CN101537016B - Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof - Google Patents
Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof Download PDFInfo
- Publication number
- CN101537016B CN101537016B CN200910083272.1A CN200910083272A CN101537016B CN 101537016 B CN101537016 B CN 101537016B CN 200910083272 A CN200910083272 A CN 200910083272A CN 101537016 B CN101537016 B CN 101537016B
- Authority
- CN
- China
- Prior art keywords
- cartilage cells
- epiphyseal plate
- microcapsule
- preparation
- epiphyseal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5036—Polysaccharides, e.g. gums, alginate; Cyclodextrin
-
- 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
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5161—Polysaccharides, e.g. alginate, chitosan, cellulose derivatives; Cyclodextrin
-
- 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
-
- 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
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0618—Cells of the nervous system
- C12N5/0623—Stem cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0655—Chondrocytes; Cartilage
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
- C12N2533/32—Polylysine, polyornithine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/54—Collagen; Gelatin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/70—Polysaccharides
- C12N2533/74—Alginate
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Engineering & Computer Science (AREA)
- Physical Education & Sports Medicine (AREA)
- Rheumatology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Cell Biology (AREA)
- Epidemiology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Nanotechnology (AREA)
- Physics & Mathematics (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Dispersion Chemistry (AREA)
- Developmental Biology & Embryology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Materials For Medical Uses (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a compound microcapsule preparation containing cartilage cells of epiphyseal plate or other cartilage cells. The compound microcapsule preparation comprises fresh cartilage cells of epiphyseal plate or immortalised cartilage cells of epiphyseal plate or other cartilage cells, a mixed carrier alginate-collagen-polylysine, a liquefying agent sodium citrate, and a curing agent calcium chloride. The number of the fresh cartilage cells of epiphyseal plate or the immortalised cartilage cells of epiphyseal plate (or other cartilage cells) is between 2*10<6> and 8*10<6>. The mixed carrier alginate-collagen-polylysine comprises the following components of 50-600 parts of alginate by weight, 10-100 parts of collagen by weight and 1-10 parts of polylysine by weight. The content of the liquefying agent sodium citrate is 2-20 parts by weight and the content of the curing agent calcium chloride is 1-10 parts by weight. The carrier material of the compound microcapsule preparation in the invention is a natural extract, has excellent biocompatibility, relatively strong mechanical strength and few side effects, and is suitable for treatment of early closure of epiphyseal injury.
Description
Technical field
The present invention relates to contain the treatment that cartilage cells of epiphyseal plate (or other chondrocytes) compound microcapsule preparation and preparation and Injury of Epiphyseal Plate thereof early close.
The carrier being particularly related to comprises sodium alginate, collagen protein, polylysine, calcium chloride, sodium citrate solution.By separated cartilage cells of epiphyseal plate or immortalized cells (or other chondrocytes) under the effect of above-mentioned carrier, the microcapsule that makes as required 100 μ m～1000 μ m by high-pressure electrostatic generator for microcapsules (can be made the microcapsule of the specification that differs in size as 100 μ m～200 μ m, 300 μ m～500 μ m etc.), be kept at transplanting use to be injected in culture fluid or normal saline, the method is not also appeared in the newspapers at present both at home and abroad.The present invention is raw materials used has good biocompatibility, and stronger mechanical strength, is a kind of safe, effective, novel cell microencapsulation preparation.The Injury of Epiphyseal Plate that can be used for treating human or animal early closes.
Background technology
Epiphyseal plate (claiming again growth plate) is the hyaline cartilage that one deck has longitudinal growth ability between long bone epiphysis and metaphysis.Be responsible for the growth of bone, its organizational structure and substrate composition are step to be changed.Physiological activity and biological behaviour are complicated, be subject to the regulation and control of whole body secretion (comprising hormone, somatomedin) drawn game radical secretion and autocrine (multiple somatomedin), dyshormonia, matrix protein gene sudden change, Nutrition and Metabolism obstacle, toxicant, physical factors etc. all can cause multiple epiphyseal plate growth promoter disease, as dyschondroplasia, dwarfism etc., Injury of Epiphyseal Plate and infection can cause epiphyseal plate partly or entirely early to close, and anamorphosis, has a strong impact on limb function and labour force.
Growth plate cartilage is the most active position of osteogenesis, and its growth is directly connected to the growth promoter of long bone.The removing of propagation, hypertrophy, substrate calcification and osteoclast that when normal, cartilage internalization must be by chondrocyte, absorbs.Along with the intrusion formation class osseous tissue of blood vessel and osteoblast, and then the ossified osseous tissue that forms of calcification.
Epiphyseal plate is divided into three parts from dissecting: 1.. the cartilage epiphyseal plate being formed by different chondrocytes; 2.. ossified metaphysis; 3.. epiphyseal plate Ranvier perichondrial ossification ditch around, wherein cartilage epiphyseal plate divides three layers, from epiphysis end, is followed successively by storage belt, propagation band and mastocyte band.Propagation band is the place of epiphyseal plate longitudinal growth, Ranvier perichondrial ossification ditch is the place of epiphyseal plate cross growth, epiphyseal plate is that blood is for abundant tissue, arrive neonatal period epiphyseal plate and just formed gradually blood fortune barrier, if due to reason damage cartilage epiphyseal plates such as fracture, inflammation, the destruction that finally causes blood fortune barrier, now epiphysis and metaphysis blood fortune are invaded epiphyseal plate.Identical traffic, this is the basic condition that hone lamella forms, and what first form is fiber vessel bridges, then slowly ossified, if Injury of Epiphyseal Plate is enough large, is the deposition of bone under permission, and bone bridge finally forms.
