Application of in-situ crosslinking hydrogel capable of intraocular injection in preparing artificial vitreous bodies
An in-situ cross-linking and hydrogel technology, applied in prosthesis, medical science, etc., can solve the problem of lack of intraocular fillings and achieve the effect of no obvious toxic reaction
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Embodiment 1
[0028] Example 1, the cell safety test of the in situ cross-linked gel of the present invention
[0029] The cell experiment steps are as follows:
[0030] 1. Dissolve 4ARM-PEG-SH and PEG-S-EMA in phosphate buffer solution with pH = 7.4 and concentration of 0.1mol / L respectively, and make solutions with a concentration of 150mg / mL, and mix the two solutions with The volume ratio was 2:1 and mixed in vitro for 3 hours to obtain gel blocks for cell experiments.
[0031] 2. Cell lines (rat retinal pigment epithelial cell D407 cell line; monkey eye retinal vascular endothelial cell RF / 6A) (CRL-1780 cell line) were routinely cultured, and the culture medium was DMEM / F12=1:1 (purchased from Hyclone, USA company), add fetal bovine serum (final concentration is 10%), and get the cells passed to 3-6 passages for experiment.
[0032] 3. Digest the cells with 0.25% trypsin, mix well to make a cell suspension, count to 50000 / ml, take a sterile 96-well plate, add 100 μl of cell suspensio...
Embodiment 2
[0040] Example 2. Safety experiment of in situ cross-linked gel filling in animal eyes of the present invention
[0041] Five Qingzilan rabbits were selected for the experiment.
[0042] The animal experiment steps are as follows:
[0043] 1. Anesthetize the Qingzilan rabbit.
[0044] 2. Cut the hemibulbar conjunctiva along the limbus, and make perfusion holes and vitrectomy holes with a 19G scleral puncture knife.
[0045] 3. Insert the perfusion head and turn on the perfusion. The formula of the perfusion solution is as follows: add 50% glucose 4ml, dexamethasone (5mg / 1ml) 1.6ml, epinephrine hydrochloride (1mg / 1ml) 0.5ml to 500ml lactated Ringer’s solution, Tobramycin (80,000 U / 1ml) 0.2ml.
[0046] 4. The vitreous cutting head extends into the vitreous cavity to remove all the vitreous.
[0047] 5. Dissolve 4ARM-PEG-SH and PEG-S-EMA respectively in phosphate buffer solution with a concentration of 0.1mol / L at pH=7.4, and prepare a solution with a concentration of 150mg / m...
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