412 results about "Decellularization" patented technology

New methods for producing tissue engineered constructs and engineered native tissues are disclosed. The methods include producing a tissue engineered construct by growing cells in vitro on a substrate and then decellularizing the construct to produce a decellularized construct consisting largely of extracellular matrix components. The construct can be used immediately or stored until needed. The decellularized construct can be used for further tissue engineering, which may include seeding the construct with cells obtained from the intended recipient of the construct. During any of the growth phases required for production of the construct, the developing construct may be subjected to various tissue engineering steps such as application of mechanical stimuli including pulsatile forces. The methods also include producing an engineered native tissue by harvesting tissue from an animal or human, performing one or more tissue engineering steps on the tissue, and subjecting the tissue to decellularization. The decellularized, engineered native tissue may then be subjected to further tissue engineering steps.

An objective of the present invention is to overcome a problem that there is an inverse relationship between the decellularization rate and the strength of tissue. This problem was solved by immersing tissue in a solution containing a non-micellar amphipathic molecule (e.g., a 1,2-epoxide polymer). Thus, the present invention provides decellularized tissue, in which the cell survival rate of the tissue is less than a level at which calcification or an immune reaction is elicited in an organism and the tissue damage rate of the tissue is suppressed to a level which permits clinical applications. Tissue prepared by the above-described treatment preferably retains a certain level of tissue strength. Further, the tissue of the present invention has an effect of performing cell replacement.

An acellular soft tissue-derived matrix includes a collagenous tissue that has been delipidated and decellularized. Adipose tissue is among the soft tissues suitable for manufacturing an acellular soft tissue-derived matrix. Exogenous tissuegenic cells and other biologically-active factors may be added to the acellular matrix. The acellular matrix may be provided as particles, a slurry, a paste, a gel, or in some other form. The acellular matrix may be provided as a three-dimensional scaffold that has been reconstituted from particles of the three-dimensional tissue. The three-dimensional scaffold may have the shape of an anatomical feature and serve as a template for tissue repair or replacement. A method of making an acellular soft tissue-derived matrix includes steps of removing lipid from the soft tissue by solvent extraction and chemical decellularization of the soft tissue.

An object of the present invention is to provide a method for producing an ADM suitable as a scaffold for a cultured skin, that is, a separation and decellularization method that enables various types of extracellular matrixes such as a basement membrane to be preserved, allows the epidermal layer to be peeled off easily, and does not cause any damage to the dermal matrix. The present invention relates to a skin separation and decellularization method comprising a step of freeze thawing harvested skin and then separating the skin into epidermis and dermis by a treatment with hypertonic saline, and a step of washing the separated dermis.

Decellularization of tissue by means of an amphipathic solvent a well-established practice. However, situations exist where the provision of enhanced decellularization is preferred. There is a demand for treating methods for coping with such situations. Thus, it is intended to provide a method for enhancing decellularization. The method comprises not only the immersing of a tissue in a solution containing an amphiphilic molecule in non-micellar form (for example, 1,2-epoxide polymer) but also performing a radical reaction (for example, treatment selected from the group consisting of exposure to gamma-ray irradiation, ultraviolet irradiation, a free radical supply source, ultrasonication, electron beam irradiation, and X-ray irradiation).

The invention described herein provides systems and apparatuses for an initial preparation of an organ or tissue scaffold comprising an extracellular matrix, and subsequent recellularization of the scaffold to ultimately form a resultant artificial organ or tissue incorporating the natural and original extracellular matrix. The techniques and equipment of the invention collectively minimize scaffold collapse, compression or physical damage to the organ as well as afford the advantages of significant maintenance of the initial natural structural and biochemical attributes of the organ or tissue. The invention is particularly useful in organ and tissue transplantation and repair.

The invention provides for methods and materials to decellularize a solid organ and to recellularize such a decellularized organ to thereby generate a solid organ.

