Acellular matrix repairing gel and new method for preparing the same

Abstract

The invention relates to the field of bio-materials, and especially relates to an acellular matrix repairing gel and a new method for preparing the same. The invention discloses the acellular matrix repairing gel and the method for preparing the same, wherein the method includes steps of acellular treatment and gelatinization treatment on tissue and organs from mammal animals to prepare the acellular matrix repairing gel. The acellular matrix repairing gel is eliminated in immunogenicity of heterologous and foreign tissue, so that activity of extracellular matrix components of the tissue is maintained as more as possible. The gel can specially repair damaged tissue and organs of human body, is strong in applicability, is suitable for requirements of various irregular-shaped repair zones and different position environments in body and has a huge clinical value.

Description

technical field

[0001] The invention relates to the preparation of tissue matrix materials, in particular to an acellular matrix repair gel and a new preparation method thereof, belonging to the field of biological materials. Background technique

[0002] The repair and reconstruction of tissues and organs refers to the process of repairing and restoring tissues and organs through regeneration by adjacent healthy cells. Due to the inherent defects in the spontaneous healing process of tissues and organs, once there is a defect or dysfunction in the tissues and organs, it will cause a certain degree of damage. Aplastic impairment or permanent defect on the Therefore, the repair and reconstruction of tissues and organs has always been the focus of related fields such as biology and medicine. With the development and in-depth research on the mechanism of tissue regeneration and repair and tissue engineering, it has become a hot spot in this field to find and develop various bi...

Examples

Embodiment 1

[0054] Example 1 Preparation of bovine sciatic nerve acellular matrix repair gel

[0055] 1. Preparation of Acellular Matrix Repair Gel

[0056] Take 20 g of bovine sciatic nerve, wash it with PBS buffer to remove blood and dirt, soak it in 200 ml of 0.1% peracetic acid solution for 0.5 h to sterilize, then cut the sciatic nerve into small pieces of about 0.1 cm, and place it in 200 ml of 2% Pull through X-100 PBS buffer solution, shake for 0.5h, then use 100ml of PBS buffer solution containing 0.1% trypsin and 1% EDTA for decellularization, shake for 4h, and then use 200ml of 5μg / ml ribonuclease and 50μg 5 g of bovine sciatic nerve decellularized matrix was obtained by removing residual nucleic acid in PBS buffer solution of / ml deoxyribonuclease, then digested with 50 ml of 0.01M hydrochloric acid solution containing 0.5% pepsin, shaken for 24 hours, and then added 5 ml of 0.1M sodium hydroxide solution and 50ml of PBS buffer, adjust pH to 7.4, and incubate at 37°C for 48h ...

Embodiment 2

[0060] Example 2 Preparation of porcine dermis acellular matrix repair gel

[0061] 1. Preparation of Acellular Matrix Repair Gel

[0062] About 100 g of pig dermis was taken, washed with PBS buffer to remove blood and dirt, soaked in 200 ml of 1% peracetic acid solution for 2 hours to sterilize, then cut the dermis into small pieces of 1 cm square and placed in 500 ml of 0.1% sodium deoxycholate In PBS buffer solution, shake for 2h, then use 500ml PBS buffer solution containing 1% trypsin and 0.1% EDTA for decellularization treatment, shake for 1h, and then use 200ml containing 50μg / ml ribonuclease and 500μg / ml DNA Enzyme in PBS buffer solution to remove residual nucleic acid to obtain 32g of pig dermis acellular matrix, then digest with 200ml PBS solution containing 0.1% papain, adjust pH to 2 after shaking for 48h to stop digestion, add 300ml PBS buffer, adjust pH To 7.4, incubate at 37℃ for 24h to obtain porcine dermal acellular matrix repair gel.

[0063] 2. Effect expe...

Embodiment 3

[0065] Example 3 Preparation of bovine tendon acellular matrix repair gel

[0066] Take 20 g of bovine tendon, wash it with PBS buffer to remove blood and dirt, soak it in 50 ml of 0.5% peracetic acid solution for 1 hour to sterilize, then cut the tendon into small pieces of about 0.5 cm, and place it in 50 ml of fat with a concentration of 0.5% In PBS buffer solution of alcohol polyoxyethylene ether (perparine), shake for 1 h, then decellularize with 60 ml of PBS buffer solution containing 0.1% trypsin and 0.1% EDTA, shake for 3 hours, and then use 40 ml of 10 μg / ml ribose Nuclease and 50μg / ml DNase in PBS buffer solution remove residual nucleic acid to obtain 5g of bovine tendon acellular matrix, which is then digested with 50ml of 0.01M hydrochloric acid solution containing 0.1% pepsin, shaken for 24h, and added with 5ml of 0.1M Sodium hydroxide solution and 50ml PBS buffer were adjusted to pH 7.4 and incubated at 37°C for 24h to obtain bovine tendon acellular matrix repair...

