202 results about "Osteoblast cell differentiation" patented technology

The present invention discloses agents and methods for inducing osteoblastic cellular differentiation, as well as the use of such agents and methods to treat patients to maintain bone mass, enhance bone formation and / orbone repair. Exemplary agents include oxysterols, alone or in combination with particular oxysterols, or other agents, such as bone morphogenic proteins, known to assist in bone formation. The invention further includes medicaments including oxysterols for the treatment of bone disorders and implants to facilitate bone repair.

The present invention discloses oxygenic oxygenic oxysterols. Also disclosed, agents and methods for protecting, blocking or rescuing marrow stromal cells from the inhibitory effects of oxidative stress on their osteoblastic cellular differentiation. Exemplary agents include oxysterols, rhBMP2, alone or in combination which are demonstrated to specifically combat oxidative stress caused by inflammatory oxidized lipids, such as xanthine / xanthine oxidase and minimally oxidized LDL. The synergistic effects of oxysterols and bone morphogenic proteins are disclosed.

The invention provides an inducing method and inducing culture medium for differentiation of bone marrow mesenchymal stem cells into osteoblasts in vitro. The inducing culture medium is composed of 1*10<-8> mol / L of dexamethasone, 50 mu mol / L of ascorbic acid and 10 mmol / L of sodium beta-glycerophosphate; and solvent is a supernatant of a sclerite complete culture medium and comprises 10% of fetal calf serum, 100 U / mL of penicillin, 100 mg / L of streptomycin, a mixture of DMEM culture fluid and F12 culture fluid and multiple growth factors secreted by bone cells in the sclerite culture process. According to the invention, bone marrow mesenchymal stem cells of a mouse are purified by replacing the cell culture fluid through an adherent cell passage method, the obtained cells of the first generation are induced, and the supernatant of the sclerite complete culture medium cultured for 72-96 hours is used as the solvent of osteoblast differentiation inducer, thereby obviously improving the in vitro osteogenic differentiation efficiency of bone marrow mesenchymal stem cells.

The invention provides a method for enhancing bone formation, inhibiting osteoclastic differentiation and / or activating osteoblastic differentiation whereby to manage, treat or achieve prophylaxis of bone disease which comprises administering to a human or animal subject suffering from, or susceptible to bone disease a therapeutically or prophylactically effective amount of a lanthanum compound.

A 3D porous scaffold material for tissue engineering to induce the generation of bone tissue is prepared from chitosan, gelatin, GPSM and calcium nitrate through sol-gel process, chemical treating and freeze drying. Its advantages are high bioactivity and degradability, proper porosity and pore diameter, and controllable properties.

The invention provides methods and compositions for differentiating stromal cells from adipose tissue into cells having osteoblastic properties, and methods for improving a subject's bone structure. The methods comprise culturing stromal cells from adipose tissue in β-glycerophosphate and ascorbic acid and / or ascorbate-2-phosphate for a time sufficient to allow differentiation of said cells into osteoblasts. Such methods and compositions are useful in the production of osteoblasts for autologous transplantation into bone at a surgical site or injury. The compositions comprise adipose stromal cells, a medium capable of supporting the growth of fibroblasts and amounts of β-(glycerophosphate and ascorbic acid and / or ascorbic-2 phosphate sufficient to induce the differentiation of said stromal cells into osteoblasts.The invention further provides methods of identifying compounds that affect osteoblast differentiation. Such compounds are useful in the study of bone development and in the treatment of bone disorders, including bone fractures and osteoporosis.

The present invention relates to an application of iridoid in the preparation of anti-osteoporosis medicines, which belongs to the medicines technical field. The invention provides an application of iridoid which is shown in a general formula I in the preparation of medicines used for treating and / or preventing osteoporosis. Experiments found that ALP activity in UMR106 cells is taken as guidance for tracing and separating an extract of iridoid possessing general formula I, the extract and each separated component are researched from the aspects of osteoblast propagation and antioxidation activity, the iridoid possessing the general formula I has the effects of substantially promoting alkaline phosphatase activity and osteoblast differentiation, so that the iridoid has anti-osteoporosis effect and can be used for preparing the medicines used for treating or preventing osteoporosis. The component of specnuezhenide and a compound G13 possesses the effects of substantially promoting the alkaline phosphatase activity and osteoblast differentiation, and the specnuezhenide has effects for obviously promoting UMR-106 cell proliferation, a better effect is obtained when the specnuezhenide is used for anti-osteoporosis effect.

