides a preparation method of the chitosan-chondroitin sulfate nano-particles, which comprises the following steps:
mixing chondroitin sulfate, a carboxyl activating agent and a buffer solution, and performing carboxyl activation to obtain carboxyl-activated chondroitin sulfate;
and mixing the carboxyl-activated chondroitin sulfate with the chitosan nanoparticles, and performing chemical grafting reaction to obtain the chitosan-chondroitin sulfate nanoparticles.
The invention mixes chondroitin sulfate, carboxyl activating agent and buffer solution to carry out carboxyl activation, thus obtaining the carboxyl-activated chondroitin sulfate. In the present invention, the carboxyl activating agent preferably includes 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and succinimide. In the invention, the mass ratio of the chondroitin sulfate to the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is preferably 1: 1-1.2, and more preferably 1: 1.1; the mass ratio of the chondroitin sulfate to the succinimide is preferably 1: 1.2-1.5, and more preferably 1: 1.3-1.4.
In the present invention, the buffer is preferably a PBS buffer, which preferably has a pH of 7.4.
In the present invention, the mixing is preferably performed under the condition of ultrasound, the power of the ultrasound is preferably 150W, and the time is preferably 20 min.
In the invention, the temperature for activating the carboxyl is preferably 25-75 ℃, and more preferably 30-50 ℃. In the invention, the activation time of the carboxyl is preferably 1-2 h, and more preferably 1.5 h. In the present invention, the activation of the carboxyl group is preferably carried out under mechanical stirring.
After the chondroitin sulfate of the activated carboxyl is obtained, the invention mixes the chondroitin sulfate of the activated carboxyl with chitosan nano-particles, and carries out chemical grafting reaction to obtain the chitosan-chondroitin sulfate nano-particles. In the present invention, the mixing is preferably performed by stirring.
In the invention, the temperature of the chemical grafting reaction is preferably 25-37 ℃, and more preferably 28-32 ℃; the time is preferably 12 to 36 hours, and more preferably 20 to 30 hours. In the present invention, the chemical grafting reaction is preferably carried out under stirring conditions, and the stirring rate is preferably 300 rpm. In the chemical grafting reaction process, carboxyl of chondroitin sulfate reacts with amino on the surface of chitosan, so that the chondroitin sulfate is grafted on the surface of chitosan nanoparticles.
After the chemical grafting reaction, the present invention preferably performs a post-treatment on the obtained chemical grafting reaction solution, and in the present invention, the post-treatment preferably includes the following steps:
and filtering the chemical grafting reaction solution, and sequentially dialyzing and freeze-drying the obtained filtrate to obtain the chitosan-chondroitin sulfate nanoparticle solid.
The present invention preferably performs the filtration using a filter head having a diameter of 200 nm. In the invention, the dialysis is preferably carried out in a PBS buffer solution with the pH value of 7.4, and the cut-off molecular weight of the dialysis is preferably 8000-12000 kDa, more preferably 9000-10000 kDa; the dialysis time is preferably 1 to 3 days, and more preferably 2 days. The present invention does not require any particular manner of freeze-drying, and any freeze-drying method known to those skilled in the art may be used.
The invention provides a drug-loaded joint lubricant, which comprises the chitosan-chondroitin sulfate nanoparticle and a drug loaded on the surface of the chitosan-chondroitin sulfate nanoparticle.
In the present invention, the drug is preferably an anti-inflammatory drug. In the invention, the medicament is preferably one or more of diclofenac sodium, isomonoside, mangiferin, fenoprofen, aspirin, loxoprofen, ibuprofen and nimesulide.
In the invention, the drug loading amount of the drug-loaded joint lubricant is 10-40 wt%, and more preferably 20-30 wt%; the load is the mass percentage of the medicine in the medicine-carrying joint lubricant.
The invention provides a preparation method of the drug-loaded joint lubricant, which comprises the following steps:
mixing chitosan-chondroitin sulfate nanoparticles, a medicament and a buffer solution, and carrying out medicament loading to obtain the medicament-loaded joint lubricant.
In the present invention, the buffer is preferably a PBS buffer, and the pH of the PBS buffer is preferably 7.4.
In the invention, the drug loading is preferably carried out under the condition of stirring, and the stirring speed is preferably 100-500 rpm, and more preferably 300-400 rpm. In the invention, the temperature of the drug loading is preferably 25-37 ℃, more preferably 28-32 ℃, and the time is preferably 12-36 h, more preferably 20-30 h.
