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PLOS One logoLink to PLOS One
. 2025 Oct 9;20(10):e0330076. doi: 10.1371/journal.pone.0330076

Shear Wave Elastography for measuring the elastic properties of the Psoas Major muscle: An intra- and inter-examiner reliability study

Gabriel Rabanal-Rodríguez 1,2, César Fernández-de-las-Peñas 3,4, Javier Álvarez-González 5,6, Alberto Roldán-Ruiz 5, Jorge Buffet-García 5,*, Juan Antonio Valera-Calero 1,2
Editor: Masatoshi Nakamura7
PMCID: PMC12510488  PMID: 41066316

Abstract

Background

The psoas major (PM) plays an important role in population with low back pain (LBP). Its evaluation considers clinical history, which can be confused with other lumbar and lower limb conditions, physical examination, with an inconsistent palpation, and imaging tests that provide inconclusive parameters in various studies. Therefore, developing reliable assessment procedures to evaluate PM elastic properties is necessary to improve diagnosis and follow-up. This study aimed to evaluate the intra and inter-examiner reliability of shear wave elastography (SWE) for calculating the PM stiffness in patients with LBP.

Design

Observational study.

Methods

Longitudinal views of the of the PM muscle using SWE were acquired bilaterally in 52 volunteers experiencing moderate LBP and disability. All measurements were performed twice, involving the assessment of shear wave speed and Young’s modulus as indicators of stiffness, with data collected by an experienced examiner and a novice examiner.

Results

Test-retest reliability showed strong consistency regardless of the examiner’s level of experience, with intraclass correlation coefficients (ICCs) exceeding 0.9 for both metrics. However, experienced examiners achieved smaller minimal detectable changes. Inter-examiner reliability was comparatively lower, with ICCs ranging from 0.854 to 0.925, and notable differences in mean values between examiners were observed (p < 0.01).

Conclusion

Excellent test-retest reliability was observed for the measurement of PM muscle stiffness in chronic LBP patients using SWE by both experienced and novice evaluators, although statistically significant differences were found between the two trials for the novice examiner. Inter-examiner reliability was lower, highlighting variability between assessors. To minimize errors and improve consistency and accuracy, if multiple examiners are involved, averaging measurements is recommended.

Introduction

The psoas major (PM) is a long muscle that originates from the transverse processes, vertebral bodies, and intervertebral discs of T12 to L5 and inserts into the lesser trochanter of the femur. Biomechanically, the PM exerts compression and shear forces on the lumbar spine, tending to produce extension moments in the upper lumbar segments and flexion moments in the lower segments. Despite its influence on lumbar spine movements, its primary function is hip flexion, with a secondary role in lumbar stabilization through force transmission during trunk and hip flexion activities [1].

From a clinical perspective, the PM muscle plays a key role in the pathogenesis of painful musculoskeletal conditions affecting the lumbar spine and the lower limbs (e.g., groin injuries, low back pain (LBP), pelvic pain, hip snapping and femoroacetabular impingement) [1] as described in numerous reports [27].

Regarding non-specific LBP, it has been established that increased PM stiffness due to continued local contraction could be a relevant etiologic factor, suggesting that tools for its evaluation, like elastography, can provide significant quantitative information for the diagnosis and assessment of these patients [8]. In this context, other authors have also noted overactivation of the PM in patients with chronic LBP [7].

The evaluation of the PM is based on the medical history, clinical examination and complementary imaging tests. Isolated assessment based on clinical history is complex and not recommended, since it may present compatible symptoms with impairments or injuries of other structures of the lumbopelvic region. In the clinical setting, the evaluation can be complemented by palpation (with evidence advising that soft tissue palpation is inconsistent in patients with LBP [9]) or by performing muscular resistance tests (with isometric contractions) or orthopedic tests including the Thomas or FADIR maneuvers [1]. As for imaging tests, the gold standard is considered to be Magnetic Resonance Imaging, although some studies have also used Computerized Tomography [5]. However, in recent years, ultrasound imaging (US) has begun to be studied as a possible alternative for the evaluation of the muscular characteristics of the PM [1012].

This tool, in particular through sonoelastography, allows to assess tissue elasticity, functioning as an adjunct to B-mode US and offering significant advantages with reduced cost, examination time and complexity. Shear wave elastography (SWE), which employs mechanical shear waves generated by the compressive acoustic waves used during B-mode imaging to measure the velocity of shear wave propagation (which correlates directly with tissue stiffness), despite competing with other elastography techniques for the assessment of musculoskeletal tissues [13], is regarded as the most reliable approach. Evaluating muscle stiffness could offer clinically significant insights, potentially identifying early disease stages when morphological changes are not yet visible on grayscale imaging, making it more informative than parameters such as size, histology, or shape descriptors.

Taking into account the available evidence on changes in the muscular characteristics of the PM in relation to LBP and disc pathology [6,1422], the uncertain physical examination with an inconclusive palpation and given the scarce published literature on the usefulness and reliability of US assessment for the determination of its properties, only including the study by Zhou et al. [23] in healthy subjects, whose results may not be extrapolated to individuals with LBP regarding their particular characteristics, the development of a study to establish valid and reliable methods for the assessment of the elastic properties of the PM is rationally justified.

