53 results about "Corneal cell" patented technology

The present invention addresses the need for improved methods and apparatuses of transplanting cells to injured or diseased cornea. The methods disclosed provide for culture of cells on the concave surface of a contact lens, followed by application of the contact lens to the eye, thereby permitting repopulation of the corneal region by the cells. A particular use for the present invention involves the transfer of genetically altered cells.

The invention discloses a preparation method for artificial corneas. The preparation method comprises the following steps of (1) collecting age-appropriate pig eyeballs; (2) scraping corneal epithelial cells and corneal endothelial cells of the pig eyeballs, reserving bowman layers, preparing lamellar corneas by using a lamellar blade, and drilling size-specific corneal slices; (3) putting the corneal slices obtained in step (2) in liquid nitrogen for 8-35 minutes, slowly rewarming the corneal slices to room temperature under normal temperature and normal pressure after the corneal slices are taken out, repeating the operation for 4-12 times; (4) putting the corneal slices which are subjected to freezing-thawing treatment in step (3) in normal saline for carrying out ultrasonic cleaning, wherein the ultrasonic working frequency is 20-250 kHz, and the ultrasonic time is 15-65 minutes; (5) putting the corneal slices which are subjected to ultrasonic treatment in a buffer system, carrying out enzymolysis on DNA (Deoxyribose Nucleic Acid) of corneal stromal cells by using endonuclease, and obtaining the artificial corneas. According to the artificial corneas prepared by the preparation method disclosed by the invention, gaps between natural collagen fibers of the artificial corneas are beneficial for inducing the growth of the corneal cells and the growth of nerve fibers of an acceptor and promoting the repairing and the reconstructing of corneas of a donor.

Described herein are methods and compositions for stimulating proliferation of cells that express adherent junctions and cease proliferation, for example, human corneal endothelial cells, by downregulation of certain cell-cell junctions. In one embodiment, downregulation is achieved using RNA interference, and contacting the cells with mitogenic growth factors and an agent that elevates intracytoplasmic cAMP. Furthermore, described herein are methods of isolating human corneal endothelial cells from keratocytes, and methods of preserving and maintaining viability of human corneal endothelial cell aggregates. Also described are surgical grafts comprising human corneal endothelial cells that have been isolated, optionally stored, and transiently contacted with an agent that downregulates expression of p 120, and a biocompatible support. The methods and compositions described herein can be used in novel therapies to help expand human corneal endothelial cells during in vitro tissue engineering and for in vivo treatment of corneal endothelial dysfunction.

The invention belongs to the field of microfluidic technology and biotechnology, and relates to a multifunctional organ chip based on a microfluidic technology, and a preparation method and an application thereof. The multifunctional organ chip based on the microfluidic technology comprises a substrate, the substrate at least comprises two structural layers, each structural layer is provided witha micro-channel and a cell culture area communicated with the micro-channel, the cell culture areas of every two adjacent structural layers are communicated, and a porous membrane is arranged at the communication position. The invention provides the multifunctional organ chip based on the microfluidic technology, which not only can stably culture human corneal cells in a culture area to form three-dimensional corneal tissues, but also can detect the integrity of an established corneal barrier, and solves the problem that in-vitro two-dimensional culture and in-vivo animal experiments are difficult to correctly reflect human corneas, and the requirements of scientific research and clinical application on an in-vitro bionic research model are met.

The present invention relates to the use of a plastically-compacted collagen gel as a substrate for the growth of corneal cells, particularly limbal corneal epithelial stem cells. Cells grown on such a substrate can be cultured to produce artificial ocular epithelia which can be used in ocular toxicity testing or for transplantation.

The invention relates to methods of modifying cells of corneal tissue to express an active agent, to modified corneal tissue, to vectors utilised in such methods and to methods of xeno- and allo-transplantation utilising the modified corneal tissue. The method of modifying cells of corneal tissue to express an active agent involves exposing harvested corneal tissue to an effective concentration for transfection of an expression vector which comprises a nucleotide sequence encoding for the active agent for a period sufficient to allow infection, such that cells of said the corneal tissue will express the active agent.

