CN102078642B - Articular cartilage restoration and regeneration stent and preparation method thereof - Google Patents
Articular cartilage restoration and regeneration stent and preparation method thereof Download PDFInfo
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- CN102078642B CN102078642B CN 201110021494 CN201110021494A CN102078642B CN 102078642 B CN102078642 B CN 102078642B CN 201110021494 CN201110021494 CN 201110021494 CN 201110021494 A CN201110021494 A CN 201110021494A CN 102078642 B CN102078642 B CN 102078642B
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Abstract
The invention discloses an articular cartilage restoration and regeneration stent and a preparation method thereof, belonging to the field of medical materials. The stent is made of decalcified spongiosa-cortical bone, and the decalcified spongiosa-cortical bone is provided with more than two holes penetrating through the decalcified spongiosa bone and the cortical bone. The holes are vertically arranged on the horizontal surface of the decalcified cortical bone and have diameter of 0.1-1mm. The width between hole centers is 0.5-5mm. The preparation method provided by the invention comprises the steps of: soaking the bone in a decalcified solution for decalcification; monitoring the decalcification process by using an atomic absorption spectrophotometer; carrying out X-ray and CT detection on a decalcified sample to prove that the bone is completely decalcified to obtain the decalcified spongiosa-cortical bone; drilling on the horizontal surface of the decalcified cortical bone by using laser in a diameter of 0.1-1mm; and cutting the decalcified cortical bone into a certain size, and freezing and storing. The articular cartilage restoration and regeneration stent is made of a natural collagen stent material without immunological rejection reaction, is easy to adsorb by cells, and has good biocompatibility and better mechanical strength.
Description
Technical field
The present invention relates to field of medical materials, particularly a kind of articular cartilage reparative regeneration support and preparation method thereof.
Background technology
Articular cartilage damage is the intractable damage in sports medical science field.Kinetic articular cartilage damage incidence rate is very high, and the survey result of Peking University's institute of sports medicine shows that the cartilage injury accounts for more than 40.5% of motion of knee joint wound.Regular meeting causes symptoms such as arthralgia behind the cartilage injury.Because the cartilaginous tissue structure is special, impassivity, blood vessel and lymphatic vessel in it, the nutrition supply of cartilage is to change by the pressure that joint motion produces, synovial fluid is exchanged between articular cavity and cartilage matrix, so the articular cartilage after the damage can not be repaired and be regenerated, and easily causes tangible dysfunction.Home and abroad repair of cartilage method is a lot, but does not still have a kind of generally acknowledged restorative procedure that improves.At present, utilize self bone marrow reparation cartilage defect, as: the little fracture technology of subchondral bone becomes one of important method of cartilage defect reparation.Because it is simple to operate, obtain good result, extensive use on clinical repair of cartilage for the cartilage defect of small size.But because little fracture is damaged by the scar piece filling of bone marrow formation, the mechanical stress of scar piece is too low, can't bear the pressure that joint motion produces.Therefore can not bear a heavy burden little 6 weeks of fracture surgery long period; Repairing the position may occur subsiding or hyperplasia; Little fracture method is unsatisfactory for the little fracture effect of the cartilage of large tracts of land and non-horizontal level, and newborn cartilage mostly is fibrous cartilage.Therefore, seek the cartilage defect filling bracket that is fit to and also become one of main direction of studying of repair of cartilage.
The repair of cartilage timbering material that uses can be divided into natural scaffold materials and man-made support material two big classes substantially at present.Natural scaffold materials is to derive from naturally occurring macromolecular material in animal and plant or the human body, has excellent biological compatibility, organizes affinity.The man-made support material is to study more material at present, mainly contains polyglycolic acid, polylactic acid and poly-anhydride etc., can adjust its physics, chemistry, biomechanics and degradation property as required, shapes easily, produces good reproducibility.
In realizing process of the present invention, the inventor finds that there is following problem at least in prior art: problems such as natural scaffold materials exists material character to change, and mechanical strength and degradation speed are restive.Natural scaffold materials commonly used has collagen protein, hydroxyapatite etc., and natural support bio-mechanical property is difficult to adjust, and is difficult to find with cartilage biomechanics similar performance.The Mechanics of Machinery of some natural biologic material support is poor, the mechanical stress that produces in the time of can't resisting joint motions, cause subsiding of mend easily, also influence the growth of cell on the material simultaneously, and poor than cartilage as hydroxyapatite rack elasticity and creep properties, the too high wearing and tearing that easily cause corresponding cartilage joint face of hardness.The biocompatibility of man-made support material is relatively poor relatively, has inflammatory reaction in various degree, and the immunogenicity problem, even the carcinogenecity problem.