Injury of Epiphyseal Plate is very serious a kind of in children's's bone injury, due to impaired limbs carrying out property growth retardation and the deformity of causing of growth plate.Epiphyseal plate is the basis of child's skeleton development, has special 26S Proteasome Structure and Function, and many reasons can cause the part or all of closure in advance of epiphyseal plate.Make limbs occur deformity and cause corresponding joint function disturbance, and along with child's growth promoter, deformity can increase the weight of gradually.The method that treatment Injury of Epiphyseal Plate part is early closed is a lot, but each method has the no thoroughness of its limitation and treatment, the implant of clinical middle application at present has autologous fat, bone is cured, cartilage, bone cement, medical silica-gel, gelfoam and autologous epiphyseal plate (comprising free vascular pedicle and band muscle flap person) though and epiphyseal plate extracellular is cultivated to transplant and is likely addressed this problem, still there are at present many problems.For example: best seed cell, between biotic environment and cartilage cells of epiphyseal plate to be related to that problem etc. has to be solved.Current molecular biology and engineered fast development, for bone induction research provides wide platform, the research of bone morphogenetic protein and somatomedin is day by day deep, for treatment Injury of Epiphyseal Plate early closes the thinking that provides new, bone marrow stromal cells in adults has differentiation potential, and is the focus that people competitively study.Injury of Epiphyseal Plate is early closed and is caused the serious deformity of limbs and dysfunction, it is common children's's bone injury disease, still lack at present the ideal material that can repair damage epiphyseal plate organizational structure and its growth function of recovery, microencapsulated cell technology is closed the treatment means that provides new damaged morning for epiphyseal plate.
Exposed cell transplantation enters in body and will run into strong immunity of organism rejection again; microcapsule can rely on the insulation blocking function of film and select permeation; guarantee that bioactive substance diffuses through; antibody or immunocyte are carried stays isolation; thereby on physiology, avoided individual immunologic rejection; become the desirable means that solve cell transplantation process immunologic rejection problem, thereby cause the interest of numerous researcheres.1997, the people such as Japan 0kada propose " cell preparation " word, utilize microencapsulated cell can resist immune attack in vivo, maintain normal physiological function, discharge continuously the medicative cellular products of tool---bioactie agent, can form long-term drug delivery system in cell gonosome.So far, first microencapsulated cell proposes as drug delivery system concept aspect pharmacy.
Sodium alginate gel has the structure similar to cartilage matrix composition proteoglycan, can make chondrocyte be grown in a kind of environment of similar physiological status, gel networks can be wrapped up chondrocyte completely simultaneously, in three dimensional structure, to grow completely, being conducive to cell and keeping phenotype stablize and secrete a large amount of substrate, is comparatively desirable cultured chondrocytes material.With nontoxic high molecular polymer, make the microcapsule with semipermeable membrane function, specific cell is wrapped in microcapsule, its principle is: the semi permeability of utilizing microcapsule membrane, the micromolecule such as oxygen, nutrient substance can see through film and supply with cell, make it to keep physiologically active, and the bioactive product of emiocytosis also can appear from microcapsule, play specific function, have immunocompetent macromolecular substances, cell etc. can not act on capsule inner cell by microcapsule membrane simultaneously, thereby play the effect of immunity isolation.
A series of closely-related extracellular matrix proteins in collagen protein family, extensively be present in mammalian body, account for 25.33% of human body protein total amount, collagen is the basic structure material of the organs such as skeleton, tendon, interosseous membrane, skin, cartilage and ligament, make it have very high tensile strength, and same cell interaction, for the adhering to of cell, transplanting, propagation, differentiation and existence provide signal of interest.Tropocollagen molecule has unique structure and aminoacid sequence to make it have two distinguishing features: (1) every 3 amino acid residues just have a glycine residue to form Gly-X-Y repetitive sequence, and the food in one's mouth propylhomoserin of (2) high-load and hydroxyl are fed propylhomoserin residue.Collagen is a kind of good tissue engineering bracket material as a kind of n cell epimatrix biopolymer, collagen has many advantages: (1) wide material sources, be easy to separation and purification from living tissue, so the price of collagen goods is relatively cheap, and supply is stable; (2) easily by body, absorbed, and there is very low antigenicity; (3) good mechanical performance, natural collagen fiber have good intensity and viscoelasticity, the most important thing is, as a kind of biomaterial, it has good biocompatibility and avirulence, and good biological plasticity and anti-fatigue ability are the essential compositions of cartilage.
Therefore, how sodium alginate and collagen protein are just become to the technical barrier that this technical field urgent need will solve as carrier parcel cartilage cells of epiphyseal plate jointly.
