136 results about "Tissue engineered skin" patented technology

A cryopreservation solution of tissue engineering products uses DMEM culture solution as a basic solvent which is added with vitamin B, vitamin C, chondroitin sulfate, beta-integrin, cromolyn sodium, cytochalasin B, L-glutamine, bovine serum albumin, fetal bovine serum, trehalose, sodium carbonate, polysucrose-70, and dimethyl sulfoxide added during freezing storage. The cryopreservation solution provided by the invention has little toxicity to cells and long storage time, and can be stored for six months under 80 DEG C below zero, and 12 to 18 months in liquid nitrogen; and the cell viability of the resuscitated cells can be over 60%. The stored tissues can be simply and conveniently used after being resuscitated for only three to five min in water bath under the temperature of 37 DEG C and being washed by sterilized saline water. The invention can be widely used, and is suitable for tissue engineering skin, tissue engineering cornea, tissue engineering blood vessel, tissue engineering nerve and the like, and also is applicable to the cryopreservation of normal tissues.

The invention provides a method for constructing tissue engineering skin. In the method, human epidermal stem cells and hypodermal fibroblasts are taken as seed cells, and a homologous acellular dermal matrix modified by a human placental type IV collagen is taken as a stent, so that active dual-layer engineering skin is compounded and constructed by using a gas-liquid interface separate culture method. The tissue engineering skin constructed by using the method is closer to the structure of natural skin; and as proved by histomorphology, the tissue engineering skin has an epidermal and hypodermal double-layer structure, wherein the hypodermal collagen stent has a complete structure, an epidermal layer is provided with a plurality of layers of cells of different differentiation degrees, and the morphological requirement of the tissue engineering skin is met.

The invention discloses a preparation method and an application of a cellulose nanocrystals (CNCs)-reinforced collagen compound substrate, which belong to the field of tissue engineering. The preparation method comprises the following steps of: preparing CNCs by adopting a method for degrading cellulose with sulfuric acid; preparing a CNCs solution and a collagen solution, and mixing the CNCs solution and the collagen solution physically; and pouring the mixed solution into a mold, and molding to obtain the CNCs-reinforced collagen compound substrate. According to the CNCs-reinforced collagen compound substrate prepared with the method, the problems of poor mechanical strength and poor hydrophilicity of the conventional collagen material are solved; and the CNCs-reinforced collagen compound substrate can be applied in the fields of wound dressings, tissue inducing regeneration membranes, soft tissue patches, tissue engineering skin, facial masks and the like. A preparation process technic provided by the invention is easy and practicable, has high repeatability, and is suitable for industrial production of CNCs-reinforced collagen compound substrates of different requirements.

The invention relates to a preparation method of tissue engineering skin containing hair follicles. In the invention, tissue engineering hair follicles are evenly inoculated in a tissue engineering dermal layer formed by compounding a bracket material and cells, then the tissue engineering dermal layer is covered by epidermal cells, and the tissue engineering skin containing the hair follicles is formed after culture; the invention overcomes the defects of little number of the hair follicles in the skin, no uniformity and uncontrollable quantity of the hair follicles caused by mixed injection formerly, and the prepared tissue engineering skin contains an epidermis, dermis and hair follicle type tissue structure, wherein the number of the hair follicles approaches to the natural number, the hair follicles are evenly distributed in artificial pores and develop continuously after in-vitro culture and transplantation, a multi-layer concentric circle structure is formed, an inner layer is hair papilla cells wrapt by micro-capsule membranes, and an outer layer is epithelium class cells; and the skin cultured in vitro or transplanted in vivo finally forms functional skin containing a hair follicle accessory structure, and the regeneration of the hair follicles after skin transplantation can be realized.

The invention relates to tissue engineering skin and a preparation method thereof. The tissue engineering skin comprises a dermis layer and an epidermis layer, and is prepared from seed cells, cell growth factors and a biological scaffold material according to a biological 3D printing technology. The tissue engineering skin is similar to natural skin, comprises the dermis layer and the epidermis layer, can be produced in batches, can be closely attached to a wound, and has high biocompatibility, the dermis layer contains blood vessels, sweat glands and hair follicles, the shape, size and thickness of the tissue engineering skin can be easily changed, the elasticity and flexibility of the skin after sound healing can be enhanced, and the tissue engineering transplantation success rate can be increased.