Summary of the invention
The objective of the invention is the defective at above-mentioned prior art, a kind of articular cartilage reparative regeneration support that has excellent biological compatibility and mechanical strength concurrently is provided.
Another object of the present invention provides the preparation method that the articular cartilage reparative regeneration is used support.
The technical scheme taked of the present invention is to achieve these goals:
The material of described support is decalcification spongiosa-cortical bone, and described decalcification spongiosa-cortical bone is provided with the aperture that runs through decalcification spongy bone and decalcification cortical bone.
Described aperture arranges perpendicular to described decalcification cortical bone horizontal surface, and the diameter of described aperture is 0.1-1mm.
Described aperture is more than two, and the centre distance of adjacent described aperture is 0.5～5mm.
The porosity of described aperture is 5%-50%.
Another technical scheme provided by the invention is: a kind of articular cartilage reparative regeneration preparation method of support may further comprise the steps:
(1) selects to comprise cortical bone and the double-deck skeleton of spongy bone;
(2) described skeleton is soaked in carries out decalcification in the decalcifying Fluid and handle;
(3) with atomic absorption spectrophotometer monitoring decalcification process;
(4) specimen after the decalcification is carried out X line and CT detection, after the complete decalcification of skeleton, obtains decalcification spongiosa-cortical bone;
(5) perpendicular to the diameter laser boring of decalcification cortical bone horizontal surface with 0.1-1mm, boring runs through the decalcification cortical bone and the decalcification spongy bone is two-layer, penetrates entire bracket;
(6) decalcification spongiosa-cortical bone is cut into the specification that use needs, refrigerated storage.
Described decalcifying Fluid is the 0.1M-0.7M ethylenediaminetetraacetic acid, pH value 6-10, temperature: 2-10 ℃.
Further, before transplanting described biological support, described biological support carries out cobalt 60 irradiations 12-72 hour.
Described skeleton is people's ilium, vertebral body bone or the long bone metaphysis of freezing.
The beneficial effect that the technical scheme that the embodiment of the invention provides is brought is:
1. using the people's bone in the ripe commercial bone bank is raw material, and the bone bank bone is to ratify by Bureau of Drugs Supervision, and is applied to clinical raw material for many years, and the abundant and safety of donor source is easy to clinical popularizing.The reparation that also can be used for the human cartilage damage is handled in a lot of animal spongiosa-cortical bone (cattle, sheep etc.) decalcifications through method of the present invention.It is many that cattle, sheep are cultured quantity, and getting bone, to do material more extensive.
2. the present invention has used protection decalcification bone supporting stand biological nature decalcification new method, has solved simultaneously with the existing problem of ethylenediaminetetraacetic acid (EDTA) decalcification.The acid of widespread (hydrochloric acid, nitric acid and formic acid etc.) decalcification method, very big to the collagen scaffold damage of bone, cause support to become fragile, the change of degradation biology and bio-mechanical property under the elastic modelling quantity, unrenewable in repair of cartilage.Use EDTA (neutral calcium ion chelating agent) (the neutral decalcifying agent of this kind is minimum to the damage of ossein support) to carry out the decalcification of people's bone among the present invention.But because the EDTA decalcifying agent can only permeate people's cortex bone surface 2mm, can't slough the calcareous of cortical bone center fully, and overlong time, can't satisfy the clinical practice needs.Therefore the present invention abandons simple decalcification cortical bone support and selects cortex-spongy bone for use, thickness of cortex of bone is only got 0.1-1mm in the support, all the other supply the damaged degree of depth with the thickness of spongy bone, because the multi-pore structure of spongy bone makes decalcifying Fluid be easy to enter the material center.Decalcification fully, and significantly shortened decalcification time, satisfy clinical practice.
3. utilize the thin layer cortical bone in the material, can use staple that repair materials is fixed in cartilage defect place, increase the firm degree of repair of cartilage support, shorten the time that patient's postoperative bears a heavy burden walking, recovers motor function.
