36 results about "Cartilaginous Differentiation" patented technology

The present invention relates to the field of biomedical materials. Particularly provided is a DNA-silk fibroin composite hydrogel microsphere. The DNA-silk fibroin composite hydrogel microsphere is prepared from DNA and silk fibroin on the basis of the micro-fluidic technology by means of combination of self-assembly physical crosslinking and photo-crosslinking. The introduction of a silk fibroin network into the composite hydrogel microsphere can provide mechanical performance for a DNA hydrogel network and reduce the production cost. RGD can remarkably improve the cell loading capability of the microsphere. The hydrogel microsphere can simulate a microenvironment of cartilage tissue by means of three-dimensional loading of high-density cells, so as to enhance substance exchange and signaling between cells, and more effectively promote the proliferation and adhesion of bone marrow mesenchymal stem cells and regulating the chondrogenic differentiation, thus promoting cartilage regeneration and repair, and providing an available material for preparing medicaments or medical instruments for cartilage defect repair.

The invention discloses composite hydrogel as well as a preparation method and application thereof. The composite hydrogel comprises a hydrogel matrix and a liposome loaded on the hydrogel matrix, the hydrogel matrix comprises a methacrylated natural protein, and the liposome is loaded with a drug; the medicine comprises a medicine for promoting cartilage differentiation and an anti-inflammatory traditional Chinese medicine monomer. According to the invention, through a liposome entrapment technology, the stability and sustained release ability of the insoluble traditional Chinese medicine monomer and the cartilage differentiation induction drug are enhanced. The particle size of the liposome is controllable, the liposome is well combined with a methacrylated natural protein network, a drug sustained release bin can be formed in the hydrogel, long-time and local precise release is achieved, the drug utilization efficiency is improved, and systematic toxic and side effects are avoided.

The invention discloses HA-PAS hydrogel for enhancing cartilage repair of early osteoarthritis and a preparation method of the HA-PAS hydrogel. According to the hydrogel, formylated hyaluronic acid, hydrazide modified low-methoxyl pectin and phenylboronic acid modified alginate form a ternary polysaccharide skeleton, a double-dynamic cross-linked network is formed through acylhydrazone bonds and boric acid ester bonds, and a triple network structure is formed by combining permanent cross-linking catalyzed by horse radish peroxidase. The hydrogel integrates functional components such as nano-selenium, a pulullan-strontium chelate and a mesenchymal stem cell source exosome, so that multiple effects of resisting oxidation, resisting inflammation, promoting cartilage differentiation and repairing subchondral bones are achieved. The compression modulus of the hydrogel is 0.8-1.5 MPa, and the hydrogel can be injected for in-situ gelling and MMP responsive degradation, shows an excellent cartilage repair effect in animal experiments, and provides a new solution for treatment of early osteoarthritis.

The present invention provides a method for producing a cartilage-like tissue from pluripotent stem cells, the method comprising a step for culturing pluripotent stem cells in a cartilage differentiation culture medium and a step for increasing the ascorbic acid concentration of the cartilage differentiation culture medium. The present invention also provides a method for regulating the hardness of induced cartilage-like tissue differentiated from pluripotent stem cells, the method comprising a step for culturing pluripotent stem cells in a cartilage differentiation culture medium and a step for increasing the ascorbic acid concentration of the cartilage differentiation culture medium.

Application methods of a nano layered double include: applying the nano layered double hydroxide in promoting chondrogenic differentiation of human umbilical cord-derived mesenchymal stem cells, and applying the nano layered double hydroxide in preparing drugs for promoting cartilage regeneration, promoting intervertebral disc repair, or treating intervertebral disc degeneration. Compared to human umbilical cord-derived mesenchymal stem cells without an addition of the nano layered double hydroxide, the human umbilical cord-derived mesenchymal stem cells with the addition of the nano layered double hydroxide have a better and faster ability to differentiate into chondrocytes and broad application prospects.