The preparation methods of chitosan-chondroitin sulfate nanoparticles and drug-loaded joint lubricants provided by the present invention are described in detail below with reference to the examples, but they should not be construed as limiting the scope of the present invention.
Example 1
(1) CS nanoparticle preparation
100mg of CS was weighed, dissolved in 50mL of 1% acetic acid solution, and then distilled water was added to 80mL, followed by addition of 40mg of sodium tripolyphosphate. Adjusting the pH value to 5.6 by using a saturated sodium carbonate solution, and reacting for 2 hours at the temperature of 4 ℃ under the condition of mechanical stirring; 2mL of glutaraldehyde aqueous solution (50%, v/v) was added dropwise, the reaction was continued for 12h with mechanical stirring at 4 ℃ and then 55mg of sodium borohydride was added and the reaction was continued for 4 h. And (3) centrifuging the solution at 10000rpm after the reaction is finished, dialyzing the precipitate obtained after centrifugation in PBS buffer solution with the pH of 7.4 for 2 days, and finally freeze-drying to obtain the CS nano-particles.
(2) CS-CHI nano-particles prepared by grafting CHI on surfaces of CS nano-particles
50mg of CHI was weighed out and dissolved in 50ml of PBS buffer (pH 7.4), and 100mg of EDC and NHS were added in this order. And after ultrasonic treatment for 20min, mechanically stirring and activating for 2h at normal temperature. After the activation reaction was completed, 75mg of CS nanoparticles was added. The mixture is placed at normal temperature and mechanically stirred to react for 1 day. And after the reaction is finished, filtering the solution by using a filter head with the diameter of 200nm, dialyzing the obtained solution in PBS buffer solution with the pH value of 7.4 for 2 days, and freeze-drying the dialyzed solution to obtain the CS-CHI nano-particles without the drug.
(3) 100mg of CS-CHI nanoparticles were weighed and dissolved in PBS buffer pH 7.4. Then 100mg of diclofenac sodium as an anti-inflammatory drug is weighed and added into the solution, and the mixture is stirred for 2 days at room temperature after being uniformly mixed. After the completion, impurities were removed by dialysis. And finally, freeze-drying the obtained solution to obtain the drug-loaded CS-CHI nano-particles.
The transmission electron micrograph of the CS nanoparticles and the CS-CHI nanoparticles is shown in FIG. 1. As can be seen from FIG. 1, compared with the blank CS nanoparticles, a layer of uniform organic matter appears on the surface of the CS-CHI nanoparticles, indicating that CHI molecules are successfully grafted on the surface of the CS nanoparticles.
Performance testing
(I) stability test
The CS-CHI nanoparticles were placed in PBS buffer at pH 7.4, PBS buffer at pH 5.4, and physiological saline, respectively, at a concentration of 2mg/mL, and the results were shown in (one), (two), and (three) of fig. 2, respectively. In FIG. 2, the dispersion conditions of ultrasonic dissolution, standing for one week and standing for two weeks are shown in the sequence from left to right in the first, second and third steps, and as can be seen from FIG. 2, the CS-CHI nanoparticles still have good stability after standing for two weeks in different media.
(II) biological toxicity study
Rat primary chondrocytes were placed in CS nanoparticle dispersion, CS-CHI nanoparticle dispersion and diclofenac sodium-loaded CS-CHI nanoparticle dispersion for incubation for different periods of time (the concentrations of the dispersions were all 1.0mg/mL), and the survival results are shown in FIG. 3. As can be seen from FIG. 3, the CS-CHI nanoparticles have good biocompatibility, and can be safely applied to the biomedical field.
(III) Friction test
Using CHI biological macromolecule and CS-CHI nano-particle lubricant as lubricant to prepare Ti for artificial joint material6Al4And performing a friction test on the surface V, wherein the mass concentration of the lubricant is 1.0mg/mL, the friction pair is PDMS (polydimethylsiloxane), the sliding frequency is 1Hz, and the load is 5N. The results are shown in FIG. 4.
As can be seen from figure 4, the CS-CHI nanoparticle lubricant has excellent water lubrication performance, which indicates that the CS-CHI nanoparticle lubricant has application potential as a lubricant of artificial joint replacement materials.
