Antibacterial and physicochemical properties are generally considered important features of the porcine acellular dermal matrix (pADM). Oxidized 2-hydroxypropyltrimethyl ammonium chloride chitosan (OHTCC) was applied to crosslink with pADM at dosages of 1, 2, 4, 8, and 16%. The properties of the crosslinked pADM (OHTCC-pADM) were evaluated. DSC and TG analysis suggested that crosslinking could promote the thermal stability, the highest T d , and T max of OHTCC-pADM (8%) was 80 C and 325 C, which has been improved by 15 C and 13 C, respectively. While FTIR and AFM tests indicated that the structure integrity of collagen could still be maintained. SEM tests demonstrated the sustained three-dimensional architecture of OHTCC-pADM with appropriate porosity. Moreover, OHTCC-pADM exhibited improved ability to resist collagenase degradation, the degradation rate of 4%-, 8%-OHTCC-pADM was <50%. The tensile strength of OHTCC-pADM proved to be superior compared to pADM. Furthermore, 8%-OHTCC-pADM exhibited nearly 90% antibacterial activity against Escherichia coli, and Staphylococcus aureus. In addition, the results of the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazoliumbromide study showed that the cytocompatibility of OHTCC-pADM decreased with the increasing the amount of OHTCC, but all relative proliferation rates were above 80%. In conclusion, our study revealed that OHTCC stabilized and functionalized pADM while preserving good cytocompatibility.