The invention provides a kit and method for inducing differentiation from bone mesenchymal stem cells to osteoblasts. The kit comprises an osteoblast inducing culture solution and an osteoblast identification solution, wherein the inducing culture solution comprises a cell culture solution A, a cell culture solution B, a cell culture solution C, a cell culture solution D and a cell culture solution E; the osteoblast identification solution comprises a cell immobilizing solution, coloring agents and a detergent; the cell culture solution A is an alpha-minimum essential medium (MEM) liquid culture medium; the cell culture solution B is fetal bovine serum; the cell culture solution C is a dexamethasone solution; the cell culture solution D is a beta-sodium glycerophosphate solution; the cell culture solution E is a vitamin C solution; the cell immobilizing solution is 4% paraformaldehyde; the coloring agents include sodium thiosulfate, silver nitrate and neutral red; and the detergent is double distilled water.

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.

The invention discloses an application of lnc (long non-coding) RNA lnc-DIF. The technical scheme is that a femur sample of an old osteoporotic mouse model and a femur sample of a hindlimb-unloaded osteoporotic mouse model are detected firstly, and high expression of the lnc RNA lnc-DIF in bone tissue of the old and hindlimb-unloaded osteoporotic mouse models is discovered. An RNA interference sequence of the lnc RNA lnc-DIF is further designed and prepared, osteogenic differentiation function experiments discover that the lnc RNA lnc-DIF inhibits osteogenic cell differentiation and can be specifically bound to osteogenesis related miR-489-3p. Accordingly, the lnc RNA lnc-DIF can be used for preparing drugs for treating osteoporosis caused by various factors as well as other diseases of a skeletal system.

Disclosed are methods and compositions comprising a novel osteoblast-specific transcription factor designated Osf2 / Cbfa1. Also disclosed are nucleic acid segments encoding this polypeptide derived from human cell lines, and the use of these polynucleotides in a variety of diagnostic and therapeutic applications. Methods, compositions, kits, and devices are also provided for identifying compounds which are inhibitors of osteoblast differentiation, and identifying Osf2 / Cbfa1 polynucleotides and polypeptides in a sample. Also disclosed are nucleic acid compositions comprising an Osf2 promoter, and the use of the promoter in heterologous and homologous gene transcription and protein production.

The invention provides an expansion in vitro purification culture method of mesenchymal stem cells and a cell culture medium used in a culture process. According to the method, mesenchymal stem cells differentiate to osteogenic stem cells. The cell culture medium comprises the following components: 8-12mu g / L of bFGF (Basic Fibroblast Growth Factor), 8-12mu g / L of EGF (Epidermal Growth Factor), 8-12mu g / L of PDGF-BB (Platelet Derived Growth Factor-BB), 10 percent of fetal calf serum, 100U / mL of penicillin and 100mg of L of streptomycin; a solvent is a basic culture medium; and the basic culture medium is a DMEM (Dulbecco Modified Eagle Medium) culture solution, an F12 culture solution or a mixture of the DMEM culture solution and the F12 culture solution. The mesenchymal stem cells obtained by adopting a method comprising the steps of early replacement of the cell culture medium, cell adherence passage and combined addition of cell factors (bFGF, EGF and PDGF-BB) in the culture solution are high in purity and strong in vitro proliferation capacity, and have good capability of differentiation to osteogenic stem cells.