And performing a friction test on the surface of the natural bovine cartilage by using the CS nano-particles and the CS-CHI nano-particles as lubricants, wherein the mass concentration of the lubricants is 1.0mg/mL, the friction pair is also the natural bovine cartilage, the sliding frequency is 1Hz, and the load is 5N. The results are shown in FIG. 5.
As can be seen from fig. 5, the CS-CHI nanoparticle lubricant has a lower coefficient of friction than the blank CS nanoparticles, indicating its potential application as an injectable joint lubricant.
(IV) Slow Release Effect test
The drug release profiles of CS nanoparticles loaded with equal amounts of diclofenac sodium and CS-CHI nanoparticle lubricant in PBS buffer are shown in figure 6. As can be seen in FIG. 6, the drug-loaded CS-CHI nanoparticle lubricant was seen to have a controlled drug release behavior, indicating its potential function as a treatment for osteoarthritis.
Example 2
Compared to example 1, the mass ratio of CHI to CS nanoparticles was 50 mg: 150mg, and the rest operations are the same, thus obtaining the CS-CHI nano-particles.
Example 3
Compared to example 1, the mass ratio of CHI to CS nanoparticles was 50 mg: 100mg, and the rest operations are the same to obtain the CS-CHI nano-particles.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A chitosan-chondroitin sulfate nanoparticle comprises a chitosan nanoparticle and chondroitin sulfate loaded on the surface of the chitosan nanoparticle.
2. The chitosan-chondroitin sulfate nanoparticle according to claim 1, wherein the chitosan nanoparticle has a particle size of 50 to 300 nm; the deacetylation degree of the chitosan nanoparticles is 75-85%, and the viscosity is 5-200 mpa.s.
3. The chitosan-chondroitin sulfate nanoparticle according to claim 1 or 2, wherein the mass ratio of the chitosan nanoparticle to chondroitin sulfate is 1.5-3: 1.
4. The method for preparing chitosan-chondroitin sulfate nanoparticles as claimed in any one of claims 1 to 3, comprising the steps of:
mixing chondroitin sulfate, a carboxyl activating agent and a buffer solution, and performing carboxyl activation to obtain carboxyl-activated chondroitin sulfate;
and mixing the carboxyl-activated chondroitin sulfate with the chitosan nanoparticles, and performing chemical grafting reaction to obtain the chitosan-chondroitin sulfate nanoparticles.
5. The production method according to claim 4, wherein the carboxyl activating agent comprises 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and succinimide;
the mass ratio of the chondroitin sulfate to the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is 1: 1-1.2;
the mass ratio of the chondroitin sulfate to the succinimide is 1: 1.2-1.5.
6. The preparation method of claim 4, wherein the temperature for activating the carboxyl group is 25 to 75 ℃ and the time is 1 to 2 hours;
the temperature of the chemical grafting reaction is 25-37 ℃, and the time is 12-36 h.
7. A drug-loaded joint lubricant, comprising the chitosan-chondroitin sulfate nanoparticle of any one of claims 1 to 3 or the chitosan-chondroitin sulfate nanoparticle prepared by the preparation method of any one of claims 4 to 6, and a drug loaded on the surface of the chitosan-chondroitin sulfate nanoparticle.
8. The drug-loaded joint lubricant of claim 7, wherein the drug is one or more of diclofenac sodium, isomonoside, mangiferin, fenoprofen, aspirin, loxoprofen, ibuprofen, and nimesulide.
9. The drug-loaded joint lubricant according to claim 7 or 8, wherein the drug loading in the drug-loaded joint lubricant is 10-40 wt%.
10. The method for preparing the drug-loaded joint lubricant of any one of claims 7 to 9, comprising the steps of:
mixing chitosan-chondroitin sulfate nanoparticles, a medicament and a buffer solution, and carrying out medicament loading to obtain the medicament-loaded joint lubricant.
