Tissue engineering natural gel stent material and preparation method and application thereof
A tissue engineering and scaffold material technology, applied in medical science, prosthesis, etc., can solve the problems of biocompatibility and mechanical strength that are difficult to meet tissue engineering, and achieve high application prospects and practical value, and the steps are simple and easy to implement. The effect of biocompatibility
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Embodiment 1
[0032] Example 1 Preparation of Natural Gel Scaffold Materials for Tissue Engineering
[0033] Take 30g of bean curd leaves and freeze them for 24 hours, put the frozen bean curd leaves and 20ml of deionized water into a mortar, mash, add water while grinding until it becomes a paste. Put the ground tofu chai juice and residue into a centrifuge tube, centrifuge at 4000 r / min for 5 minutes, then suck out the supernatant, put the residue into a bag, seal it and store it in a freezer at -4°C for later use. Add the supernatant obtained from the crude extraction into 50ml of 90% ethanol, oscillate and mix evenly, then put it into a centrifuge tube and centrifuge at a speed of 5000 r / min for 10min, then suck out the supernatant, and take 3ml of the supernatant obtained in the previous step The supernatant was added dropwise to 1mol / L CaCl 2 Test the principle of gel in the solution. If there is no obvious gel phenomenon, dissolve the obtained precipitate in water and heat it at 6...
Embodiment 2
[0034] Example 2 Preparation of tissue engineered grafts
[0035] Remove excess fat and muscle from the fresh pork hind legs, retain the knee cartilage and femur, cut off the cruciate ligament, carefully open the patella, expose the joint cavity, and continue to separate the tissue until the articular cartilage surface is fully exposed, taking care not to damage the articular cartilage. Carefully cut out the square cartilage tissue, hold it with tweezers and soak it in 10ml PBS for cleaning. After the samples were collected, the cartilage tissue was washed 3 times with double antibodies, and the supernatant was discarded. Transfer the cartilage tissue to a 10ml petri dish, fix the cartilage with tweezers, and remove the liquid with a pipette. Add a small amount of PBS to the petri dish, just cover the tissue, chop the cartilage tissue, use a gun tip to move the chopped tissue into a 50ml centrifuge tube, shake it well for 2-3 times, and pour off the liquid , add about 20ml...
Embodiment 3
[0037] Example 3 Tissue Engineering Gel Scaffold Material Used in the Detection of Cytotoxic MTT in Chondrocyte Culture
[0038] The concentration of the chondrocyte obtained in embodiment 2 is made into 1×10 4Cell / mL, inoculate in 96 well plate by 100uL every hole, change into the tissue engineering natural gel support that contains 1 mg / ml obtained by embodiment 1 after cultivating 24 hours (the gel support after the steam sterilization that obtains) granule), the simple DMEM culture medium is used as a negative control, and the DMEM culture fluid containing 649g / L phenol is used as a positive control, and each sample is repeated 8 samples at each time point. After culturing for 2, 4 and 7 days respectively, 100 μL of 0.5 mg / ml 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution was added to each well plate. After incubating at 37°C for 4 hours, aspirate the culture medium in each well plate and add 150 μL of DMSO (dimethyl sulfoxide). After the 9...
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