Decellularized black carp skin biomimetic substrate for regeneration restoration as well as preparation method and application thereof
A decellularized herring technology, applied in the field of decellularized herring skin biomimetic matrix for regenerative repair and its preparation, can solve the problems of delaying conventional wound healing and no degreasing treatment, so as to improve biological safety and compatibility, cell ablation The effect of high removal rate and avoiding religious barriers
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Embodiment 1
[0041] Example 1 Preparation of decellularized herring skin biomimetic matrix for regenerative repair
[0042] (1) Pretreatment of herring skin: fresh herring skin is scraped off residual scales and redundant muscle tissues, rinsed with deionized water at 4°C several times to remove residual blood; cut into small pieces of 30×50mm according to different anatomical parts, and quickly Freeze at -40°C, take out and thaw under running water at room temperature before use;
[0043] (2) Remove mucous membrane / liquid and degrease: at 4°C, stir with 3.5% NaCl solution with a material-to-liquid ratio of 1:20 (w / w) for 1 hour, then remove the waste liquid, and roughly clean the herring skin to remove part of the redundancy; above The steps are repeated 3 times;
[0044] (3) Microstructure protection: at 4°C, stir with PBS buffer solution (0.1M, pH7.4) with a solid-to-liquid ratio of 1:20 (w / w) for 2 hours and then remove the waste liquid; repeat the above steps twice;
[0045] (4) Cle...
Embodiment 2
[0052] Example 2 Preparation of decellularized herring skin biomimetic matrix for regenerative repair
[0053] (1) herring skin pretreatment: with embodiment 1.
[0054] (2) Remove mucous membrane / liquid and degrease: at 4°C, stir with 5% NaCl solution with a material-to-liquid ratio of 1:10 (w / w) for 1 hour, then remove the waste liquid, and roughly clean the herring skin to remove part of the redundancy; above The steps are repeated 2 times.
[0055] (3) Microstructure protection: at 4°C, stir with PBS buffer solution (0.2M, pH7.0) with a solid-to-liquid ratio of 1:10 (w / w) for 2 hours, then remove the waste liquid; repeat the above steps once.
[0056] (4) Cleaning: at 4°C, stir for 4 hours with a 5% NaCl solution with a solid-to-liquid ratio of 1:10 (w / w), and then remove the waste liquid.
[0057] (5) Initial decellularization treatment: at 4°C, 2% sodium deoxycholate (containing 50 μg / ml gentamicin sulfate) with a solid-liquid ratio of 1:10 (w / w) was stirred for 18 hou...
Embodiment 3
[0063] Example 3 Preparation of Decellularized Herring Skin Biomimetic Matrix for Regeneration and Repair
[0064] (1) herring skin pretreatment: with embodiment 1;
[0065] (2) remove mucous membrane / fluid and degrease: with embodiment 2;
[0066] (3) Microstructure protection: at 4°C, stir for 2 hours with D-Hank’s buffer solution (pH 7.2) with a solid-to-liquid ratio of 1:10 (w / w), and then remove the waste liquid; repeat the above steps twice.
[0067] (4) Cleaning: at 4°C, stir with a 4% NaCl solution with a material-to-liquid ratio of 1:10 (w / w) for 4 hours and then remove the waste liquid;
[0068] (5) Initial decellularization treatment: at 4°C, 0.5% sodium dodecylsulfonate (containing gentamicin sulfate 50 μg / ml) with a solid-liquid ratio of 1:10 (w / w) was stirred for 12 hours. The herring skin was rearranged to remove part of the redundant tissue formed in the above treatment process; at 4°C, it was washed 3 times with Hank's buffer solution with a solid-to-liquid ...
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