2.2 Evaluation of decellularization
2.2.1 Histological and immunofluorescent evaluation
For confirmation of decellularization, DP samples were fixed in 4%
paraformaldehyde for 24 h and embedded in paraffin. After dehydration
and paraffin embedding, the DP samples were cut into 5-μm-thick sections
for histological examination using hematoxylin and eosin (H&E)
staining. The stained sections were visualized and photographed on an
optical microscope (Leica, DM3000, Germany). Residual nuclei within the
DP were observed by DAPI staining viewed on an Olympus BX53 microscope
(Olympus, Japan).
2.2.2 Quantification of DNA, glycosaminoglycan (GAG) and collagen
content
The DNA content in DP samples was determined using a Genomic DNA
Extraction Kit (Takara, China) following the manufacturer’s
instructions. The GAG content in DP samples was evaluated using a DMMB
Colorimetry Kit (GenMed, USA). A hydroxyproline assay kit (Abcam, USA)
was utilized to quantify the collagen content of DP samples according to
the manufacturer’s instructions.