67 results about "Chondrocyte cell" patented technology

A production method for producing cell colony forming units in vitro from a mammalian tissue explant, the method comprises the steps of a) growing a piece of the mammalian tissue explant in a growth medium to obtain cell colony forming units from immature cells from the piece of explant, and b) harvesting cells from one or more of the cell colony forming units for use in Autologous Cell Implantation / transplantation methods. The mammalian tissue explant is selected from the group consisting of cartilage; bone such as, e.g., bone marrow; connective tissue; muscle tissue such as, e.g., smooth muscle tissue, heart tissue, liver tissue and skeletal muscle tissue; skin tissue such as, e.g., periosteum; mucosal tissue; brain tissue, pancreas tissue and blood vessels. In particularly, the mammalian tissue explant is cartilage, such as elastic, fibro, hyalin or articular hyalin cartilage. The cells obtained are suitable for use in autologous implantation / transplantation methods. In a specific embodiment, the cell obtained are chondroytes, especially for use in autologous chondrocyte implantation (ACI) methods.

The invention relates to a culture method for inducing adipose tissue-derived stromal cells to differentiate to chondrocyte, and aims to solve the problem that the prior art is low in differentiation rate. The culture method comprises the following steps: 1) performing separation of primary cells of adipose tissue-derived stromal cells; 2) performing amplification and passage of the adipose tissue-derived stromal cells; and 3) performing differentiation culture on P5-generation adipose-derived stem cells to chondrocyte, namely, adding the adipose tissue-derived stromal cells into a condition culture medium, namely, an adipose tissue-derived stromal cell chondrocyte differentiation culture medium, after P5 generation of passage, performing chondrocyte differentiation culture, replacing the medium of the cells every 3 days, observing the morphological change of the cells, and after 16 days of induction, performing Alcian blue dyeing, and identifying the chondrocyte differentiation situation of the adipose tissue-derived stromal cells. The result of Alcian blue dyeing identification shows that the chondrocyte can be formed, and the Alcian blue dyeing is positive. The culture method has the characteristics of being simple and feasible, short in induction time as the induction culture medium is a serum-free culture system, good in test repeatability and high in osteoblast differentiation rate.

An aseptic / sterile medical kits are comprising a cartilage regeneration kit, a bone regeneration kit or an umbilical cord blood storage kit in a configuration that each process performs according to functionally-specialized kit sets for each step, via division of overall processes into corresponding steps for isolation, culture, collection and storage of cells, and implantation of desired cells into target sites of the body. The cartilage is regenerated by cartilage tissue collection; chondrocyte isolation; chondrocyte medium change and subculture; preparation of chondrocyte therapy product; media for isolation / culture / preparation / cryopreservation of cells; and media for isolation / culture / cryopreservation of cells, using the cartilage regeneration kit. The bone is regenerated by bone marrow collection; osteoblast isolation; osteoblast medium change and subculture; and preparation of osteoblast therapy product, using the bone regeneration kit. Additionally, the umbilical cord blood is stored by umbilical cord blood collection; hematopoietic stem cell isolation; and cryopreservation of hematopoietic stem cells, using the umbilical cord blood storage kit.

A process for culturing chondrocytes to form constructs which contain higher percentages of cells that retain the chondrocytic phenotype are disclosed. The tissue engineered constructs may be formed into neocartilage-containing compositions for a variety of in vitro and in vivo purposes. Methods of treating individuals in need of articular cartilage growth by implanting a new composition are disclosed.

The invention relates to an application of autologous adipose-derived mesenchymal stem cells in preparation of medicines for treating joint diseases. The application is characterized in that adipose-derived mesenchymal stem cells (Ad-MSCs) separated and extracted from autologous adipose tissues of the patient are injected to the diseased articular cavity in a large dose once. The adopted mesenchymal stem cells have immunophenotyping and representing biological marks of Ad-MSC through flow cytometry authentication, and can be directionally induced and differentiated to osteoblast and chondrocyte. Primary cells are directly injected to the articular cavity of the diseased joint to treat various joint diseases. With the adoption of the autologous adipose-derived mesenchymal stem cells as the primary cells, the cells have better differentiative potential and the ability of inhibiting inflammation and immunoregulation ability. The adipose-derived mesenchymal stem cell transplant provides a novel effective means for treating osteoarticular diseases. The method is simple and feasible, and the quantity of cells collected is great and the trauma is small. The autologous mesenchymal stem cells of the patient are applied without invading related ethics, so that the risk of immunological rejection, infectious diseases and the like is avoided.

