This research explores a novel steel fiber made by twisting together two straight steel wires. The pull-out behavior of the twin-twisted steel fiber was investigated, proving its slip-hardening ability in cement-based matrices. The number of twists per unit length, fiber embedment length, matrix compressive strength, and fiber equivalent diameter were the experimental variables. Hooked-end fibers were also investigated for comparison. Pull-out test results of twin-twisted steel fibers quantified the mechanisms of frictional bonding and mechanical untwisting and their effects on the pull-out response and slip hardening of steel fibers. The maximum pull-out load increased with the increase of fiber twists per unit length, which increased the likelihood of damage to the matrix tunnel and had a negative effect on the energy absorption capacity. Defects at interfaces and scratches on fiber surfaces were examined using SEM imaging. Twin-twisted steel fibers reached an enhanced hardening response and energy absorption capacity compared to hooked fibers yet showed lower maximum bond strength. New bond strength and energy dissipation indexes were introduced and could effectively be used for fiber optimization.