Background: Cultural and spiritual beliefs have preserved the use of traditional practices, including al hijama (wet cupping), which is widely recognized for its purported therapeutic value and historical significance. Despite its popularity, the practice often lacks formal training and is frequently administered by unqualified practitioners, leading to false beliefs and potentially unsafe practices. Objective: To investigate the scientific knowledge and beliefs of hijama practitioners of Karachi, Pakistan, with a focus on identifying misconceptions, variations in practice, and the influence of cultural and religious beliefs on the practice of hijama therapy. Methodology: A cross-sectional exploratory study was conducted involving 114 hijama practitioners from Karachi, who were recruited through various WhatsApp groups and enrolled in a seminar held in January 2023. The participants were required to complete a 37-item questionnaire covering their qualifications, practice methods, hygiene standards, and beliefs about the hijama therapy. Data were analyzed using spss version 20, employing mean, standard deviation, chi-square tests, and frequency distribution. Results: Most participants (97.4%) were female, with only 39.5% possessing a license to practice. Many practitioners (89.5%) were motivated by the religious significance of hijama. The knowledge regarding the scientific theories and mechanism of action of hijama therapy was known by 56.4%. Practices varied widely regarding hygiene, blade type, and cup placement. 38.1% did not consistently check for infectious diseases. Additionally, the practice of reciting Qur’anic verses was widespread, believed to enhance the therapy's effectiveness which shows strong religious connotation. Conclusion: The study reveals a substantial gap in formal training among hijama practitioners, leading to variations in practice and potential safety concerns. There is a pressing need for standardized training and regulations to ensure the safe and effective practice of hijama therapy. Further research is essential to establish evidence-based guidelines which improve the public and physician awareness towards this therapy’s risks and benefits.