The invention belongs to the technical field of medicine; in order to solve the problem that no efficient and safe cartilage siRNA delivery system is provided for limiting application of RNAi in treating cartilage diseases, the invention provides a cartilage siRNA liposome delivery system and a preparation method of the delivery system. The delivery system consists of lipidosome, a lipidosome modifier and a small nucleic acid drug, wherein the lipidosome is phosphatidylcholine, cholesterol or Dlin-KC2-DMA; the lipidosome modifier is a polyethylene glycol lipid conjugate; and the small nucleic acid drug is Ihh siRNA. According to the invention, a liposome nano based cartilage RNA interference delivery system is modified, and the formed composite has positive charges, is safe and small in volume, and can be distributed in a full-lamellar cartilage tissue. Carrier experiments verify that siRNA can be delivered into cartilage cells to successful knock out corresponding genes inside the cartilage cells, therefore a bottleneck of gene knockout in the cartilage tissue is broken through; and the siRNA, as a target gene, can be used for treating PTOA (post-traumatic osteoarthritis).

The invention relates to a method to prepare and preserve homogeneous bioactive bone tissue, its steps: 1, obtaining: on the aseptic condition, obtaining homogeneous bone tissue; 2, finishing: eliminating soft tissue and periosteum, eliminating bone marrow tissue, and reserving partial joint bursa, ligament and adhesive point of muscle; 3, defatting: dipping in chloroformic methanol, washing with physiological saline and then packaging; 4, gradient temperature reducing: placing the preprocessed bone tissue in program temperature reducer to make gradient temperature reduction up to -40 deg.C--60 deg.C for 20-80 min.; 5, preserving: putting the bone tissue in the -60 deg.C - 80 deg. C refrigerator. It can keep the integrality of bone ultramicro structure, bone inducing activity, bioforce intensity and maintain a certain quantity of survival soft bone cells.

The invention provides a preparation method of cross-linking microspheres based on microfluidics. The preparation method comprises the following steps of pushing an injector of a water phase solutionand an oil phase solution which are prepared in advance through a pushing and injecting pump device, enabling a water-oil two-phase solution to form water-in-oil type emulsion liquid drops at an exitchannel of a micro-fluidic chip, and performing illumination at an ultra-violet curing lamp to prepare PEGDA photo-crosslinking microspheres; and enabling the PEGDA photo-crosslinking microspheres tobe charged into a sodium hydroxide solution so as to obtain PEGDA-chitosan cross-linking microspheres, wherein the diameter range of the cross-linking microspheres is 240-440[mu]m, and is controlled through the flow velocity and the flow velocity ratio of the water phase solution and the oil phase solution. The preparation method is flexible and easy to operate. A preparation method of an injecting type chondrocyte composite body comprises the following steps of A1, preparing the cross-linking microspheres; and A2, enabling the cross-linking microspheres to carry chondrocyte. Biological cellsare carried on the cross-linking microspheres having a dual-network hydrogel system, and through proliferation and disintegration of the biological cells, the injecting type cell composite body can beprepared, and is a novel tissue repair material which is widely applied.

The invention relates to the field of miRNAs and exosomes, in particular to an application of miRNA in the preparation of a medicine for preventing and treating osteoarthritis, an exosome with the high expression of the miRNA, and an application of the exosome. The invention provides an application of miR-155-5p in the preparation of a medicine for preventing and / or treating osteoarthritis, and anucleotide sequence of the miR-155-5p is as shown in SEQ ID NO: 1. The application provided by the invention can effectively improve the expression of the miR-155-5p, and the miR-155-5p can promote cartilage cell proliferation, migration and extracellular matrix secretion and inhibit cell apoptosis by targeting Runx2, and thereby achieves the effect of preventing or treating osteoarthritis.

