A kind of decellularized biological amniotic membrane comprising loose layer and preparation method thereof
A technology of decellularization and loose layer, which is applied in the field of biological amnion and its preparation technology, can solve the problems of large loss of cell proliferation and anti-inflammation, reduced repair and anti-inflammatory effect, easy to be destroyed and removed, and achieve the improvement of repair and anti-inflammatory function , reduce loss, and ensure safety
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[0022] A method for preparing the decellularized biological amniotic membrane comprising a loose layer as described above, comprising the following steps:
[0023] Carry out virus inactivation to the washed biological amniotic membrane with peracetic acid-sodium chloride solution to obtain inactivated amniotic membrane;
[0024] Wash the inactivated amniotic membrane with normal saline and shake it, and wash it again with sodium bicarbonate solution to obtain the rewashed amniotic membrane;
[0025] The protease solution and the rewashed amnion are mixed and incubated, and the purified water is shaken and washed, and after being shaken and washed with a detergent, the decellularized biological amnion is prepared.
[0026] Further, in a preferred embodiment of the present invention, before virus inactivation, the freshly stripped amniotic membrane is washed with physiological saline, and then purified with peracetic acid-sodium chloride solution for 12-16 minutes at room temper...
Embodiment 1
[0040] This embodiment provides a method for preparing the decellularized bio-amnion that contains the loose layer. The method can prepare the decellularized bio-amnion that retains the loose layer, including the following steps:
[0041] 1.1 Wash the freshly stripped amniotic membrane twice with normal saline at room temperature;
[0042] 1.2 After cleaning, use 1.7mL / cm 2 Add a peracetic acid-sodium chloride solution with a peracetic acid content of 0.1% and a sodium chloride content of 0.8mol / L as a purification solution for virus inactivation, and purify by shaking at room temperature for 16 minutes;
[0043] 1.3 After the virus is inactivated, wash twice with normal saline at room temperature and rewash with 0.5% sodium bicarbonate solution to obtain rewashed amnion;
[0044] 1.4 Use 0.25% trypsin solution for 0.17mL / cm 2 The dosage is mixed with the rewashed amniotic membrane and incubated at 35°C for 150 minutes, and washed with pure water for 3 times;
[0045] 1.5 t...
Embodiment 2
[0047] This embodiment provides a method for preparing the decellularized bio-amnion comprising a loose layer, through which the acellular bio-amnion retaining the structure of the loose layer can be prepared, including the following steps:
[0048] 1.1 Wash the freshly stripped amniotic membrane with saline at room temperature for 3 times;
[0049] 1.2 After cleaning, use 1.7mL / cm 2 Add peracetic acid-sodium chloride solution with a peracetic acid content of 0.1% and a sodium chloride content of 1.2mol / L as a purification solution for virus inactivation, and shake and purify at room temperature for 12 minutes;
[0050] 1.3 After the virus is inactivated, wash with normal saline at room temperature for 3 times, and wash again with 10% sodium bicarbonate solution to obtain rewashed amniotic membrane;
[0051] 1.4 Use 0.4% trypsin solution for 0.24mL / cm 2 The amount of the amnion was mixed with the rewashed amnion and incubated at 39°C for 220 minutes, and washed with pure wat...
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