CN110772356B - Implanted auricular cartilage composite support - Google Patents
Implanted auricular cartilage composite support Download PDFInfo
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- CN110772356B CN110772356B CN201911116229.0A CN201911116229A CN110772356B CN 110772356 B CN110772356 B CN 110772356B CN 201911116229 A CN201911116229 A CN 201911116229A CN 110772356 B CN110772356 B CN 110772356B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/18—Internal ear or nose parts, e.g. ear-drums
- A61F2/186—Nose parts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/30—Joints
- A61F2/30756—Cartilage endoprostheses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/30—Joints
- A61F2002/30001—Additional features of subject-matter classified in A61F2/28, A61F2/30 and subgroups thereof
- A61F2002/30108—Shapes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/30—Joints
- A61F2/30756—Cartilage endoprostheses
- A61F2002/30764—Cartilage harvest sites
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- Health & Medical Sciences (AREA)
- Heart & Thoracic Surgery (AREA)
- Animal Behavior & Ethology (AREA)
- Cardiology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Otolaryngology (AREA)
- Veterinary Medicine (AREA)
- Vascular Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Rheumatology (AREA)
- Pulmonology (AREA)
- Materials For Medical Uses (AREA)
- Prostheses (AREA)
Abstract
The invention discloses an implanted auricular cartilage composite bracket; comprises a first bracket and a second bracket; one ends of the first support and the second support are both connected with a connecting frame, and the first support and the second support form a 'human' structure through the connecting frames; the first bracket, the second bracket and the connecting bracket respectively comprise wire carving line fiber cores and ear cartilage bracket layers, and the ear cartilage bracket layers are wrapped outside the wire carving line fiber cores; according to the invention, the soft ear bone tissue is used as a support, and the thread carving fiber core is implanted in the center between the soft ear bone tissue and the soft ear bone tissue, so that the supporting force of the soft ear bone support is increased, the strength and hardness of the soft ear bone support are increased, and the stability of the soft ear bone support is further improved; through the ' human ' type structure that sets up, ride and stride in the septum of nose, increase its mechanical properties, further increase cartilage support's stability.
Description
Technical Field
The invention belongs to the technical field of medical biomaterials, and particularly relates to an implantable soft-ear bone composite scaffold.
Background
The clinical application of the cartilage support is commonly used in plastic surgery and orthopaedics, but with the increase of the application, a plurality of defects also exist, for example, in plastic surgery, the auricular cartilage cushion nose tip is used for taking out the cartilage of an auricle and is cushioned at the position of the nose tip so as to achieve the purpose of shaping the nose tip. The ear cartilage cushion pen point mainly comprises two methods, one is that a nose tip support is built, then the nose tip edge is lifted and upwarps, and the other is that the ear cartilage is made into a shield shape and is cushioned below the skin of the nose tip, so that the purpose of shaping the nose tip is achieved.
The soft ear bone support is adopted for supporting at the present stage, the support supporting force of the soft ear bone alone is insufficient, the nasal tip prosthesis cannot be supported for a long time, the traditional cartilage support is poor in stability and irregular in appearance, the physical microstructure is not neat and uniform enough, adhesion, growth and appreciation of cells are not facilitated, and postoperative healing is not facilitated.
The prior chinese patent application 201780016117.0 discloses a multi-phase osteochondral scaffold for osteochondral defect repair, the scaffold comprising a bone phase and a cartilage phase, wherein the bone phase comprises a support matrix, the cartilage phase comprises a polymer matrix, and the scaffold comprises a non-porous layer between the bone phase and the cartilage phase. The present invention also describes a multi-phase osteochondral scaffold for osteochondral defect repair, the scaffold comprising a bone phase and a cartilage phase, wherein the bone phase comprises a support matrix, the cartilage phase comprising a polymer matrix, and wherein the support matrix is tapered such that the support matrix is smaller in size at a lower end of the support matrix than at an upper end of the support matrix.