4. the loose and porous structure of spongy bone in the support, can hold bone marrow and derived from bone marrow stem cell that more subchondral bones discharge, have that broader space allows bone marrow stem cell adheres to, the exchange of migration and nutrient substance, accelerate propagation and the differentiation of stem cell, the reparation of cartilage defect.
5. the natural collagen timbering material that adopts of the present invention, the problem of no immunological rejection is easy to cell and adsorbs good biocompatibility.
6. self be rich in bone morphogenetic protein (BMPs) at decalcification spongiosa-cortical bone support, can promote that stem cell or the marrow stromal cell in the middle of the bone marrow in the induction rack changes into chondrocyte or osteocyte, promote the secretion of stem cell secretion, cartilage or bone matrix.To the startup of repair of cartilage, growth, regulation and control and mouldingly all play an important role.
7. easy and simple to handle, do not need second operation, saved in the operation and drawn materials the time, be easy to clinical practice.Decalcification spongiosa-cortical-bone articular cartilage the bracket for regenerating of vertical boring of the present invention, utilize little fracture technology to discharge the bone marrow stem cell of self during use, make it diffuse to decalcification cancellous bone scaffolds inside by the vertical boring on the support, the decalcification cancellous bone scaffolds supports the growth of autologous bone marrow stem cell 3 D stereo, and the bone morphogenetic protein (BMPs) on the support induces bone marrow stem cell to bone, cartilage-derived growth, differentiation simultaneously.The present invention has avoided external support cultured cell in the organizational project, and exogenous factor is induced, and the step that product Hui Zhi etc. are complicated need not second operation, more is applicable to clinical practice.
Description of drawings
Fig. 1 is the articular cartilage reparative regeneration support sketch map that the embodiment of the invention provides;
Fig. 2 is the cutaway view of Fig. 1;
Fig. 3 is the articular cartilage reparative regeneration that provides of the embodiment of the invention with the preparation flow figure of support;
Fig. 4 is the articular cartilage reparative regeneration support scanning electron microscope structure chart that the embodiment of the invention provides;
Fig. 5 is that people's decalcification cortex-cancellous bone scaffolds that the embodiment of the invention provides is repaired the damaged operation figure of knee cartilage;
Fig. 6 is that the damaged back observation knee cartilage defect repair place that drew materials in 12 weeks of people's decalcification cortex-cancellous bone scaffolds reparation knee cartilage that the embodiment of the invention provides schemes substantially;
Fig. 7 is that people's decalcification cortex-cancellous bone scaffolds that the embodiment of the invention provides is repaired knee cartilage damaged 12 all HE colored graphs;
Fig. 8 is that people's decalcification cortex-cancellous bone scaffolds that the embodiment of the invention provides is repaired knee cartilage damaged 12 all toluidine blue cartilage specificity colored graphs;
Fig. 9 is that people's decalcification cortex-cancellous bone scaffolds that the embodiment of the invention provides is repaired knee cartilage damaged 12 all II Collagen Type VI immunohistochemical staining figure.
The specific embodiment
For making the purpose, technical solutions and advantages of the present invention clearer, embodiment of the present invention is described further in detail below in conjunction with accompanying drawing.
Referring to Fig. 1 and Fig. 2, a kind of articular cartilage reparative regeneration support, the material of described support is decalcification spongiosa-cortical bone, and decalcification cortical bone 1 is connecting decalcification spongy bone 3, and described decalcification spongiosa-cortical bone is provided with the aperture 2 that runs through decalcification spongy bone 3 and decalcification cortical bone 1.
As a kind of preferred, aperture 2 is perpendicular to the 1 horizontal surface setting of decalcification cortical bone, and the diameter of aperture 2 is 0.1-1mm, can be 0.1mm, 0.5mm, 1mm.Aperture 2 is more than two, and the distance at the center, hole of adjacent aperture 2 is 0.5～5mm, and hole centre distance can be 0.5mm, 1.2mm, 3.4mm, 5mm.The porosity of aperture 2 is 5%-20%, and the support general thickness is greater than 2mm.