Summary of the invention
One of object of the present invention is to provide a kind of good biocompatibility, and mechanical strength is high, and toxic and side effects is little, has and fills and secretion bioactie agent, discharges continuously the medicative cellular products of tool (microencapsulated cell preparation).
Above-mentioned purpose of the present invention reaches by the following technical programs:
Containing cartilage cells of epiphyseal plate (or other a chondrocytes) compound microcapsule preparation, it is characterized in that: comprise fresh cartilage cells of epiphyseal plate or immortalization cartilage cells of epiphyseal plate (or other chondrocytes); Comprise mixed carrier alginate-collagen protein-polylysine; Comprise liquefier sodium citrate; Comprise firming agent calcium chloride.
Describedly containing fresh cartilage cells of epiphyseal plate or immortalization cartilage cells of epiphyseal plate (or other chondrocytes) number, be: 2～8 * 10
6.
Consisting of of described mixed carrier: alginate 50 to 600 weight portions, collagen protein 10 to 100 weight portions, polylysine 1 to 10 weight portion.
Consisting of of described liquefier sodium citrate: sodium citrate 2 to 20 weight portions.
Consisting of of described firming agent calcium chloride: 1 to 10 weight portion.
Described cartilage cells of epiphyseal plate (or other chondrocytes) compound microcapsule that contains can be the microcapsule being stored in the middle of culture fluid or in normal saline.
Described be stored in the microcapsule in culture fluid or the Microcapsules Size scope in normal saline of being stored at 100～1000 μ m.
The particle size range of described microcapsule also can be made different magnitude range as required, as 100～200 μ m; 200～300 μ m; 300～550 μ m; 500～750 μ m or 700～950 μ m etc.
The cartilage cells of epiphyseal plate of described purification is cartilage cells of epiphyseal plate or cryopreservation resuscitation cartilage cells of epiphyseal plate or the immortalization cartilage cells of epiphyseal plate of fresh separated.
Two of object of the present invention is to provide the preparation method of a kind of cartilage cells of epiphyseal plate or other chondrocyte compound microcapsules.
Containing a preparation method for cartilage cells of epiphyseal plate or other chondrocyte compound microcapsule preparations, its step is as follows:
(1) sodium alginate of 50 to 600 weight portions is weighed, dissolve; Obtain microcapsule solution;
(2) collagen protein of 10 to 100 weight portions is weighed, dissolve, obtain another kind of microcapsule solution;
(3) polylysine of 1 to 10 weight portion is weighed, dissolve, obtain another kind of microcapsule solution;
(4) sodium citrate of 2 to 20 weight portions is weighed, dissolve, obtain another kind of microcapsule liquefaction solution;
(5) calcium chloride of 1 to 10 weight portion or barium chloride are weighed, dissolve, obtain another kind of microcapsule solid-to-liquid ratio solution;
(6) cartilage cells of epiphyseal plate of purification is pressed to 2～8 * 10
6quantity, obtains Cell sap;
(7) by gained Cell sap and sodium alginate and collagen solution mixing, and react with described consolidation liquid by high-pressure electrostatic bull microcapsule drop generating device, be solidified into the micro gel bead of circle or similar round, the micro gel bead that micro gel bead must be strengthened in step (3) gained polylysine solution effects, micro gel bead, again in the effect of step (4) gained sodium citrate, must contain cartilage cells of epiphyseal plate compound microcapsule preparation after liquefaction.
Described high-pressure electrostatic bull microcapsule drop generating device comprises: an electrostatic generator, on described electrostatic generator, there are positive and negative polarities, positive pole is connected with the syringe needle of microinjection apparatus, negative pole is connected with the rustless steel steel ring being immersed in described consolidation liquid, mixed carrier solution and cartilage cells of epiphyseal plate are housed in injection device, splash in described consolidation liquid and form micro gel bead, gained cartilage cells of epiphyseal plate compound recipe micro gel bead is deposited to the bottom (each step is all in aseptic lower operation) of container, after the liquefaction of solidify-sodium citrate solution of polylysine, must contain cartilage cells of epiphyseal plate (or other chondrocytes) compound microcapsule again, it can be the microcapsule being stored in the middle of culture fluid or in normal saline.
The cartilage cells of epiphyseal plate of described purification is cartilage cells of epiphyseal plate or cryopreservation resuscitation cartilage cells of epiphyseal plate or the immortalization cartilage cells of epiphyseal plate of fresh separated.
The purification (amplification) of described cartilage cells of epiphyseal plate (or other chondrocytes): get rabbit childhood, under aseptic condition, get distal femur growth plate cartilage piece, be cut into the piece of 1mm * 1mm * 1mm size, after one's mother's sister's enzymic digestion, centrifugal, washing, hyaluronic acid enzymic digestion; Centrifugal, washing, type i collagen enzymic digestion; Centrifugal, washing, extract in cartilage cells of epiphyseal plate .DMEM culture medium and carry out primitive cell culture, with one's mother's sister's enzyme, primary cell digested again, cultivates to making cell confluency 70% left and right, uses SV40LTAg gene transfecting cell, and positive cell clone, cultivates and go down to posterity.