The existing Chinese patent application 201510789943.1 discloses a collagen-based cartilage scaffold, and the preparation method comprises the following steps: (a) freezing and molding the laid collagen solution, soaking and dehydrating the collagen solution by a hypertonic buffer solution, and extruding the collagen solution to form a film; (b) spreading a layer of collagen solution on the surface of the membrane, and freeze-drying; (c) and (3) integrally crosslinking by using a crosslinking agent, washing with water, and freeze-drying to obtain the collagen-based cartilage scaffold.
However, in the above technical solutions of the prior art, the cartilage tissue is simply used as the scaffold, which is prone to cause insufficient supporting force, resulting in the technical problems of low strength, poor hardness and poor stability of the cartilage scaffold.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide an implanted soft-ear bone composite support, wherein soft-ear bone tissues are used as supports, and a thread carving fiber core is implanted in the center between the soft-ear bone tissues, so that the supporting force of a soft-ear bone support is increased, the strength and hardness of the soft-ear bone support are increased, and the stability of the soft-ear bone support is further improved; through the ' human ' type structure that sets up, ride and stride in the septum of nose, increase its mechanical properties, further increase cartilage support's stability.
The invention provides the following technical scheme:
an implantable auricular cartilage composite scaffold; comprises a first bracket and a second bracket; one ends of the first support and the second support are both connected with a connecting frame, and the first support and the second support form a 'human' structure through the connecting frames; first support, second support and link all include line carving line fibre core and ear cartilage support layer, the parcel of line carving line fibre core outside has ear cartilage support layer.
Preferably; the thread carving thread fiber core is of a spiral structure.
Preferably, the acute included angle of the human-shaped structure of the first bracket and the second bracket ranges from 35 degrees to 50 degrees.
Preferably, the preparation method of the implanted auricular cartilage composite scaffold comprises the following steps:
s1: adding the wire carving line into an N, N-dimethylformamide solvent, stirring, carrying out ultrasonic treatment, standing for a period of time, adding an alkaline solution for solidification, stretching while solidifying, washing, and drying to obtain a wire carving line fiber core;
s2: taking transparent ear cartilage tissue, cutting into pieces, adding active agent and protease inhibitor for centrifugal treatment, removing supernatant, and retaining precipitation solution;
s3: adding buffer solution of an active agent and a protease inhibitor into the precipitation solution obtained in S2, shaking and uniformly mixing, centrifuging, and repeatedly washing the precipitation part to obtain cartilage particles for later use;
s4: uniformly spreading the cartilage particles obtained in the step S3 in a culture dish, and then carrying out vacuum freeze-drying in a freeze dryer to obtain cartilage extracellular matrix powder;
s5: freezing the cartilage extracellular matrix powder in the S4 in liquid nitrogen, then thoroughly grinding the powder by a high-speed machine, and screening the powder by a screen to obtain more fine cartilage extracellular matrix powder;
s6: crosslinking with crosslinking agent, washing with water, and freeze drying to obtain ear cartilage scaffold; and implanting the thread carving thread fiber core into the soft ear bone bracket to obtain the soft ear bone composite bracket.
Preferably, the PH value of the buffer solution in step S3 is 7-8, and the buffer solution is tris buffer solution, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, or polyethylene glycol octyl phenyl ether buffer solution.
Preferably, in step S4, the cartilage particles are uniformly spread in a culture dish to a thickness of 2-6 mm.
Preferably, in step S4, the vacuum freezing time is 20-25 h.
Preferably, the crosslinking agent is carbodiimide and N-hydroxysuccinimide.
Preferably, the freezing time T1 in step S6 is 25-46.5h, and the crosslinking time T2 is 45-50 h. In order to improve the strength of the composite scaffold, the freezing time T1 and the crosslinking time T2 satisfy T1. T2 is 1135 is 2350.
