77 results about "RUNX2" patented technology

This invention provides a method for obtaining a gene involved in regulation of cartilage differentiation, in which a transcription factor, preferably Runx2 / Cbfa1, is forcedly expressed in a cell that is deficient such transcription factor, preferably in a Runx2 / Cbfa1-deficient chondrocyte, and the gene, the expression of which is thereby induced, is selected using DNA chip analysis, subtraction, or other means as well as a Runx2 / Cbfa1-deficient chondrocyte useful for carrying out such method. The invention also provides a polynucleotide obtained by such method, a polypeptide encoded by such polynucleotide, an antibody against such polypeptide, a recombinant vector comprising such polynucleotide, a transformant comprising such recombinant DNA vector, a cell expressing such polypeptide, a transgenic animal of such polynucleotide, an animal model of a bone and / or joint disease (preferably osteoarthritis), a method for screening for a therapeutic agent and / or prophylactic agent for a bone and / or joint disease (preferably osteoarthritis) using the aforementioned objects, a candidate compound for a therapeutic agent and / or prophylactic agent selected by such method, a pharmaceutical composition for a bone and / or joint disease (preferably osteoarthritis), and a method for diagnosing such disease.

A plasmid, viral or linear DNA molecule containing a nucleic acid sequence derived from the promoter region of the hCol1α2 gene, which is selectively transported into the nuclei of cells in the osteoblast lineage. The sequence can be used independently as a nuclear entry sequence only, and / or as a nuclear entry sequence without regard to position, in a vector or linear DNA that directs gene expression and nuclear entry. The disclosure further includes a chimeric DNA sequence derived by the addition of osteoblast-specific enhancer sequences to the nuclear entry sequence / promoter sequence, to increase osteoblast-specific expression while retaining osteoblast-specific nuclear import. An enhancer sequence is derived from the promoter region of the human Core Binding Factor alpha 1 (Cbfa1 / Runx2) gene. The Cbfa1 / Runx2 promoter can be added to the sequence derived from, or alternatively, comprising the promoter region of the hCol1α2 gene. Also provided are methods of use of the novel sequences.

The invention discloses a biological agent for promoting osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSC), and a pharmaceutical composition for prevention and / or treatment ofadolescent idiopathic scoliosis (AIS). The inventor finds that lncAIS is key lncRNA involved in progress of AIS, wherein the lncAIS interacts with NF90 to promote HOXD8mRNA stability, thus transcription of RUNX2 in the BM-MSC is enhanced, and thus osteogenic differentiation of the normal BM-MSC is caused. By contrast, the NF90 cannot be collected due to down-regulating of the lncAIS in the BM-MSCof an AIS patient , thus the HOXD8mRNA stability is eliminated, and in this way, transcription of the RUNX2 for osteogenic differentiation is hindered. Thus, the BM-MSC of overexpression lncAIS, HOXD8or the RUNX2 can be used for preparing the biological agent for promoting osteogenic differentiation of the BM-MSC and the pharmaceutical composition for prevention and / or treatment of the AIS, and thus the new means is provided for prevention and / or treatment of the AIS.

The invention discloses application of vibration stimulation in regulation of in vitro osteogenesis and adipogenic differentiation of bone marrow-derived mesenchymal stem cells and belongs to the technical field of cell biotechnology and bone tissue engineering. Osteogenesis differentiation culture and adipogenic differentiation culture are respectively carried out on bone marrow-derived mesenchymal stem cells of people in vitro, vibration stimulation is carried out once a day and every time is 20-45 minutes, and the in vitro osteogenesis differentiation and adipogenic differentiation of bone marrow-derived mesenchymal stem cells are facilitated to be promoted. Vibration stimulation is applied to the bone marrow-derived mesenchymal stem cells, formation of calcium of extracellular matrix and expression of osteogenesis differentiation marker genes of RUNX2 and COL1A1 in mRNA level can be upwards regulated, generation of lipid of epimatrix and expression of adipogenic differentiation marker genes of PPARG2 and CEBPA mRNA can be restrained, and a safe and efficient seed cell source is provided for promoting stem cell osteogenesis differentiation to generate more functional osteoblasts.