Described be stored in the microcapsule in culture fluid or the Microcapsules Size scope in normal saline of being stored between 100～1000 μ m.The particle size range of described microcapsule can be made different magnitude range as required.(as 100～200 μ m; 200～300 μ m; 300～550 μ m; 500～750 μ m; Or 700～950 μ m etc.).Gained cartilage cells of epiphyseal plate compound microcapsule preparation early closes treatment for Injury of Epiphyseal Plate.
Three of object of the present invention is to provide the application containing cartilage cells of epiphyseal plate or other chondrocyte compound microcapsule preparations, is used for the treatment of Injury of Epiphyseal Plate, for the filling after epiphyseal plate excision and effectively secretion.
Immunity isolation is the core of microcapsule technology, adopt nontoxic high molecular polymer to make the little sacculus with semipermeable membrane function, to treat that transplanted cells is wrapped in this sacculus, capsule inner cell can see through this microcapsule membrane and obtain nutrition, secreted bioactive substance also can be discharged by microcapsule, there is immunocompetent macromolecular substances simultaneously and can not pass through this film as immunocyte, immunoglobulin etc., thereby play immunologic barrier effect.This artificial cell is transplanted in host, can be prevented host immune system attack and long-term surviving.
Because collagen protein has, prevent ossified ability and for the regeneration of chondrocyte provides substrate, thereby further synthetic II Collagen Type VI recovers epiphyseal plate and keeps opening.Cartilage cells of epiphyseal plate is after alginate-collagen protein-polylysine-sodium citrate and calcium chloride parcel, without In vitro culture implantable epiphyseal plate defective region immediately, should be more suitable for the filling of epiphyseal plate defective region and reparation, and more effectively stop blood vessel to be grown into and prevent that bone bridge from forming.Under local physiological environment, reconstruction is normal epiphyseal plate organizational structure, and can recover preferably the growth function of epiphyseal plate, and the treatment of closing early for epiphyseal plate disease damage provides the required graft with good function activity.
Due to Injury of Epiphyseal Plate part, early to close the origin cause of formation more, the treatment of Injury of Epiphyseal Plate is a difficult problem of Orthopedic Clinical always, Clinical Processing method is various, after the damaged generation of bone, the physiology of bone is seriously damaged continuously, when utilizing exogenous cell therapy, must have applicable carrier, by carrier, make the growth factor release of cell generation to bony defect position, induce new bone formation to reach therapeutic purposes.The method that can obtain cell derived and immortalized cells by autologous or allogeneic is set up frozen reserve cell storehouse, obtains desirable cell derived, increases application prospect clinically.
After having had good cell, select alginate and collagen protein as main carriers, adopt high-pressure electrostatic generator for microcapsules that cartilage cells of epiphyseal plate is wrapped in microcapsule, after being implanted to bone bridge defective region, cell can obtain permanently effective existence, secretion somatomedin, promote cartilage-derived growth, the difficult problems such as solution is easily aging to the histiocyte after transplanting both at home and abroad at present, and effect is bad.Implant after utilizing the rear microencapsulation cartilage cells of epiphyseal plate of transplanting as Bar excision, and can effectively overcome immunological rejection, the microencapsulation cartilage cells of epiphyseal plate that has a physiological property by implantation after Bar excision extends the epiphyseal plate open hour, solve after Bar excision suffering limb energy for growth still lower than normal and a tender difficult problem that is just being less than 20 degree angulation deformities.According to the size of epiphyseal plate defective region determine microencapsulated cell amount number, control the microencapsulated cell amount of implanting, reach and grasp the epiphyseal plate speed of growth.
The artificial cell of syringeability microencapsulation is a kind of less invasive techniques, have easy and simple to handle, the advantage such as plastotype is random, and wound is little.Therefore due to microencapsulation cartilage cells of epiphyseal plate itself and natural growth plate cartilage, organize very similarly, after transplanting, can replace original growth plate cartilage, become and repair the comparatively ideal graft of Injury of Epiphyseal Plate, have great clinical application potential.
Below by embodiment, the invention will be further described, but do not mean that limiting the scope of the invention.
The specific embodiment
Embodiment 1
The preparation of the cartilage cells of epiphyseal plate of fresh separated (or other chondrocytes) compound microcapsule
1, the preparation before parcel:
1. the processing of glass drying oven and rustless steel apparatus:
By the glass drying oven cleaning up and rustless steel apparatus airing, be placed on and under 180～260 degrees Celsius, toast 3 hours (degerming reduce phlegm and internal heat source) in high temperature roaster.
2. the preparation of cartilage cells of epiphyseal plate separation solution:
A. the preparation of one's mother's sister's protein enzyme solution:
Take 5 grams of commercially available one's mother's sister's protease, be placed in above-mentioned glass drying oven, add sterile water for injection until all dissolve, obtain one's mother's sister's protein enzyme solution.
B. the preparation of hyaluronic acid enzymatic solution:
Take 2 grams of commercially available hyaluronidases, be placed in above-mentioned glass drying oven, add sterile water for injection until all dissolve, obtain hyaluronic acid enzymatic solution.
The preparation of C.I Collagenase Type:
Take 1 gram of commercially available type i collagen enzyme, be placed in above-mentioned glass drying oven, add sterile water for injection until all dissolve; Obtain type i collagen enzymatic solution.
