Objective: The aim of this in vitro study was to evaluate the effect of human plasma, compared to physiologic saline, on microtensile bond strengths of 5 self-conditioning dentin adhesives when used as a dentin perfusion medium. Method and Materials: One hundred and fifty extracted human third molars were prepared in a special manner allowing the simulation of intrapulpal pressure and dentin perfusion. The specimens were randomly divided into 10 experimental groups. Five groups were perfused with physiologic saline, while the others were perfused with diluted human plasma, each under constant hydrostatic pressure of 30 cm H2O. One saline group and 1 human plasma group was then assigned to each of the following dentin adhesives: Clearfil SE Bond (Kuraray), Clearfil Protect Bond (Kuraray), Xeno 3 (Dentsply), Futurabond NR (Voco), and iBond (Heraeus Kulzer). The microtensile bond strength for each group was evaluated using a Zwick universal testing machine 10 minutes after light curing. Results: Pairwise comparison showed a significant increase of bond strength in all human plasma-perfused subgroups (P .05, Tukey's test). The influence of the different dentin adhesives was significant (P .001, analysis of variance). The highest significant values were observed for Futurabond NR and iBond with human plasma (P .05, closed test procedure). Conclusion: The use of human plasma might be a suitable alternative to imitate perfused dentin conditions in testing devices and might eliminate the discrepancy between different in vitro investigations focusing on bond strength of dentin adhesives. Keywords: dentin, dentin bonding agents, dentin perfusion, human plasma, microtensile bond strength, saline, self-conditioning