Transcriptomic Profiling and Histological Validation of Preputial Fibrosis in Hypospadias

<b>Background:</b> Hypospadias is often associated with abnormal prepuce development. Investigating the differences between the inner prepuce of hypospadias patients and normal controls at the transcriptomic level and histological characteristics helps to reveal the causes of its developmental abnormalities or implement targeted treatments. <b>Materials and Methods:</b> Dorsal and ventral inner preputial tissues were collected from 31 hypospadias patients and 21 phimosis children (controls). Differences in gene expression between the two groups were studied via transcriptomic sequencing and enrichment analysis. Corresponding histological features were further validated by histological staining. <b>Results:</b> Transcriptomic sequencing results showed that, compared to the control group, the dorsal inner prepuce of the hypospadias group had 97 upregulated and 10 downregulated genes; the ventral prepuce had 140 upregulated and 99 downregulated genes. Among all upregulated genes, 44 were closely related to fibrosis. Other significantly enriched terms included cornified envelope formation, efferocytosis, C-type lectin receptor signaling pathway, and complement and coagulation cascades. Histological validation revealed that the dorsal inner prepuce of hypospadias children contained more collagen fibers, a higher ratio of type I/III collagen, and lower microvessel density, showing some correlation with the severity of hypospadias. <b>Conclusions:</b> This study demonstrated a hyper-fibrotic state in the inner prepuce of hypospadias, which may significantly impact post-operative wound healing and complications.