The composition for repairing the meniscus comprises a growth differentiation factor and emodin, and is used for improving inflammation and oxidative stress in a microenvironment and promoting a cell chondrogenic differentiation process. Through verification, according to the core-shell co-delivery nano-carrier provided by the invention, two bioactive compounds, namely emodin and a growth differentiation factor, are wrapped by mesoporous silicon dioxide nano-particles, and the two bioactive compounds are used for promoting fibrocartilage differentiation. The double-drug nano delivery system provides continuous release of emodin and GDF, synergistically regulates severe inflammation and oxidation environments, and promotes in-vitro and in-vivo fibrous cartilage regeneration. Benefited from this, according to the invention, a three-dimensionally printed meniscus-shaped PCL framework is combined with the composition by using cartilage-specific matrix hydrogel, a stability-enhanced meniscus replacement model is constructed, a cartilage immune microenvironment is synergistically adjusted, and successful repair of animal (rabbit) full meniscus replacement is realized.

The use of a polylactic-co-glycolic acid scaffold in promoting cartilage differentiation and regeneration belongs to the technical field of biomedical materials. The present invention includes the use of a polylactic-co-glycolic acid scaffold for non-disease diagnosis and treatment purposes in promoting cartilage differentiation and regeneration. The polylactic-co-glycolic acid copolymer is PLGA (50:50). The polylactic-co-glycolic acid scaffold can promote the differentiation of cartilage precursor cells into cartilage, and increase the formation of cartilage collagen matrix and cartilage proteoglycans. The degradation release liquid of the PLGA (50:50) scaffold material can effectively promote the differentiation of cartilage precursor cells into cartilage, and increase the formation of cartilage collagen matrix and cartilage proteoglycans. The release liquid of the PLGA (50:50) scaffold can be changed into a cartilage microenvironment, thereby playing a role in promoting cartilage regeneration.

The invention discloses a metal gel composite osteochondral repair porous scaffold material based on a heterogeneous double-layer bonding technology and a preparation method of the metal gel composite osteochondral repair porous scaffold material. The specific process comprises the following steps: 1) obtaining an inorganic magnesium alloy porous lower-layer bracket by adopting a metal 3D printing technology; (2) mixing a cartilage differentiation promoting growth factor and a nano-carrier loaded with the cartilage differentiation promoting growth factor into methacrylated I-type collagen serving as a matrix, and carrying out 3D printing to obtain an organic porous upper-layer bracket; (3) carrying out polarization treatment on the to-be-bonded surface of the lower-layer bracket by using a plasma surface activation technology, and pre-infiltrating the to-be-bonded surfaces of the upper-layer bracket and the lower-layer bracket by using a water-absorbing self-curing adhesive; and (4) bonding an upper-layer scaffold and a lower-layer scaffold, and aging the adhesive to obtain the metal gel composite osteochondral repair porous scaffold material based on the heterogeneous double-layer bonding technology. The repairing porous scaffold material has an inorganic lower-layer structure and an organic upper-layer structure which are organically combined by using a bonding technology, so that the problem of simultaneously repairing bones and cartilages is solved in a targeted manner.

The present invention relates to a novel peptide having various physiological activities and a use thereof. The novel peptide, according to the present invention, may induce the increase of constituent components of joints in chondrocytes, increase chondrogenic differentiation of mesenchymal stem cells and extracellular matrix generation, and also inhibit the increase of inflammatory cytokines or inflammatory proteins, and inhibit osteoclast differentiation and formation, and thus may be usefully employed as an active ingredient for a pharmaceutical composition for cartilage regeneration or prevention or treatment of inflammatory disease or osteoporosis, or a health functional food for alleviating such symptoms.

The present invention generally relates to the field of cell culture technology, and specifically, to a culture medium and a culture method for infrapatellar fat pad stem cells. The infrapatellar fat pad stem cell culture medium comprises a basal culture medium, and working concentrations of 3v / v%-7v / v% platelet lysate, 10ng / mL-30ng / mL TGF-β1, 0.5μmol / L-1.5μmol / L dexamethasone, and 0.1μg / mL-1μg / mL non-cytokine proinflammatory factors. The culture medium can promote the growth of IPFP-ASCs and has a significant promoting effect on the in vitro cartilage differentiation ability of IPFP-ASCs.

