Quantitative characterization of the colorectal cancer in a rabbit model using high-frequency endoscopic ultrasound

Purpose: Colonoscopy accompanied with biopsy works as the routine endoscopic strategy for the diagnosis of colorectal cancer (CRC) in clinic; however, the colonoscopy is limited to the tissue surface. During the last decades, enabling technologies are emerging to complement with the colonoscopy for better administration of CRC. The conventional low-frequency (<12 MHz) endoscopic ultrasound (EUS) guided fine-needle aspiration (FNA) has been widely used to assess the lesion penetration. With the high-frequency ultrasound transducer (>20 MHz), EUS allows more precise visualization of the colorectal abnormalities. In order to achieve the accurate detection or in situ characterization of the colorectal lesions, the EUS diagnosis needs more patho-physiological related information in the micro-structural or molecular level. Quantitative ultrasound (QUS) technique, which could extract the micro-structural information from the ultrasound radio-frequency (RF) signal, is promising for the non-invasive tissue characterization. To date, the knowledge of the high-frequency endoscopic QUS for the CRC characterization has not been fully determined. Methods: In this work, to our best knowledge, it is the first application of the QUS technique based on a customized high-frequency EUS system (30.5 MHz center frequency) to characterize the colorectal malignancies in a VX2 rabbit CRC model. To eliminate the response from the ultrasound electronic system and transducer, the ultrasound signals from colon tissue were calibrated. And, the resulting quasi-liner ultrasound spectra were fit by the linear regression test. As a result, three spectral parameters, including the slope (k), intercept (I) and Midband Fit (M), were obtained from the best-fit line. The three spectral parameters were compared between the malignant tissue regions and adjacent normal tissue regions of the colon tissue specimen ex vivo. The independent t-test was conducted between the three parameters from the normal and malignant group. The statistical method of Fisher Linear Discriminant (FLD) was used to explore the linear combinations of the three parameters, so as to provide more tissue micro-structural features than the single parameter alone. The three FLD values were derived from three different combinations among k, I and M. The threshold was selected from the statistical analysis to optimize the differentiation criteria between the malignant and the normal tissues. The color-coded images were used to display the local FLD values and combined with the EUS B-mode image. Results and Conclusions: The mean Midband Fit (M) and intercept (I) showed significant differences between the malignant and normal tissue regions. The statistical analysis showed that there were significant differences in all the mean FLD values of the spectral parameter combinations (kI, kM and IM) (t test, P < 0.05). And, the combined image result from the B-mode image and color-coded image could visually correlate with the histology result. In conclusion, the high-frequency endoscopic QUS technique was potential to be used as a complementary method to distinguish the colorectal malignancies by leveraging its morphological and micro-structural ultrasound information.