Microdialysis has shown intratendinous glutamate levels to be significantly higher in Achilles tendons with painful tendinosis than in normal pain-free tendons, and treatment with eccentric training has shown good clinical results with diminished tendon pain during activity. In six patients with chronic painful Achilles tendinosis we performed microdialysis for 2 h, before and after the 12-week eccentric training program. The treatment was successful in all six patients, and the mean VAS score (amount of pain during Achilles tendon loading) decreased from 69 before treatment to 17 after treatment. There was no significant difference between the intratendinous glutamate levels before and after treatment. Our results offer no obvious neurophysiological explanation but showed that successful treatment with eccentric training was not associated with lowered intratendinous glutamate levels.