A preparation method of decellularized biological amniotic membrane with controllable product performance
A decellularized amniotic membrane technology, which is applied in the field of preparation of decellularized biological amnion, can solve the problems of the influence of material structure and performance, the decrease of the thickness and toughness of biological amniotic membrane, and the inability of reagents to remove cells, etc.
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preparation example Construction
[0037]The preparation method includes: putting the sterilized amnion tissue into two different decellularized solutions successively for treatment; the concentration of the decellularized solution, the time and temperature of the treatment are according to the product requirements of the decellularized biological amnion The tensile strength and degradation time are calculated and determined by the above formula.
[0038] In a specific embodiment: the decellularized solution is selected from acid-base, hypertonic saline solution, hypotonic solution, sodium lauryl sulfate, trypsin, etc.;
[0039] Described acid-base is selected from hydrochloric acid, acetic acid, EDTA, sodium hydroxide solution, potassium hydroxide solution, ammoniacal liquor etc.;
[0040] The hypertonic saline solution is selected from 1-3 mol / L sodium chloride or potassium chloride solution, and the hypotonic solution is selected from 0.9% sodium chloride or potassium chloride solution or purified water.
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experiment example 1
[0057] The verification of experimental example 1 prediction formula of the present invention
[0058] Table 1 prediction formula of the present invention
[0059]
[0060] The above prediction formula is based on the mass concentration of the decellularized solution (X 1 , %), processing time (X 2 , h), and temperature (X 3 , ℃) 3 factors as independent variables, the degradation time of amniotic membrane (Y 1 , days), tensile strength (Y 2 , Mpa), as the response value, the prediction formula was used to prepare the amnion product and to verify the performance of the final amnion product. The specific experimental design and results are shown in Table 2 and Table 3 below.
[0061] The specific operation method is: estimate the degradation time of the product at 38-65 days, estimate the tensile strength at 6.7-13.5Mpa, calculate the three factors that need to be controlled in the preparation process through the above prediction formula, and get as follows: Independent...
experiment example 2
[0076] Verification of Experimental Example 2 Optimal Conditions
[0077]Under the optimal process, the predicted values corresponding to each response variable were estimated, and three batches of amnion samples were re-produced, and 1 L of trypsin solution with a concentration of 0.20% was prepared, and the above-mentioned cleaned amnion was soaked in trypsin solution for 12.7 hours at 4°C; prepare SDS solution with a concentration of 0.20%, soak the above amniotic membrane in SDS solution for 20min, shake frequency 110r / min, 4 times in total. The above-mentioned amnion is washed 4-8 times, freeze-dried, irradiated and sterilized for preservation.
[0078] Table 5 The predicted value and experimental value of the response variable under the optimal process
[0079]
[0080] The predicted values and experimental values of each response variable under the optimal process are shown in Table 5. It can be seen that the predicted values of the response variables are ve...
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