The invention discloses a ply tissue engineering corneal frame and a manufacturing method and an application thereof, relating to a ply tissue engineering corneal frame of the biomedicine. Compared with the existing materials, the invention provides a ply tissue engineering corneal frame and a manufacturing method and an application thereof, which has abundant resources, high transparency, good biocompatibility, thorough decellularization and strong safety, and the performance approaches to that of a fresh cornea, so that the ply tissue engineering corneal frame can be accepted by majority of patients and applied clinically for a long term. The ply tissue engineering corneal frame is an animal derived decellularized ply corneal stroma sheet and does not contain cellular constituents; collagenous fibers are tidily arranged, and gaps are regular; corneal light transmittance is 80-95%, and tensile strength is 2-5N/mm<2>. The ply tissue engineering corneal frame can be served as the substitute for various donor materials for corneal transplantation and can be used for treating a series of diseases of corneal trauma and chemical burn, corneal tumor and a series of hyperplastic diseases, a series of diseases of corneal neovascularisation and scar, corneal immune diseases, a series of diseases caused by corneal transplantation exclusive reaction and other keratopathy.

The invention discloses a method for preparing human body or animal accellular tissues. According to the method, the decellularized tissue obtained by treating human or animal tissues, such as skin, pleuroperitoneal membrane, esophagus mucous membrane, small intestine mucous membrane, pericardium, vessel, nervus, cardiac valves, bone, cartilage, muscle tendon, and the like through processes of inactivation of virus, derosination, decellularization, and the like is an acellular three-dimensional network structure tissue, and extracellular matrix components are kept. The tissue prepared by the method can be used as an implantable repair material for a human body and also can be used as a support material of tissue engineering cell culture. The skin, the pleuroperitoneal membrane, the esophagus mucous membrane, the small intestine mucous membrane, the pericardium, and the like also can be used as a wound covering material.

The invention relates to a decellularization cornea preparation method, which adopts steps of corneal epithelium layer removing, ultraviolet cross-linking, viral inactivation, decellularization treatment, gradient dehydration, radiation protection and sterilization. According to the present invention, the prepared decellularization cornea has characteristics of complete antigen removing, no excitation of host acquired immunity reaction, good biocompatibility, low damage on nature corneal stroma, maintaining of structure characteristics of the nature corneal, and maintaining of effective components of the nature corneal so as to provide physical and chemical properties similar to the nature corneal; and after the prepared decellularization cornea is transplanted, animal experiment results show that characteristics of transparent transplanted cornea, no scar formation, no melting generation and no neovascularization are provided, the transplanted cornea and the recipient bed are completely integrated after transplanting a month, the transplanted cornea is subjected to complete epithelization after three months, corneal stroma cells migrate toward the decellularization cornea graft so as to prove that the tissue just takes place slow reconstruction, and the transplanted cornea after 6 months does not show significant difference in histology and appearance detection compared with the nature cornea.

The invention provides a repair material for biological derivative tendon. The material takes homogeneous or heterogeneous tendon as a raw material and is formed through disinfection, degreasing, deproteinization, decellularization, freeze drying, depolymerization, devillication recombination and secondary disinfection. A method for preparing a tendon scaffold material comprises the following steps that: on one hand, a periphery structure of a tendon tissue is destructed through treatment processes of removing soft tissues, the disinfecting, the degreasing and the decellularization, and the repellency of a transplantation receipt to the tendon is reduced; on the other hand, a basic mechanism, partial enzyme activity of the tendon tissue and the continuity of the tendon tissue are further maintained through a freeze drying process so as to provide a good scaffold structure for the revascularization and cell tissue of the tendon.

The invention discloses scaffolds of umbilical cord decellularized Wharton jelly for tissue engineering and a preparation method thereof. Umbilical cords are employed as the raw material and their outer membranes and vascular tissues are peeled off. And the rest part of the umbilical cords is subjected to hypotonic freeze-thaw, mechanical pulverization, differential centrifugation, enzymatic digestion for decellularization. Then the umbilical cord Wharton jelly is collected and injected into a mold. After freeze drying and crosslinking, multiple three dimensional porous sponge scaffolds and composite scaffolds can be obtained. The method of the invention has the advantages of wide material source, low cost, simple technology. And the prepared scaffolds are characterized by controllable fine structure, appropriate degradation rate, good biocompatibility, and biomechanical strength, which are in favor of cell adhesion and the uniform distribution of seed cells within the scaffolds, as well as seed cell multiplication, migration and growth. Thus, the scaffolds of umbilical cord decellularized Wharton jelly in the invention can be widely applied in the tissue engineering field such ascartilage, bone, skin and nerve, with a favorable clinical application prospect.