The invention relates to the technical field of hydrogel and preparation thereof, and discloses chitosan-based gradient hydrogel as well as a preparation method and application of the chitosan-based gradient hydrogel. Thiolated chitosan and curcumin-loaded multi-vesicle liposome are dissolved in an alpha-ketoglutaric acid aqueous solution; and dissolving the N-hydroxysuccinimide acrylate and the hydroxyapatite in another deionized water, mixing the two solutions, and adjusting the pH value of the system to be neutral to obtain the bone layer gel. According to the method for preparing the gel, hydroxyapatite is replaced by cartogenin to obtain the cartilage layer gel. And placing the bone layer gel on the bottom layer and the cartilage layer gel on the top layer, and integrally preparing to obtain the chitosan-based gradient hydrogel. The chitosan-based osteochondral gradient hydrogel constructed by the invention can better simulate an osteochondral gradient structure and an osteochondral interface thereof, the cartilage layer gel promotes cartilage differentiation, the bone layer gel promotes osteogenic differentiation, and slow release of curcumin regulates inflammation microenvironment and maintains phenotype.

The application relates to an anti-inflammatory and cartilage regeneration promoting material, a preparation method and application thereof, and belongs to the technical field of biomedical materials. The anti-inflammatory and cartilage regeneration promoting material comprises a PLGA (50:50) polymer material, and a huperzine A derivative dimer loaded on the PLGA (50:50) polymer material through 3D printing. The application also provides the use of the anti-inflammatory and cartilage regeneration promoting material in the preparation of anti-inflammatory, cartilage protection or cartilage regeneration promoting drugs. The PLGA / A10E scaffold has good biocompatibility, the PLGA / A10E scaffold degradation release liquid can effectively promote the differentiation of cartilage precursor cells into cartilage, and increase the formation of cartilage collagen matrix and cartilage proteoglycan. The in-vitro degradation liquid of the PLGA / A10E scaffold can promote the expression of SOX9 under non-inflammatory conditions; under LPS-mediated inflammatory conditions, the expression of MMP-13 can be inhibited, thereby promoting the synthesis of cartilage matrix and inhibiting the degradation of cartilage matrix.

The application discloses a kind of biomimetic composite scaffold with periosteum-cortical bone-cancellous bone structure and its preparation method and application.The scaffold is assembled by independent periosteum biomimetic scaffold module and independent bone trunk biomimetic module.The periosteum biomimetic scaffold module is cylindrical tube structure hydrogel, loaded with zinc silicate particles and periosteum related cells, with the biological functions of promoting cartilage differentiation, secreting nerve and vascular factors and synthesizing periosteum matrix proteins.The bone trunk biomimetic scaffold module is cylindrical bioceramic, with multiple vertical pipes embedded in the dense layer of outer wall to simulate cortical bone, and with staggered structure inside to simulate cancellous bone.Animal experiments show that the biomimetic composite scaffold can effectively promote the integrated regeneration of bone trunk and periosteum in rabbit radial critical size defect model.Therefore, the application provides a new solution for the biomimetic design and regeneration repair of bone tissue engineering.

The application provides a method for improving the content of cartilage glycosaminoglycan by using cationic polymer, and the method promotes the content of glycosaminoglycan in cartilage tissue to be obviously increased by treating cartilage with a preparation containing polybrene, creates a good microenvironment for cartilage repair, and promotes the cartilage to be better repaired and regenerated; the method can improve the content of glycosaminoglycan in the differentiated tissue of mesenchymal stem cells by culturing the mesenchymal stem cells in an induction medium containing polybrene, promotes the mesenchymal stem cells to be better differentiated into cartilage, and has a wide clinical application prospect.

The present invention is to provide a 3D cartilage organoid block prepared by differentiating mesenchymal stem cells into 3D spheroid cartilage tissues, a basic unit for the 3D cartilage spheroid block. The inventors found that both the amount of GAG matrix and the expression of the collagen type2 increased. Therefore, the method of this invention provides clinically applicable cartilage tissues by effectively enhancing the function of the cartilage differentiation constructs according to 2D culture. The 3D cartilage organoid block can be usefully applied to the area, such as, articular cartilage regeneration and plastic surgery, where cartilage tissues restoration is required.