The present invention discloses a 3D printing composite magnetic metal scaffold preparation method, which comprises: establishing a three-dimensional model with a porous structure, printing a three-dimensionally-penetrating porous metal scaffold by using a 3D printing technology, and modifying the porous metal scaffold with poly dopamine and Fe3O4@PDA composite magnetic nanoparticles through a one-step method so as to obtain the 3D printing composite magnetic metal scaffold. According to the present invention, the 3D printing composite magnetic metal scaffold can meet the requirement of the bone repair material on the mechanical strength, and further has advantages of advantages of good biocompatibility, good hydrophilicity and reduced cytotoxicity, wherein the coated material has weak magnetism, such that the osteoblast differentiation can be easily achieved, and the osteogenesis is easily achieved; and the preparation method has advantages of simple and easy operation, rapidness, high efficiency and easy production.

The invention discloses a two-layer composite periodontal defect repair material based on photo-crosslinking hydrogel and a preparing method of the material. The periodontal defect repair material isprepared with biomacromolecules GelMa and synthetic macromolecules PEGDMA which both have photo-crosslinking features. The two-layer composite hydrogel material can integrate the advantages of a GelMAfiber hydrogel layer and the advantages of a PEGDMA hydrogel layer, so that the two-layer composite hydrogel material has the shapes of three-dimensional fibers and communicated pores, good biocompatibility, high hydrophilicity and excellent mechanical performance and stability. The GelMA fiber hydrogel layer can regulate and control differentiation of periodontal ligament stem cells towards osteoblasts through fiber size on the condition that no growth factor is added, and tissue regeneration of the alveolar bone is effectively promoted; the PEGDMA hydrogel layer can effectively stop soft tissue (gingival tissue) from invading a defect area, and an effective space is provided for the tissue regeneration of the alveolar bone; in the meanwhile, the transportation of nutrient substances isnot influenced while the stopping effect is achieved.

It is intended to provide an osteogenesis promoter capable of promoting osteogenesis by promoting the differentiation of osteoblasts, and foods, drinks, drugs or feeds for promoting osteogenesis. Namely, an osteogenesis promoter capable of promoting osteogenesis by promoting the differentiation of osteoblasts which comprises, as the active ingredient, lactoperoxidase and / or a digestion product obtained by digesting lactoperoxidase with a protease such as trypsin; and foods, drinks, drugs or feeds for promoting osteogenesis containing lactoperoxidase and / or its digestion product.

The invention provides a method for inducing bone mesenchymal stem cells to differentiate into osteoblast. The method is characterized in that little osteoblast is added in a bone mesenchymal stem cell culture system to form an osteogenesis micro-environment; the quantity of the osteoblast is more than 30% of the total cell quantity preferably. The method firstly uses the osteogenesis micro-environment provided by little osteoblast, induces the BMSCs to form mature bone tissue in vitro by a co-culture method and has a wide prospect in the field of bone tissue restoration and reconstruction.

The present invention provides a method of inducing undifferentiated cells into induced osteoblasts. The method includes: A) a step of providing an induced osteoblast differentiation inducing agent obtained by culturing chondrocytes capable of hypertrophication in a differentiation agent producing medium containing dexamethasone, β-glycerophosphate, ascorbic acid and a serum component; and B) a step of culturing the undifferentiated cells in an undifferentiated cell culture medium containing the induced osteoblast differentiation inducing agent and a medium component, to differentiate the undifferentiated cells into the induced osteoblasts.

The invention relates to prevention and treatment of bone diseases caused by glucocorticoid medicines. Concretely, the invention relates to application of Wnt signal pathway to prevention and treatment of bone diseases caused by glucocorticoid medicines. Expression of C / EBPalpha factor is down regulated at a later period that BMP2 induces a mouse mesenchymal stem cell line C3H10T1 / 2 to be differentiated into osteoblast, and overexpression of the factor can inhibit differentiation of osteoblast. The reason causing down-regulated expression of C / EBPalpha is that distal promoters (-1286bp / -1056bp) of C / EBPalpha are subjected to hypermethylation at the differentiation later-period of osteoblast and are silenced. Interruption of the methylation process promotes high expression of C / EBPalpha and leads to osteogenic / adipogenic differentiation unbalance. A Dex-induced osteoporosis model proves that Wnt / beta-catenin signal pathway has important regulation and control effects on expression and methylation of C / EBPalpha in differentiation of osteoblast. A stimulant / activator of the pathway is capable of rescuing osteogenic / adipogenic differentiation unbalance caused by Dex.