The invention discloses an experimental method for researching anti-articular cartilage cell apoptosis of resveratrol. The ability of resveratrol to inhibit articular cartilage cell apoptosis is verified by establishing crowd research, in vivo experiments and in vitro experiments, and the mechanism of action of resveratrol to inhibit articular cartilage cell apoptosis is discussed to verify the 'effect axis hypothesis' of resveratrol-p38 MAPK signal pathway down-regulation-anti-apoptosis, which proves that the potential action targets of resveratrol in the screened p38 MAPK pathway have a targeting relationship, and further defines the overall regulation effect of resveratrol on a plurality of targets in the pathway. Experiments prove that resveratrol has the ability to inhibit articular cartilage cell apoptosis, and provides experimental basis for clinical practice of resveratrol in treating osteoarthritis.

The invention relates to a trachea support and its application. The trachea stent is provided with support rings and support links, each of the support rings is C-shaped, and each support ring is connected in series by support links, so that each support ring and support links are fixed into one. The present invention also provides a tissue engineered trachea, which is made by wrapping the chondrocyte-gelatin / polycaprolactone electrospun membrane composite on the outside of the trachea stent. Using the tracheal stent or tissue engineered trachea of ​​the present invention, the tracheal stenosis can be repaired by the extratracheal support suspension method, and a new C-shaped cartilage ring can be formed after implantation, finally making it have the potential to grow.

The invention provides a composition containing a ghrelin active agent and a method of using the composition to induce stem cells differentiated to cartilage cells. The invention specifically providesa method of using the composition to treat osteoarthritis and a method of using the composition to treat cartilage injury.

The invention belongs to the field of articular cartilage substitute materials, and provides a preparation method of polyvinyl alcohol-acrylamide-agarose hydrogel with high mechanical strength. Polyvinyl alcohol and acrylamide are used as matrix materials, and a small amount of agarose is added. Acrylamide forms a first network through a chemical cross-linking agent, and polyvinyl alcohol is physically cross-linked in a refrigeration cycle to form a second network. Agarose can form hydrogen bonds with molecular chains of polyvinyl alcohol and acrylamide, so that the mechanical strength of the hydrogel is further improved. In addition, due to the introduction of agarose, the hydrogel can adhere to cartilage cells and promote cartilage cell proliferation. The hydrogel prepared by the method is expected to be applied to articular cartilage replacement.

The invention discloses a multicellular biological composite scaffold, a preparation method and application thereof. The scaffold comprises a lower-layer structure for forming a circular permutation mode of (A-B-A)m and an upper-layer structure for forming a circular permutation mode of (C-D-C)n, and A is a lower-layer composite material of bioactive ceramic powder containing three elements of Li,Mg and Si and an organic material with biocompatibility, B is stem cells with osteogenic differentiation potential, C is an upper-layer organic material containing an organic material with biocompatibility, D is chondrocyte, m and n are positive integers between 1 and 100. The multicellular biological composite scaffold can simulate the physiological structure of osteochondral complex tissue andefficiently promote the integrated repair of osteochondral defect.

The invention discloses a preparation method of a zin-ion-containing tissue engineering scaffold with an anti-inflammatory function. The method comprises the following steps: separating Wharton's jelly from a human umbilical cord; preparing a pure Wharton's jelly scaffold through the processes of repeated low-temperature crushing, freeze thawing, gradient centrifugation, pancreatin digestion and elution, vacuum freeze compression drying and the like; performing induced deposition on the pure Wharton jelly scaffold in simulated body fluid containing zinc ions; putting the scaffold in an ammoniawater solution in a hydrothermal kettle and performing a gas-phase hydrothermal reaction; and then performing vacuum freeze drying to obtain the cartilage tissue engineering scaffold containing zincions. The tissue engineering scaffold prepared by the method not only has high histocompatibility but also can successfully load zinc ions and release the zinc ions through a slow release effect, so that macrophage polarization is regulated and an anti-inflammatory effect on cartilage injury in an inflammatory environment is realized; and stem cells can be induced to differentiate into chondrocytes; and a good internal immune environment is provided for cartilage repair.