The invention discloses a dual-layer collagen-chitosan sponge scaffold and a preparation method thereof, and relates to a tissue engineering scaffold. The invention provides the dual-layer collagen-chitosan sponge scaffold for the tissue engineering skin and the preparation method thereof. The scaffold is a dual-layer mesh structure, the upper and the lower layers are collagen and chitosan composite films, and the hole diameters of the layered and hole-shaped structure are mutually penetrated. The collagen hydration solution is dissolved in acetic acid for preparing the collagen dispersion solution with the concentration of 0.2 percent to 0.5 percent and 0.5 percent to 0.8 percent; the chitosan is dissolved in the acetic acid for preparing the chitosan dispersion solution with the concentration of 0.2 percent to 0.5 percent and 0.5 percent to 0.8 percent; the two dispersion solutions are mixed to obtain two collagen-chitosan solutions; the collagen-chitosan solution with the concentration of 0.5 percent to 0.8 percent is frozen overnight, the sponge-like white scaffold is obtained by drying, and the single-layer scaffold is obtained by cross-linking after the film pressing, cleaning, freezing overnight and drying; the collagen-chitosan solution is poured in for freezing overnight and drying to obtain the dual-layer scaffold, and the cross-linking, the cleaning, the freezing overnight, the freezing and the drying are carried out.

The present invention is a tissue engineering skin containing the peripheral blood stem cell and the preparation method. The present invention puts the peripheral blood stem cell and the skin fibroblast cell inside the biological frame. The active tissue engineering skin containing the peripheral blood stem cell is built by planting the epidermal cell on the surface. The tissue engineering skin is similar to the natural skin tissue in structure. Compared with the present skin replacing product, the present invention has properties of improving the blood supply in the wound and promoting the wound healing. The present invention can improve the elastic and toughness properties of the skin as well as the mechanical wear tolerance. The present invention can also reduce the scar proliferation and improve the healing quality. The present invention can improve the success rate of the artificial skin planting and can be effectively applied in repairing the skin defect, which is hard to heal.

The invention relates to the technical fields of tissue engineering and medical wound repair. At present, a living skin substitute constructed by using the materials of polylactic acid, polyglycolic acid, collagen, hyaluronic acid, and the like as a dermic bracket has the defects that on one hand, host materials are difficult to extract, the living skin substitute has complicated manufacturing technology and is expensive in cost and difficult to widely popularize and apply clinically; and on the other hand, the living skin substitute does not have a skin basilar membrane structure so that healed skin does not resist pressure and wear, and the living skin has unfirm adhesion to the epidermis and is easy to shed and break or form water blisters so that the structural and morphological development of the normal epidermis is influenced; and allogeneic acellular dermis is taken from cadaver skin and is limited in sources and expensive in cost, and thus clinical application is limited. The invention aims at providing a skin substitute which uses surface-finished and modified amnion as the basilar membrane and blood plasma as stroma, which has the advantages of wide material sources, low cost and simple preparation method. An animal experiment proves that the complete basilar membrane and hemidesmosomes can be retained in in-vivo transplantation, and the formation of an epidermal structural form is accelerated and promoted.

The invention relates to in-vitro inducing differentiation of umbilical cord mesenchymal stem cells into skin tissue engineering seed cells. The invention discloses a new research direction for human umbilical cord mesenchymal stem cells, i.e. in-vitro inducting differentiation into skin tissue engineering seed cells. The invention also discloses an experimental method thereof, which comprises the following steps: collecting the caesarean umbilical cords in aseptic environment; culturing original-generation umbilical cord mesenchymal stem cells (UCMSCs) by an enzyme digestion or tissue-piece adherent method; after purifying and subculturing to the third generation, inducing by using different in-vitro condition induction liquids to differentiate into epidermis stem cells and fibroblasts; and inducing and differentiating epidermis stem cells and fibroblasts into sudoriferous gland cells by using coculture mode with the normal sudoriferous cells. After the induced cells are digested, liquid nitrogen is stored for later use. The invention provides seed cells for skin tissue engineering, and has the advantages of strong multiplication capacity, easy inducing differentiation, low immunogenicity, abundant sources and easy obtainment.