Referring to Fig. 3, a kind of articular cartilage reparative regeneration preparation method of support may further comprise the steps:
Step 101: select to have under the cortical bone skeleton of spongy bone, at 0.5-3mm, decalcifying Fluid is the 0.1M-0.7M ethylenediaminetetraacetic acid, pH value 6-10, temperature: 2-10 ℃ with the thickness of cortex of bone finishing;
Step 102: skeleton is soaked in carries out decalcification in the decalcifying Fluid and handle;
Step 103: with atomic absorption spectrophotometer monitoring decalcification process;
Step 104: the specimen after the decalcification carries out the x line and CT detects, and with the complete decalcification of proof skeleton, obtains decalcification spongiosa-cortical bone;
Step 105: in the diameter laser boring of decalcification cortical bone horizontal surface with 0.1-1mm;
Step 106: detect decalcification spongiosa-cortical bone support each several part nano-elastic modulus by nano impress biomechanics tester, filter out the decalcification spongiosa-cortical bone similar with the normal cartilage elastic modelling quantity as articular cartilage reparative regeneration support, decalcification spongiosa-cortical bone is cut into the specification that use needs, refrigerated storage;
Step 107: before transplanting described biological support, described biological support carries out cobalt 60 irradiations 12-24 hour.
The articular cartilage reparative regeneration of employing present embodiment with the restorative procedure key step of support is:
At first, dig in the osteochondral defect zone and scrape off except fiber becomes, irregular pathological changes cartilage and subchondral bone.Then, carry out the subchondral bone grinding, osteochondral defect is accomplished the shape of comparison rule.The boring of carrying out subchondral bone then allows to discharge the bone marrow of self.
Then, the decalcification bone supporting stand with laser drill is handled is trimmed to the shape that meets osteochondral defect, inserts damaged place, and cortical bone partly is bottom.Subchondral bone can infiltrate through by the aperture on the support cortical bone in the decalcification spongy bone owing to self the bone marrow that boring discharges, and then reparation osteochondral defect place.Self bone marrow stem cell that discharges has strengthened repair ability.Self bone marrow that simple subchondral bone boring discharges, no support can not be born stress and can't be according to the reparation of ad hoc structure standard, and the articular cartilage of generation lacks normal structure.And adopted decalcification bone articular cartilage bracket for regenerating of the present invention, the bone marrow of release is infiltrated in the decalcification spongy bone through the decalcification cortical bone duct in the support, both increased the mechanical strength of repairing early stage mend, the mechanical stress of mend has can be by the regulation and control bone marrow repair of damaged shape 3 D stereo.
The articular cartilage reparative regeneration preparation method of support:
Preparation flow figure purchases the frozen people's ilium of profound hypothermia or vertebral body bone as shown in Figure 3, is soaked in the 0.1M-1MEDTA decalcifying Fluid, and the temperature of decalcifying Fluid is in 6 ℃, and pH is 8, utilizes chelating to slough calcium in the cortical bone, and decalcifying Fluid is changed every day.With deionized water preparation EDTA decalcifying Fluid, carry with the plastic containers Sheng.The decalcifying Fluid of changing every day detects calcium ion concentration with monitoring decalcification process with atomic absorption spectrophotometer, and the Ca chelate concentration＜0.5g/L in decalcifying Fluid is complete decalcification, and the specimen after the decalcification carries out the x line and CT detects, and proves complete decalcification.Because decalcification cortical bone surface texture is too fine and close, is unsuitable for growing into of cell.Therefore the present invention by the laser drill technology as shown in Figure 2, with the diameter vertical drilling of decalcification cortical bone horizontal surface with 0.5mm, laser drill is adopted in boring, (different with clinical patient application aperture on the experimental rabbit animal model, because the joint of people, rabbit varies in size, so the aperture is all different with pitch-row), relatively hole interbody spacer 2mm is set in the back, then, and as shown in Figure 3, laser drill decalcification spongiosa-cortical bone support is cut into certain size specification, refrigerated storage.Before transplanting, the decalcification of the laser drill of preparation spongiosa-cortical bone support carries out cobalt 60 irradiations sterilization in 18 hours.
Referring to Fig. 4, the structure scanning electron microscope (SEM) photograph of laser drill decalcification spongiosa-cortical bone cartilage joint bracket for regenerating spongy bone part.As seen collagen fiber are loose in the decalcification spongy bone is staggered, and is full of the space between collagen fiber, and pore structure is staggered to communicate, and is conducive to the infiltration of bone marrow matrix and the factor and adhering to of cell.The natural lamella sample of decalcification cortical bone, some advantage is arranged: 1. collagen structure can resist vertical pull strength and pressure, is conducive to the regenerating bone or cartilage support and is fixed in cartilage defect.2. the gap of sheet interlayer is conducive to the infiltration of bone marrow matrix and the factor and adhering to of cell; Be conducive to the exchange of cytotrophy material.
Repair the operation process of cartilage defect experiment at animal model
For verifying that articular cartilage reparative regeneration of the present invention with the effect of support in method for repairing and mending, has carried out the sham operated experiment at animal model.Select New Zealand's large ear rabbit of 2.5-3.0kg, anesthesia, preserved skin, sterilization, as shown in Figure 5, the otch knee joint is exposed the femur coaster, does the 4mm diameter at the femur coaster, dark damaged of 1.5mm, after subchondral bone is holed little fracture processing, accomplish the 4mm diameter at the laser drill decalcification spongiosa-cortical bone support that has disinfected, the cylinder of height 1.5mm is packed into damaged.The layering sew up wound, 12 weeks of postoperative draw materials, and carry out histological observation.
Fig. 6 carries out the substantially figure of repair of cartilage after 12 weeks for New Zealand's large ear rabbit uses laser drill decalcification spongiosa-cortical-bone articular cartilage reparative regeneration with support.The repair place of visible arrow indication on the cardinal principle specimen has presented the cartilage sample and changed, and is very similar with normal surrounding tissue, not obvious with the intersection of normal cartilage, the slit do not occur.Do not see obvious immune inflammation reaction.
New Zealand's large ear rabbit laser drill decalcification spongiosa-cortical-bone articular cartilage bracket for regenerating referring to Fig. 7, Fig. 8 and Fig. 9 carries out the Histological section dyeing of repair of cartilage after 12 weeks.As seen the decalcification of the repair place of arrow indication spongiosa-cortical bone support is substituted by new life's hyaline cartilage fully, and subsiding does not appear in the repair place, with the intersection fluid-tight engagement of the normal cartilage on figure right side, blur boundary.Because the early stage subchondral bone of observing is not also substituted by bone, presents chondroid tissue.Do not see inflammatory cell infiltrations such as macrophage, do not see obvious immune inflammation reaction.Cambium HE colored graph such as Fig. 7; Cambium Toluidine blue staining (specificity is at mucopolysaccharide composition in the cartilage matrix) is positive in Fig. 8; Cambium II Collagen Type VI immunohistochemical staining is positive in Fig. 9, proves that cambium is rich in the hyaline cartilage that the II Collagen Type VI is similar normal articular cartilage.
The decalcification bone articular cartilage bracket for regenerating of laser drill of the present invention is to use cortical bone and the spongy bone of allogeneic people's decalcification to prepare, when decalcification spongiosa-cortical-bone articular cartilage bracket for regenerating of employing laser drill is repaired, easy and simple to handle, be easy to clinical practice.And use articular cartilage reparative regeneration of the present invention can solve the shortcoming of heteroplastic transplantation rejection with support, autologous bone marrow not only provides autologous stem cells during reparation, the microenvironment of cells survival is provided simultaneously, than simple chondrocyte or stem cell transplantation higher cells survival rate is arranged.The inventive method uses allochthonous decalcification people bone support to solve the biocompatibility issues of synthetic support.The mechanical stress of the decalcification bone supporting stand by biomechanics test shows preparation and elastic modelling quantity are more close to cartilage, it is poor to have solved natural biologic material rack mechanical mechanics such as silkworm silk, collagen protein, fibrin, people's amniotic membrane, the problem of the mechanical stress that produces in the time of can't resisting joint motions.Can not cause subsiding of mend, but the also growth of cell on the support material simultaneously.And decalcification spongiosa-cortical-bone articular cartilage bracket for regenerating is to use decalcification spongiosa-cortical bone of people, itself be rich in bone morphogenetic protein (BMPs), can promote stem cell or marrow stromal cell in the middle of the bone marrow in the induction rack to change into chondrocyte or osteocyte, promote the secretion of stem cell secretion cartilage or bone matrix.To the startup of repair of cartilage, growth, regulation and control and mouldingly all play an important role.