The invention disclosed relates to the culturing of mammalian e.g. human cells, and in particular to the culturing and expansion of substantially undifferentiated human epidermal keratinocytes exhibiting stem cell-like characteristics, and in co-culture with human dermal fibroblasts utilizing a low calcium serum free and animal by-product free medium derived from a commercially available medium. The medium consists of a commercially available basal medium, and a single fibroblast growth factor (FGF) or a mimic thereof e.g. FGF7/KGF. A method is also disclosed for treating mammalian e.g. human skin wound injuries, by applying to the wound an effective amount of substantially pure mammalian e.g. human epidermal stem cell-like keratinocytes in a substantially undifferentiated state, and optionally additionally, dermal fibroblasts e.g. human dermal fibroblasts. Both cell types can be grown separately or in a co-culture for application purposes.

The invention provides a collagen-based composite cornea substitute with bioactivity and a preparation method thereof. The composite cornea substitute is prepared by the following steps: taking collagen and 3-(methacrylamide)propyl-dimethyl(3-sulfopropyl)amide (MPDSAH) as raw materials and water as a solvent, crosslinking the collagen with zero-length cross linker carbodiimide (EDC); taking Irgacure 2959 as a photo initiator and polyethyleneglycol diacrylate (PEGDA) as a crosslinking agent to perform initiated polymerization and crosslinking for the MPDSAH; and after even mixing, injecting a die with the mixture for cure forming. After freeze drying, the composite cornea constitute has a three-dimensional porous structure, can adsorb a large amount of cornea cell growth factors, can be stored for a long time, and can recover the primary shape and performance like before freeze drying after water absorption and expansion. The prepared composite cornea substitute has good histocompatibility, can induce and promote the cornea regeneration, and can be biodegraded along with the cornea regeneration, and the introduction of the growth factors can promote the enrichment of self growth factors and reduce the occurrence of postoperative complications.

The invention discloses a sodium hyaluronate eyedrop containing deproteinized calf blood extractive. The components comprise deproteinized calf blood extractive, sodium hyaluronate, pH buffering agent, metal chelating agent, osmotic pressure regulation agent and injection water. The preparation method has the following steps: weighing the sodium hyaluronate, adding the injection water to resolve, and obtaining sodium hyaluronate solution after overnight swelling; fetching the deproteinized calf blood extractive, the pH buffering agent, the metal chelating agent and the osmotic pressure regulation agent, adding the injection water to resolve, mixing with the sodium hyaluronate solution after resolving, and agitating uniformly after supplementing the injection water to full dose; filtering and sterilizing after adjusting the pH of the solution, embedding to single dose packaging containers after being qualified by testing, and sealing and obtaining the product. The eyedrop disclosed by the invention is used for curing corneal wounds and xerophthalmia, can promote the corneal cells to incept and utilize the energy, and improves the recovery rate of the corneal wounds. The preparation method of the eyedrop avoids the stimulus to the surface cells of eyes, and prevents the contamination caused by microorganisms in tears and in air during use and storage.

The invention belongs to the field of biomedical materials, and discloses a collagen film with 3D micro-patterns as well as a preparation method and application thereof. The collagen film with the 3Dmicro-patterns is prepared by the following steps: forming 3D micro-patterns on a silicon wafer by adopting a photolithography technology by using a mask with 3D micro-patterns; pouring PDMS prepolymers onto the silicon wafer so as to be subjected to cross-linking and curing, thereby obtaining a PDMS base with the 3D micro-patterns; dissolving collagen in an acidic solution, adding water, and carrying out uniform stirring so as to obtain a collagen solution; adding a cross-linking agent into the collagen solution so as to be subjected to cross-linking, thereby obtaining cross-linking reactionproducts; and then, removing bubbles from the cross-linking reaction products, pouring the bubble-removed cross-linking reaction products onto the PDMS base, carrying out air-drying so as to have a film formed, and carrying out stripping so as to obtain the collagen film with the 3D micro-patterns. The preparation technology of the collagen film with the 3D micro-patterns is simple in processes and relatively low in cost; and the prepared film combines advantages of collagen and patterned surface, thereby having good physicochemical properties and biocompatibility. The collagen film with the 3D micro-patterns can be used in corneal tissue engineering so as to induce corneal tissue regeneration; moreover, the collagen film with the 3D micro-patterns is capable of performing regulation on corneal cell behavior, thereby forming a structure similar to native tissues.