The invention relates to a method for culturing seed cells of tissue engineering. The method for amplifying the seed cells of the tissue engineering comprises the following steps of: (1) adding a macroporous microcarrier into a bioreactor, hydrating and sterilizing and then adding 50 to 100ml of fibroblast culture medium, and adding fibroblasts for culturing; (2) inhibiting the fibroblasts from amplifying when the fibroblasts are attached to the microcarrier and the growth area is near 50 percent of surface area of the microcarrier, and standing and then pouring out culture solution; and (3) washing the obtained carrier by using sterile, calcium-free and magnesium-free phosphate buffered solution (PBS), then adding an epidermal cell culture medium and epidermal cells, and continuing to stir and culture until the cells overgrow the carrier to obtain the seed cells of skin tissue engineering. The quantity and function of the cells cultured by the method can both be apparently increased,the damage of the cells is reduced, the cells can be directly used as seed cells for skin construction of the tissue engineering, and the effect is superior to those of methods using epidermal cell suspension and epidermal cell patches.

The invention relates to a preparation method of tissue engineering skin containing appendant organs. In the invention, the preparation method comprises the following steps of: inoculating amniotic mesenchyme stem cells, amniotic mesenchyme stem cells induced to the directions of hair papillae, amniotic epithelial cells and amniotic epithelial cells induced to the directions of the epithelia of the sweat gland into a gel solution, then inoculating the amniotic epithelial cells and the amniotic epithelial cells induced by keratinocyte in a warp direction on the surface of a structure of the hair follicle and the sweat gland after culturing by a dermal layer by induced culture forming the structure of the hair follicle and the sweat gland, and obtaining the tissue engineering skin with the structure of the hair follicle and the sweat gland after culturing. The skin has elasticity, toughness, vigorous cellular metabolism, strong multiplication capacity, sufficient secretion of extracellular matrices, tight linkage of the cuticular layer, the inophragma and the cells of the dermal layer and little possibility of falling off, enhances the success rate of transplantation, can promote the healing of a wound surface, enhances the effects of restoration and replacement and has the function of regulating the immunological rejection of a receptor; by the contained structure of the hair follicle and the sweat gland, the function of the skin is more complete; the adopted amniotic tissues are postpartum wastes and have extensive source, strong cell multiplication capability and multiplepassage frequency; and the invention is beneficial to the mass production of seed cells and lowers the industrialization cost.

The invention relates to tissue-engineered skin containing blood vessels and hair follicle structures based on 3D printing and a preparation method thereof. The tissue-engineered skin is composed of an epidermal layer, an acellular dermal scaffold and a dermis layer, wherein in the epidermal layer, epidermal stem cells are used seed cells, the seed cells are printed on the upper surface of the acellular dermal scaffold through a 3D printer after being compounded with a hydrogel carrier to differentiate and form a normal epidermal structure, in the dermis layer, mesenchymal stem cells, vascularendothelial cells, dermal papilla cells and adipose-derived stem cells are used as seed cells, the seed cells and a hydrogel carrier are compounded, through the 3D printer, gelatin slow-release micropheres compounded with cytokines are printed on the lower surface of the acellular dermal scaffold, and meanwhile, the hydrogel compound of the seed cells is printed in the gelatin slow-release micropheres to form a dermis structure with a three-dimensional spatial structure.