technical field

[0001] The invention relates to the preparation of tissue matrix materials, in particular to an acellular matrix repair gel and a new preparation method thereof, belonging to the field of biological materials. Background technique

[0002] The repair and reconstruction of tissues and organs refers to the process of repairing and restoring tissues and organs through regeneration by adjacent healthy cells. Due to the inherent defects in the spontaneous healing process of tissues and organs, once there is a defect or dysfunction in the tissues and organs, it will cause a certain degree of damage. Aplastic impairment or permanent defect on the Therefore, the repair and reconstruction of tissues and organs has always been the focus of related fields such as biology and medicine. With the development and in-depth research on the mechanism of tissue regeneration and repair and tissue engineering, it has become a hot spot in this field to find and develop various bi...

Examples

Embodiment 1

[0054] Example 1 Preparation of bovine sciatic nerve acellular matrix repair gel

[0055] 1. Preparation of Acellular Matrix Repair Gel

[0056] Take 20 g of bovine sciatic nerve, wash it with PBS buffer to remove blood and dirt, soak it in 200 ml of 0.1% peracetic acid solution for 0.5 h to sterilize, then cut the sciatic nerve into small pieces of about 0.1 cm, and place it in 200 ml of 2% Pull through X-100 PBS buffer solution, shake for 0.5h, then use 100ml of PBS buffer solution containing 0.1% trypsin and 1% EDTA for decellularization, shake for 4h, and then use 200ml of 5μg / ml ribonuclease and 50μg 5 g of bovine sciatic nerve decellularized matrix was obtained by removing residual nucleic acid in PBS buffer solution of / ml deoxyribonuclease, then digested with 50 ml of 0.01M hydrochloric acid solution containing 0.5% pepsin, shaken for 24 hours, and then added 5 ml of 0.1M sodium hydroxide solution and 50ml of PBS buffer, adjust pH to 7.4, and incubate at 37°C for 48h ...

Embodiment 2

[0060] Example 2 Preparation of porcine dermis acellular matrix repair gel

[0061] 1. Preparation of Acellular Matrix Repair Gel

[0062] About 100 g of pig dermis was taken, washed with PBS buffer to remove blood and dirt, soaked in 200 ml of 1% peracetic acid solution for 2 hours to sterilize, then cut the dermis into small pieces of 1 cm square and placed in 500 ml of 0.1% sodium deoxycholate In PBS buffer solution, shake for 2h, then use 500ml PBS buffer solution containing 1% trypsin and 0.1% EDTA for decellularization treatment, shake for 1h, and then use 200ml containing 50μg / ml ribonuclease and 500μg / ml DNA Enzyme in PBS buffer solution to remove residual nucleic acid to obtain 32g of pig dermis acellular matrix, then digest with 200ml PBS solution containing 0.1% papain, adjust pH to 2 after shaking for 48h to stop digestion, add 300ml PBS buffer, adjust pH To 7.4, incubate at 37℃ for 24h to obtain porcine dermal acellular matrix repair gel.

[0063] 2. Effect expe...

Embodiment 3

[0065] Example 3 Preparation of bovine tendon acellular matrix repair gel

[0066] Take 20 g of bovine tendon, wash it with PBS buffer to remove blood and dirt, soak it in 50 ml of 0.5% peracetic acid solution for 1 hour to sterilize, then cut the tendon into small pieces of about 0.5 cm, and place it in 50 ml of fat with a concentration of 0.5% In PBS buffer solution of alcohol polyoxyethylene ether (perparine), shake for 1 h, then decellularize with 60 ml of PBS buffer solution containing 0.1% trypsin and 0.1% EDTA, shake for 3 hours, and then use 40 ml of 10 μg / ml ribose Nuclease and 50μg / ml DNase in PBS buffer solution remove residual nucleic acid to obtain 5g of bovine tendon acellular matrix, which is then digested with 50ml of 0.01M hydrochloric acid solution containing 0.1% pepsin, shaken for 24h, and added with 5ml of 0.1M Sodium hydroxide solution and 50ml PBS buffer were adjusted to pH 7.4 and incubated at 37°C for 24h to obtain bovine tendon acellular matrix repair...

Images