Objective       To determine the effect of osmotic shock combined with sonication and detergent treatment on the histology and bioactive components of newborn bovine dermis.       Methods       The reticular dermis was prepared from neonatal male bovine skin (n=8)

and sequentially treated with osmotic shock

sonication and Triton X-100. HE staining

DAPI staining and nucleic acid electrophoresis were used to evaluate cellular debris. The collagen bundle structure and cellular components were observed by scanning electron microscopy (SEM). PicoGreen

DMMB and BCA were applied to assay the contents of DNA

sulfate polysaccharide (sGAG) and protein in the dermis before and after decellularization. The contents of TGF-β1

EGF

bFGF and KGF in the dermis were measured by ELISA.       Results        Morphological observation displayed that the decellularized dermis showed the integral three-dimensional structure

interspersed collagen bundles

and satisfying removal of cells

and no obvious cell or cellular debris were seen. Compared with natural dermis

the treated sample showed significant decrease in the contents of DNA (2 516.1±324.2 vs 249.5±53.8 ng/mg

P=0.000)

sGAG (5.92±0.50 vs 2.48±0.24 μg/mg

P=0.000)

TGF-β1 (478.8±196.1 vs 180.3±111.0 pg/g

P=0.009) and EGF (10.52± 2.78 pg/g vs undetectable). The content of bFGF (788.6±333.8 pg/g vs 364.8±294.8 pg/g

P=0.424) was reduced by 53.7%. No significant change was found in the content of KGF (0.033±0.000 vs 0.033±0.000 pg/g

P=0.433).       Conclusion       Osmotic shock combined with sonication and detergent treatment effectively achieves acellular extracellular matrix with integral histological structure and partially preserved major bioactive components of the newborn bovine dermis.