The invention discloses a preparation method of a natural-tissue-derived decellularized and decalcified bone material. According to the method, any bone tissue of a mammal is treated with a protease inhibitor-containing normal saline buffer, an organic solvent, Tirton X-containing PBS (phosphate buffer saline), SDS (sodium dodecyl sulfonate)-containing PBS, pancreatin-containing PBS, deoxyribonuclease-containing PBS, an EDTA (ethylene diamine tetraacetic acid) isotonic solution and ultrasonic waves, and the decellularized and decalcified bone material is obtained. Cancellous bones and cortical bones can be decellularized completely and simultaneously, the conditions are mild, damage to an ECM (extracellular matrix) is avoided, rapidness and stability are realized, and the obtained decellularized and decalcified bone material has the advantages of good biocompatibility, high plasticity, high biomechanical strength and the like and can be used for clinically repairing bone regeneration and repair disorder such as bone defects, nonunion and the like caused by various causes.

The invention provides a biological tissue matrix material, and a preparation method and a purpose thereof. The biological tissue matrix material is characterized by comprising an extracellular matrix, wherein the extracellular matrix comprises collagenous fiber, growth factors and fibronectin. The biological tissue matrix material is prepared from small intestinal submucosa matrix materials. The preparation method is characterized by comprising the following steps that decellularization is performed: decellularized liquid contains trypsin and PBS solution, and also contains EDTA, EDTA-2Na or EDTA-4Na; the decellularized liquid is used for treatment in multi-frequency ultrasonic environment for decellularization; the dual-frequency ultrasound at least contains two ultrasonic frequencies with different frequencies. The decellularization process technology is improved; the DNA residual quantity of the obtained product is lower; the immunogenicity is lower; the anti-infection capability is higher; the restoration capability is higher.

The invention belongs to the field of tissue engineering, and discloses a vascularization support for tissue engineering and a construction method of the vascularization support. The construction method includes (1), pre-treating an acellular matrix to remove impurities possibly introduced in decellularization and storage processes; (2), producing an acellular matrix/composite material compound support according to a tissue engineering support forming method; (3), post-treating the acellular matrix/composite material compound support obtained in the step (2), and removing residual substances in the production process to obtain the vascularization support for tissue engineering. The construction method has the advantages that the method is based on basic principles in organism vascularization processes and is conformity to migration and multiplication features of endothelial cells, microenvironments for organism vascularization are simulated to the greatest extent, the cellular matrix/composite material compound support is construed to develop advantages of both the acellular matrix and a composite material, and the support product capable of meeting tissue engineering vascularization requirements is constructed.

The invention relates to a scaffold material for tissue engineering construction and a preparation method thereof, and particularly relates to a compound tissue engineering scaffold containing a PLGA (poly(lactic-co-glycolic acid) strengthening net, and a preparation method and application thereof. The compound tissue engineering scaffold containing a PLGA strengthening net comprises a porous scaffold having good biocompatibility and a PLGA strengthening net, wherein the PLGA strengthening net is formed by weaving PLGA fibers, and the PLGA strengthening net is tightly combined with the porous scaffold. The compound tissue engineering scaffold containing a PLGA strengthening net has mechanical property similar to that of the acellular dermal matrix, and is more beneficial to the maintenance of three-dimensional porous structures of the scaffold, thereby being more beneficial to the ingrowth of cells, blood vessels and tissues; and because of the good biocompatibility of collagen, chitosan and other naturally-derived macromolecules, the biological performance of the compound scaffold is maintained, thereby being beneficial to the adhesion, propagation and secretion of cells. Thus, the compound tissue engineering scaffold is a good renewable material applicable to the construction of tissues and organs of tissue engineering.