The invention discloses tissue engineering skin which comprises seed cells and an acellular allogenic dermal matrix, wherein the seed cells and the acellular allogenic dermal matrix form a skin appendage; the seed cells are arranged, spread flat and printed at corresponding positions on the front side of the acellular allogenic dermal matrix according to an anatomical structure of normal skin by using a 3D (Three-dimensional) printer. The invention also discloses a method for preparing the tissue engineering skin. The invention provides perfect tissue engineering skin, and the tissue engineering skin can be rapidly constructed in a short time in an operating room (the operation is finished). Meanwhile, the tissue engineering skin comprises the seed cells and dermal matrix, the seed cells are basal layer cells, have the activities of multipotential stem cells and multipotential differentiation functions and are arranged and printed in the corresponding positions according to the anatomical structure of the normal skin, the skin appendage is easily formed, the cure rate is accelerated after the operation, and functions of the skin are restored.

A tissue-engineered artificial skin able to regulate the pigment secretion is configured through externally culturing the ceratin cells, fibroblasts and melanocyte of skin, proportionally mixing by tissue engineering method, and forming artificial skin.

A tissue-engineered skin containing vascular structure features that it contains skin fibroblasts, keratinous cells and vascular endothelial cells. Its configuring method includes separating said three kinds of cells from eath other, external culture, proportionally mixing, and configuring the artificial skin with vascular structure. Its advantages are high survival rate and high resistance to infection.

The invention discloses a low-temperature preserved solid culture medium used for a tissue engineering skin model and a preservation method of the low-temperature preserved solid culture medium. The low-temperature preserved solid culture medium is prepared by the following steps: adding a low-temperature protection agent into a common culture medium for epidermis cells to prepare a basic culture solution; and mixing the basic culture solution with agarose gel and condensing to form the solid culture medium suitable for the skin model. The tissue engineering skin model is embedded into the culture medium, is preserved at 4-8 DEG C for 24-72 hours, and then is resuscitated by a common epidermis cell culture solution. The solid culture medium can be used for reducing tissue activity reduction and structure damages of cells and skin tissues, caused by a low-temperature preservation condition, to the greatest extent.

The invention relates to a method for constructing tissue-engineering skin. The construction method adopts an autocrine extracellular matrix of fibroblast as a biological scaffold of the tissue-engineering skin. The method utilizes the extracellular matrix excreted by the cultured fibroblast as the biological scaffold and human epidermal cells as seed cells, and the cultured tissue-engineering skin is closer to the structure of natural skin. The method has the advantages that: firstly, the method is simple and convenient, and favorable for synthesis and mass production of products because thematerials take the form of liquid; secondly, the obtained product has the structure which is closer to the natural skin, and the method is favorable for improving the sensibility and the precision ofprediction of the toxicity of compounds; thirdly, the product does not contain additional biomaterials, so that the risk of cross infection of zoonosis cannot occur; and fourthly, the obtained product is convenient for quality control, so that the variation between batches is reduced.

The present invention is a tissue engineering skin containing the fat layer and the preparation method. The present invention is characterized in that the present invention is manufactured by the fat cell, the skin fibroblast cell, the epidermal cell and the biological frame. The fat cell and the skin fibroblast cell are mixed inside the biological frame. The active tissue engineering skin containing the fat layer is built by planting the epidermal cell on the surface. The prepared issue engineering skin containing the fat layer not only has a certain degree of elastic and toughness properties, but also has short incubation time and no significant immunological rejection. The tissue has similar structure as the natural skin and the subcutaneous adipose tissue. The present invention has multiple effects in aspects of anti infection, homeostasis and inhabiting the scar formation.

The invention discloses a vascularized full-thickness tissue-engineered skin layer-by-layer assembled by hydrogel, nanofiber scaffolds and skin cells and a preparation method thereof, and belongs to the technical fields of macromolecule materials and biomedical materials. Artificial tissue engineered skin consists of an epidermal layer and a corium layer, wherein the epidermal layer is formed by alternately stacking nanofiber scaffolds located above the corium layer with seed cells. The corium layer consists of lower-layer nanofiber scaffolds, upper-layer hydrogel scaffolds and three kinds ofseed cells, and the seed cells are distributed on the surface of the nanofiber scaffolds, inside the hydrogel and on the surface of the hydrogel. The vascularized full-thickness tissue-engineered skinis prepared by combination of an electrospinning technology and a macromolecular complexation technology with a fiber / cell layer-gel layer-fiber / cell layer-by-layer self-assembly technology. The artificial tissue-engineered skin with biological function can be used for regeneration and repair of various tissues, especially for wound healing, angiogenesis, scar formation reduction and the like.

