Extracellular matrix freeze-drying protection liquid and application method thereof

Abstract

The invention relates to an extracellular matrix freeze-drying protection liquid. The extracellular matrix freeze-drying protection liquid takes normal saline as a solvent and comprises the following components: chondroitin sulfate, vitamin C, polyethylene glycol, trehalose, glucan, serine and threonine. The invention has the following advantages: the protection liquid has high penetrability, can be vitrified under a low-temperature condition, is rich in negative polar groups and hydroxyl groups, and can be efficiently bonded with collagen molecules; the stable and uniform extracellular matrix freeze-drying effect can be ensured; the protection liquid can be formed into the vitreous state at low temperature, thereby preventing ice crystals from damaging the protein structure; and a protection film is formed at the outer layer of the matrix in stead of hydrogen bonds formed by water molecules and collagen, thereby protecting the collagen function and the stability of the three-dimensional structure. The extracellular matrix freeze-drying protection liquid has low cytotoxicity, and can be used immediately after being rinsed with normal saline or sterilized water; the freeze-dried extracellular matrix can be preserved at normal temperature for more than three years when being packaged in vacuum; and the extracellular matrix freeze-drying protection liquid is suitable for the preservation of purified extracellular matrix such as collagen, elastin, glucoprotein and the like, and is also suitable for various biologic materials constructed by extracellular matrix materials.

Description

technical field [0001] The invention belongs to the technical field of medical biological material protection, and in particular relates to a protective solution for freeze-drying of extracellular matrix (abbreviated as freeze-drying) and a use method thereof. Background technique [0002] Extracellular matrix is ​​secreted by cells and can initiate intracellular signaling pathways through specific surface receptors to regulate cell differentiation, proliferation, migration, adhesion and other functions. It is derived from natural tissues and has good biocompatibility. The extracellular matrix is ​​mainly composed of collagen, laminin, fibronectin and other components; it is obtained by removing cellular components in biomaterials by physical and chemical means. This type of biomaterials is used in clinical repair and tissue engineering tissue construction. It has obvious curative effects and advantages; currently there are dozens of products on the market based on extracel...

AI Technical Summary

Problems solved by technology

Moreover, the extracellular matrix is ​​insoluble in water and cannot form a eutectic point with the lyoprotectant, so the protective agent and the extracellular matrix cannot form a stable co-state during the freezing process. Therefore, the lyoprotectant for soluble small molecular proteins is used in When the extracellular matrix is ​​removed, the generated ice crystals will damage the three-dimensional structure of the matrix, resulting in changes in the physical and chemical properties of the matrix

In addition, the surface of collagen molecules has a large number of positive charges, while the electronegativity of carbohydrates is weak, so the adhesion to collagen is weak, and the ability to compete with water molecules to replace hydrogen bonds is also weak, and the protection of collagen molecules is insufficient. , easy to collapse the collagen structure and decrease its activity