D.DMEM/Ham F12 culture fluid, hyclone, is all purchased from U.S. Gibico company.
The purification of the new fresh cell of described growth plate cartilage (or other chondrocytes): get rabbit childhood, under aseptic condition, get distal femur growth plate cartilage piece, be cut into the piece of 1mm * 1mm * 1mm size, after one's mother's sister's enzymic digestion, centrifugal, washing, hyaluronic acid enzymic digestion, centrifugal, washing, type i collagen enzymic digestion, centrifugal, washing, extract cartilage cells of epiphyseal plate, in DMEM culture medium, carry out primitive cell culture and go down to posterity.
3. the preparation of sodium alginate soln:
Take 2 kilograms of commercially available sodium alginates, be placed in glass drying oven, stir on one side, add normal saline on one side, until sodium alginate all dissolves, obtain sodium alginate soln.
4. the preparation of collagen solution:
Take 0.5 gram of commercially available collagen protein, be placed in glass drying oven, add normal saline to all dissolving, obtain collagen solution.
5. the preparation of calcium chloride solution:
Take 1 kilogram of commercially available calcium chloride, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain calcium chloride solution;
6. the preparation of polylysine solution:
Take 1 gram of commercially available import polylysine, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain polylysine solution;
7. the preparation of sodium citrate solution:
Take 2 grams of commercially available sodium citrates, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain sodium citrate solution;
8. the cartilage cells of epiphyseal plate of above-mentioned collagen solution, sodium alginate soln and fresh separated (or other chondrocytes) is mixed, obtain mixed solution; With disposable sterilized injector, draw above-mentioned mixed liquor, by high-pressure electrostatic bull microcapsule drop generating device, splash in above-mentioned calcium chloride solution, with Ga
2+be cross-linked to form rapidly cancellated calcium alginate collagen type microcapsule, Na in consolidation liquid
+can with Ca in calcium alginate
2+crosslinked, make calcium alginate collagen surface of microcapsule all the time with a considerable amount of negative charges, add positively charged polylysine solution, then intermolecular cross-linking occurs, the formed network structure of this intermolecular cross-linking is more close compared with the network structure of calcium alginate intramolecular crosslinking.Gained microcapsule sinks to below container, sucts after clear liquid and adds polylysine solution to make strengthening, and then through sodium citrate liquefaction, must need the microcapsule of scope.By microcapsule add culture fluid or normal saline stand-by.
By the upper solution decant of said vesse, microcapsule is below washed three times with normal saline, with large size needle applicator, inhale the microcapsule with a small amount of normal saline, change suitable syringe needle and be injected directly into Injury of Epiphyseal Plate position.
9. the preparation of part defect model inside young rabbit proximal ends of tibia epiphyseal plate:
Select the large ear rabbit of health in 5 week age, 1% the capable intraperitoneal anesthesia of pentobarbital sodium, conventional sterile working, bilateral tibial upper end inner incision is got in operation, appear proximal ends of tibia growth plate cartilage, excision epiphyseal plate inside part approximately 1/3～1/2, layer-by-layer suture periosteum, subcutaneous and skin.Then the cartilage cells of epiphyseal plate compound microcapsule of preparation is injected to Injury of Epiphyseal Plate place, left side, does not transplant as a control group at proximal tibia Injury of Epiphyseal Plate place, right side, raises routine observation in postoperative cage.
Embodiment 2
The preparation of cryopreservation resuscitation cartilage cells of epiphyseal plate (or other chondrocytes) compound microcapsule
1, the preparation before parcel:
1. the processing of glass drying oven:
By the glass drying oven airing cleaning up, be placed on and under 180～260 degrees Celsius, toast 3 hours (degerming reduce phlegm and internal heat source) in high temperature roaster.
2. the processing of recovery cartilage cells of epiphyseal plate:
From liquid nitrogen container, take out cryopreservation tube, drop into rapidly in 37 ℃ of warm water, and frequently shake, make it melt as early as possible.From water-bath, take out cryopreservation tube, sucking-off cell suspension, injects centrifuge tube and drips 10 times of above culture fluid, and low-speed centrifugal after mixing, washs 3 times, with Trypan Blue, detects cell survival rate, and the cartilage cells of epiphyseal plate of purification is pressed to 2～8 * 10
6cell number is wrapped up, and makes the rear cartilage cells of epiphyseal plate compound microcapsule of recovery stand-by.
3. the preparation of sodium alginate soln:
Take 2 kilograms of commercially available sodium alginates, be placed in glass drying oven, stir on one side, add normal saline on one side, until sodium alginate all dissolves, obtain sodium alginate soln.
4. the preparation of collagen solution:
Take 0.5 gram of commercially available collagen protein, be placed in glass drying oven, add normal saline to all dissolving, obtain collagen solution.