The invention provides an effect of long-chain non-coding RNA in adipogenic differentiation and osteogenic differentiation of stem cells, and belongs to the technical field of stem cells. Experimentsprove that in the adipogenic differentiation process of mesenchymal stem cells, the expression quantity of the LINC02075 is obviously increased, inhibition of LINC02075 can obviously inhibit mRNA andprotein expression of adipogenic differentiation related genes PPAR gamma, C / EBP alpha and AP2, thereby inhibiting the adipogenic differentiation of the mesenchymal stem cells. Meanwhile, the invention proves that inhibition of LINC02075 can significantly promote mRNA and protein expression of osteogenic differentiation related genes RUNX2, ALP and OCN to promote osteogenic differentiation of themesenchymal stem cells

The present invention relates to a method to enhance the activity of Runx2, a major target protein of Bone Mophogenetic Protein (BMP), by Runx2 acetylation, more precisely, a method to activate BMP-mediated bone formation pathway by protecting Runx2 from ubiquitination, indicating that Runx2 is protected from degradation by the increase of Runx2 acetylation making the protein more stable. The method to enhance Runx2 activity of the present invention can be utilized for the prevention and the treatment of bone disease such as osteoporosis, osteogenesis imperfecta, periodontal disease and fracture, by inducing bone formation by inhibiting Runx2 degradation.

The invention discloses application of aloin in improvement of osteogenic capability of a bone mesenchymal stem cell (BMSC). According to the application of the invention, the BMSC is treated by aloins with different concentrations, and cell viability of the BMSC is detected; influence of the aloin on osteoblast differentiation of the BMSC is evaluated by adopting alkaline phosphatase activity andmineralization experiments; real-time fluorescent quantitative PCR detection is carried out for detecting expression situation of osteogenesis-related genes such as Runx2, ALP, OCN and OPN; variationof an ERK1 / 2-Runx2 signal channel is detected by virtue of a Western blotting process and an immumofluorescence process; and results show that the aloin promotes the osteoblast differentiation of theBMSCs by activating the ERK1 / 2-Runx2 signal channel, the osteogenic capability of the BMSCs is improved, and the research results provide data support for research and development of a new drug usedfor preventing and treating osteoporosis, so that the aloin has a greater application prospect.

The invention relates to a method for improving adipose-derived stem cell differentiation efficiency through gene editing. After the optimal gRNA sequence is obtained through screening and optimization, miR-221 is knocked out to obtain transgenic adipose-derived stem cells, a specific monoclonal antibody is obtained through RUNX2 screening, the monoclonal antibody can inhibit the activity of RUNX2, and after the monoclonal antibody is added into an induction medium, the adipose-derived stem cells can be specifically promoted to be differentiated into chondrocytes, and good induction efficiencyand application value are realized.

The invention encompasses compositions and methods for effectively interfering, reducing and preventing conversion of vascular smooth muscle cells (VSMCs) and circulating stem cells to osteoblastic bone-like cells, thereby reducing and / or preventing vascular calcification (VC) or calcium mineral (hydroxyapatite) deposition in the vasculature. The severity and extent of calcification in the major arteries reflect atherosclerotic plaque burden and strongly predict cardiovascular morbidity and mortality. The present inventive compositions used for administration to human and other mammalian subjects comprise select actives that inhibit, interfere or regulate the biochemical processes leading to such calcification and include (1) at least one agent that modulates expression and / or activity of peroxisome activated protein receptor gamma (PPAR-γ); (2) at least one agent that inhibits expression and / or suppresses activity of one or more of the osteogenic transcription factors (Cbfα1 / Runx2, Osterix, Msx2) and β-catenin signaling; (3) at least one agent that inhibits expression and / or suppresses activity of one or more of bone morphogenetic proteins (BMPs: BMP 2 and 4), alkaline phosphatase (ALP), and osteocalcin; (4) at least one agent that inhibits the activity of Reactive Oxygen Species (ROS); and (5) at least one agent that suppresses one or more of inflammatory mediators including interleukins IL-1α, IL-1β, IL-6, NF-κB, TNF-α, matrix metalloproteinases (MMPs) and prostaglandin E2 (PGE2). The compositions may further comprise at least one agent that promotes expression and / or carboxylation of matrix Gla protein (MGP). Advantageously, these select actives include materials such as phytonutrients, vitamins and minerals that have been broadly used in food and drink products and are safe for human and pet / animal consumption. Compositions with such combinations have the ability to prevent, treat and reverse VC not only in coronary arteries but also in other tissues capable of undergoing undesirable calcification. In addition the present compositions are effective against associated conditions or contributory factors / inducers to VC, including diabetes, obesity, hypertension, inflammation, oxidative stress, osteoporosis and arthritis.

