The purpose of this investigation was to examine the changes in unipennate muscle architecture after distraction osteogenesis. Nine adult goats underwent 20% tibial lengthening in one of the hind limbs. Immediately after distraction, lengthened and contralateral (untreated) tibialis caudalis (TC) muscles were harvested. Lengths of the muscle belly, muscle fiber (FL), sarcomere (SL), tendon (TL), and superficial aponeurosis, as well as muscle mass, pennation angle (PA), and physiological cross-sectional area (PCSA), were compared between the treated and contralateral sides. Lengthened TC muscle demonstrated 20.8% increase in belly length, 4.39% increase in TL, and 36.7% increase in FL, while PA decreased by 37.2% (P ¼ 008). Muscle length increase was mainly due to lengthening of muscle belly, which resulted both from FL increase and 15.3% length increase in the aponeurosis component of muscle belly, without significant effect of the PA decrease. The FL increase was due to SL increase, not to sarcomere neogenesis, while mass and PCSA did not change. We concluded that although muscle architecture can be adversely affected by distraction because of deficient sarcomere neogenesis, PCSA can remain unchanged, giving false impression of preserved function. Change in PA plays only minimal role in muscle adaptation to distraction. Anat Rec, 290:461-467, 2007Rec, 290:461-467, . 2007 Wiley-Liss, Inc.