The invention provides a dura graft and a preparation method thereof. The dura graft is prepared from an acellular dermal matrix. The acellular dermal matrix is obtained from allogenic skin by crosslinking treatment, digestion treatment with a protease solution, decellularization with a hypertonic saline solution containing a surfactant, DNA degradation, amino acid nutrition treatment and freeze drying. The invention also provides applications of the dura graft in preparing repair materials used in the treatment of dural damage. The acellular dermal matrix is obtained from human skin tissue and undergoes physicochemical and biochemical treatment, so that all the components that may cause immunological rejection are removed. A fibrous stereoscopic scaffold structure of the original tissue is completely reserved, and new vessels and fibroblasts rapidly grow in the dura graft after implantation. Therefore, the dura graft has good clinical effects.

The invention discloses a microenvironment construction method for three dimensional culture of cells based on a specificity extracellular matrix and an application. A specificity extracellular matrix extraction method, the specificity extracellular matrix and a photocuring material are combined to prepare hydrogel, and the invention relates to an application of the hydrogel to in vitro three dimensional culture and particularly an organ chip technique. According to the technical scheme, in accordance with tissue or organs of different density, surfactants having different decellularization capacity are selected, through combination of the manner that enzyme treatment is selected and an ammonia solution is used as a solvent, particularly-compact tissue or organs are treated, and the preferred combination of the enzyme and surfactants comprises Trypsin and a Triton X-100 solution, and the solvent is a 0.05%-0.5% ammonia solution. The extracellular matrix and a light-initiating colloid-forming material are compounded for preparing the hydrogel so as to construct cell culture microenvironment. The method has the advantages of being high in colloid-forming speed, simple in colloid-forming condition, controllable in hydrogel mechanical property and the like.

The invention relates to a biological amnion and a preparation method thereof. The biological amnion has three layers including a slow release layer, an amnion layer and a collagen layer from top to bottom, wherein the slow release layer consists of collagen and biological active factors, the amnion layer consists of an amnion subjected to decellularization treatment, and the collagen layer is formed by freeze-drying and compounding collagen. The biological amnion is prepared through the steps of raw material pretreatment, virus inactivation, decellularization treatm. The biological amnion prepared by the invention has the characteristics of an effect of slowly releasing the active factors, low antigenicity on removing epithelial cells, convenience in product operation, difficulty in curling, good adhesiveness with surrounding tissues, difficulty in sliding and the like. Meanwhile, by using a process for performing the decellularization treatment on the amnion, disclosed by the invention, a natural compact collagen structure in the amnion can be effectively retained, the biological amnion can fully achieve a physical barrier effect after being applied to tenorrhaphy, and an animal experiment proves that the biological amnion can effectively achieve an effect of preventing tissue adhesion.

The invention discloses a method for removing cells from a human amnion. The method is characterized by comprising the following steps of: obtaining a clean semitransparent amnion, rinsing the semitransparent amnion by using a PBS (Phosphate Buffer Solution), soaking the semitransparent amnion in a comprehensive PBS liquid, shocking, rinsing the amnion by using the PBS after the amnion is taken out, successively soaking the amnion in steapsin/PBS liquid and deoxyribonuclease/PBS liquid for treatment, rinsing the amnion by using double-antibody/PBS liquid, taking out the decellularized amnion and putting the decellularized amnion in the PBS for preservation and standby. The method has the advantages that the cells are removed from the amnion in a manner that a surfactant is combined with steapsin and deoxyribonuclease, the cells can be prompted to be completely and efficiently removed from the amnion due to the combined use of the steapsin and the deoxyribonuclease, and meanwhile, the extracellular matrix is reserved to the maximum and is enabled to be free from damage.

Disclosed is a method for processing porcine cornea using an aqueous NaCl solution and an aqueous trypsin/EDTA solution to decellularize enucleated porcine cornea. The porcine cornea processed by the method causes neither inflammation nor immune rejection. The porcine corneal stroma decellularized by the method can be recellularized together with host keratocytes after transplantation.