The invention belongs to the technical field of tissue engineering and gene delivery, in particular to a double-gene activated bone-cartilage compound transplant, a preparation method and application thereof. In the method, seed cells are respectively planted on two layers of bracket materials which are activated by different genes and have the same main constituents, and the seed cells of the upper layer and the lower layer are simultaneously induced to respectively differentiate to chondrocyte and osteoblast by utilizing an in situ gene transfection principle so as to form a tissue engineering bone-cartilage compound transplant in vitro, wherein the seed cells are bone mesenchymal stem cells; the main constituents of the bracket materials are chitosan and gelatin; and in the two different genes, one is a gene differentiated from any known induced stem cell to the chondrocyte, while the other one is a gene differentiated from any known induced stem cell to the osteoblast. By utilizing the method, the bone-cartilage compound transplant is prepared in vitro and further used for restoring bone-cartilage.

The invention provides methods and compositions for differentiating stromal cells from adipose tissue into cells having osteoblastic properties, and methods for improving a subject's bone structure. The methods comprise culturing stromal cells from adipose tissue in β-glycerophosphate and ascorbic acid and / or ascorbate-2-phosphate for a time sufficient to allow differentiation of said cells into osteoblasts. Such methods and compositions are useful in the production of osteoblasts for autologous transplantation into bone at a surgical site or injury. The compositions comprise adipose stromal cells, a medium capable of supporting the growth of fibroblasts and amounts of β-glycerophosphate and ascorbic acid and / or ascorbic-2 phosphate sufficient to induce the differentiation of said stromal cells into osteoblasts. The invention further provides methods of identifying compounds that affect osteoblast differentiation. Such compounds are useful in the study of bone development and in the treatment of bone disorders, including bone fractures and osteoporosis.

The invention discloses a micro RNA which can induce differentiation from stem cells to osseous cells and an application thereof. The micro RNA disclosed by the invention is one of the following RNAs, namely a double-stranded RNA of a1) or a single-stranded RNA of b1): a1) the nucleotide sequence of one strand is a sequence 1-9 in a sequence table while that of the other strand is a reverse complement to the sequence 1-9 in the sequence table; b1) a recombinant RNA molecule is obtained by connecting a single-stranded RNA with the nucleotide sequence which is the sequence 1-9 in the sequence table and a single-stranded RNA with the nucleotide sequence which is a reverse complement to the sequence 1-9 in the sequence table by a connection sequence; and the connection sequence is TTCAAGAGA. The micro RNA can be used for preparation of medicaments for the treatment of orthopedic disorders and the medicaments can be adopted for the treatment of the orthopedic disorders of osteoporosis, bone fracture, hairline fracture, femoral head necrosis and the like.

The invention belongs to the technical field of cytobiology and bone tissue engineering, relates to a compound used for treating osteoporosis, and especially relates to an application of raloxifene in mesenchymal stem cell in-vitro osteoblast differentiation. According to the invention, raloxifene is added during a mesenchymal stem cell in-vitro osteoblast differentiation culturing process. With the application, mRNA and protein level expressions of osteoblast differentiation marker genes ALP and BMP2, and calcium ion sedimentation can be increased; and mesenchymal stem cell differentiation towards osteoblast can be promoted. The invention relates to the application of raloxifene in mesenchymal stem cell in-vitro osteoblast differentiation, and in treatments of diseases such as fractures,bone defects, and bone wounds.

The invention discloses application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis. According to the technical scheme, the detection of thighbone tissue samples of elderly patients with osteoporosis shows that miR-489-3p is low in expression in human osteoporosis tissue samples and can be used as a marker gene for diagnosing osteoporosis; an analogue and an antagonist of miR-489-3p are designed and prepared, then the analogue and the antagonist of the miR-489-3p are transfected into mice pre-osteoblasts MC3T3-E1 for carrying out a mineralization-alizarin red s dyeing experiment and an ALP dyeing experiment; technical detection of qPCR and Western blot shows that miR-489-3p can accelerate osteocyte differentiation and also can carry out targeted regulation on non-receptor protein-tyrosine-phosphatase6 (PTPN6) molecules. The application of miR-489-3p to preparation of medicines for diagnosing and treating human osteoporosis shows that miR-489-3p can regulate PTPN6 molecules to be used as target sites of action of medicines for treating skeleton system diseases such as osteoporosis.