5. the preparation of calcium chloride solution:
Take 1 kilogram of commercially available calcium chloride, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain calcium chloride solution;
6. the preparation of polylysine solution:
Take 1 gram of commercially available import polylysine, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain polylysine solution;
7. the preparation of sodium citrate solution:
Take 2 grams of commercially available sodium citrates, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain sodium citrate solution;
8. above-mentioned collagen solution, sodium alginate soln and cartilage cells of epiphyseal plate are mixed, obtain mixed solution; With disposable sterilized injector, draw above-mentioned mixed liquor, by high-pressure electrostatic bull microcapsule drop generating device, splash in above-mentioned calcium chloride solution, gained microcapsule sinks to below container, after sucting clear liquid, add polylysine solution to make strengthening, and then through sodium citrate liquefaction, must need the microcapsule of scope.By microcapsule add culture fluid or normal saline stand-by.
By the upper solution decant of said vesse, microcapsule is below washed three times with normal saline, with large size needle applicator, inhale the microcapsule with a small amount of normal saline, change suitable syringe needle and be injected directly into Injury of Epiphyseal Plate position.
9. the preparation of part defect model inside young rabbit proximal ends of tibia epiphyseal plate:
Select the large ear rabbit of health in 5 week age, 1% the capable intraperitoneal anesthesia of pentobarbital sodium, conventional sterile working, bilateral tibial upper end inner incision is got in operation, appear proximal ends of tibia growth plate cartilage, excision epiphyseal plate inside part approximately 1/3～1/2, layer-by-layer suture periosteum, subcutaneous and skin.Then the cartilage cells of epiphyseal plate compound microcapsule of preparation is injected to Injury of Epiphyseal Plate place, left side, does not transplant as a control group at proximal tibia Injury of Epiphyseal Plate place, right side, raises routine observation in postoperative cage.
Embodiment 3
The preparation of immortalization cartilage cells of epiphyseal plate (or other chondrocytes) compound microcapsule
1, the preparation before parcel:
1. the processing of glass drying oven:
By the glass drying oven airing cleaning up, be placed on and under 180～260 degrees Celsius, toast 3 hours (degerming reduce phlegm and internal heat source) in high temperature roaster.
2. amplification and the frozen processing of immortalization cartilage cells of epiphyseal plate (or other chondrocytes):
The amplification of described immortalization cartilage cells of epiphyseal plate: get rabbit childhood, get under aseptic condition after piece → one's mother's sister enzymic digestion of distal femur growth plate cartilage piece → be cut into 1mm * 1mm * 1mm size → centrifugal, washing → hyaluronic acid enzymic digestion → centrifugal, washing → type i collagen enzymic digestion → centrifugal, washing → extract and carry out primitive cell culture → primary cell is digested again → cultivate to making cell confluency 70% left and right → with SV40LTAg gene transfecting cell → positive cell clone → cultivate and go down to posterity (or frozen) with one's mother's sister's enzyme in cartilage cells of epiphyseal plate → DMEM culture medium.
From liquid nitrogen container, take out cryopreservation tube, drop into rapidly in 37 ℃ of warm water, and frequently shake, make it melt as early as possible.From water-bath, take out cryopreservation tube, sucking-off cell suspension, injects centrifuge tube and drips 10 times of above culture fluid, and low-speed centrifugal after mixing, washs 3 times, with Trypan Blue, detects cell survival rate, and the cartilage cells of epiphyseal plate of purification is pressed to 2～8 * 10
6wrap up, make immortalization cartilage cells of epiphyseal plate (or other chondrocytes) compound microcapsule stand-by.
3. the preparation of sodium alginate soln:
Take 2 kilograms of commercially available sodium alginates, be placed in glass drying oven, stir on one side, add normal saline on one side, until sodium alginate all dissolves, obtain sodium alginate soln.
4. the preparation of collagen solution:
Take 0.5 gram of commercially available collagen protein, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain collagen solution.
5. the preparation of calcium chloride solution:
Take 1 kilogram of commercially available calcium chloride, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain calcium chloride solution;
6. the preparation of polylysine solution:
Take 1 gram of commercially available import polylysine, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain polylysine solution;
7. the preparation of sodium citrate solution:
Take 2 grams of commercially available sodium citrates, be placed in glass drying oven, add sterile water for injection to all dissolving, obtain sodium citrate solution;
8. above-mentioned collagen solution, sodium alginate soln and immortalization cartilage cells of epiphyseal plate (or other chondrocytes) are mixed, obtain mixed solution; With disposable sterilized injector, draw above-mentioned mixed liquor, by high-pressure electrostatic bull microcapsule drop generating device, splash in above-mentioned calcium chloride solution, gained microcapsule sinks to below container, after sucting clear liquid, add polylysine solution to make strengthening, and then through sodium citrate liquefaction, must need the microcapsule of scope.By microcapsule add culture fluid or normal saline stand-by.
By the upper solution decant of said vesse, microcapsule is below washed three times with normal saline, with large size needle applicator, inhale the microcapsule with a small amount of normal saline, change suitable syringe needle and be injected directly into Injury of Epiphyseal Plate position.
9. the preparation of part defect model inside young rabbit proximal ends of tibia epiphyseal plate:
Select the large ear rabbit of health in 5 week age, 1% the capable intraperitoneal anesthesia of pentobarbital sodium, conventional sterile working, bilateral tibial upper end inner incision is got in operation, appear proximal ends of tibia growth plate cartilage, excision epiphyseal plate inside part approximately 1/3～1/2, layer-by-layer suture periosteum, subcutaneous and skin.Then the immortalization cartilage cells of epiphyseal plate compound microcapsule of preparation is injected to Injury of Epiphyseal Plate place, left side, does not transplant as a control group at proximal tibia Injury of Epiphyseal Plate place, right side, raises routine observation in postoperative cage.