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| CN202210291118.9A CN114558176B (en) | 2022-03-23 | 2022-03-23 | Chitosan-chondroitin sulfate nanoparticle and drug-loaded joint lubricant |
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Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008119824A1 (en) * | 2007-04-03 | 2008-10-09 | Bioiberica, S.A. | Nanoparticulate composition of chitosan and chondroitin sulfate |
| US20100092572A1 (en) * | 2007-01-29 | 2010-04-15 | Peter Kaeuper | Chitosan-based colloidal particles for rna delivery |
| CN102145174A (en) * | 2011-04-06 | 2011-08-10 | 西安交通大学 | Chondroitin sulfate nano-selenium and preparation method thereof |
| US8114842B1 (en) * | 2004-10-05 | 2012-02-14 | Gp Medical, Inc. | Nanoparticles for drug delivery |
| US20130244972A1 (en) * | 2006-12-11 | 2013-09-19 | Mor Research Applications Ltd. | Injectable chitosan mixtures forming hydrogels |
| CN107802888A (en) * | 2017-10-31 | 2018-03-16 | 无锡中科光远生物材料有限公司 | A kind of preparation method of the nano fiber scaffold of promotion regenerating bone or cartilage |
| CN108938597A (en) * | 2018-08-24 | 2018-12-07 | 东华大学 | A kind of chitosan graft polymer medicine-carrying nanoparticles and its preparation and application |
| CN108976317A (en) * | 2018-06-29 | 2018-12-11 | 中国科学院兰州化学物理研究所 | A kind of chitosan biological macromolecular and its preparation method and application |
| CN109789223A (en) * | 2016-08-25 | 2019-05-21 | 医药研究产品有限公司 | Joint cavity injection composition comprising nucleic acid and chitosan |
| US20190224132A1 (en) * | 2016-10-04 | 2019-07-25 | University Of Florida Research Foundation, Inc. | Chondroprotective nanoparticles for the treatment of osteoarthritis |
| CN114080399A (en) * | 2019-05-24 | 2022-02-22 | 考麦德制药公司 | Chitosan and application thereof |
-
2022
- 2022-03-23 CN CN202210291118.9A patent/CN114558176B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8114842B1 (en) * | 2004-10-05 | 2012-02-14 | Gp Medical, Inc. | Nanoparticles for drug delivery |
| US20130244972A1 (en) * | 2006-12-11 | 2013-09-19 | Mor Research Applications Ltd. | Injectable chitosan mixtures forming hydrogels |
| US20100092572A1 (en) * | 2007-01-29 | 2010-04-15 | Peter Kaeuper | Chitosan-based colloidal particles for rna delivery |
| WO2008119824A1 (en) * | 2007-04-03 | 2008-10-09 | Bioiberica, S.A. | Nanoparticulate composition of chitosan and chondroitin sulfate |
| CN102145174A (en) * | 2011-04-06 | 2011-08-10 | 西安交通大学 | Chondroitin sulfate nano-selenium and preparation method thereof |
| CN109789223A (en) * | 2016-08-25 | 2019-05-21 | 医药研究产品有限公司 | Joint cavity injection composition comprising nucleic acid and chitosan |
| US20190224132A1 (en) * | 2016-10-04 | 2019-07-25 | University Of Florida Research Foundation, Inc. | Chondroprotective nanoparticles for the treatment of osteoarthritis |
| CN107802888A (en) * | 2017-10-31 | 2018-03-16 | 无锡中科光远生物材料有限公司 | A kind of preparation method of the nano fiber scaffold of promotion regenerating bone or cartilage |
| CN108976317A (en) * | 2018-06-29 | 2018-12-11 | 中国科学院兰州化学物理研究所 | A kind of chitosan biological macromolecular and its preparation method and application |
| CN108938597A (en) * | 2018-08-24 | 2018-12-07 | 东华大学 | A kind of chitosan graft polymer medicine-carrying nanoparticles and its preparation and application |
| CN114080399A (en) * | 2019-05-24 | 2022-02-22 | 考麦德制药公司 | Chitosan and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| RABIA GUL等: "Biodegradable Ingredient-Based Emulgel Loaded with Ketoprofen Nanoparticles", 《AAPS PHARMSCITECH》 * |
| TAGHIPOUR-SABZEVAR V等: "Targeted delivery of a short antimicrobial peptide against CD44-overexpressing tumor cells using hyaluronic acid-coated chitosan nanoparticles: An in vitro study", 《JOURNAL OF NANOPARTICLE RESEARCH》 * |
| 朱笑微等: "《平衡医学 平衡针灸与平衡药物 21世纪人类健康指南》", 31 August 2002, 中国科学技术出版社 * |
| 李澜 等: "用于软骨修复的组织工程水凝胶", 《生命科学》 * |
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