The purpose of this study was to characterize microneurovascular (MNV) muscle transplants immunohistochemically up to 10 years after transfer. Histological data was related to long-term functional outcome. The study comprised 17 patients with a mean age of 41 years suffering from complete unilateral long-lasting facial paralysis. A two-stage procedure was performed between 1986 and 2001. The gracilis, latissimus dorsi, and serratus muscles were used in four, eight, and five patients, respectively. Eighteen biopsy samples were taken from MNV muscle grafts during secondary refinement procedures. In one patient, the tissue samples were collected at two different time points. Immunohistochemistry testing revealed muscle fiber type distribution (anti-myosin fast), proliferating satellite cells (Ki-67), and reinnervation (S-100). Muscle atrophy was assessed histomorphometrically. In a recent study, patient characteristics and the function of the flap were evaluated. Histological data were compared with clinical data and long-term functional outcomes of the patients. In biopsy samples taken 1-10 (mean 31 months) years after MNV muscle transfer, the mean muscle fiber diameter was 38 (range 14-70) microm, indicating a 40% decrease compared with control values. Muscle atrophy was not type-specific and the mean percentage of type II fibers was not altered. Individual variation was, however, considerable. Proliferative activity of satellite cells was seen in 60% of the samples but it tended to decline with an increase in follow-up time. All samples showed intramuscular reinnervation. In statistical analysis severe atrophy correlated with prolonged intraoperative ischemia (P=0.04). The good long-term functional outcome correlated with dominance of fast fibers in muscle grafts (P=0.03). Atrophy tended to be more pronounced in the serratus than in the other muscles (ns). In summary, despite dense muscle reinnervation, morphology of the muscle is not fully restored after muscle transfer. Ischemia time affects muscle morphology. Adaptation of the graft to fast-twitch muscle activity favors better mimic function. The proliferative activity of satellite cells declines with prolonged follow-up time.