Objective To assess the antimicrobial elution characteristics, toxicity, and antimicrobial activity of amikacin-impregnated ferric-hyaluronate implants (AI-FeHAI) for amikacin delivery to the tarsocrural joint of horses. Study Design Experimental study. Sample Population AI-FeHAI implants, equine cartilage, and synovium, and horses (n=6). Methods In vitro study: Five AI-FeHAI were placed in saline solution with daily replacement until implant degradation. Eluent was tested for amikacin concentration and bioactivity. Synovial and cartilage explants were incubated in the presence or absence of AI-FeHAI for 72 hours and subsequently assessed for morphology, viability, and composition. Synovial explants were incubated with Staphylococcus aureus in the presence or absence of AI-FeHAI. Spent medium was cultured daily and explants were assessed for morphology and viability after 96 hours. In vivo study: AI-FeHAI were placed in 6 tarsocrural joints. Standard cytologic analysis and amikacin concentration (SFAC) were determined in synovia obtained regularly for 28 days thereafter. Similar analyses were conducted after a single intra-articular injection of amikacin 6 months later. Results In vitro study: Amikacin concentrations exceeded 16 g/mL and inhibited S. aureus growth for 8 days. AI-FeHAI had no effect on cartilage explants. AI-FeHAI eliminated bacteria from synovial explants. In vitro study: After AI-FeHAI placement, SFAC was highest (140.78+63.81 g/mL) at first sampling time. By 24 hours SFAC was <16 g/mL. After intra-articular injection, SFAC was the highest (377.91 ± 40.15 g/mL) at first sampling time. By 48 hours SFAC was <16 g/mL. Conclusions A single intra-articular amikacin injection demonstrated superior pharmacokinetics than AI-FeHAI prepared as described. Clinical Relevance AI-FeHAI cannot be recommended for clinical use.