A kind of bladder repair material and its use and preparation method
A technology for repairing materials and antibodies, applied in the fields of medical tissue engineering and bladder tissue repair, can solve problems such as difficulty in effectively promoting muscle regeneration and few smooth muscle cells, and achieve excellent repair performance, good recovery effect, and the effect of promoting regeneration.
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Embodiment 1
[0048] Embodiment 1 Preparation of a new tissue repair material
[0049] 1. Preparation of SIS
[0050] (1) Cleaning the intestines: take a pair of fresh pig small intestines, remove excess chyme in the intestines, turn the small intestines inside out, and clean the inner surface repeatedly with normal saline, then cut the small intestines longitudinally with scissors, and finally cut them into long lengths Intestine segment about 15cm.
[0051] (2) Physical curettage: use a spatula to scrape off the serosa layer and muscle layer on the intestinal segment to obtain a preliminary SIS. This process needs to be repeatedly scraped off until the milky white translucent film is separated, and washed 2-3 times with normal saline.
[0052] (3) Degreasing: After repeatedly rinsing with deionized water, wrap the SIS membrane with gauze, squeeze out part of the water, and remove excess water with filter paper. After mixing the chloroform-methanol mixture at 1:1 v / v, soak the dried SIS...
Embodiment 2
[0065] Embodiment 2 Preparation of a tissue repair material
[0066] 1. Preparation of SIS
[0067] With embodiment 1 step 1.
[0068] 2. Thiolation of SIS Membranes
[0069] (1) Cut the SIS material into discs with d = 15.6 mm (24-well plate size), and place 10 SIS discs in a glass dish with d = 60 mm;
[0070] (2) Prepare 2ml of Traut's solution of 5mg / ml with PBS;
[0071] (3) Completely soak the materials (10 tablets / dish) in the step (1) in Traut's solution, and react at room temperature for 1.5 hours. After the reaction, wash with PBS 2-3 times.
[0072] 3. Coupling of Antibody to SIS Membrane
[0073] This example takes the antibody CD29 as an example to illustrate the coupling process of the antibody to the SIS membrane. 4-(N-Maleimidomethyl)cyclohexane-1-carboxylic acid sulfosuccinimidyl ester sodium salt (Sulfo-SMCC; sigma) was prepared at a concentration of 0.5 mg / ml in 100 μl of PBS React with 10 μl (1 mg / ml) CD29 antibody in a glass dish at room temperature...
Embodiment 3
[0076] Embodiment 3 Preparation of a tissue repair material
[0077] 1. Preparation of SIS
[0078] With embodiment 1 step 1.
[0079] 2. Thiolation of SIS Membranes
[0080] (1) Cut the SIS material into discs with d = 15.6 mm (24-well plate size), and place 10 SIS discs in a glass dish with d = 60 mm;
[0081] (2) Prepare 2ml of Traut's solution of 10mg / ml with PBS;
[0082] (3) Completely soak the materials (10 tablets / dish) in the step (1) in Traut's solution, and react at room temperature for 1.5 hours. After the reaction, wash with PBS 2-3 times.
[0083] 3. Coupling of Antibody to SIS Membrane
[0084] This example takes the antibody CD29 as an example to illustrate the coupling process of the antibody to the SIS membrane. With 100 μl, the 0.5mg / ml 4-(N-maleimidomethyl)cyclohexane-1-carboxylic acid sulfosuccinimide sodium salt (Sulfo-SMCC; sigma) prepared in PBS was mixed with 10 μl (1 mg / ml) of CD29 antibody was reacted in a glass dish for 1.5 hours at room temp...
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