The present invention relates to a composition which is for preventing or treating bone diseases and includes CCR2 as an active ingredient. It was confirmed that a C—C chemokine receptor type 2 (CCR2) protein, a polynucleotide encoding the CCR2 protein, or a mesenchymal stem cell transduced with the CCR2 according to the present invention neutralize bone disease factor MCP-1, thereby reducing the collagen epitope (CTX-II) and collagen metabolism factors (MMP1 and MMP3) relating to collagen absorption in the body. In addition, it was confirmed that the expression of the SOX9 gene and anti-inflammatory cytokines (TGF-β and IL-10) related to cartilage differentiation was significantly increased. Thus, the present invention has excellent regenerative ability against osteoarthritis and excellent pain suppression and alleviation effects, and thus can be effectively used for preventing or treating bone diseases such as osteoarthritis.

The present application relates to the field of biomedicine technology, and in particular to an induced pluripotent stem cell with dual reporter genes. The induced pluripotent stem cell with dual reporter genes comprises an endogenous COL2A1 locus and a COL10A1 locus in its genome, wherein the COL2A1 locus comprises a first reporter gene and an endogenous COL2A1 gene, and the COL10A1 locus comprises a second reporter gene and an endogenous COL10A1 gene, wherein the first reporter gene and the second reporter gene encode different proteins, respectively. The induced pluripotent stem cell with dual reporter genes has the ability to differentiate into chondrocytes and further induce the generation of hypertrophic cartilage organoids. The cell line of the above-mentioned cells can be used to construct human hypertrophic cartilage organoids in vitro, to track cartilage differentiation and hypertrophy in real time in vitro, and can be used in the fields of disease modeling, drug screening, tissue transplantation, etc.

The present invention relates to a complex for promoting cartilage differentiation comprising stem cells or a culture thereof and a cartilage cell-free crush, a method for fabricating cartilage by using the complex, cartilage fabricated via the method, a pharmaceutical composition comprising the complex for preventing or treating arthropathy, a method for preventing or treating arthropathy using the composition, a composition for inducing cartilage cell differentiation comprising the prepared cartilage cells, and a method for fabricating cartilage using the prepared cartilage cells. The complex for promoting cartilage differentiation comprising the stem cells or the culture thereof and the cartilage cell-free crush provided in the present invention is not only differentiated into cartilage cells without side effects even in vivo, but also promotes differentiation of inherent stem cells in vivo into cartilage cells via a paracrine effect exhibited in the differentiated cartilage cells to effectively regenerate the damaged cartilage, and thus the complex may be widely used for treatment of arthropathy involving the cartilage damage.

The invention discloses a preparation method and application of an injectable hydrogel-loaded acellular matrix, belongs to the technical field of biological materials and biomedical engineering, and solves the problem that in the prior art, the effect of applying BMSCs to cartilage injury repair is not ideal enough. The method comprises the following steps: under the action of fluid shear force, inducing mesenchymal stem cells to carry out chondrogenic differentiation by using TGF-beta3, and carrying out decellularization treatment on a cell aggregate to obtain a decellularized matrix; the modified hyaluronic acid and chitosan are cross-linked, and HPCC hydrogel is obtained; and mixing the acellular matrix with the HPCC hydrogel to obtain the injectable hydrogel loaded acellular matrix. According to the invention, in-vitro directional induction of cartilage differentiation of BMSCs is carried out through combination of fluid shear force and TGF-beta3, and the curative effect is promoted.

Use of a milk fruit extract for the preparation of a composition for improving bone quality and promoting chondrocyte differentiation. The milk fruit extract is extracted from early-harvested fruits of Chrysophyllum cainito using water as a solvent.

