As we know different methods and materials of decellularization have different effects on ECM. Which one is better for cartilage?
Hello
Treatment with 2% SDS appears to be the most promising method for decellularization . Is divided into two Phase .
Decellularization Phase I
At t=4 wks, self-assembled constructs (n=6/group) were removed from culture and exposed to one of five decellularization treatments, for either 1 h or 8 h. The decellularization treatments included:
1% SDS
2% SDS
2% TnBP
2% Triton X-100
Hypotonic/Hypertonic Solution (half-time of each)
Hypotonic: 10 mM Tris HCl, 5 mM EDTA, 1 μM PMSF
Hypertonic: 50 mM Tris HCl, 1 M NaCl, 10 mM EDTA, 1 μM PMSF
All treatments included 0.5 mg/ml DNase Type I, 50 μg/ml RNase, 0.02% EDTA, and 1% P/S/F, in PBS. Both 1 h control and 8 h control groups were exposed to this same solution without detergent treatments. These treatments were applied at 37°C with agitation. Following the 1 h or 8 h treatment, the constructs were washed for 2 h in PBS at 37°C with agitation. Additionally, an untreated control was assessed immediately following construct removal from culture, without the treatment or wash steps.
Decellularization Phase II
At t=4 wks, self-assembled constructs (n=6/group) were removed from culture and exposed to 2% SDS for 1, 2, 4, 6, or 8 h. As in phase I, all treatments included 0.5 mg/ml DNase Type I, 50 μg/ml RNase, 0.02% EDTA, and 1% P/S/F, in PBS. These treatments were applied at 37°C with agitation. Following the SDS treatment, the constructs were washed for 2 h in PBS at 37°C with agitation. Additionally, an untreated control was assessed immediately following construct removal from culture, without the treatment or wash steps.
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i hope help you , Good Luck .
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