The invention belongs to the biotechnical field and in particular to tissue engineering skin containing appendage and a preparation method thereof. The preparation method comprises the following steps: performing transfection of a urine cell to induce to generate a multifunctional stem cell; continuously inducing the multifunctional stem cell to generate a mesenchymal stem cell; differentiating the mesenchymal stem cell to form an epidermis cell, a fibroblast and a hair follicle cell; and constructing tissue engineering double-layered skin containing the appendage with a biological stent material amniotic membrane. The seed cell of the tissue engineering skin prepared by the preparation method is the urine cell which is convenient to obtain and short in cultivation period. The hair follicle structure which is contains facilitates union of a wound, and the tissue engineering skin containing the appendage provided by the invention lays a foundation for follow-up development of the tissue engineering skin containing the appendage.

The invention discloses a culture medium that is used in transportation of tissue engineered skin. A basic culture fluid is added with additional hydrocortisone, epidermal growth factors, isoproterenol, insulin, transferrin, triiodothyronine, adenine, vitamin C, cholera toxin, L-serine, L-novain, palmic acid, linoleic acid, arachidonic acid, bovine pituitary extract, bovine serum albumin, penicillin, streptomycin and fungizone B. The obtained culture medium has strong stability and long shelf life of about 12 months, completely meet the demands of large-range transportation of the tissue engineered skin products, can effectively maintain the structure and functions of the tissue engineered skin in the transportation process, becomes solid after being cooled, and consequently has convenient carry in the transportation process.

The invention discloses a novel human epidermal culture method. The method includes the following steps of (1), skin sample treatment and cell separation; (2), culture of epidermal cells; (3), passage of the epidermal cells and stripping of tissue engineered skin; (4), quality inspection of the tissue engineered skin. A preparation method of a KMSFM culture medium includes: taking DSFM of Gibco as a basis; using low calcium ion concentration; adding bovine pituitary extract (BPE), bovine insulin, bovine transferrin, bFGF, cortisol, heparin and endothelin at the same time. By the novel human epidermal culture method, bovine serum is removed, mixing of non-human-derived ingredients can be reduced greatly, and cost and unsafe factors are reduced; a culture system coated by I-type collagen is used, cell growth slowness caused by removing of nourishing layer cells is avoided, and the novel human epidermal culture method can be used for treating skin pigment diseases such as leukoderma and giant pigmented nevus.

The invention discloses an xenogenic dermal reticular layer scaffold with basilar membrane removal and no cells, which is taken from an dermal reticular layer of a mammal, and has a natural three-dimension structure with larger pores of the dermal reticular layer; the dermal scaffold removes a basilar membrane and has no cells; and the thickness of the dermal scaffold is between 0.2 and 0.8mm. The scaffold is prepared by performing sterile dermis getting, then removing subcutaneous tissue, epidermis, the basilar membrane and a dermal papillary layer, then orderly using distilled water to soak and using trypsinase and an EDTA.Na2 digestive juice to digest, using a phosphate buffer solution to wash after the persistent oscillation, then placing into a Triton X-100 solution for soaking and performing the persistent oscillation, and then using the phosphate buffer solution to wash. The scaffold completely removes the source of a main immunogenic substance in dermis, and reduces the immunogenicity; the scaffold has strong nutrition permeability, quick vascularization speed, high survival rate and good effect when used for the repair of skin wounds, the construction of tissue engineering skin, and tissue filling; and the scaffold has the advantages of rich dermis sources, simple preparation technology, short production period, and low cost.