The present invention discloses an artificial auricle material with a dual framework of vascularization and chondrogenicity, and a preparation method thereof. The method comprises: 1) performing 3D modeling as needed to obtain an artificial auricle scaffold model with a large pore, and pre-setting a vascularization differentiation framework on the scaffold; 2) using multi-material multi-nozzle light-curing 3D printing technology, simultaneously printing the artificial auricle scaffold with a GelMA solution and printing a vascularization framework within the scaffold using a GelMA solution containing mesenchymal stem cells (MSCs); and culturing the vascularization differentiation to obtain an intermediate-form artificial auricle with the vascularization framework; 3) combining a multiple-immersion light-curing method, introducing a ColMA solution containing MSCs into the pores of the intermediate-form artificial auricle; and culturing the chondrogenic differentiation to obtain an artificial auricle material with both a vascularization and chondrogenicity framework. The simultaneous provision of vascularization and chondrogenicity frameworks within the scaffold provides a design paradigm for an artificial auricle with complete physiological functions.

The invention discloses a composite gel artificial auricle material based on a fiber web substrate strengthening strategy and a preparation method thereof. The preparation method comprises the following steps: (1) designing and preparing a transparent mold of the fiber net substrate and a normal auricle shape; (2) preparing a polylactic acid-glycolic acid copolymer (PLGA) fiber web through electrospinning, cutting the PLGA fiber web into a required shape, and fixing the PLGA fiber web in a mold after heat setting; (3) pouring a ColMA solution containing cartilage promoting factors and mesenchymal stem cells, and carrying out light curing to obtain a composite gel artificial auricle inner layer; and (4) carrying out chondrogenic differentiation culture on the composite material for 4-8 weeks, then fixing the composite material in the mold again, and carrying out photocuring on a ColMA gel protective layer on the surface layer of the composite material to obtain the composite gel artificial auricle material based on the fiber web substrate strengthening strategy. The gel artificial auricle material has a bionic internal structure and an attractive appearance, and a pre-model is provided for research, development and practical application of artificial auricle products.

The application discloses a composition for repairing meniscus, which comprises a growth differentiation factor and emodin, improves inflammation and oxidative stress in a microenvironment, and promotes cell chondrogenic differentiation process. It is verified that the "core-shell" co-delivery nano carrier provided by the application uses mesoporous silica nanoparticles to wrap two bioactive compounds, emodin and a growth differentiation factor, and the latter is used for promoting fibrocartilage differentiation. The dual-drug nano delivery system provides sustained release of emodin and GDF, synergistically regulates a harsh inflammatory and oxidative environment, and promotes fibrocartilage regeneration in vitro and in vivo. Benefited from this, the application also combines a three-dimensionally printed meniscus-shaped PCL frame with the composition by using a cartilage-specific matrix hydrogel, constructs a reinforced stability meniscus replacement model, has a synergistic regulation on a cartilage immune microenvironment, and realizes successful repair of a whole meniscus replacement operation of an animal (rabbit).

The present invention provides an MFAP5-positive synovial lining layer stem cell for repairing articular cartilage injury and an identification method thereof. By performing single-cell sequencing on synovial samples in animal model samples with cartilage defects and the regenerated cartilage-like tissue that has a physical connection with the synovial samples, and through bioinformatics analysis means, a cell subset that plays a key role in the regeneration of cartilage-like tissue, namely the MFAP5-positive subset, is found in synovial interstitial cells, and an identification method for this cell subset is clarified. This cell subset can be used as a more effective cell source for promoting cartilage regeneration, is expected to greatly shorten the in vitro cartilage differentiation time, and can be used to efficiently achieve in-situ repair of cartilage defects in vivo, having broad clinical application prospects in cartilage repair and regeneration.

The invention belongs to the technical field of biotechnology and tissue engineering, and particularly relates to application of pilose antler bone matrix vesicles in preparation of a medicine for treating bone injury, in the pilose antler bone matrix vesicles prepared through an enzymolysis method, the pilose antler cartilage matrix vesicles can remarkably promote cartilage differentiation, the osteogenesis promoting capacity of the pilose antler osteogenesis matrix vesicles is extremely high, and the pilose antler cartilage matrix vesicles can be used for preparing medicines for treating bone injury. The pilose antler bone matrix vesicle can improve bone mineral density, bone volume fraction, bone trabecula number, bone trabecula thickness, bone trabecula separation degree and cortical bone mineral density, can play a significant role in the aspect of osteoporosis resistance, and has good application in the aspect of bone injury treatment.