The present invention relates to a Eupatorium spp. extract having anti-obesity effects as a result of decreasing adipocytes and increasing osteoblasts, as well as the effects of preventing bone disease or fractures by increasing osteoblasts and of preventing osteoporosis by decreasing adipocytes and increasing osteoblasts by the same proportion in mesenchymal stem cells. DCM fraction layers of a Eupatorium spp. stem extract collected on a monthly basis and Eupatorium spp. stem extract collected only in September may inhibit the activity of PPARγ, AP2, CD36, adiponectin C / EBPα, and LPL, which serve as significant factors for adipocyte differentiation in C3H10T1 / 2 cells and primary mesenchymal stem cells, which are pluripotent stem cell lines, and may increase the activity of ALP, osterix, CO1I and RUNX2, which serve as significant factors for osteoblast differentiation. The Eupatorium spp. extract of the present invention exhibits the effects of increasing bone mineral density (BMD) and decreasing adipocytes in bone marrow in an osteoporosis animal model experiment involving an ovariectomy, and therefore may be used as a useful material for preventing and treating osteoporosis.

The invention relates to a method for improving osteogenic differentiation efficiency of human amniotic mesenchymal stem cells. The method adds an induction composition in the human amniotic mesenchymal stem cells, wherein the composition comprises 0.1-2g / L of hyaluronic acid, 0-200mg / L of prostaglandin receptor-2 selective agonist, 1-100nmol / L of dexamethasone, 0-10nmol / L of beta-glycerophosphate and 1-100mg / L of ascorbic acid. Compared with a conventional inducing method, the osteogenic differentiation efficiency of the human amniotic mesenchymal stem cells can be significantly improved, expression and activity of alkaline phosphatase are promoted, and expression of osteogenesis-related genes of RunX2, Osx, BSP, Ocn, ALP, Col1a1 and the like and formation of mineralized calcium nodules are enhanced. The method for improving the osteogenic differentiation efficiency of the human amniotic mesenchymal stem cells can be applied in osteogenic differentiation of the human mesenchymal stem cells, and applied in bone diseases of bone defects, bone trauma and the like.

The invention relates to a method for co-expression of Runx2 and Osterix in mesenchymal stem cells or other non-osteoblasts by a special mode that the expression level of Runx2 is less than the expression level of Osterix, thus speeding up the induction of osteoblast differentiation. The invention provides a pharmaceutical composition for prevention or treatment of osteoblast differentiation related diseases. The pharmaceutical composition contains Runx2 protein and Osterix protein, wherein the Runx2 protein content is smaller than the Osterix protein content. The invention also provides application of the Runx2 protein and the Osterix protein in preparation of the pharmaceutical composition for prevention or treatment of osteoblast differentiation related diseases, and in the pharmaceutical composition, the content of the Runx2 protein is less than that of the Osterix protein. The pharmaceutical composition and the special co-expression mode of Runx2 and Osterix provided in the invention can be used for treatment of osteoporosis, osteogenesis imperfecta, periodontal diseases, fractures and other bone diseases, and also can be used for research, development and production of tissue-engineered bones. The invention also provides a method for screening drugs preventing and treating bone diseases.

The present invention discloses a new medical use of a growth hormone release hormone (GHRH) agonist, particularly applications of a growth hormone release hormone agonist in preparation of anti-vascular calcification drugs. According to the present invention, GHRH-A can increase the cAMP level and the protein kinase A (PKA) activity in smooth muscle cells, can decrease the expression and the activity of NAPDH oxidase in smooth muscle cells, can decrease the ROS level in smooth muscle cells, can decrease the expression and the activity of Runx2 in smooth muscle cells, can decrease the expression and the activity of alkaline phosphatase (ALP) in cells, and can decrease the inorganic phosphorus content in blood so as to reduce the calcium deposition and the phosphorus deposition in aorta, reduce the activity of ALP in aorta, and achieve the purposes of treatment or prevention of vascular calcification and arteriosclerosis.

