Extracellular matrix composite hyaluronic acid gel and preparation method, application and biological material thereof
A technology of hyaluronic acid and biomaterials, applied in tissue regeneration, drug delivery, pharmaceutical formulations, etc., can solve the problems of secondary tissue damage and limit clinical application, and achieve the reduction of secondary damage, good biocompatibility, good The effect of histocompatibility
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[0052] The present invention does not limit the preparation method of the extracellular matrix. Optionally, the extracellular matrix is derived from commercially available products; alternatively, the extracellular matrix can be prepared by conventional methods in the art. The conventional methods in the art include but are not limited to It is used in combination with one or more of repeated freeze-thaw methods, chemical detergent methods, osmotic solution methods or enzyme digestion methods; wherein chemical detergents include but are not limited to SDS, LiDS, sodium cholate, EDTA, TritonX-100, TritonX-114, NP-40 or neoglucoside, etc.
[0053] In some preferred embodiments, the extracellular matrix is derived from an extracellular matrix gel obtained by dissolving acellular matrix material. Extracellular matrix gel is a polymer network system composed of natural proteins and other components in the extracellular matrix obtained by dissolving the decellularized extracellu...
Embodiment 1
[0073] Example 1 Preparation of extracellular matrix gel
[0074] The preparation of extracellular matrix gel using porcine heart extracellular matrix as raw material comprises the following steps:
[0075] (1) Preparation of porcine heart extracellular matrix: take fresh porcine heart, wash it three times with PBS buffer, cut the myocardium into small pieces of 2cm×2cm×0.5cm, incubate with 2% SDS on a shaker at room temperature for 72 hours with an interval of 12 hours The medium was changed once; 1% Triton X100 was incubated on a shaker at room temperature for 72 hours, and the medium was changed every 12 hours; the PBS buffer was washed on a shaker at room temperature for 96 hours, and the medium was changed once every 6 hours, and lyophilized for 12 hours to obtain White blocky myocardial decellularized matrix material.
[0076] (2) Preparation of cardiac extracellular matrix gel: The cardiac acellular matrix material obtained in (1) was ground into a powder using a freez...
Embodiment 2
[0077] Example 2 Preparation of Extracellular Matrix Composite Hyaluronic Acid Gel
[0078] (a) React 2-iminothiophene with extracellular matrix gel at room temperature for 2 hours to generate extracellular matrix-2-iminothiophene compound. The extracellular matrix gel is prepared according to the method provided in Example 1.
[0079] (b) Simultaneously take Sulfo-SMCC reagent and incubate with hyaluronic acid for 1 hour to generate hyaluronic acid-Sulfo-SMCC compound.
[0080] (c) then mix the extracellular matrix-2-iminothiophene compound and the hyaluronic acid-Sulfo-SMCC compound obtained in the above steps and incubate at room temperature for 1 hour to covalently crosslink the hyaluronic acid to the extracellular matrix material, Extracellular matrix complex hyaluronic acid gel, such as figure 1 shown.
[0081] The dosage and concentration of each substance in the above-mentioned covalent crosslinking step are preferably: 2-iminothiophene, Sulfo-SMCC and hyaluronic aci...
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