The present invention provides a medical kit and a method using the same. For this purpose, an aseptic / sterile medical kit is provided which comprises a cartilage regeneration kit 10, a bone regeneration kit 20 or an umbilical cord blood storage kit 30, in a configuration that each process can be performed according to the functionally-specialized kit sets for each step, via division of the overall processes into the corresponding steps for isolation, culture, collection and storage of cells, and implantation of the desired cells into the target sites of the body. Therefore, the cartilage is regenerated by cartilage tissue collection; chondrocyte isolation; chondrocyte medium change and subculture; preparation of a chondrocyte therapy product; media for isolation / culture / preparation / cryopreservation of cells; and media for isolation / culture / cryopreservation of cells, using the cartilage regeneration kit 10. The bone is regenerated by bone marrow collection; osteoblast isolation; osteoblast medium change and subculture; and preparation of an osteoblast therapy product, using the bone regeneration kit 20. In addition, the umbilical cord blood is stored by umbilical cord blood collection; hematopoietic stem cell isolation; and cryopreservation of hematopoietic stem cells, using the umbilical cord blood storage kit 30.

The methods described herein are for conserving highly expanded cells that have functional properties such as potential for use in neotissue constructs. For example, highly expanded chondrocytes that can be used to construct neocartilage exhibiting functional properties similar to native articular cartilage. The methods and systems feature processes that form functional, human cartilage using cells that have been expanded to at least 1.5×105 times or P3 or greater. This enables a large quantity of engineered cartilage implants to be produced from few cells.

A reinforced matrix membrane containing one or more scaffold-forming proteins suitable for cell growth and for use in chondrocyte cell transplantation, and method of making same. The scaffold is incubated with the collagen matrix in solutions, colloidal dispersions, or suspensions of stabilizing proteins. The reinforced matrix may be used in tissue engineering, cartilage transplantation, bone and cartilage grafting, healing, joint repair and the prevention of arthritic pathologies.

The invention relates to a zuogui pill nanoparticle drug delivery system and a preparation method thereof. The zuogui pill nanoparticle drug delivery system comprises a zuogui pill extract and a polybutylcyanoacrylate nano carrier, wherein the mass ratio of the zuogui pill extract to the polybutylcyanoacrylate nano carrier is 1 to (0.1-10). Due to the technical scheme, with polybutylcyanoacrylate as a raw material, the zuogui pill nanoparticle drug delivery system is prepared by an improved emulsion polymerization method. The zuogui pill nanoparticle drug delivery system used by people has the advantages of continuity, high efficiency, controllability and the like, is capable of continuously and efficiently releasing the zuogui pill effective components, promoting chondrocyte proliferation and differentiation, and is natural, free of toxicity and high in efficiency.

The invention provides a micro-nano hydrogel microsphere for targeted adjustment of a mitochondrial respiratory chain as well as preparation and application of the micro-nano hydrogel microsphere. According to the invention, hyaluronic acid microspheres are prepared through a microfluidic technology, and SS-31 peptide and Wyrgrl peptide modified resveratrol-loaded long-circulating liposome are connected in a microsphere nano network through a non-covalent bond, so that a micro-nano hydrogel microsphere system with a targeted cell MRC adjustment function is constructed. The system has efficient cellular uptake efficiency and mitochondrial targeting, and can significantly improve the MRC function, reduce proton leakage, protect mitochondria, down-regulate ROS expression, and promote generation of cartilage extracellular matrix. Meanwhile, the system can effectively slow down the progress of osteoarthritis in a rat osteoarthritis model. The micro-nano hydrogel microsphere system for the targeted adjustment and control of the cell MRC function, provided by the invention, has great potential in treating various degenerative diseases related to MRC dysfunction.

