Myofibroblasts: A New Factor Affecting the Hyperlipidemia-Induced Elastic Abnormality of Corpus Cavernosum in Rabbits Detected by 2-D Shear Wave Elastography.

OBJECTIVE: Two-dimensional shear wave elastography (2-D SWE) has been proven to detect hyperlipidemia-induced elastic abnormality in the corpus cavernosum. This study investigated cytological factors affecting the elasticity of the corpus cavernosum in rabbits with hyperlipidemia using single-cell RNA sequencing (scRNA-seq). METHODS: Male New Zealand white rabbits were randomly divided into a hyperlipidemia group (high-cholesterol diet) and a control group (standard diet). Penile 2-D SWE was performed to detect the elastic abnormality in the corpus cavernosum. ScRNA-seq was performed to observe cellular changes in the corpus cavernosum of rabbits with hyperlipidemia. Immunohistochemistry, immunofluorescence and histological examinations were conducted to verify the results of scRNA-seq. RESULTS: Two-dimensional SWE revealed that the Young's modulus of the corpus cavernosum was significantly greater in the hyperlipidemia group than that in the control group (p < 0.001). Histological findings revealed extracellular matrix accumulation within the corpus cavernosum, with stronger staining of collagen types I and Ⅲ. ScRNA-seq revealed that fibroblasts, smooth muscle cells, and endothelial cells were the major cell types in the corpus cavernosum. A novel subtype of fibroblasts (myofibroblast) was discovered in the hyperlipidemia group, which was verified by immunofluorescence staining and gene ontology analysis. Fibroblasts, smooth muscle cells and endothelial cells were three cellular sources for myofibroblasts. CONCLUSION: Myofibroblasts are activated and proliferate and secrete large amounts of collagen fibers in the corpus cavernosum during hyperlipidemia, leading to abnormal Young's modulus detected by 2-D SWE and their recognition as a new factor affecting the hyperlipidemia-induced elastic abnormality of the corpus cavernosum.

Factors affecting the shear wave elastic quantitative measurement of penile tissue in rats.

Background: As a new ultrasound technology, 2-dimensional shear wave elastography (2D-SWE) can evaluate the elastic characteristics of penile tissue. However, no studies have reported the main factors affecting the shear wave elastic quantitative measurement (SWQ) in penile tissue. Objectives: To analyze the main factors affecting the SWQ reflecting the elastic characteristics of penile tissue by 2D-SWE. Methods: Twenty healthy male Sprague-Dawley rats (5-60 weeks old) were selected for this study. We performed the 2D-SWE examination on the penis using the Aixplorer ultrasound system, with SWQ as the measurement index. We performed penile immunohistochemistry analysis with the positive area proportion (PAP) of alpha-smooth muscle actin (PAPS) and type III collagen fiber (PAPC) as the outcomes. Then, we conducted multiple linear regression analysis to explore the correlation of SWQ with PAPS and PAPC and established the regression equation. Results: The multiple linear regression analysis showed that the linear regression equation (SWQ = 10.376 - 0.05 PAPS - 0.07 PAPC) was statistically significant (F = 21.153, P < 0.001). The content of smooth muscle cells (SMCs) and collagen fibers was negatively correlated with SWQ, affecting 42.6% of the total variation in SWQ (R2 = 0.426). Conclusions: SMCs and collagen fibers are the main factors affecting the SWQ value of penile tissue and the primary tissue components determining the SWQ when using 2D-SWE to quantitatively evaluate the elastic characteristics of penile tissue.

Experimental investigation of early assessment of corpora cavernosa fibrosis with two-dimensional shear wave elastography.

This study explored the usefulness of two-dimensional shear wave elastography (2D-SWE) in the early assessment of corpora cavernosa fibrosis (CCF). New Zealand male rabbits were randomly assigned to an experimental group or a control group. Recombinant human transforming growth factor beta 1 (TGF-ß1) was injected into the dorsal penis tissue of rabbits in the experimental group. Conventional ultrasound and 2D-SWE examinations were performed before and 20 days after injection. Penile histological analysis was performed by hematoxylin-eosin staining, sirius red staining, and immunohistochemistry. Measurement of 2D-SWE examination results was performed using shear wave elastography quantitative measurement (SWQ). Histological analysis outcomes were the proportion of smooth muscle cells (SMCs), collagen fibers (CFs), collagen type I (Col I), and collagen type III (Col III), as well as the SMCs/CFs ratio, measured by sirius red staining. Other histological analysis outcomes were the positive area proportion (PAP) of TGF-ß1 (PAPT), fibronectin (PAPF), and Col III (PAPC), measured by immunohistochemistry. After recombinant human TGF-ß1 injection, SWQ was higher in the experimental group than that in the control group (P < 0.001); however, there were no differences in conventional ultrasound results. There were significant differences in histological outcomes between the two groups (all P < 0.05). These results indicated that 2D-SWE was superior for identifying early histological changes in CCF.