Claims (8)
1. containing a cartilage cells of epiphyseal plate compound microcapsule preparation, it is characterized in that: comprise fresh cartilage cells of epiphyseal plate or immortalization cartilage cells of epiphyseal plate; Comprise mixed carrier alginate-collagen protein-polylysine; Comprise liquefier sodium citrate; Comprise firming agent calcium chloride; Described containing fresh cartilage cells of epiphyseal plate or immortalization cartilage cells of epiphyseal plate number 2 * 10
6～8 * 10
6; Consisting of of described mixed carrier alginate-collagen protein-polylysine: alginate 50 to 600 weight portions, collagen protein 10 to 100 weight portions, polylysine 1 to 10 weight portion; The amount of described liquefier sodium citrate is 2 to 20 weight portions; Described firming agent calcium chloride is 1 to 10 weight portion.
2. according to claim 1 containing cartilage cells of epiphyseal plate compound microcapsule preparation, it is characterized in that: the described cartilage cells of epiphyseal plate compound microcapsule that contains is stored in the middle of culture fluid or in normal saline.
3. according to claim 2 containing cartilage cells of epiphyseal plate compound microcapsule preparation, it is characterized in that: the cartilage cells of epiphyseal plate that described fresh cartilage cells of epiphyseal plate is fresh separated, or cryopreservation resuscitation cartilage cells of epiphyseal plate.
4. containing a preparation method for cartilage cells of epiphyseal plate compound microcapsule preparation, its step is as follows:
(1) sodium alginate of 50 to 600 weight portions is weighed, dissolve; Obtain microcapsule solution;
(2) collagen protein of 10 to 100 weight portions is weighed, dissolve, obtain another kind of microcapsule solution;
(3) polylysine of 1 to 10 weight portion is weighed, dissolve, obtain another kind of microcapsule solution;
(4) sodium citrate of 2 to 20 weight portions is weighed, dissolve, obtain another kind of microcapsule liquefaction solution;
(5) calcium chloride of 1 to 10 weight portion or barium chloride are weighed, dissolve, obtain consolidation liquid;
(6) cartilage cells of epiphyseal plate of purification is pressed to 2 * 10
6～8 * 10
6quantity, obtains Cell sap; Described cartilage cells of epiphyseal plate is fresh cartilage cells of epiphyseal plate or immortalization cartilage cells of epiphyseal plate;
(7) by gained Cell sap and sodium alginate and collagen solution mixing, and mix with described consolidation liquid by high-pressure electrostatic bull microcapsule drop generating device, be solidified into the micro gel bead of circle or similar round, the micro gel bead that micro gel bead and step (3) gained polylysine solution effects must be strengthened, micro gel bead again with the effect of step (4) gained sodium citrate, after liquefaction, must contain cartilage cells of epiphyseal plate compound microcapsule preparation.
5. the preparation method containing cartilage cells of epiphyseal plate compound microcapsule preparation according to claim 4, it is characterized in that: described high-pressure electrostatic bull microcapsule drop generating device comprises: an electrostatic generator, on described electrostatic generator, there are positive and negative polarities, positive pole is connected with the syringe needle of microinjection apparatus, negative pole is connected with the rustless steel steel ring being immersed in described consolidation liquid, mixed carrier solution and cartilage cells of epiphyseal plate are housed in injection device, splash in described consolidation liquid and form micro gel bead, gained cartilage cells of epiphyseal plate compound recipe micro gel bead is deposited to the bottom of container, through polylysine, solidify again, after sodium citrate solution liquefaction, must contain cartilage cells of epiphyseal plate compound microcapsule, the described cartilage cells of epiphyseal plate compound microcapsule that contains is stored in the middle of culture fluid or in normal saline.
6. the preparation method containing cartilage cells of epiphyseal plate compound microcapsule preparation according to claim 5, is characterized in that: the cartilage cells of epiphyseal plate that described fresh cartilage cells of epiphyseal plate is fresh separated, or cryopreservation resuscitation cartilage cells of epiphyseal plate.
7. the preparation method containing cartilage cells of epiphyseal plate compound microcapsule preparation according to claim 6, it is characterized in that: the preparation method of described cryopreservation resuscitation cartilage cells of epiphyseal plate is: from liquid nitrogen container, take out cryopreservation tube, drop in 37 ℃ of warm water rapidly, and frequently shake, make it melt as early as possible; From water-bath, take out cryopreservation tube, sucking-off cell suspension, injects centrifuge tube and drips 10 times of above culture fluid, and low-speed centrifugal after mixing, washs 3 times, with Trypan Blue, detects cell survival rate, and the cartilage cells of epiphyseal plate of purification is pressed to 2 * 10
6～8 * 10
6wrap up, make the rear cartilage cells of epiphyseal plate compound microcapsule of recovery stand-by.