The above only is preferred embodiment of the present invention, and is in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, is equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (5)
1. an articular cartilage reparative regeneration support is characterized in that, the material of described support is decalcification spongiosa-cortical bone, and described decalcification spongiosa-cortical bone is provided with the aperture that runs through decalcification spongy bone and decalcification cortical bone; Described aperture be perpendicular to decalcification cortical bone horizontal surface by laser boring, boring runs through described decalcification cortical bone and described decalcification spongy bone is two-layer, penetrates entire bracket and makes.
2. support according to claim 1 is characterized in that, described aperture arranges perpendicular to described decalcification cortical bone horizontal surface, and the diameter of described aperture is 0.1-1mm.
3. support according to claim 2 is characterized in that, described aperture is more than two, and the centre distance of adjacent described aperture is 0.5～5mm.
4. support according to claim 1 is characterized in that, the porosity of described aperture is 5%-50%.
5. an articular cartilage reparative regeneration may further comprise the steps with the preparation method of support:
(1) selects to comprise that cortical bone and the double-deck skeleton of spongy bone, described skeleton are people's ilium, vertebral body bone or the long bone metaphysis of freezing;
(2) described skeleton is soaked in carries out decalcification in the decalcifying Fluid and handle, described decalcifying Fluid is the 0.1M-0.7M ethylenediaminetetraacetic acid, pH value 6-10, temperature: 2-10 ℃;
(3) with atomic absorption spectrophotometer monitoring decalcification process;
(4) specimen after the decalcification is carried out X line and CT detection, after the complete decalcification of skeleton, obtains decalcification spongiosa-cortical bone;
(5) perpendicular to the diameter laser boring of decalcification cortical bone horizontal surface with 0.1-1mm, boring runs through the decalcification cortical bone and the decalcification spongy bone is two-layer, penetrates entire bracket;
(6) decalcification spongiosa-cortical bone is cut into the specification that use needs, refrigerated storage; Before transplanting described biological support, described biological support carries out cobalt 60 irradiations 12-72 hour.
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| CN103202724B (en) * | 2013-03-03 | 2015-09-23 | 北京工业大学 | A kind of method utilizing laser technology to carry out cartilage punching |
| CN103301505B (en) * | 2013-06-07 | 2014-11-19 | 钟春燕 | Method for preparing bacterial cellulose three-dimensional exhibition microporous bracket |
| CN105854082B (en) * | 2015-01-20 | 2019-03-22 | 北京大学第三医院 | A kind of Meniscus scaffold and preparation method thereof |
| EP3773298A1 (en) | 2018-04-03 | 2021-02-17 | Convergent Dental, Inc. | Laser system for surgical applications |
| CN109528358B (en) * | 2019-02-01 | 2023-11-24 | 北京爱康宜诚医疗器材有限公司 | cartilage substitute |
| CN114942169B (en) * | 2022-06-02 | 2023-06-23 | 四川大学华西医院 | Method for manufacturing complete bone joint pathological section of mouse |
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| CN101332313A (en) * | 2008-08-06 | 2008-12-31 | 北京大学第三医院 | Bracket for regenerating decalcification cortical-bone articular cartilage with vertical drill and preparation method thereof |
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| US7815926B2 (en) * | 2005-07-11 | 2010-10-19 | Musculoskeletal Transplant Foundation | Implant for articular cartilage repair |
| US20070276506A1 (en) * | 2006-05-25 | 2007-11-29 | Biomet Manufacturing Corp. | Demineralized osteochondral plug |
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| CN101332313A (en) * | 2008-08-06 | 2008-12-31 | 北京大学第三医院 | Bracket for regenerating decalcification cortical-bone articular cartilage with vertical drill and preparation method thereof |
| CN101816806A (en) * | 2010-02-05 | 2010-09-01 | 余方圆 | Cartilage complex of tissue engineering bone and method for preparing same |
| CN201949388U (en) * | 2011-01-19 | 2011-08-31 | 北京大学第三医院 | Joint cartilage bone repair and regeneration support |
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Effective date of registration: 20180821 Address after: 100089 Beijing Daxing Zhongguancun Science Park Daxing Biomedical Industry Base No.26 Yongwang West Road Hospital 8-2 Building 401 Patentee after: Beijing Wan Jie Tianyuan medical instruments Limited by Share Ltd Address before: 100191 49 Garden Road North, Haidian District, Beijing. Co-patentee before: Ao Yingfang Patentee before: The Third Affiliated Hospital of Peking University |
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