Use of hijab can influence the scalp’s condition, including its microbiome. To date there has been no study comparing scalp microbiome in women wearing hijab to that in women not wearing hijab. This was a cross-sectional study conducted from August 2019 to April 2021. Healthy women aged 18 years old or older who had not undergone menopause were recruited. Those in the hijab group should wear hijab minimum 8 h a day for at least 5 years. After wash-out period, the sample was collected from the subject’s scalp. Next Generation Sequencing (NGS) was performed with primer V3-V4 region of 16S rRNA and ITS1 DNA for bacteria and fungi, respectively. Alpha diversity and beta diversity were identified, along with functional analysis. Actinobacteria and Ascomycota were the most dominant phyla on the scalp. S. capitis was more prominent in the hijab group while S. cohnii was more prominent in non-hijab group. Additionally, M. restricta was more common in hijab group while M. globosa was more common in non-hijab group. This study emphasizes the difference of scalp microbiome in women wearing hijab compared to women not wearing hijab, which indicated that women wearing hijab are more prone to seborrheic dermatitis.