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Cell culture Cell lines were maintained in RPMI 1640 (Thermo Fisher, 11875119) with 5% FBS (Sigma-Aldrich, SH30396.03) and 1% penicillin and streptomycin (Thermo Fisher, 15070063)

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demonstrated in 1999 that topical or intraperitoneal administration increased re-epithelialization by 42 percent over saline controls at day 4 and by 61 percent at day 7 in a rat wound model.18 Allan Goldstein at George Washington University, who pioneered the isolation of thymosins, described thymosin beta-4 as a multi-functional regenerative peptide with applications from cardiac repair to corneal healing.19 Relevance to Endurance Athletes The endurance rationale for TB-500 centers on three effects: angiogenesis promotion (new blood vessel formation improving tissue perfusion), inflammation modulation (calming excessive responses that delay recovery), and anti-fibrotic activity (reducing scar tissue that causes stiffness and re-injury risk).20 In preclinical models, accelerated healing of connective tissue and improved muscle recovery after endurance exercise have been reported