The present invention identifies genes whose expression pattern is altered when pre-osteoblastic cells undergo differentiation into mature osteoblasts. The genes identified may be used as markers for the differentiation process. The present invention also provides methods to screen agents that are capable of modulating the differentiation process. The present invention also provides methods of identifying therapeutic agents that stimulate bone formation by analyzing the expression of one or more of the genes identified.

The invention belongs to the field of biomedical engineering, and concretely relates to a method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes. A simple, ecologic, safe and effective method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes is lacked in the prior art. The method for inducing oriented differentiation bone marrow mesenchymal stem cells into osteocytes, provided by the invention, comprises the following steps: 1, constructing a porous calcium phosphate scaffold; 2, compounding collagen to the scaffold obtained in step 1 to form a fiber network structure; and 3, inoculating the fiber network with bone marrow mesenchymal stem cells, pre-culturing the bone marrow mesenchymal stem cells for 4-48 h, and culturing the bone marrow mesenchymal stem cells in an ossification induction medium. The calcium phosphate-collagen scaffold constructed in the invention provides a suitable microenvironment, and the collagen is compounded to the calcium phosphate scaffold to realize the cascade amplification of a signal among mediated cells; and the bone marrow mesenchymal stem cells are cultured in the ossification induction medium to enhance the expression of ossification associated transcription factors and promote the differentiation of the bone marrow mesenchymal stem cells into the osteocytes.

The invention belongs to the field of biomedical engineering, and particularly relates to a method for inducing mesenchymal stem cells to be directionally differentiated into osteoblasts. The method for inducing the mesenchymal stem cells to be directionally differentiated into the osteoblasts has the advantages that the problem of deficiency of a simple, convenient, economical, safe and effectivemethod for inducing mesenchymal stem cells to be differentiated into osteoblasts in the prior art can be solved; macromolecular three-dimensional structures are constructed, appropriate micro-environments are provided, collagen composition and mediation intercellular signal cascade amplification are adopted, culture is carried out on the mesenchymal stem cells in osteogenic induction culture media, accordingly, expression of osteogenesis-related transcription factors can be strengthened, and the mesenchymal stem cells can be promoted to be differentiated into the osteoblasts.

The invention relates to a method of in-vitro amplification and purification culture of mesenchymal stem cells in differentiation from mesenchymal stem cells to osteoblasts. A used cell culture medium comprises the following components: 5-20% of PL platelet lysate, 2-5U / ml of heparin, 5-10microgram / ml of doxycycline, and a basic culture medium DMEM or a basic culture medium AMEM. According to the method disclosed by the invention, the method comprising steps of separation and extraction of mesenchymal stem cells, cell inoculation, cell culture, cell culture fluid replacement and cell propagation are adopted, and the obtained mesenchymal stem cells are high in purity, high in in-vitro amplification capacity, and good in the capacity of differentiation to osteoblasts.

The invention discloses a BMP-2 loaded strontium substituted hydroxyapatite / silk fibroin / heparin composite scaffold and a preparation method and an application of the BMP-2 loaded strontium substituted hydroxyapatite / silk fibroin / heparin composite scaffold. The advantage that the silk fibroin and the strontium substituted hydroxyapatite are used in bone repair is taken; the heparin is combined onthe composite scaffold to slowly release a growth factor; the BMP-2 is then adsorbed, the growth factor is slowly released to promote the in vitro proliferation of a marrow stroma mesenchymal cell andinduce the marrow stroma mesenchymal cell to differentiate into a bone precursor cell, meanwhile, the doping of the strontium element effectively improves the bone induction capability, the mesenchymal stem cell in the body is collected to differentiate to a bone cell, and the composite scaffold is expected to be widely applied in the field of biological materials.