The present invention relates to diagnosis and / or prognosis of cardiovascular disease and cardiovascular events. A prognostic risk score in relation to the cardiovascular events, more particular after intervention, can be determined by counting the number of deregulated genes (or derived proteins) in their isolated monocytes. Deregulation means low expression of COX1 and / or COX4I1, and / or TFAM, and / or RUNX2.

The application relates to flavonoid micromolecules, and in particular relates to an application of chrysin and naringenin in promotion of bone formation and differentiation process which are regulated via Runx2, Osx, Col1A1, OCN, OPN, ALP, Fra1 and NF-kB1 and in treatment of diseases related to inflammatory factor TNF-alpha mediated inflammatory bone resorption.

The invention relates to application of adipose-derived stem cells edited by a gene editing technology to improvement of differentiation efficiency. After the optimal gRNA sequence is obtained throughscreening and optimization, miR-221 is knocked out to obtain transgenic adipose-derived stem cells, a specific monoclonal antibody is obtained through RUNX2 screening, the monoclonal antibody can inhibit the activity of RUNX2, and after the monoclonal antibody is added into an induction medium, the adipose-derived stem cells can be specifically promoted to be differentiated into chondrocytes, andgood induction efficiency and application values are realized.

The invention discloses an application of miR-296 and a simulant thereof in promoting osteogenic differentiation of bone marrow mesenchymal stem cells and bone formation. The regulation effect of themiR-296 on osteogenic differentiation of the bone marrow mesenchymal stem cells and bone formation is explored through in-vitro experiments. In-vivo experiments prove that the formation of new bones can be promoted through the transfection of the miR-296, and the in-vitro experiments prove that the miR-296 can improve osteogenic differentiation ability of the bone marrow mesenchymal stem cells andpromote in-vivo bone formation by promoting the formation of osteoblast calcium nodules and the activity of a marker enzyme ALP, as well as promoting the expression of Osterix and Runx2 which are important regulatory transcription factors in the process of osteogenic differentiation. The invention provides a new technical means for the treatment of osteoporosis.

The invention encompasses compositions and methods for effectively treating and / or preventing the development and / or progression of osteoporosis and related disorders such as osteoarthritis and rheumatoid arthritis, and for promoting overall bone and joint health. This is accomplished by addressing multiple key mechanisms that lead to such disorders. The invention includes compositions comprising a combination of agents having biological activities that effectively suppress, regulate or interfere with the various key biochemical processes and mechanisms that increase the risk for development and / or progression of osteoporosis. The present compositions and methods simultaneously promote bone formation and reduce bone resorption by (a) stimulating osteoblast formation and osteogenesis; (b) suppressing adipocyte differentiation; (c) inhibiting osteoclast formation; and (d) increasing apoptosis of osteoclasts. The inventive compositions used for administration to human and other mammalian subjects having or at risk for development of osteoporosis comprise (1) at least one agent capable of modulating expression and / or activity of one or more of peroxisome activated protein receptor gamma (PPAR-γ), CAAT / enhancer binding protein-α (C / EBPα) and Sterol Regulatory Element-Binding Protein (SREBP-1); (2) at least one agent that activates expression and / or activity of one or more of the osteogenic transcription factors (Runx2 / Cbfα1, Dlx5, Osterix, Msx2); (3) at least one agent that activates expression and / or activity of one or more of bone morphogenetic proteins (BMPs: BMP 2 and 4), alkaline phosphatase (ALP), and osteocalcin; (4) at least one agent capable of activating Wnt / β-catenin signaling pathway; (5) at least one agent that inhibits the activity of pro-oxidants including reactive nitrogen species and reactive oxygen species (ROS); (6) at least one agent that suppresses one or more of inflammatory mediators including interleukins IL-1α, IL-1β, IL-6, NF-κB, TNF-α, matrix metalloproteinases (MMPs) and prostaglandin E2 (PGE2); and (7) at least one agent that induces the expression of and / or activates one or more of adenosine monophosphate-activated protein kinase (AMPK), sirtuin (SIRT1) and adiponectin (AP).