Preferably, in order to further increase the stability of the ear cartilage scaffold, the specific surface area A of the ear cartilage cell particles is 850-1920cm2Per gram; the pH value is 7-8 to increase the adhesiveness of the ear cartilage cells and increase the contact area with the cross-linking agent, thereby being beneficial to the proliferation and growth of the ear cartilage cells. The specific surface area A and the pH value of the ear osteocyte particles meet the following requirements: a & PH is 5950 or more and 15360 or less.
Preferably, the cell content M of the ear cartilage extracellular matrix, the specific surface area A of the ear cartilage cells and the pH value satisfy the following relation:
A＝μ·(PH)3/M1/2；
in the above formula, mu is a specific surface area coefficient, and the value range is 3.24-6.65; only numerical operations are performed here.
Preferably, in order to improve the bioactivity and strength of the composite scaffold, the PH value, the freezing time T1 and the crosslinking time T2 of the auricular chondrocytes satisfy the following relationship:
T2/T1＝λ·PH1/2；
wherein, the lambda is a relation coefficient and the value range is 0.35 to 0.74.
In addition, in the process of preparing the ear cartilage scaffold, the taken ear cartilage tissue is cut up, washed three times by phosphate buffer solution at 4 ℃, then placed in 10mM Tris buffer solution containing protease inhibitor, repeatedly centrifuged for 3 times by a centrifuge at 3000rpm, placed in 10mM Tris buffer solution containing protease inhibitor, fully mixed, centrifuged at 7000rpm, the supernatant is removed, and the precipitation solution is reserved; adding 1% of polyethylene glycol octyl phenyl ether buffer solution and tris buffer solution into the precipitate, shaking for 8h on a constant temperature bed at 4 ℃, centrifuging, and washing for three times by using phosphate buffer solution; and (3) keeping the washed cartilage particles in a digestive enzyme mixed solution at 37 ℃ for digestion for 12h, continuously shaking, centrifuging and repeatedly washing.
The cartilage particles which are thoroughly cleaned are placed in a culture dish and evenly paved, and are frozen in a low-temperature refrigerator at the temperature of 80 ℃ below zero for 1h, and then are frozen in a freeze drying agent under vacuum for 24h to obtain the cartilage extracellular matrix powder.
Prefreezing cartilage extracellular matrix powder in liquid nitrogen for 20s, and grinding at 60Hz frequency for 1min in a high-speed grinder; repeatedly grinding for multiple times to ensure that the cartilage extracellular matrix is fully ground, removing the ground powder, and repeatedly screening by using a 100-micrometer screen to obtain more fine cartilage extracellular matrix powder;
preparing the finally obtained fine cartilage extracellular matrix powder into suspension in the process of crosslinking the ear cartilage scaffold, and preparing the ear cartilage scaffold by a freezing method; firstly, freezing the suspension at-20 ℃ for 1h, freezing at-80 ℃ for 1h, and then putting the suspension into a refrigerator for freezing for 48 h; then putting carbodiimide and N-hydroxysuccinimide for crosslinking for 48h at 4 ℃, washing excessive crosslinking agent by using a phosphate-hydrochloric acid buffer solution, and then putting the ear cartilage scaffold at-80 ℃ for freeze-drying and dehydrating again to obtain the ear cartilage scaffold.
Compared with the prior art, the invention has the following beneficial effects:
(1) according to the implanted type auricular cartilage composite stent, the auricular cartilage tissues are used as the stent, and the thread carving thread fiber core is implanted in the center between the auricular cartilage tissues, so that the supporting force of the cartilage stent is increased, the strength and hardness of the auricular cartilage stent are increased, and the stability of the auricular cartilage stent is further improved.
(2) The invention relates to an implanted type ear cartilage composite bracket; through the ' human ' type structure that sets up, ride and stride in the septum of nose, increase its mechanical properties, further increase cartilage support's stability.
(3) The implanted type ear cartilage composite scaffold improves the ear cartilage scaffold prepared in the prior art, has regular shape and more regular and uniform physical microstructure, is favorable for adhesion, growth and proliferation of cells and growth and healing.
