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Preparation and application of bioengineering decellularized dermal matrix

A decellularized dermis and bioengineering technology, applied in tissue regeneration, medical science, prosthesis, etc., can solve the problems of losing the basic biological functions of decellularized tissues, destroying cells, and destroying the internal structure of tissues

Active Publication Date: 2016-08-10
拜欧迪赛尔成都生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing acellular dermis manufacturing technologies all use chemical reagents to destroy the cells, and combine glutaraldehyde and other cross-linking agents to cross-link to reduce immunogenicity, which has the disadvantages of destroying the collagen structure and remaining biological toxicity.
In recent years, the method of removing heterogeneous skin cells by physical means has also been proposed, hoping to solve the shortcomings of the preparation of dermal materials by chemical reagent treatment. However, these decellularization methods often lead to damage to the internal structure of the tissue and lose the original structure of the decellularized tissue. basic biological functions

Method used

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  • Preparation and application of bioengineering decellularized dermal matrix
  • Preparation and application of bioengineering decellularized dermal matrix
  • Preparation and application of bioengineering decellularized dermal matrix

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Pig back skin is taken, and after cleaning and hair removal, medium-thick dermis skin slices with a thickness of 0.3-0.5 mm and a width of 1-5 cm are reserved for future use.

[0024] 1. Protect the dermal tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 40g / L, hyaluronic acid 20g / L, neomycin 5mg / L, adjust the pH value to 7.2 , the osmotic pressure is 380mOsm;

[0025] 2. Seal the dermis in a plastic bag filled with protective liquid;

[0026] 3. Under the condition of 400MPa high static pressure, treat 8 times, each time is 5 minutes;

[0027] 4. After taking out the dermis, place it in a protective solution containing 2% Pluronic F68+2000U / ml DNase at a temperature of 30°C, set the shaker speed to 100 rpm, and treat for 4 hours;

[0028] 5. After the detergent and enzyme treatment, take out the conjunctiva and rinse it in the protective solution for 3 hours.

Embodiment 2

[0030] Pig back skin is taken, and after cleaning and hair removal, medium-thick dermis skin slices with a thickness of 0.3-0.5 mm and a width of 1-5 cm are reserved for future use.

[0031] 1. Protect the conjunctival tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 50g / L, glycerin 200g / L, neomycin 5mg / L, adjust the pH value to 7.2, The osmotic pressure is 400mOsm;

[0032] 2. Seal the dermis in a plastic bag filled with protective liquid;

[0033] 3. Under the condition of 600MPa high static pressure, treat 5 times, each time is 3 minutes;

[0034] 4. After taking out the dermis, place it in a protective solution containing 5% sodium lauryl sarcosine + 4000U / ml DNase at a temperature of 25°C, set the speed of the shaker at 100 rpm, and treat for 3 hours ;

[0035] 5. After the detergent and enzyme treatment, take out the conjunctiva and rinse it in the protective solution for...

Embodiment 3

[0037] Pig back skin is taken, and after cleaning and hair removal, medium-thick dermis skin slices with a thickness of 0.3-0.5 mm and a width of 1-5 cm are reserved for future use.

[0038] 1. Protect the conjunctival tissue with protective solution throughout the whole process. The formula of the protective solution is: RPMI-1640 medium, add chondroitin sulfate 30g / L, hyaluronic acid 50g / L, neomycin 5mg / L, adjust the pH value to 7.2, the osmotic pressure is 450mOsm;

[0039] 2. Seal the dermis in a plastic bag filled with protective liquid;

[0040] 3. Under the condition of 800MPa high static pressure, treat 5 times, each time is 2 minutes;

[0041] 4. After taking out the dermis, place it in a protective solution containing 2% sodium dodecylsulfonate + 2000U / ml DNase at a temperature of 30°C, set the shaker speed to 100 rpm, and treat for 5 hours;

[0042] 5. After the detergent and enzyme treatment, take out the conjunctiva and rinse it in the protective solution for 8 ...

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Abstract

The invention discloses a novel bioengineering decellularized dermal matrix and a preparing method thereof. The preparing method includes the steps that porcine skin is pretreated to obtain dermal layer skin graft, and a mixed protective solution is used in all the dermal matrix decellularization process; a high hydrostatic pressure technology is used for preseparating cells in dermal tissue; residual cell nucleuses are digested through a composite nuclease method; detergent is used for removing loose smudge cell sheets; the mixed protective solution is used for rinsing. Suitable and efficient physical means are used and supplemented with a small amount of enzymes and detergent, cell components in the dermis are fully removed, under whole-process protection of the protective solution, it can be ensured that immunogenicity of the dermal matrix is reduced to the maximum on the basis of ensuring complete removal of the cell nucleuses, a normal three-dimensional collagenous fiber structure and array polarity of the dermal matrix are reserved, and therefore the prepared decellularized dermal matrix is high in stability and biocompatibility.

Description

technical field [0001] The invention relates to the field of preparation of bioengineering medical materials, in particular to a method for preparing decellularized dermal matrix Background technique [0002] Patients with severe trauma, extensive burns, and skin ulcers have huge wounds with skin loss or are difficult to heal, and autologous skin transplantation is basically the main method for treating defect wounds, but the source of autologous skin is often tight, especially when large areas of skin defects Obtaining autologous skin is even more difficult. On the other hand, taking autologous skin is a new trauma to the patient; therefore, in the treatment of these diseases, it has become a clinical need to use skin substitutes to cover the wound temporarily or for a long time. [0003] Natural dermal material is the first choice for skin grafts due to its similarity to human skin. Among them, heterogeneous skin is cheap and comes from a wide range of sources, but there...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/36
CPCA61L27/362A61L27/3687A61L27/3691A61L27/60A61L2430/40
Inventor 史真史伟云
Owner 拜欧迪赛尔成都生物科技有限公司
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