The invention discloses a method for the directional induction of the differentiation of MSCs. The method for the directional induction of the differentiation of MSCs comprises the following steps: inoculating and adsorbing in-vitro induction cells to the lower surface of a polycarbonate film, inoculating in-vitro MSCs to the upper surface of the polycarbonate film, and co-culturing the induction cells and the MSCs to promote the differentiation of the MSCs to target cells, wherein each of the MSCs and the induction cells cannot pass through the holes of the polycarbonate film. The differentiation of UCMSCs to cuticle cells has a beneficial effect on the clinic skin regeneration research progress in the future. The method develops ideas for the increase of the induction method for the multi-directional differentiation of the MSCs, and provides bases for the construction of tissue engineering skins through adopting the UCMSCs as seed cells.

The invention discloses a method for fast constructing and repairing skin defects by tissue engineering skins and belongs to the field of medical methods. The method can be used for solving the defects of low survival rate of skin grafts and long period of the traditional medical method, wherein the method is formed by a razor-thin graft and a cell suspension in combination with a composite skin grafting method. The method for fast constructing and repairing the skin defects by the tissue engineering skins has good effects of promoting wound healing, enhancing the survival rate of the razor-thin graft, and improving the color and texture of skins, and can effectively treat skin defects resulted from various reasons such as a trauma and a surgery.

The invention relates to a regeneration material of artificial dermis substitution and a preparation method thereof. In the regeneration material of a dermis substitution for tissue engineering skin for loading rhGM-CSF, the regeneration material is formed by a heparinizing collagen-chitosan bracket loading the rhGM-CSF solution. The invention further discloses a preparation method of the regeneration material. The invention provides an artificial dermis substitution with good performance for curing the whole skin coloboma of trauma, burning, chronic skin ulcer, thereby being capable of remarkably quickening the vascularization process of the regeneration material of artificial dermis substitution, decreasing the infection risk, promoting the healing of the surface of a wound and reducing the hyperplasia of scar, and thus easing the pain of patients. The invention can be widely applied for the aspects of trauma, burning, surgery reshaping, and the like. The preparing method is simple, the source of the material is wide, the production efficiency is high, and the invention is suitable for industrial production.

A vascularized full thickness tissue engineered skin assembled by hydrogel, nanofibrous scaffolds and skin cell layers and a preparation method thereof relate to a technical field of polymer materials and biomedical materials. The artificial tissue engineered skin includes an epidermis layer and a dermis layer. The epidermal layer is formed by alternately stacking upper nanofibrous scaffolds located above the dermis layer and a kind of seed cells. The dermis layer is formed by lower nanofibrous scaffolds, the hydrogel layer above the lower nanofibrous scaffolds, and three kinds of seed cells distributed on surfaces of the lower nanofibrous scaffolds as well as inside and on a surface of the hydrogel layer. The artificial tissue engineered skin is prepared by a combination of electrospinning technology, polymer complexation technology and fiber / cell layer-gel layer-fiber / cell layer self-assembly technology. The bio-functional artificial tissue engineered skin can be used for regeneration and repair of various tissues.

The invention relates to a method for producing dual-layer organism skin with basilar membrane. Wherein, it is formed by preparing organism skin cultivate base, preparing gel solution, cultivating breeding box and cultivating screen support; it adjusts the calcium density of cultivate base and adds external laminin to accelerate the basilar membrane formation, to strengthen the connection between organism protective skin and corium. The invention has simple process while all materials are liquid and better compatible, without hurting transplanted one; and the skin will not be separated easily; and the product has better compatibility.

The invention discloses a method for building tissue engineered skin by composite DED (de-epidermidalized dermis) of amnion endothelial cells. The hAECs (human amnion endothelial cells) are used as seed cells; human DED is used as a stent material; the seed cells are planted onto the stent material to build the hAECs DED; the seed cells are planted onto the stent materials to build the hAECs-DED tissue engineered skin. The method comprises the following steps of (1) hAECs separation culture; (2) FB (fibroblast) separation culture; (3) DED preparation; (4) hAECs-DED tissue engineered skin building. The amnion endothelial cells are used as the seed cells; the human DED is used as a stent; the seed cells and the stent are all from the human body and have the homeology; the built tissue engineered skin has the tissue structures and functions similar to the normal skin, and also has some features of normal skin proliferation and differentiation.