The invention relates to the field of cells, in particular to a composition, differentiation inducing culturing liquid containing the composition and an inducing method. Selected dental pulp stem cells (dental pulp stem cell, DPSCs) are same with MSCs of other sources, have self-renewing and multi-directional differentiation capacity and have the advantages that cell proliferation in vitro is easy, and the immunogenicity of the cells is low, and the selected dental pulp stem cells can be taken as seed cells to promote repairing and regeneration of injured tissue. Thedifferentiation inducing culturing liquid can effectively promote specificity differentiation, towards cartilage cells, of DPSCs. Based on the differentiation inducing culturing liquid, the provided method ofchondrogenesisdifferentiation for the DPSCs subjected to in-vitro inducing culturingcan successively achieve chondrogenesis differentiation inducing on the DPSCs.

The invention discloses a culture dish for chondrocyte cell sheets. The culture dish comprises a cell culture dish container body made of polystyrene, an upper end cover made of polystyrene is arranged on the cell culture dish container body, an arc-shaped cell suspension adding opening is formed in the side wall of the cell culture dish container body, a sealing end cover is arranged on the arc-shaped cell suspension adding opening, and a polycarbonate semipermeable membrane capable of filtering molecules with 5Kb molecular weight is arranged in the middle of the interior of the cell culturedish container body. The polycarbonate semipermeable membrane provided by the invention allows free passage of metabolic waste and nutrient components, and does not allow passage of a factor corresponding to the chondrogenesis promotion. During liquid change every time, only the liquid above the semipermeable membrane is changed, the chondrogenesis promotion factor does not need to be additionallyadded, the autocrine factor is reserved, and the production cost is reduced. The product can contain more liquid, the operation of changing the liquid every day is avoided, and the pollution possibility and the heavy work are reduced. Meanwhile, the culture dish is reasonable and simple in structural design, easy to implement and suitable for application and popularization.

The invention discloses a method for establishing an osteoarthritis osteoclast-like cell model of mesenchymal stem cells after directional cartilage differentiation through a two-step method. The method comprises the steps that firstly, the mesenchymal stem cells are induced to be subjected to directional differentiation into articular cartilage cells, and then the articular cartilage cells are induced by adopting IL-1beta in-vitro induction to establish the osteoarthritis model. According to the method, the two-step method is used for establishing the osteoarthritis model after directional cartilage differentiation for the first time, and by exploring the cartilage cell morphology and expression change of phenotype genes aggrecan and Co12alpha1, the optimal use concentration and cell induction time of a solution are determined. The method is easy to operate and low in cost and can be applied to related studies such as cell quality evaluation of the mesenchymal stem cells after directional cartilage differentiation, early warning of easy infection of adult osteoarthritis, verification of osteoarthritis drug action targets and screening of drug toxicity / drug functions.

The invention discloses application of atler stem cell exosome in preparation of products for improving or treating osteoarthritis and delaying cell senescence. The invention particularly discloses application of a velvet antler-derived stem cell exosome in preparation of products for preventing, improving or treating individual osteoarthritis and application thereof in preparation of products for delaying cell senescence. The cell exosome has a stable treatment effect, and can inhibit or reduce the expression of inflammatory factors; the grip strength of arthritis mice is enhanced, and the motion function is recovered; the bone density of a joint area is enhanced, so that the mechanical capacity of a skeleton structure is enhanced, and the symptom of osteoarthritis is improved; cartilage injury in the joint area is relieved; the growth and differentiation of cartilage cells are effectively promoted, and the damage and injury of soft tissues are relieved; the composition can effectively improve and treat osteoarthritis, has the effect of delaying senescence of human mesenchymal stem cells, and has wide application prospects in the fields of osteoarthritis treatment and cosmetics.