8. according to the application in the medicine of the filling for the preparation of after Injury of Epiphyseal Plate or epiphyseal plate excision and effectively secretion containing cartilage cells of epiphyseal plate compound microcapsule preparation described in any one in claim 1-3.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910083272.1A CN101537016B (en) | 2009-04-30 | 2009-04-30 | Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof |
| PCT/CN2010/072395 WO2010124655A1 (en) | 2009-04-30 | 2010-04-30 | Complex microcapsule comprising epiphyseal plate chondrocytes and preparation method and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910083272.1A CN101537016B (en) | 2009-04-30 | 2009-04-30 | Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101537016A CN101537016A (en) | 2009-09-23 |
| CN101537016B true CN101537016B (en) | 2014-03-26 |
Family
ID=41120557
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910083272.1A Active CN101537016B (en) | 2009-04-30 | 2009-04-30 | Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN101537016B (en) |
| WO (1) | WO2010124655A1 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101537016B (en) * | 2009-04-30 | 2014-03-26 | 北京宏医耀科技发展有限公司 | Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof |
| CN105457100B (en) * | 2015-12-21 | 2018-09-25 | 上海赛立维生物科技有限公司 | Artificial liver cell microfluidic body microcapsule preparation method and its microfluid microcapsule generating apparatus |
| CN111617318B (en) * | 2020-05-26 | 2021-12-10 | 南京市儿童医院 | Preparation method of injectable epiphyseal plate regeneration hydrogel for children |
| CN112778544B (en) * | 2021-01-21 | 2021-12-03 | 江南大学 | Crosslinked hydrogel for muscle stem cell culture and preparation method and application thereof |
| CN113813448B (en) * | 2021-10-08 | 2022-12-06 | 大连大学附属中山医院 | Hardness-adjustable hydrogel support containing cartilage-like pitted structure |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100355456C (en) * | 2003-08-15 | 2007-12-19 | 华侨大学 | Calcium alginate/poly arginine-chitin glycan composite medicine release -controlled microcap sule and preparing method thereof |
| JP2006051157A (en) * | 2004-08-11 | 2006-02-23 | Olympus Corp | Chondrifying capsule |
| CN101537016B (en) * | 2009-04-30 | 2014-03-26 | 北京宏医耀科技发展有限公司 | Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof |
-
2009
- 2009-04-30 CN CN200910083272.1A patent/CN101537016B/en active Active
-
2010
- 2010-04-30 WO PCT/CN2010/072395 patent/WO2010124655A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| CN101537016A (en) | 2009-09-23 |
| WO2010124655A1 (en) | 2010-11-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102188748B (en) | Novel chondrocyte epimatrix membrane and preparation method thereof | |
| Zhao et al. | Osteogenic media and rhBMP-2-induced differentiation of umbilical cord mesenchymal stem cells encapsulated in alginate microbeads and integrated in an injectable calcium phosphate-chitosan fibrous scaffold | |
| US20140227235A1 (en) | Cartilage cell treatment comprising collagen, hyaluronic acid derivative, and stem cell derived from mammal umbilical cord | |
| US20070104692A1 (en) | Breast tissue regeneration | |
| Marycz et al. | Adipose stem cell combined with plasma-based implant bone tissue differentiation in vitro and in a horse with a phalanx digitalis distalis fracture: a case report. | |
| JPS59192364A (en) | Soft bone and repairing method thereof | |
| CN105998067A (en) | Process, tube and device for the preparation of wound healant composition | |
| CN101537016B (en) | Compound microcapsule preparation containing cartilage cells of epiphyseal plate, preparation method and application thereof | |
| CN108865986B (en) | Mesenchymal stem cell preparation for repairing articular cartilage damage/defect and preparation method and application thereof | |
| CN102387824A (en) | Rapid preparation and use of engineered tissue and scaffolds as individual implants | |
| CN105358188A (en) | Lipofilling with ex-vivo expanded adipose tissue-derived stem cells for cosmetic breast filling or for facial filling and/or rejuvenation | |
| CN108478863A (en) | The preparation method and products thereof of compound small-caliber artificial blood vessel | |
| CN105969726A (en) | Method for preparing adipose-derived stem cells by means of extraction | |
| CN1836034A (en) | Methods of producing neurons | |
| CN110585484A (en) | Composite bone powder for bone tissue and preparation method and application thereof | |
| CN107137763A (en) | A kind of study of vascularized tissue engineering bone and preparation method thereof | |
| CN106421917A (en) | Method for preparing composition for repairing cartilage injuries | |
| JP7350344B2 (en) | Method for producing chondrocyte pellets from human induced pluripotent stem cells and its uses | |
| JP2007037718A (en) | Artificial cartilage tissue and its production method | |
| CN106474156A (en) | Purposes in preparing medicine for the compositionss | |
| CN110237302A (en) | A kind of preparation method of articular cartilage repair materials-autologous platelet rich plasma combination hyaluronic acid gel | |
| WO2010125722A1 (en) | Culture substrate for cartilage comprising artificial collagen, and cartilage regeneration therapy method using the culture substrate | |
| CN111407785A (en) | Composite acellular matrix injection for treating femoral head necrosis and using method | |
| CN103874763A (en) | Preparation of parental cell bank from foetal tissue | |
| AU2006200600A1 (en) | Cartilage composition for transplantation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |