35 results about "Cells bone" patented technology

The invention discloses a tissue engineering bone / cartilage double-layer scaffold and a construction method and application in medical science. The method adopts natural cartilage and bone as materials, respectively removes the antigenicity of the cartilage and the bone through de-cell, and adopts a freezing and drying method to prepare the double-layer scaffold simultaneously suitable for growthof the two issues of the bone and the cartilage. A preparation method comprises that: acellular bone is soaked in suspension of cartilage acellular matrix microfilaments, and the double-layer scaffoldof which the upper layer is a cartilage acellular matrix three-dimensional porous spongy scaffold and the lower layer is an acellular bone scaffold is prepared by the freezing and drying method and acrosslinking method. The cartilage acellular matrix spongy scaffold part of the double-layer scaffold can be introduced with factors promoting the formation of the cartilage, and the acellular bone part can be introduced with factors promoting the formation of the bone, and the factors can be respectively inoculated with cells for forming the cartilage or the bone to construct a living tissue engineering bone / cartilage complex, which is used for repairing bone and cartilage complex defect or total joint defect clinically.

A cell-removed heterogeneous bone matrix is prepared from the rib and extremity bone of pig through physical and chemical processing, and removing cells and the heterogeneous protein from tissue by use of proteolytic enzyme and Triton-X100. Its advantages are natural components and biologic chemical strength, low antigenicity, high compatibility and bone conductivity, and low cost.

Disclosed is a construction method of sclerotomal cell idiosyncratic transcription factor - 2 gene decorated tissue engineered bone through employing a natural biologically derived material (acellular bone matrix) and seed cells (osteoblast specific transcription factor-2 gene modified mesen chymal stem cells), carrying out extracorporal dynamic culture and restoration for large segment long bone defection. The method can realize the clinic cicatrisation of bone defections.

The invention discloses an osteochondral stent as well as a preparation method and application thereof. The osteochondral stent comprises a cartilage layer, an adhesion layer and a bone layer, wherein two sides of the adhesion layer are respectively connected with the cartilage layer and the bone layer; one or more of the cartilage layer, the adhesion layer and the bone layer are of a porous structure. The osteochondral stent can be used for repairing osteochondral defects, can fully capture cells, has a strong osteochondral injury repairing function, and is simple and convenient to operate.

The described invention provides soft tissue grafts, hard tissue grafts, and composite soft / hard tissue grafts and methods of producing such grafts. The grafts comprise a three-dimensional carrier matrix, a growth factor composition comprising an autologous platelet-rich fibrin and a cell culture composition comprising a culture medium, a population of cells suspended in the culture medium, and cells impregnated on or in a surface of osteoconductive particles.

The invention relates to an acellular bone matrix composite with a partially anti-freezing function and a cell capturing function and a preparation method thereof, and the bone matrix material is formed by assembling sulfuric acid polysaccharide with an anti-freezing function and chitosan or fibronectin onto the surfaces of an acelluar bone matrix. The preparation method of the acellular bone matrix composite comprises the following steps: firstly, preparing the acellular bone matrix material; and then assembling the sulfuric acid polysaccharide and other materials with the anti-freezing function and the chitosan or the fibronectin onto the surfaces ( the inner surface and the outer surface) of the acelluar bone matrix in a layer upon layer mode under an intermolecular electrostatic interaction or ligand interaction method so as to form a controlled-release coating. The composite retains the natural structure, biological characteristics and low immunogenicity of bone tissues, and simultaneously the composite coating has favorable anti-freezing function and seeded cell capturing function. The composite serving as a material for filling bone defects or a tissue engineering bone bracket has obvious function of promoting vascularization, and has scientific, reasonable, simple, convenient and feasible processing and manufacturing.

The invention discloses a method for culture of urine-derived pluripotent stem cells by virtue of in vitro small molecule induction. The method comprises the following steps: obtaining urine-derived cells from a human urine sample; carrying out in vitro small molecule induction and culturing on the obtained primary cells to obtain P1 generation hUSCs clones growing with adherence; picking up the hUSCs clones in good growing state to inoculate; and continuously carrying out continuous cell culture to obtain hUSCs with the cells which are elongated and good in state. The hUSCs prepared by the method disclosed by the invention, by virtue of induction, can be directionally differentiated to osteoblasts, adipose cells, skeletal muscle cells, nerve cells, fibroblasts or smooth muscle cells. The hUSCs are transplanted into a diabetic lower extremity ischemia model, and research results show that the hUSCs can remarkably enhance blood flow recovery and angiogenesis in an ischemia part. The hUSCs prepared by the method can be applied to treating and repairing diseased tissues and organs of diabetes and diabetic complications, cardiovascular, cerebrovascular and renovascular diseases, Alzheimer's disease, osteoporosis, arthritis and the like as well as early-warning tumors of the urinary system and screening cell drugs.

The invention discloses acellular bone matrix hydrogel with reserved natural hydroxyapatite and a preparation method of the acellular bone matrix hydrogel, and belongs to the field of bone regeneration and repair materials in tissue engineering. According to the organic-inorganic composite hydrogel, cortical bones or cancellous bones from the same source or different sources are used as raw materials, natural bone minerals of different degrees are reserved through degreasing, decellularization and decalcification treatment, then enzymolysis digestion and purification treatment are conducted, the pH and salt concentration of a solution are adjusted to physiological conditions, and then a pre-gel solution is obtained; and when in use, the temperature is raised to 37 + / -1 DEG C, and the hydrogel is formed through self-assembly. According to the bone tissue decellularized extracellular matrix hydrogel, natural hydroxyapatite components in bone matrixes are effectively reserved, hydroxyapatite and collagenous fibers are well combined and uniformly dispersed, and the bone tissue decellularized extracellular matrix hydrogel can be used as temperature-sensitive injectable hydrogel to be applied to bone tissue engineering and has bright application prospects.

The present invention proposes formative agent of protein complex, in which a polyphenol is useful component, and the agent is useful as gene complex, cell adhesion inhibitor or immune tolerogen. The polyphenol of forming the agent is selected from catechin group consisting of epigallocatechin-gallate, tannic acids, or proanto-dianisidine, a protein of the protein complex is selected from proteins consisting of animal proteins composed of polypeptide chain of peptide-combined amino acids, vegetative proteins, nucleus proteins, glycogen proteins, lipo-proteins and metal proteins, the gene complex comprises by compositing genes by polyphenol catechins in order to introduce genes to cells of animals or human bodies, a cell composed of the cell adhesion inhibitor is selected from cells consisting of an animal cell including a stem cell, skin cell, mucosa cell, hepatocyte, islet cell, neural cell, cartilage cell, endothelial cell, epidermal cell, osteocyte or muscle cell isolated from human or animal organism, or sperm, ovum or fertilized egg of domestic animals or fishes and a tissue or an organ for transplantation of the immune tolerogen is selected from the tissue or the organ consisting of skin, blood vessel, cornea, kidney, heart, liver, umbilical cord, bowels, nerve, lung, placenta or pancreas.

The invention discloses a method for fluorescence labeling of gymnosperm pollen tube fibril bone and its application and is to provide a method for fluorescence labeling of gymnosperm pollen tube fibril bone and the application of this method in microscope inspection of gymnosperm pollen tube fibril bone. The said labelling method includes the following steps: 1) fixing the gymnosperm pollen tube with 40-60mM Pipes cushion solution containing 2-4 % cavaform; 2) washing the pollen tube; 3) placing the pollen tube in 0.2-0.3 % glycerin to infiltrate for 30-60min; 4) washing the pollen tube; 5) dyeing the pollen tube and avoiding the sun with 150-250nM phalloidine by fluorescence labeling for 0.5-1.5h and fluorescence labeling the fibril bone. The invention is of simple operation and is quick, has small damage to the cells, and has good labeling effect, which will play an important role in study gymnosperm pollen tube cell bone and have a high actual application value.

The invention belongs to the field of biological materials for tissue engineering repair and tissue regeneration promotion, and particularly relates to a preparation method and application of a blended hydrogel biological scaffold material with biological activity. The blended hydrogel biological scaffold material is prepared by sequentially mixing an acellular bone meal matrix and methacrylate-modified gelatin according to a certain proportion. The preparation method of the scaffold material provided by the invention is safe, non-toxic and easy to operate, the acellular bone meal with low immunogenicity is obtained by performing acellular treatment on bones of alpha-1,3-galactoside gene knockout pigs, and the acellular bone meal has the characteristic of low immunogenicity after being implanted into a living body. The tissue engineering scaffold material is obtained by blending the acellular bone meal with biological activity and low immunogenicity with the GelMA hydrogel, the hydrogel material mixed with the acellular bone meal reserves the extracellular matrix of a natural material, the hydrogel can also provide a good supporting effect for the scaffold material, the biocompatibility is good, and thereby the scaffold material can be used as a bone tissue engineering material to be applied to hard bone defects in vivo, skull repair and the like. Compared with a traditional treatment method, the scaffold material has wide application prospects.

The invention relates to an active scaffold for multi-cell printing and nervating bone regeneration as well as a preparation method and application of the active scaffold. The multi-cell printing nervous bone regeneration active scaffold is a multi-cell printing nervous bone regeneration active scaffold containing nerve cells. The multi-cell printing nervous bone regeneration active scaffold has an upper / lower layer structure, wherein the upper layer structure is at least one layer containing a bioactive inorganic material; the upper layer structure comprises an organic hydrogel material with good biocompatibility and an upper layer support of nerve-related cells, and the lower layer structure comprises at least one layer of lower layer support comprising a bioactive inorganic material, the organic hydrogel material with good biocompatibility and bone-related cells.

A 3D decellularized bone scaffold seeded with cancer cells, such as prostate cancer cells or Ewing's sarcoma is provided. The three-dimensional includes Ewing's sarcoma (ES) tumor cells; and an engineered human bone scaffold. The engineered human bone scaffold further includes osteoblasts that secrete substance of the human bone, and osteoclasts that absorb bone tissue during growth and healing. The engineered human bone scaffold includes the tissue engineered three-dimensional model which recapitulates the osteolytic process. The engineered human bone scaffold is engineered by co-culturing of osteoblasts and osteoclasts. The osteoblast is produced by cell differentiation process from mesenchymal stem cells. The osteoclast is produced by cell differentiation from human monocytes, wherein the human monocytes are isolated from buffy coats. The scaffold can be used with cancer cell lines to identify therapeutic targets to slow, stop, and reverse tumor growth and progression as well as to predict the efficacy of potential therapeutics.

A gradient mineralized cancellous bone matrix material and a preparation method thereof are provided, and the preparation method includes: processing naturally-derived bone tissue with an immunogenicity removal treatment for decellularization, and processing an obtained decellularzed bone with a gradient demineralization treatment to obtain the gradient mineralized cancellous bone matrix material. The present invention expands a porosity of the bone matrix material and a collagen exposure degree on a surface thereof, which effectively releases growth factors and improves adhesion of the material to the cells, so as to up-regulate genes and proteins related to cell regeneration. The present invention not only retains the biomechanical properties and three-dimensional microstructure of natural bone ECM scaffolds, but also plays an active role for osteogenesis, angiogenesis and collagen mineralization in the early stage of fracture, thereby increasing engraftment adhesion of cells and promoting differentiation induction of cells.

The invention discloses a novel dolastatin 10 cyclopeptide derivative, a preparation method thereof and application thereof in antitumor drugs. The novel dolastatin 10 derivative with cyclopeptide structure may act as a novel antitumor compound, having good inhibitory effect against tumor cells, especially human colon cancer cells, leukemia cells, osteosarcoma cells and the like. The novel dolastatin 10 cyclopeptide derivative is prepared by converting a linear-chain pentapeptide intermediate, adding an amide condensing agent and carrying out liquid synthetic cyclization. The preparation method provided herein is low in cost, convenient to operate and high in efficiency. The novel dolastatin 10 cyclopeptide derivative has good inhibitory effect against cancers, especially HCT-116 human colon cancer cells, and has high stability and low toxicity; basis is laid for the development of anticancer drugs to treat colon cancer and other cancers.

The invention discloses an artificial periosteum compounded with an acellular periosteum matrix as well as a preparation method and application of the artificial periosteum. The polyvinyl alcohol / sodium carboxymethyl cellulose is adopted as a gel substrate material of the artificial periosteum, and silk fibroin and acellular periosteum matrix hydrogel are added to form a gel film material with a uniform structure. The artificial periosteum prepared by the invention is close to the physical and biological characteristics of a natural periosteum, has better biocompatibility, provides a highly hydrated 3D network structure similar to an extracellular matrix and a rough surface for cells, is beneficial to the adhesion and growth of the cells, is beneficial to the proliferation and differentiation of osteoblasts and osteogenic cells. The artificial periosteum can be used for further promoting the formation of new bones, has similar performances to periosteum in mechanics, also has good anti-hemolysis and anti-coagulation effects, is a very potential ideal material for repairing soft tissues, and has a very wide application prospect in the field of bone tissue engineering.

The invention relates to the technical field of cell culture, in particular to a stent-free periodontal ligament stem cell sphere as well as a preparation method and application thereof. The stent-free periodontal ligament stem cell spheres are prepared by utilizing a cell sphere culture plate three-dimensional culture technology and are applied to tissue engineering related to osteogenic differentiation. The stent-free periodontal ligament stem cell sphere disclosed by the invention has good stability and biological activity and relatively strong osteogenic differentiation efficiency, and can be used as a cell bone substitute material in the fields of periodontal tissue engineering and bone tissue engineering or used in combination with a scaffold material to improve the osteogenic efficiency. The stent-free periodontal ligament stem cell spheres cultured and collected by the method are uniform and consistent in size, and the cell activity can be maintained for a long time; and meanwhile, the preparation method disclosed by the invention is repeatable, simple, convenient and feasible, does not need to depend on large-scale equipment or devices, and is beneficial to quickly and effectively obtaining the cell sphere structure.

The invention relates to the technical field of bone tissue engineering, and discloses a preparation method of a functional bone repair composite scaffold. The method comprises the following steps: preparation of nano decellularized bone meal, preparation of a functional porous nano bone meal-gelatin-chitosan-dexamethasone composite scaffold, and crosslinking. The invention further discloses a nano bone meal-gelatin-chitosan-dexamethasone functional bone repair composite scaffold and application thereof. According to the invention, the functional bone repair composite scaffold has bone matrix components, cell adhesion sites, sufficient mechanical strength and a three-dimensional structure suitable for cell growth and proliferation and nutrient metabolism, the composite scaffold created by combining bone meal with gelatin and chitosan has good connectivity, the nano bone meal is uniformly dispersed in the pore wall, pores and skeleton of the scaffold, has a relatively high surface area, and facilitates the adhesion, growth and proliferation of osteoblasts, and due to introduction of the dexamethasone, the problems of tissue fibrosis and the like caused by inflammatory reaction caused by implantation of the allogenic scaffold can be effectively reduced, and the dexamethasone has an obvious promoting effect on proliferation and differentiation of the osteoblasts.

The present invention relates to: a method of preparing an osteoblast from a somatic cell of a mammal, the method including introducing a bone-related gene or an expression product thereof and a reprogramming-related gene or an expression product thereof, or introducing a reprogramming-related gene or an expression product thereof independently into the somatic cell, the bone-related gene including at least one kind selected from the group consisting of Runx2 (R), Osterix (O), and Dlx5 (D), the reprogramming-related gene including at least one kind selected from the group consisting of Oct family, c-Myc (M), L-Myc (L), Klf family, Lin-28, and Sox2; and an osteoblast prepared by the method.

The invention discloses a decellularized scaffold and a preparation method thereof.The preparation method comprises the steps that decellularized bone powder is prepared, the decellularized bone powder, hydroxyl calcium polyphosphate and PLGA are mixed and then subjected to electrostatic spinning, and a carrier scaffold is obtained; soaking the carrier scaffold in a basic fibroblast growth factor solution to obtain a basic fibroblast growth factor-loaded carrier scaffold; and inoculating osteoblasts on the carrier scaffold loaded with the basic fibroblast growth factors to obtain the decellularized scaffold. According to the preparation method, cells and fat can be efficiently removed through a specific decellularization method, then decellularized bone particle powder, hydroxyl calcium polyphosphate and PLGA are subjected to electrostatic spinning to prepare the biological scaffold, the scaffold is made to have good biocompatibility, pore diameter structure and mechanical strength, and then the biological scaffold can be used for preparing the biological scaffold. And basic fibroblast growth factors and osteoblasts are loaded, so that proliferation and differentiation of the osteoblasts can be promoted, and an osteogenesis promoting effect is achieved.

The invention pertains to the use of mesenchymal stromal cells (MSC) in the treatment of bone disorders or injuries. The invention provides MSC and preparations of specifically pooled MSC for use in the manufacturing of bone graft material for implanting into or attaching to bones in order to enhance bone regeneration after surgery or injury, or to treat various bone disorders, such as osteonecrosis. The invention provides bone graft material, a method for its production, bone graft implants, and medical methods and uses of the inventive products.

The invention discloses a preparation method of an acellular periosteum matrix gel material derived from natural tissues. The invention uses the acellular periosteum matrix derived from mammals to be freeze-dried to maximize the retention of biologically active molecules; and grinds it into powder, Control the particle size of the powder, increase the contact area of ​​the solution to accelerate digestion, and obtain a standardized and uniform acellular periosteum matrix gel with good osteogenic activity potential. In addition, cross-linking growth factors through EDC has a good effect on promoting the excellent regeneration and vascularization of new bone tissue. It can be used to repair bone defects caused by various etiologies in clinic, bone nonunion and other bone regeneration disorders.

A cell-removed heterogeneous bone matrix is prepared from the rib and extremity bone of pig through physical and chemical processing, and removing cells and the heterogeneous protein from tissue by use of proteolytic enzyme and Triton-X100. Its advantages are natural components and biologic chemical strength, low antigenicity, high compatibility and bone conductivity, and low cost.

The invention belongs to the technical field of bone-tendon interface injury repair, and specifically relates to regional stem cell-induced activity decellularized bone-tendon interface leaf support, specifically comprising: step 1, obtaining hBMP-2, hTGF-β3 and hGDF-7, and preparing 100ng / ml working solution of hBMP-2, hTGF-β3 and hGDF-7; Step 2, preparation of hBMP-2, hTGF-β3 and hGDF-7 slow-release colostrum; Step 3, preparation of double bionic decellularized bone tendon interface leaf scaffold ; Step 4, preparation of regional directional induced slow-release decellularized bone tendon interface leaf scaffold; step 5, preparation of regional stem cell-induced activity decellularized bone tendon interface leaf scaffold; providing a natural and natural shape for clinical bone tendon interface reconstruction A new type of tissue engineering graft with reliable mechanics and excellent activity can achieve rapid and high-quality healing after bone-tendon interface injury.

The invention discloses a sawing-punching integrated preparation device and method for a decellularized bone regeneration and repair material. The sawing-punching integrated preparation device comprises a pneumatic quick-change clamping device, a two-shaft linkage workbench, a bone sawing machine, a middle conveying device, a punching machine and a bone grain screening device, wherein the pneumatic quick-change clamping device adapts to clamping of large bones of animals with different shapes and sizes, and the two-axis linkage workbench drives the pneumatic quick-change clamping device to perform two-dimensional translation; the bone sawing machine is used for sawing animal big bones into animal bone slices, and adjusting the thickness of the bone slices; the middle conveying device conveys the animal bone slices to the punching machine for punching, and has a function of screening qualified animal bone slices; the punching machine punches the animal bone slices into bone grains and conveys the bone grains to the bone grain screening device; and the bone grain screening device screens qualified bone grains. According to the sawing-punching integrated preparation device, automatic control integrating two-shaft linkage work, animal big bone sawing, intermediate transportation, multi-die punching and screening collection is achieved, so that the whole logistics process of processing animal big bones into finished bone grains is achieved, and the processing efficiency of the animal big bone grains is greatly improved.

The invention discloses a novel dolastatin 10-cyclic peptide derivative, its preparation method and its application in antitumor drugs. The dolastatin 10 derivative with a cyclic peptide structure of the present invention can be used as a new type of anti-tumor compound, and has a good inhibitory effect on tumor cells, especially human colon cancer cells, leukemia cells, osteosarcoma cells and the like. The dolastatin 10-cyclic peptide derivative of the present invention is obtained by transforming a straight-chain pentapeptide intermediate, adding an amide condensation agent, and obtaining through liquid-phase synthesis and cyclization. The preparation method provided by the invention has low cost, convenient operation and high efficiency. The compound of the invention has a good inhibitory effect on cancer, especially HCT-116 human colon cancer cells, has high stability and low toxicity, and lays the foundation for the development of anticancer drugs for treating colon cancer or other types of cancer.

The invention relates to the field of biomedical materials, in particular to an oral guided bone regeneration membrane prepared from amniotic periosteal acellular matrix and a preparation method thereof. The present invention adopts amniotic periosteum tissue to prepare and obtain amniotic periosteal acellular matrix guided bone regeneration membrane through raw material treatment, decellularization, sample disinfection and other processes, which is used in guided bone regeneration for repairing alveolar bone or jaw bone defect. The amniotic periosteum-derived acellular matrix guided bone regeneration membrane effectively removes the cellular components in the tissue, eliminates the immunogenicity of the xenogeneic tissue, retains the active components of the amniotic periosteal extracellular matrix, and provides a good microenvironment for bone repair , to promote the repair and reconstruction of oral bone defects. In addition, the method for preparing the sheep acellular periosteum of the present invention has low cost, and compared with other mammals, the thickness and mechanical properties of the sheep periosteum material are more suitable for the application of the oral guided bone regeneration technology.

The invention discloses a bone filling adhesive which has good biocompatibility and is convenient to implant, and a preparation method of the bone filling adhesive as well as application of the bone filling adhesive. The bone filling adhesive is characterized by being made from PLGA (poly(lactic-co-glycolic acid)), ethyl acetate, biological glue, a phosphate buffer and distilled water serving as raw materials according to the ratio, wherein ethyl acetate can be replaced by acetone or dichloromethane, the bone filling adhesive is prepared by the steps of dissolving PLGA intoethyl acetate or acetone or dichloromethane, and dissolving the biological glue into the phosphate buffer. The bone filling adhesive is added into a solvent, and after being dissolved, the bone filling adhesive is implanted in an injection manner. The bone filling adhesive has the beneficial effects that when bone fractures are treated, the bone filling adhesive is added into the solvent and the mixture is implanted in an injection manner by utilizing good biodegradability and biocompatibility of the PLGA and the biological glue, and the implantation manner is accurate, convenient and fast; the bone filling adhesive is capable of promoting and adjusting growth and differentiation of subperiosteal peripheral cells, accelerating formation of bone matrixes and a mineralizing biological mechanism, quickly healing the bone fractures and quickly fixing and supporting bone fracture parts; and the bone filling adhesive can be proliferated and disintegrated into bone generating cells, has good bone conduction property and is absorbed into bone fracture areas to form calluses and further repair the bone fractures.

The present invention relates to the identification, isolation, expansion and characterization of a specific type of multipotent adult cardiac stem cell. These adult stem cells are characterized in that they naturally express a specific pattern of markers, which can be used to assist with their isolation and expansion. In particular, the cells express SOX17 and GATA4, but do not express Oct4, Nanog, c-kit and telomerase reverse transcriptase. These cells are able to differentiate into one or more of the following cell types: adipocytes, osteocytes, endothelial cells and / or smooth muscle cells. They also display an unprecedented capacity for immunoregulation as well as providing, activating and / or inducing repair of damaged cardiac tissue. These adult stem cells may be used as therapeutic agents including, without limitation, for the regeneration of tissue, particularly for regeneration of damaged cardiac tissue, such as myocardium.

The invention discloses a method for fluorescence labeling of gymnosperm pollen tube fibril bone and its application and is to provide a method for fluorescence labeling of gymnosperm pollen tube fibril bone and the application of this method in microscope inspection of gymnosperm pollen tube fibril bone. The said labelling method includes the following steps: 1) fixing the gymnosperm pollen tube with 40-60mM Pipes cushion solution containing 2-4 % cavaform; 2) washing the pollen tube; 3) placing the pollen tube in 0.2-0.3 % glycerin to infiltrate for 30-60min; 4) washing the pollen tube; 5) dyeing the pollen tube and avoiding the sun with 150-250nM phalloidine by fluorescence labeling for 0.5-1.5h and fluorescence labeling the fibril bone. The invention is of simple operation and is quick, has small damage to the cells, and has good labeling effect, which will play an important role in study gymnosperm pollen tube cell bone and have a high actual application value.