Background: Online toxicity, encompassing behaviors such as harassment, bullying, hate speech, and the dissemination of misinformation, has become a pressing social concern in the digital age. The 2022 Mpox outbreak, initially termed "Monkeypox" but subsequently renamed to mitigate associated stigmas and societal concerns, serves as a poignant backdrop to this issue. Objective: In this research, we undertake a comprehensive analysis of the toxic online discourse surrounding the 2022 Mpox outbreak. Our objective is to dissect its origins, characterize its nature and content, trace its dissemination patterns, and assess its broader societal implications, with the goal of providing insights that can inform strategies to mitigate such toxicity in future crises. Methods: We collected more than 1.6 million unique tweets and analyzed them from five dimensions, including context, extent, content, speaker, and intent. Utilizing BERT-based topic modeling and social network community clustering, we delineated the toxic dynamics on Twitter. Results: We identified five high-level topic categories in the toxic online discourse on Twitter, including disease (46.6%), health policy and healthcare (19.3%), homophobia (23.9%), politics (6.0%), and racism (4.1%). Through the toxicity diffusion networks of mentions, retweets, and the top users, we found that retweets of toxic content were widespread, while influential users rarely engaged with or countered this toxicity through retweets. Conclusions: By tracking topical dynamics, we can track the changing popularity of toxic content online, providing a better understanding of societal challenges. Network dynamics spotlight key social media influencers and their intents, indicating that addressing these central figures in toxic discourse can enhance crisis communication and inform policy-making.