The invention provides an MFAP5 positive synovial lining layer stem cell for articular cartilage injury repair and an identification method thereof, which are characterized in that a synovial sample in a cartilage defect animal model sample and a regenerated cartilage-like tissue in physical connection with the synovial sample are subjected to single-cell sequencing, and a bioinformatics analysis means is adopted to identify the MFAP5 positive synovial lining layer stem cell. According to the method, a cell subset, namely an MFAP5 positive subset, which plays a key role in cartilage-like tissue regeneration in synovial interstitial cells is found, an identification method of the cell subset is defined, and the cell subset can be used as a more effective source cell for promoting cartilage regeneration, so that the in-vitro cartilage differentiation time is expected to be greatly shortened, and the cartilage regeneration can be promoted. The method can be used for efficiently realizing in-vivo cartilage defect in-situ repair and has a wide clinical application prospect in cartilage repair and regeneration.

The invention provides an intervertebral disc regeneration model, human nucleus pulposus cells (iHNPCs) and human fibrous ring cells (iHAFCs) reversibly immortalized by hTERT (human telomerase reverse transcriptase) are successfully constructed, and based on the two cells, after stimulation of chondrogenic differentiation factors and osteogenic factors, an iHAFCs / PPCNg mixture and an iHNPCs / GelMA hydrogel mixture are respectively loaded to the periphery and the central area of a 3D intervertebral disc-like structure of a POC support, so that the 3D intervertebral disc-like structure of the POC support is formed. After the intervertebral disc model is implanted under the skin of a nude mouse for 42 days, cartilage-like tissues exist in the central area of the intervertebral disc model, and mature bone tissues exist in the peripheral area of the intervertebral disc model. The intervertebral disc regeneration model simulates the structure of a natural intervertebral disc, provides an important biomedical engineering material for IVD tissue engineering, has potential application value in intervertebral disc degenerative diseases, and also provides an important means for researching intervertebral disc cell functions and developing an effective IVD tissue engineering method.

The present invention relates to a method for selecting stem cells having enhanced ability to differentiate cartilage, a cell therapy product comprising stem cells selected by said selection method, and a method for producing the same.

The invention discloses application of KDM6B protein in enhancing cartilage differentiation capacity of apical papilla stem cells. Experiments prove that KDM6B knocks down and inhibits cartilage differentiation of SCAPs, and overexpresses KDM6B to enhance cartilage differentiation of SCAPs; it is further proved that KDM6B can promote SCAPs mediated cartilage repair, and the effectiveness of KDM6B in cartilage tissue and subosseous bone repair and regeneration is proved in vivo. The invention lays an important foundation for research on treatment of cartilage injury by using the odontogenic stem cells in the future, and has an important application prospect.

The invention belongs to the technical field of bone tissue engineering and regenerative medicine, and particularly discloses a piezoelectric-photo-thermal dual-mode composite stent for articular cartilage repair and a preparation method of the piezoelectric-photo-thermal dual-mode composite stent. The preparation method comprises the three steps of MXene preparation, electrostatic spinning and paper folding structure construction, poly-L-lactic acid and MXene nanosheets are compounded to prepare a fiber membrane, and then the fiber membrane is subjected to hot pressing through a corrugated mold to form the three-dimensional stent with the periodic creased structure. The scaffold has piezoelectric property and photo-thermal property at the same time, promotes cartilage differentiation through the piezoelectric effect, inhibits subchondral bone angiogenesis through the photo-thermal effect, maintains a low-oxygen microenvironment, realizes integral repair of cartilage-subchondral bone functional units, can be used for treatment of cartilage defects, osteochondral defects and osteoarthritis, and has a wide application prospect. The problem that the cartilage steady state is damaged due to the fact that a traditional piezoelectric material promotes bone vascularization is solved.