The invention relates to an ocular surface in-situ drug and a preparation method thereof. The ocular surface in-situ drug comprises a hydrogel system which takes PluronicF127, PluronicF68 and methacrylic acid-hyaluronic acid (HA-MA) as main components and has a temperature response characteristic, and a nano drug-loaded microsphere which contains an active drug or wraps the active drug, in-situ gel is formed at the ocular surface temperature, and the drug release time is prolonged. By combining the hyaluronic acid derivative HA-MA with high biocompatibility and low cytotoxicity, a brand-new temperature-triggered hydrogel sustained-release system formula for eyes is provided, so that drug sustained release on the ocular surface is realized, and the treatment on bacterial/fungal keratitis and conjunctivitis is achieved; and meanwhile, the toxicity of the material is reduced, and the repair of corneal cells can be promoted during treatment.

The invention relates to limbal stem cells and a preparation method thereof. The preparation method comprises the following steps: obtaining corneal tissues, and pre-treating the to-be-used corneal tissues; adding a cell cryopreservation solution containing a cell inducer into the pre-treated corneal tissues; freezing the corneal tissues with the cell cryopreservation solution, wherein the cell inducer is a reagent which can induce inflammation or stress response of part of cells in the corneal tissues, but does not cause cell death directly. The method can achieve screening of the limbal stemcells and various kinds of maturely differentiated corneal cells in the corneal tissues through simple low-temperature refrigeration operation before cell culture, can achieve long-term cryopreservation of the high-activity limbal stem cells at the same time, does not need specific instruments or equipment, is simple to operate, high in screening efficiency and high in purity of screened cells, and can improve the ratio and activity of the limbal stem cells.

The invention discloses a moxifloxacin eye drop having both a bionic function and a targeting function and a preparation method of the moxifloxacin eye drop. Specifically, the moxifloxacin eye drop comprises the following components according to mass concentration: 5-6g/L of moxifloxacin hydrochloride, 3-5g/L of xanthan gum, 1-4g/L of sodium hyaluronate, 6-8g/L of an isoosmotic adjusting agent, and the balance of distilled water; the pH value is regulated through a pH regulator to 6.0-8.0. A xanthan gum/sodium hyaluronate gel system is creatively applied compositely to achieve both the bionic function of tears and the targeting function of a cornea; the xanthan gum has the bionic rheological property similar to that of the tear, so that higher viscosity is achieved when the shearing force is low, and the xanthan gum is evenly coated on the surface of an eyeball when the shearing force is high; the sodium hyaluronate acts on a receptor CD44 of a corneal cell, so that the cell intake is increased, and the targeting absorption is achieved. Compared with a common moxifloxacin eye drop, the moxifloxacin eye drop having both the bionic function and the targeting function has the characteristics of compatibility with the tear, no foreign-matter irritation, long residence time, higher bioavailability and the like.

Methods for cell-based delivery of collagen VII to treat epidermis bullosa and corneal erosion are provided. The present disclosure also provides compositions and pharmaceutical compositions comprising, or consisting essentially of, or further consisting of, a keratinocyte sheet or a corneal cell sheet.

The invention discloses a microwave electrodeless ultraviolet light source, a system and application, and belongs to the technical field of ultraviolet sterilization. According to the ultraviolet light source, inert gas and Zn and/or a Zn compound are filled into a quartz lamp tube, and the inert gas in the quartz lamp tube is excited by a microwave device to generate plasma. Then, the Zn and/or the Zn compound are heated and evaporated, the Zn atoms collide with the electrons or the inert gas atoms in the excited state, transition from the ground state to the excited state is achieved, and then ultraviolet light with the wavelength of 213 nm is radiated when the Zn atoms return to the ground state. Compared with the prior art, the ultraviolet light source provided by the invention has the advantages that Zn and/or a Zn compound is used for replacing mercury as a luminous substance, the raw material is cleaner, and microwave excitation is used for replacing a metal electrode, so that the problem that the service life of the light source is shortened due to electrode consumption is avoided. In addition, the dominant wavelength of the ultraviolet light source is 213nm, so that the skin and corneal cells of a human body are not damaged while bacteria are effectively killed, and the application range is wider.