The invention discloses a micro-fluidic chip-based bionic in-vivo articular cartilage microenvironment structure, and relates to the technical field of microfluidics, the micro-fluidic chip-based bionic in-vivo articular cartilage microenvironment structure comprises an upper layer, a middle layer and a lower layer which are stacked in sequence, the upper layer and the lower layer are both provided with pressure systems, and the middle layer is provided with a concentration gradient generator and a plurality of stem cell culture units. The concentration gradient generator is communicated with the cell culture unit, and the pressure system corresponds to the cell culture unit. According to the method, a physical microenvironment and a chemical microenvironment are integrated, high-throughput screening can be carried out on a cartilage cell in-vitro amplification environment by utilizing a small amount of cells, and the optimal factor concentration and compressive stress stimulation intensity suitable for cartilage cell proliferation and maintaining phenotypes are selected in an optimized manner; the structure proves the feasibility of the micro-fluidic chip technology in cartilage tissue engineering application, simulates the in-vivo mechanical stimulation microenvironment, and provides a scheme for in-vitro rapid amplification of cartilage cells, thereby improving the quality of seed cells and improving the autologous cartilage cell transplantation treatment effect.

The invention discloses an effect of copper wire stem root in preparation of a medicine for treating osteoarthritis. According to the invention, chemical components contained in the copper wire stem root are separated and represented, and it is found for the first time that the copper wire stem root contains various anti-osteoarthritis active components, including 6-hydroxy-2-isopropyl-7-methyl-1, 4-naphthoquinone, protocatechualdehyde, (E)-3, 3, 4, 5, 6-tetramethyl-1, 3, 4, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, 5-tetramethyl-1, 3, the compound is prepared from 2, 4-dihydroxybenzylidene acetone, 3-chloro-4-hydroxy-benzoic acid, kaempferol-3-O-rutinoside, p-hydroxymethylene benzyl acetone and the like. According to the application disclosed by the invention, the (E)-3, 4-dihydroxybenzylidene acetone, the 3-chloro-4-hydroxy-benzoic acid and the p-hydroxymethylene benzyl acetone are found for the first time, so that not only can the content of MMP-13 in a cartilage cell culture supernatant be remarkably reduced, but also the survival rate of the cartilage cells after IL-1beta intervention can be remarkably improved; therefore, the copper wire stem root and the three compounds can be used for preparing the medicine for treating osteoarthritis.

The invention relates to application of an umbilical cord stem cell preparation in preparation of a medicine for treating osteoarthritis. The umbilical cord stem cell preparation comprises umbilical cord mesenchymal stem cells, a tissue extracting solution and isoliquiritigenin. The umbilical cord stem cells can reduce inflammatory response and improve expression of cartilage cell marker genes to a certain extent, the tissue extracting solution is added, and the isoliquiritigenin and the umbilical cord stem cells are mixed for use, so that the repair function of the umbilical cord stem cells can be effectively improved, and the efficacy of the umbilical cord stem cells for treating osteoarthritis is exerted.

The present invention discloses a method for in vitro proliferation of chondrocytes. The method comprises primary cell culture and cell passage culture. Growth factors of bFGF, PDGF, etc. are added ina primary culture medium to promote the primary cell culture, the growth factors of the bFGF, the PDGF, IGF-I, etc. having synergistic effects are added in a passage culture medium, so that the cellnumber is proliferated and at the same time cell hypertrophy can be effectively avoided; and at the same time, primary cells are subjected to a two-dimensional culture to enable the chondrocytes to beappropriately proliferated, and besides, preferably passage cells are subjected to a three-dimensional culture to restore phenotypes of degenerated cartilage tissues, so that the method proliferatesthe number of the chondrocytes and can also main the normal tissue phenotypes.

The invention relates to the technical field of biology, in particular to an application of alpha-ketoglutaric acid in preparation of drugs, and the drugs are used for 1) treating osteoarthritis and related diseases; and / or, 2) inhibiting the decomposition phenotype of cartilage cells; and / or 3) promoting synthetic phenotypes of cartilage cells; and / or 4) promoting skin hair follicle regeneration. The pharmaceutical composition provided by the invention can protect cartilage tissues and prevent the progress of osteoarthritis, and has great potential application value for clinical treatment of osteoarthritis. In addition, the medicine provided by the invention can promote obvious increase of skin hair follicles, and also has great potential application value in treatment and improvement of alopecia.