(4) According to the implanted auricular cartilage composite scaffold, the particle size of the finely ground auricular cartilage extracellular matrix is smaller, the area of the contacted chemical cross-linking agent is larger, the degradation rate of the auricular cartilage composite scaffold is delayed, the contact probability of internal cytokines and seed cells is increased, and the stability is further improved.
(5) The implanted type auricular cartilage composite scaffold further increases the stability of the auricular cartilage scaffold and the adhesiveness of auricular cartilage cells by limiting M, A and PH, increases the contact area with a cross-linking agent, is beneficial to the proliferation and growth of auricular cartilage cells, and is beneficial to growth and healing.
(6) The strength of the composite scaffold is improved by setting the freezing time T1 and the cross-linking time T2.
(7) The pH value of the auricular chondrocytes, the freezing time T1 and the cross-linking time T2 are set to meet the requirements, so that the biological activity and the strength of the composite scaffold are improved.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
Fig. 1 is a front view of the present invention.
Fig. 2 is a top view of the present invention.
Fig. 3 is a right side view of the present invention.
Fig. 4 is a schematic view of the internal structure of the stent of the present invention.
In the figure: 1. a first bracket; 2. a second bracket; 3. a connecting frame; 4. thread carving thread fiber core; 5. ear cartilage framework layer.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described in detail and completely with reference to the accompanying drawings. It is to be understood that the described embodiments are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
The first embodiment is as follows:
as shown in fig. 1-4, an implantable auricular cartilage composite scaffold; comprises a first bracket 1 and a second bracket 2; one ends of the first support 1 and the second support 2 are both connected with a connecting frame 3, and the first support 1 and the second support 2 form a 'human' shaped structure through the connecting frame 3; first support 1, second support 2 and link 3 all include line carving line fibre core 4 and ear cartilage support layer 5, 4 outside parcels of line carving line fibre core have ear cartilage support layer 5.
The thread carving thread fiber core 4 is of a spiral structure. The value range of the acute included angle of the human-shaped structures of the first bracket 1 and the second bracket 2 is 35-50 degrees.
In addition, in the process of preparing the ear cartilage scaffold, the taken ear cartilage tissue is cut up, washed three times by phosphate buffer solution at 4 ℃, then placed in 10mM Tris buffer solution containing protease inhibitor, repeatedly centrifuged for 3 times by a centrifuge at 3000rpm, placed in 10mM Tris buffer solution containing protease inhibitor, fully mixed, centrifuged at 7000rpm, the supernatant is removed, and the precipitation solution is reserved; adding 1% of polyethylene glycol octyl phenyl ether buffer solution and tris buffer solution into the precipitate, shaking for 8h on a constant temperature bed at 4 ℃, centrifuging, and washing for three times by using phosphate buffer solution; and (3) keeping the washed cartilage particles in a digestive enzyme mixed solution at 37 ℃ for digestion for 12h, continuously shaking, centrifuging and repeatedly washing.
The cartilage particles which are thoroughly cleaned are placed in a culture dish and evenly paved, and are frozen in a low-temperature refrigerator at the temperature of 80 ℃ below zero for 1h, and then are frozen in a freeze drying agent under vacuum for 24h to obtain the cartilage extracellular matrix powder.
Prefreezing cartilage extracellular matrix powder in liquid nitrogen for 20s, and grinding at 60Hz frequency for 1min in a high-speed grinder; repeatedly grinding for multiple times to ensure that the cartilage extracellular matrix is fully ground, removing the ground powder, and repeatedly screening by using a 100-micrometer screen to obtain more fine cartilage extracellular matrix powder;
preparing the finally obtained fine cartilage extracellular matrix powder into suspension in the process of crosslinking the ear cartilage scaffold, and preparing the ear cartilage scaffold by a freezing method; firstly, freezing the suspension at-20 ℃ for 1h, freezing at-80 ℃ for 1h, and then putting the suspension into a refrigerator for freezing for 48 h; then putting carbodiimide and N-hydroxysuccinimide for crosslinking for 48h at 4 ℃, washing excessive crosslinking agent by using a phosphate-hydrochloric acid buffer solution, and then putting the ear cartilage scaffold at-80 ℃ for freeze-drying and dehydrating again to obtain the ear cartilage scaffold.
Example two:
the preparation method of the implanted auricular cartilage composite scaffold comprises the following steps:
s1: adding the thread carving thread into an N, N-dimethylformamide solvent, stirring, carrying out ultrasonic treatment, standing for a period of time, adding an alkaline solution for solidification, stretching while solidifying, washing, and drying to obtain a thread carving thread fiber core 4;
s2: taking transparent ear cartilage tissue, cutting into pieces, adding active agent and protease inhibitor for centrifugal treatment, removing supernatant, and retaining precipitation solution;
s3: adding buffer solution of an active agent and a protease inhibitor into the precipitation solution obtained in S2, shaking and uniformly mixing, centrifuging, and repeatedly washing the precipitation part to obtain cartilage particles for later use;
s4: uniformly spreading the cartilage particles obtained in the step S3 in a culture dish, and then carrying out vacuum freeze-drying in a freeze dryer to obtain cartilage extracellular matrix powder;
s5: freezing the cartilage extracellular matrix powder in the S4 in liquid nitrogen, then thoroughly grinding the powder by a high-speed machine, and screening the powder by a screen to obtain more fine cartilage extracellular matrix powder;
s6: crosslinking with crosslinking agent, washing with water, and freeze drying to obtain ear cartilage scaffold; and implanting the wire carving wire fiber core 4 into the soft ear bone bracket to obtain the soft ear bone composite bracket.
In the step S3, the pH value of the buffer solution is 7-8, and the buffer solution is a tris buffer solution, a disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution or a polyethylene glycol octyl phenyl ether buffer solution.
In step S4, the cartilage particles are uniformly spread in a culture dish to a thickness of 2-6 mm.
In step S4, the vacuum freezing time is 20-25 h.
The cross-linking agent is carbodiimide and N-hydroxysuccinimide.
In step S6, the freezing time T1 is 25-46.5h, and the crosslinking time T2 is 45-50 h. In order to improve the strength of the composite scaffold, the freezing time T1 and the crosslinking time T2 satisfy T1. T2 is 1135 is 2350.
EXAMPLE III
The difference between the first and second embodiments is that in order to further increase the stability of the ear cartilage scaffold, the specific surface area A of the ear cartilage cell particles is 850-1920cm2Per gram; the pH value is 7-8 to increase the adhesiveness of the ear cartilage cells and increase the contact area with the cross-linking agent, thereby being beneficial to the proliferation and growth of the ear cartilage cells. The specific surface area A and the pH value of the ear osteocyte particles meet the following requirements: a & PH is 5950 or more and 15360 or less.
The cell content M of the ear cartilage extracellular matrix, the specific surface area A of the ear cartilage cells and the pH value satisfy the following relation:
A＝μ·(PH)3/M1/2；
in the above formula, mu is a specific surface area coefficient, and the value range is 3.24-6.65; only numerical operations are performed here.
In order to improve the bioactivity and strength of the composite scaffold, the pH value, the freezing time T1 and the crosslinking time T2 of the auricular chondrocytes satisfy the following relationship:
T2/T1＝λ·PH1/2；
wherein, the lambda is a relation coefficient and the value range is 0.35 to 0.74.