This information could enable researchers to continue with a line of knowledge on the association between SWE-obtained parameters and LBP-related variables as suggested on clinical practice guidelines [24], as well as deepening into SWE precision for the discrimination between LBP and non-LBP populations.

Thereby, the primary objective of this study is to evaluate the intra and inter-examiner reliability of a SWE technique for assessing the stiffness of the PM muscle in a cohort of patients experiencing LBP.

Methods

Study design

Between 29th February and 31st May 2024, a longitudinal observational study was carried out to evaluate the intra and inter-examiner reliability estimates of a SWE procedure. The study was conducted in a physiotherapy laboratory within the Faculty of Health Sciences of the Francisco de Vitoria University. To ensure the quality of the report, the guidelines outlined in the Reporting Reliability and Agreement Studies (GRRAS) [25] and Enhancing the QUAlity and Transparency Of health Research (EQUATOR) frameworks [26] were followed. Furthermore, participants’ rights were safeguarded in compliance with the Declaration of Helsinki, and the study protocol was reviewed and approved by the Ethics Committee of the Francisco de Vitoria University (15/2024) prior to data collection.

Participants

A sample of patients with LBP was recruited through local announcements posted at the campus of the Francisco de Vitoria University. The inclusion criteria required participants to be between 18 and 65 years old and to have experienced at least one clinically significant episode of LBP within the past year, without neurological signs, along with mild-to-moderate pain intensity (minimum 3 out of 10 on the Visual Analogue Scale [27]) and associated disability (at least 12 out of 100 on the Oswestry Disability Index [28]) at the time of data collection.

Individuals were excluded if they were undergoing pharmacological treatment that could influence muscle tone, had a history of spinal surgeries, neuropathies (e.g., radiculopathies or myelopathies), serious medical conditions (e.g., tumors, fractures, neurological disorders, or systemic diseases), clinically relevant asymmetries, or widespread musculoskeletal conditions such as fibromyalgia.

Participants who met these criteria were asked to read, understand, and sign a written informed consent form before being scheduled for data collection.

Sample size calculation

The minimum sample size required for this study was calculated using the methodology outlined by Walter et al. [29] for reliability studies involving intraclass correlation coefficients (ICCs). Since no previous studies had evaluated intra or inter-examiner reliability estimates in clinical populations, data from asymptomatic subjects were used as a reference. Zhou et al. [23] reported test-retest reliability ICCs ranging from 0.89 to 0.92 in a sample of 52 subjects.

Based on these references, the following parameters were applied: the minimum acceptable ICC (ρ0) was set at 0.7 (the lower limit of “good reliability” as defined in the literature [30]), the expected ICC (ρ1) at 0.92, the significance level (α) at 0.05, the statistical power (1 – β) at 0.95, with 2 raters (k) and an anticipated dropout rate of 10% due to the longitudinal nature of the study. Consequently, a total of 29 participants were required to ensure adequate statistical power.

Examiners

The study involved two examiners: one experienced examiner with over 10 years of expertise in musculoskeletal US and clinical management of musculoskeletal conditions, and one novice examiner with less than 1 year of experience in both areas who had completed 20 hours of US training.

To enhance methodological rigor (as assessing the left and right side from the same participant cannot be fully considered as independent datapoints), imaging acquisition procedures were randomized for both the order in which volunteers participated and the initially examined side. For each measurement, the entire positioning process of the participant and the probe was restarted to minimize the potential influence of prior alignment or muscle relaxation. The study was conducted in two separate sessions: the first in the morning, between 9:00 and 11:00, and the second in the afternoon, from 13:00–15:00. A strict isolation protocol was followed, with the two examiners working on alternating days to eliminate any possibility of communication or agreement between them.

Participants were asked to attend twice a day for two consecutive days (24-hour interval). The first two visits were in day one (in the morning with examiner A and in the afternoon with examiner B) and the second two visits were in day two (in the morning with examiner B and in the afternoon with examiner A). A single image was obtained in each session. Subsequently, a third investigator coded all the images. Finally, each examiner measured their own images in a randomized order, ensuring blinding to both the participant’s identity and the evaluated side.

US acquisition protocol

The US device used for image acquisition was a Canon Aplio A, equipped with a convex transducer 8C1 (Canon Medical Corp, 1385 Shimoishigami, Otawara, Tochigi 324–8550, Japan). Standard console settings were applied for all acquisitions (Frequency: 5 MHz, Gain: 80 dB, Dynamic Range: 60, Depth: 12 cm).

The imaging acquisition protocol adhered to the guidelines established by Zhou et al. [23]. Participants were positioned in a lateral decubitus posture to ensure spinal and lower limb neutrality. A wedge-shaped cushion was placed posterior to the upper thoracic region to maintain the torso perpendicular to the examination table. Additionally, a square cushion was inserted between the thighs to maintain neutral hip alignment, and a towel was used when necessary to support a neutral lumbar spine position. Participants were instructed to relax their muscles during the procedure to prevent morphological bias caused by muscle contractions [31].

