This study is a monocentric, non-interventional feasibility study. 40 women with sonographically visible early stage (cT1c-cT2, cN0-cN1) invasive breast cancer who were scheduled to undergo primarily BCS in our center were included. Diagnosis of invasive breast cancer was made by preoperative core biopsy. Specimens of patients who had received neoadjuvant chemotherapy, mastectomy, specimens of re-excision surgery, status post radiation therapy of the affected breast or preoperatively diagnosed DCIS were excluded. We did not exclude patients with postoperatively diagnosed DCIS accompanying the preoperatively known invasive breast cancer. Therapy planning was according to national guidelines [15]. No changes on the standard surgery or therapy procedure were conducted. The tumor was marked preoperatively by sonographic fine needle insertion. After excision of the tumor the specimen was fixed on a KliniTray™ (KLINIKA-Medical GmbH, Germany). Intraoperative specimen ultrasound was performed using hand-held ultrasound (Voluson™ S8, GE Healthcare GmbH, Linear-Array-Sonde ML6-15, 4.0 MHz-15.0 MHz), so additional tissue could be excised if the tumor excision appeared incomplete. This procedure corresponds to the standard of care in our breast cancer center for sonographically detectable tumor masses that are primarily treated surgically. Axillary surgery was undertaken in the same session, according to national guidelines. A multidisciplinary team reviewed all cases preoperatively and postoperatively, and adjuvant radiotherapy or systemic therapy was administered according to institutional and national guidelines. The surgery was not interrupted or prolonged by the following additional specimen examination. After the hand-held specimen sonography, specimen sonography was carried out using ABUS. For this, the specimen stayed fixed on the KliniTray™ and was put in a corresponding plastic box. Pieces of base lead were placed on the exposed part of the KliniTray™ to weigh the tray down, without changing the fixation of the preparation. The plastic box was completely filled with isotonic saline solution. The use of this solution should prevent any possible influence on the cells. An ABUS membrane was then placed on the box and a plastic bag filled with ultrasound gel was placed on the ABUS membrane. This bag was additionally coated with ultrasound gel for lubrication to ease the movement of the transducer and the ABUS transducer was attached. The construction is shown in Fig. 1. The ABUS recording of the specimen was then carried out by one of the trained medical assistants according to the standard ABUS procedure. The transducer was placed with low compression. In order to avoid the risk of damage to the ABUS system, the ABUS transducer had no contact with the isotonic water solution in this arrangement (see Fig. 2). Evaluation of the created images was carried out by two independent physicians trained on ABUS evaluation. The first evaluation consisted of the quality of the images. The images were classified as “easy to evaluate” or “difficult to evaluate”. High image contrast and quality which enable a clear differentiation of the tumor from the surrounding area were necessary to classify an image as “easy to evaluate”. Further assessment was conducted as follows exclusively for the images which had been classified as “easy to evaluate”: The size of the tumor was measured in three dimensions (horizontally, vertically and ventro-dorsally). The safety margins, i.e. the distance between the tumor and the edge of the specimen, were evaluated in six directions: ventrally, dorsally, caudally, cranially, laterally and medially. The findings were compared to the report of the HHUS and to the histopathological report, where the size of the tumor and the radial distances from the edge of the tumor to the edge of the specimen in six directions were described. Furthermore, the histopathological report included hormone status, HER2 status, proliferation index Ki 67, the type of breast cancer (such as lobular carcinoma and non specific type) and the need for re-excision. In order to ensure correct correlation with the histopathologic examination of the margins and their directions, the orientation of the specimen was preserved on the KliniTray™ throughout every step of the procedures described above. ABUS measurements and HHUS measurements of the specimen were documented in millimeter in the specified dimensions. Image interpretations of ABUS were carried out without knowledge of the corresponding HHUS and without knowledge of pathological results.
Statistical analysis
The ability of ABUS and HHUS examination to predict tumor size and safety margins was assessed by comparing the safety margins and tumor size measurements as predicted by the imaging modalities with the histopathologic report. Microsoft Office Excel 365 (Microsoft Corporation, Redmond, WA) was used for data collection. Statistical analysis and graphical representation were performed using IBM SPSS Statistic 25 (IBM Corporation, Armonk, NY) and validated by the authors. Concordance between the tumor sizes measured with ABUS and HHUS was calculated with Excel 365. A paired t-test was used for to assess the differences between the tumor sizes measured with HHUS and ABUS, respectively. These measured tumor sizes were also compared with final pathologic tumor size. Statistical significance was assumed for a P value of less than 0.05 for all tests.
Ethical and legal aspects
This study was carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki). The study was registered by the local ethical committee (reference No 21-2309-104). All members of the research team committed themselves to the confidentiality of the information provided as well as to data protection and are subject to medical confidentiality. The data is not accessible to anyone outside the research team.