The invention belongs to the technical field of stem cells, and particularly relates to an accelerant for promoting mesenchymal stem cell osteogenic differentiation. Fluorescent quantitative PCR experiments, Western Blot experiments and alizarin red staining experiments find that bullacta exarata polypeptide can obviously promote the expression of the mRNA level and the protein level of mesenchymal stem cell osteogenic differentiation related genes ALP, BGLAP and RUNX2, and can obviously promote the mineralization level of bone marrow mesenchymal stem cells, so that the bullacta exarata polypeptide can be used as the accelerant of the mesenchymal stem cell osteogenic differentiation to promote osteogenic differentiation of the mesenchymal stem cells, and more osteoblasts are provided for treatment of osteoporosis, bone injury and other diseases.

The invention relates to application of vaccarin in the preparation of a drug for preventing angiosteosis, belonging to the field of traditional Chinese medicine preparations. Vaccarin is applied to the development of the drug for preventing the angiosteosis so as to well treat angiosteosis. A cell experiment result of the application presents that vaccarin (VAC) is capable of remarkably decreasing deposition of calcium ions of vascular smooth muscle cells (VSMCs) caused by a calcification induction medium, inhibiting the increase of alkaline phosphatase (ALP) and simultaneously inhibiting the expression of a Runt-related transcription factor 2 (runt-related transcription factor 2, Runx2) of a calcification-related protein induced by the calcification induction medium and a bone morphogenetic protein 2 (Bone morphogenetic protein 2, BMP-2); and an in-vivo animal experiment further proves that VAC is capable of effectively treating a calcification model induced by vitamin D3 and nicotine and effectively improving the angiosteosis, is a traditional Chinese medicine for treating the angiosteosis and can be taken as novel drugs, health products and nutrition products for treating the angiosteosis, and a new path and measure are provided for treating angiosteosis-related drugs.

The invention relates to the field of prevention and / or treatment of CAVD. The inventor finds that caffeic acid phenethyl ester (CAPE) can effectively down-regulate the expression of osteogenic differentiation marker genes RUNX2 and ALP in valvular interstitial cells to inhibit the calcification of the valvular interstitial cells, thereby suppressing or delaying the valve calcification; and thus application of caffeic acid phenethyl ester in preparation of the medicine for preventing and / or treating the individual CAVD is provided. Therefore, novel selection is provided for treating the CAVD by using a non-surgical method.

The invention provides application of astragalus saponin I to preparing dug for treating osteoporosis. Monomer compounds of astragalus saponin I (namely As-I) which can promote osteoblast differentiation are screened from traditional Chinese medicine astragalus, the osteogenic differentiation promoting function of astragalus saponin I is comprehensively verified, it is definite that astragalus saponin I has the effect of improving the expression level of bone formation key factors of Runx2, BMP-2, BGP and beta-catenin through a Wnt pathway, and therefore osteoblast differentiation is promoted; astragalus saponin I can down-regulate the RANKL / OPG expression ratio so that osteoclast differentiation can be inhibited, the purpose of treating osteoporosis is further achieved, and astragalus saponin I has great significance in osteoporosis treatment. Astragalus saponin I serves as a basis and is compounded with pharmaceutic adjuvants to form medicine for treating osteoporosis, and a new idea is provided for treating the disease.

The invention provides application of long chain non-coded RNA-NKILA in bone tissue damage repair. Experiments verify that overexpressed NKILA can promote calcium ion deposition in a mesenchymal stemcell obviously, enhance the activity of alkaline phosphatase in the mesenchymal stem cell obviously and improve expression quantities of osteogenic differentiation markers RUNX2, SP7 and SPP1 genes inthe mesenchymal stem cell obviously.

The invention discloses the application of KGN to preparation of drugs for improving the osteogenic differentiation capability of BM-MSCs, belonging to the field of cell medicine. Experimental resultsshow that KGN can promote the osteogenic differentiation of BM-MSC and improve the gene expression levels of SIRT1, ALP, COL1A1, RUNX2 and BGLAP and the expression contents of corresponding proteins.Therefore, the application of invention proves that KGN can promote the osteogenic differentiation potential of stem cells. The application KGN to preparation of drugs for improving the osteogenic differentiation capability of BM-MSCs in the invention can provide a brand-new bone-healing-capability-promoting scheme for medical research, tissue engineering research and the like.

