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Acellular dermal matrix and preparation method and application thereof

A decellularized dermis and matrix technology, applied in the field of medical biomaterials, can solve the problems that the acellular dermal matrix is ​​not suitable for corneal repair materials, the removal of intracellular substances is not complete, and the preparation method is not described in detail.

Inactive Publication Date: 2015-01-21
李学民 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] Existing patents disclose various preparation methods of acellular dermal matrix. For example, the patent with the announcement number CN201350162Y and the invention name of an acellular allogeneic dermal matrix tissue patch discloses a kind of acellular allogeneic dermal matrix as a basement membrane. , immerse it in a methoxil traditional Chinese medicine solution, so that at least one side of the basement membrane has a methoxil traditional Chinese medicine layer, but this patent does not describe in detail the preparation method of the acellular allogeneic dermal basement membrane; the publication number is CN1775189, and the name is a kind of acellular The patent application for the dermal matrix uses mammalian skin to be repeatedly treated with sodium hydroxide solution to remove the DNA of the cellular genetic material in the tissue, and then use a detergent to remove the lipids in the cell membrane. Although this method can produce an acellular dermal matrix, but Its preparation process is cumbersome and cannot effectively remove cell debris components; the publication number is CN1569260, and the patent application titled a preparation method of acellular dermal matrix material uses healthy animal skin as raw material, and uses a compound enzyme skin collagen treatment agent and alkali, etc. Acellular dermal matrix is ​​prepared by treating with various chemical substances, but this method easily leads to residues of various chemical substances, which affects the growth of fibrovascular tissue after implantation, and the preparation process is complicated and takes a long time; the publication number is CN1343523, the name The patent application for microporous allogeneic (species) acellular dermal substitute discloses that human cadaver skin is soaked in traditional enzyme-digested detergent to prepare acellular dermal matrix, but this method does not completely remove intracellular substances and is easy to cause immunogenic residues
However, none of these acellular dermal matrices are suitable as corneal repair materials

Method used

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  • Acellular dermal matrix and preparation method and application thereof
  • Acellular dermal matrix and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0051] Example 1 Preparation of acellular dermal matrix

[0052] 1) Obtain the split skin including the epidermis and dermis from the donated corpse; the corpse is purchased from Peking University Health Science Center;

[0053] 2) Processing the cut skin obtained in step 1) into a freeze-dried sheet at a temperature of -15 to -80°C, then removing the 50-200 μm epidermal layer to expose the papillary layer below, and cutting the 800 μm papillary layer to obtain a dermal matrix;

[0054] 3) Use 0.01M sodium phosphate buffer fixative containing 1.5g / 100ml glutaraldehyde to fix the dermal matrix obtained in step 2), the fixation time is 2 hours, and wash with distilled water after fixing;

[0055] 4) using NaOH with an equivalent concentration of 1.5N to remove cellular components in the dermal matrix, and the reaction time is 10 hours to obtain an acellular dermal matrix;

[0056] 5) washing the acellular dermal matrix with 0.01M sodium phosphate aqueous solution for 10 minut...

Embodiment 2

[0059] Example 2 Preparation and use of acellular dermal matrix

[0060] 1) Obtain the split skin including the epidermis and dermis from the corpse;

[0061] 2) Use 0.01M sodium phosphate buffer solution containing 1.5g / 100ml glutaraldehyde to fix the sectioned skin obtained in step 1) for 2 hours, and wash with distilled water after fixing;

[0062] 3) Use KOH alkaline solution with an equivalent concentration of 2.0N to remove the cellular components in the skin, and the reaction time is 60 hours to obtain decellularized split skin;

[0063] 4) washing the decellularized skin with distilled water for 50 minutes;

[0064] 5) Process the decellularized skin section obtained in step 4) into a freeze-dried sheet at a temperature of -27°C, then remove the 50-200 μm epidermal layer to expose the underlying papillary layer, and cut the 100 μm papillary layer to obtain an acellular dermal matrix ;

[0065] 6) Incubate the acellular dermal matrix obtained in step 5) in a 0.1M g...

Embodiment 3

[0068] Example 3 Corneal Transplantation Test

[0069] The acellular dermal matrix with a size of 1 cm×1 cm prepared in Example 1 was cut into small pieces of 2 mm×5 mm, and implanted into the right corneas of 15 New Zealand white rabbits to evaluate the biocompatibility and transparency of the carrier. Under general anesthesia, a 3×6 mm corneal stroma incision was made in the temporal peripheral region along the superior-inferior direction with a depth of half the thickness of the cornea. The ADM was inserted into this incision with forceps without suturing. The unoperated fellow eye served as a positive control. Gentamicin was injected subconjunctivally on the day of surgery. Tobramycin and dexamethasone eye drops were used for the first week after surgery. Subsequent clinical evaluation of corneal light permeability, neovascularization, and rejection was performed through slit lamp examination.

[0070] experiment such as image 3 Shown: The observation period was 24...

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Abstract

The invention provides an acellular dermal matrix which can be used as a cornea repair material. The acellular dermal matrix contains a papillary layer with the thickness of 100-800 microns. The invention also provides a preparation method of the acellular dermal matrix and a processing method of the acellular dermal matrix before its application. On the other hand, the invention provides an application of the acellular dermal matrix in preparation of a cornea repair material. The acellular dermal matrix has transparent cornea pertinency. The inventor compares sections of ultrathin acellular papillary layer's skin patch, human full-thickness skin and full-thickness cornea sections. The result shows that cells in the ultrathin acellular papillary layer's skin patch are completely removed, and reserved extracellular matrix is arranged in order and the arrangement of the extracellular matrix is close to that of cornea fiber. Thus, the acellular dermal matrix can be used as the cornea repair material to be used in treating and / or improving eye diseases such as cornea thinning and the like.

Description

technical field [0001] The invention belongs to the field of medical biomaterials, in particular to an acellular dermal matrix for repairing transparent cornea and a preparation method thereof. Background technique [0002] Acellular Dermal Matrix (ADM) is obtained by removing cellular components in the epidermis and dermis from animal skin or human cadaver skin through a series of physical and chemical methods, and retaining dermal matrix components with extremely low immunogenicity. In 1995, American Life Cell Company first reported the preparation method of acellular dermal matrix, and at the same time carried out research on its clinical application. Acellular dermal matrix was first used as a dermal substitute to replace and repair dermal defects caused by burns and trauma, but later studies found that acellular dermal matrix can be used not only for burns and trauma wounds, but also for mucosal repair and soft tissue filling. . [0003] Acellular dermal matrix is ​​d...

Claims

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Application Information

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IPC IPC(8): A61L27/36
Inventor 杨淑珍
Owner 李学民
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