The technical scheme realizes an implanted soft-ear bone composite bracket, the soft-ear bone tissue is used as the bracket, and the wire carving fiber core 4 is implanted in the center between the soft-ear bone tissue and the bracket, so that the supporting force of the soft-ear bone bracket is increased, the strength and the hardness of the soft-ear bone bracket are increased, and the stability of the soft-ear bone bracket is further improved; the human-shaped structure is arranged to straddle the nasal septum, so that the mechanical property of the human-shaped structure is improved, the stability of the cartilage support is further improved, and the ear cartilage support prepared in the prior art is improved, so that the shape of the ear cartilage support is regular, the physical microstructure is more uniform, the adhesion, growth and proliferation of cells are facilitated, and the postoperative healing is facilitated; the particle size of the finely ground ear cartilage extracellular matrix is smaller, the contact area of the chemical cross-linking agent is larger, the degradation rate of the ear cartilage scaffold is delayed, the contact probability of internal cell factors and seed cells is increased, and the stability is further improved; the M, A and PH are limited, so that the stability of the auricular cartilage scaffold is further increased, the adhesiveness of the auricular cartilage cells is increased, and meanwhile, the contact area with the cross-linking agent is increased, so that the proliferation and growth of the auricular cartilage cells are facilitated, and the healing is facilitated.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention; any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. The preparation method of the implanted auricular cartilage composite stent is characterized in that the implanted auricular cartilage composite stent comprises a first stent (1) and a second stent (2); one ends of the first support (1) and the second support (2) are connected with a connecting frame (3), and the first support (1) and the second support (2) form a human-shaped structure through the connecting frame (3); the first bracket (1), the second bracket (2) and the connecting bracket (3) respectively comprise a wire carving wire fiber core (4) and an ear cartilage bracket layer (5), and the ear cartilage bracket layer (5) is wrapped on the outer side of the wire carving wire fiber core (4);
the specific surface area A of the auricular chondrocyte particles is 850-1920cm2Per gram; the PH value is 7-8 to increase the adhesiveness of the ear cartilage cells and increase the contact area with the cross-linking agent, thereby being beneficial to the proliferation and growth of the ear cartilage cells;
the preparation method comprises the following steps:
s1: adding the thread carving thread into an N, N-dimethylformamide solvent, stirring, carrying out ultrasonic treatment, standing for a period of time, adding an alkaline solution for solidification, stretching while solidifying, washing, and drying to obtain a thread carving thread fiber core (4);
s2: taking transparent ear cartilage tissue, cutting into pieces, adding active agent and protease inhibitor for centrifugal treatment, removing supernatant, and retaining precipitation solution;
s3: adding buffer solution of an active agent and a protease inhibitor into the precipitation solution obtained in S2, shaking and uniformly mixing, centrifuging, and repeatedly washing the precipitation part to obtain cartilage particles for later use;
s4: uniformly spreading the cartilage particles obtained in the step S3 in a culture dish, and then carrying out vacuum freeze-drying in a freeze dryer to obtain cartilage extracellular matrix powder;
s5: freezing the cartilage extracellular matrix powder in the S4 in liquid nitrogen, then thoroughly grinding the powder by a high-speed machine, and screening the powder by a screen to obtain more fine cartilage extracellular matrix powder;
s6: crosslinking with crosslinking agent, washing with water, and freeze drying to obtain ear cartilage scaffold; and implanting the thread carving thread fiber core (4) into the ear cartilage support to obtain the ear cartilage composite support.
2. The method for preparing an implantable auricular cartilage composite scaffold according to claim 1, wherein; the thread carving thread fiber core (4) is of a spiral structure.
3. The method for preparing an implantable soft-ear-bone composite scaffold according to claim 1, wherein the acute included angle of the "human" shaped structure of the first scaffold (1) and the second scaffold (2) ranges from 35 ° to 50 °.
4. The method of claim 1, wherein the buffer solution in step S3 has a pH value ranging from 7 to 8, and is selected from the group consisting of Tris buffer solution, disodium hydrogen phosphate-potassium dihydrogen phosphate buffer solution, and polyethylene glycol octylphenyl ether buffer solution.
5. The method of claim 1, wherein in step S4, the cartilage particles are uniformly spread in a culture dish to a thickness of 2-6 mm.
6. The method of claim 1, wherein the vacuum freezing is performed for 20-25 hours in step S4.
7. The method for preparing an implantable auricular cartilage composite scaffold according to claim 1, wherein the cross-linking agent is carbodiimide-N-hydroxysuccinimide.
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