Active scaffold material capable of promoting bone regeneration as well as preparation method and application of active scaffold material
A scaffold material and bone regeneration technology, applied in the field of biomedical materials, can solve the problems of low concentration, chemical cross-linked molecules entering the body, easy dispersion, etc., to achieve improved local concentration, good biocompatibility and biodegradability, good stability
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[0074] A method for preparing the demineralized bone matrix scaffold material for promoting bone regeneration provided by the embodiments of the present invention includes:
[0075] Provides a demineralized bone matrix with a porous internal structure;
[0076] The decalcified bone matrix is contacted with a chemokine solution, so that the decalcified bone matrix is loaded with chemokines to form the decalcified bone matrix scaffold material.
[0077] Further, the preparation method includes: combining the collagen binding region with the stromal cell-derived factor by adopting point gene fusion expression method, so as to form the chemokine.
[0078] Further, the preparation method includes: slowly applying the chemokine solution on the decalcified bone matrix, so that the decalcified bone matrix is loaded with chemokines.
[0079] Further, the concentration of the chemokine solution is above 10 nM.
[0080] Further, the preparation method includes: providing allogene...
Embodiment 1
[0089] (1) Preparation method of demineralized bone matrix scaffold material modified with chemokines
[0090] 1) Firstly, the Escherichia coli expression vector of the chemokine is constructed by using the genetic engineering method known in the industry, and the high-purity chemokine is purified and expressed, wherein the chemokine uses SDF1α;
[0091] 2) Secondly, the decalcified bone matrix material is treated with an acid-base method. Specifically, the long bone tissue of the allogeneic source is provided, cut into a suitable size, and then treated in acetone for 48 hours to remove the soft tissue, fat and other substances on the surface. Then wash with double distilled water several times and then process decalcification with 0.6M HCI solution, freeze-dry after final double distilled water washing; Standby after irradiation sterilization again, the decalcified bone matrix that obtains has the internal structure of loose hole (such as Figure 2A-Figure 2B As shown), it ca...
Embodiment 2
[0105] 1. Preparation of demineralized bone matrix scaffold material modified by CBD-SDF1α and characterization of its macroscopic and microscopic structures
[0106] First, the Escherichia coli expression vector of CBD-SDF1α / NAT-SDF1α was constructed by genetic engineering method, and high-purity CBD-SDF1α / NAT-SDF1α was purified and expressed. Secondly, it is to utilize the acid-base method to obtain the decalcified bone matrix material (DMB stent), and it is ready for use after irradiation sterilization (the same as in Example 1, and the appearance can be referred to Figure 2A-Figure 2B ). Under sterile conditions, dissolve 10nM CBD-SDF1α / NAT-SDF1α with 20μl of PBS, and add dropwise and multiple points onto the sheared material. Note that this step should be performed slowly to ensure sufficient biological activity factors. loaded onto demineralized bone matrix scaffold materials such as Figure 7A The loading of CBD-SDF1α / NAT-SDF1α to the demineralized bone matrix scaffo...
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