According to the present invention, based on the exogenous introduction method of bone tissue cell specific expressing transcription factor Runx2 into human skin fibroblasts, a set of culture system for promoting differentiation of fibroblasts at various stages is developed so as to effectively promote transformation into osteoblasts, and the prevent invention belongs to the field of cell biology and tissue engineering. The system comprises three sets of culture liquids, wherein the culture liquids respectively act on three key stages in the transdifferentiation process, the high survival rate and the high proliferation of the RUNX2 positive cells are maintained in the stage a, the expression of the RUNX2 within a specific time is regulated in the stage b, and the transformation efficiency from the cells to the osteoblasts is improved and the transformation time is shortened after the cells are re-programmed so as to achieve the transdifferentiation of the human skin fibroblasts to the osteoblasts. According to the present invention, the three sets of the culture liquids are matched, and the respective acting time can be flexibly regulated according to the final use purpose of the cells so as to culture the large-scale inductive osteoblasts; the culture system is used for cell therapies of bone metabolic diseases, fractures and the like; and the pharmaceutical composition of the present invention can further be used for development and research and production of the tissue engineering bonds, provides convenience for basic research, and provides prospects for clinical applications.

The invention discloses application of bortezomib in preparation of medicine for treating osteosarcoma. A new function of the proteasome inhibitor bortezomib is provided, that is, the bortezomib can be used for directionally inducing osteogenic differentiation of osteosarcoma and can be used for treating the osteosarcoma. Through experimental verification, while bortezomib of medium and high concentrations induces apoptosis of osteosarcoma cells, bortezomib of low concentration can further induce osteogenic differentiation of the osteosarcoma cells, specifically, calcific nodules in the osteosarcoma cells F5M2 and U-2 OS are increased by the bortezomib; alkaline phosphatase activity is increased; and expression of osteogenic differentiation markers (ALP, OPN, Runx2) is increased.

The invention provides a method for improving the osteogenesis ability of mesenchymal stem cells, which comprises the following steps: inoculating the mesenchymal stem cells into an alpha-MEM culturemedium containing 10% of fetal calf serum, culturing, digesting and subculturing by 0.2% of pancreatin when the fusion degree of the mesenchymal stem cells is 80-85%, inoculating the obtained mesenchymal stem cells into an alpha-MEM culture medium containing 10% of fetal calf serum for culture, and adding osteogenesis induction liquid and a metformin aqueous solution for induction when the densityof the mesenchymal stem cells is greater than 90% for induction for 14-21 days. The metformin acts on the mesenchymal stem cells, alizarin red staining results show that the metformin can significantly improve the calcium mineralization nodule degree of the cells, and it is indicated that the metformin has a significant induction effect on mesenchymal stem cell osteogenic differentiation, the metformin can significantly improve the expression of the mesenchymal stem cell osteogenic differentiation related genes SIRT1, ALP, COL1A1, RUNX2 and BGLAP, and the improvement of the osteogenic differentiation capacity of the mesenchymal stem cells is of great significance to stem cell transplantation and bone biology research.

The present invention relates to the field of prevention and / or treatment of calcific aortic valve disease. Researches show that andrographolide can effectively down-regulate expressions of osteogenicdifferentiation marker genes RUNX2 and ALP in valvular interstitial cells, inhibit valvular interstitial cell calcification, and thus inhibit or delay valve calcification, thereby providing a use ofthe andrographolide in preparation of medicines for preventing and / or treating the individual calcific aortic valve disease. A new option for using a non-surgical method to treat the calcific aortic valve disease is provided.

The invention provides an application of a soybean peptide in promoting osteogenic activity, which comprises the following steps: detecting the proliferation level of osteoblasts, the activity of a marker ALP in the early stage of differentiation and a marker OCN in the later stage of differentiation, and detecting that the soybean peptide CBP promotes the proliferation and differentiation of the osteoblasts on the biochemical level; in the process of promoting osteoblast differentiation by the soybean peptide CBP, three classical signal pathways, namely ERK, JNK and p38 bypasses, of an MAPK signal pathway participate in the process and a key signal transduction step; the soybean peptide CBP promotes gene expression and phosphorylation level of a key factor Runx2 of an MAPK signal channel in osteoblasts.