In order to study the effect of biological regenerant on the interaction between aged asphalt and aggregate, based on molecular dynamics, the interface models of bio-oil recycled asphalt-weak alkaline aggregate under dry and wet conditions were established respectively. The interaction energy at the interface was calculated. The results show that, among the three bio-oil rejuvenators, cashew nut shell oil has the best effect on relieving the agglomeration of asphaltene molecules. The van der Waals force between aged asphalt and aggregate will be weakened by three kinds of regenerant. forming hydrogen bonds on the surface of aggregate.The hydrogen bonds on the surface of aggregate can be formed by cashew nut shell oil and tall oil, improving the adhesion of aged asphalt and and reducing the humidity sensitivity of aged asphalt. When the asphalt regenerant will be selected, the hydrogen bonding between aggregate and regenerant should be considered, which is helpful to improve the adhesion between recycled asphalt and aggregate.