Patients with chronic diseases like HIV and TB often seek treatment from both traditional and biomedicine healthcare providers. Collaboration with traditional medicine practitioners (TMPs) has been advocated by UNAIDS in Sub-Saharan Africa and was promising in some countries. However, no formal collaboration has been established in Ethiopia, and the perspectives of healthcare professionals (HCPs) on such collaboration remain unexplored. In this study, descriptive qualitative design was used to explore HCPs' experiences and perspectives on collaborating with TMPs in HIV and tuberculosis care in Metekel Zone, Northwest Ethiopia. HCPs working directly in HIV and TB clinics in five health facilities were purposively included. Data were collected between March 5 and April 30, 2022, through in-depth interviews using semi-structured questionnaires. Interviews were audio-recorded, transcribed verbatim, inductively coded, and thematically analyzed using MAXQDA 2020. The study included 25 key informant HCPs. They reported varied experiences with TMPs, including no interaction, occasional informal social interactions, and rare cases of informal referrals. While some HCPs expressed skepticism about the efficacy and safety of TMP practices, most acknowledged potential benefits if TMPs were limited to managing minor conditions and opportunistic infections. The major reported challenges to collaboration were unstandardized herbal practices, the secrecy of TMPs, differing treatment philosophies, the absence of legal frameworks, and logistical barriers. To establish collaboration, informants recommended legal frameworks, registering genuine healers, protecting intellectual property, organizing regular forums for TMP engagement, fostering research partnerships, and providing financial and tailored training programs for both groups. In conclusion, this study revealed HCPs had mixed experiences and perspectives on collaboration with TMPs in Metekel Zone. While some informants recognize the potential benefits of collaboration, others remain skeptical about role of TMPs. Therefore, strong government support, clear legal and research frameworks, and trainings and mutual understanding were recommended to foster collaboration.