Acoustic coupling gel was applied to the transducer, which was initially positioned superior to the iliac crest along the mid-axillary line. The cranial end of the transducer was then tilted posteriorly by approximately 20° to center the L4 vertebra in the imaging field using B-mode US (Fig 1A). At this location, the PM muscle was identified as the structure overlying the vertebral bodies (Fig 1B).

Fig 1. Transducer positioning and an illustrative example of shear-wave elastography image. (A) Transducer positioning; (B) Shear wave imaging identifying the most relevant musculoskeletal structures: quadratus lumborum, psoas major and lumbar spine; (C) Quantitative measurement of shear wave imaging: Young’s modulus (kPa) and shear wave speed (m/s).

Fig 1

To capture images of the PM, the transducer’s orientation was adjusted to align parallel to the muscle fibers’ long axis, ensuring the central portion of the transducer was perpendicular to the muscle fibers. Once aligned, the SWE mode was activated, positioning the region of interest in the center of the muscle and covering at least 50% of its area to capture the image, adapting the dimensions to each participant depending on their characteristics. Finally, images were measured by outlining a freely drawn quantification box, ensuring it did not overlap the muscle edges and excluded the muscle fasciae (Fig 1C). All SWE images were acquired using the built-in software of the Canon Aplio A system, which automatically applies standard post-processing algorithms to optimize image quality and measurement accuracy. These include real-time smoothing filters to reduce noise and enhance the visibility of shear wave propagation, as well as temporal averaging to stabilize the color-coded elastograms. The system also applies quality maps to guide the placement of the region of interest, ensuring that measurements are taken only from areas with sufficient signal stability. No manual or external image processing was performed after acquisition.

After contouring the region of interest, the US device automatically provides the shear wave speed and automatically calculates the Young’s modulus. The relationship between these two parameters is based on the physical equation E = 3ρ·SWS2 where E is Young’s modulus (in kilopascals), ρ is the tissue density (usually assumed to be ~ 1000 kg/m3 for soft tissue), and SWS is the shear wave speed (in meters per second). This formula assumes the tissue behaves as a linear, isotropic, and purely elastic medium. Although biological tissues are more complex, this approximation allows the ultrasound system to express stiffness in both m/s (direct measure of SWS) and kPa (converted Young’s modulus), depending on clinical preference or research requirements.

The InBody 770 bioimpedance device (Biospace, Urbandale, IA, USA) was used to assess body composition parameters, collecting values for water volume, body weight, body mass index (BMI) and muscle and body fat mass.

Statistical analysis

All data processing and analysis were performed using the Statistical Package for the Social Sciences (SPSS) v.29.1.1 (Armonk, NY, USA) for Mac OS. All tests were two-tailed, with a significance threshold set at p < 0.05. The distribution of continuous variables was initially evaluated using histograms and Shapiro-Wilk tests.

The demographic and clinical characteristics of the sample were described using descriptive statistics. For categorical variables, frequencies and percentages were reported. Continuous variables were described using measures of central tendency (mean or median) and dispersion (standard deviation or interquartile range), depending on whether the data followed a normal distribution. Gender differences were analyzed using Student’s T-tests for independent samples, reporting the mean difference, 95% confidence intervals, and p-values.

For the reliability analyses, the following metrics were calculated: 1) The mean and standard deviation for each US metric obtained by both operators; 2) The measurement disagreement between examiners, computed as the difference between trials for intra-examiner reliability and as the difference between examiners for single measures and the average of two attempts for inter-examiner reliability, along with the absolute error to avoid underestimating errors; 3) ICCs (ICC3,1 for intra-examiner reliability and ICC3,2 for inter-examiner reliability) using a two-way mixed-effects model for consistency; 4) The standard error of measurement (SEM), calculated as the standard deviation of the mean multiplied by the square root of 1 minus the ICC; 5) The minimal detectable change (MDC), determined as 1.96 times the square root of 2 times the SEM [30]; and 6) the coefficient of variation, calculated to express the relative variability of the measurement error with respect to the average value of the parameter assessed. It was calculated by dividing the absolute error by the mean of the corresponding variable. The result was then multiplied by 100 to express the coefficient of variation as a percentage.

Classification tresholds for ICCs were established following the general guidelines proposed by Koo et al. [30], assigning poor reliability for values under 0.5, moderate reliability for values between 0.5 and 0.75, good reliability for values between 0.75 and 0.9 and excellent reliability for values exceeding 0.9.

Finally, Bland-Altman plots were generated for both shear wave speed and Young’s modulus across examiners. For each parameter, the difference between Trial 1 and Trial 2 values was plotted against their mean. The plots include the mean bias (solid red line) and the 95% limits of agreement (mean difference ±1.96 × standard deviation of the differences; dashed lines). This approach allows for visual inspection of the consistency and spread of repeated measurements. Additionally, to assess the presence of systematic error, we examined whether the mean difference significantly deviated from zero using paired-sample t-tests. Proportional bias was evaluated by performing linear regression analyses between the differences and the means of the two trials; a significant regression slope would indicate proportional error. These analyses were conducted separately for novice and experienced examiners, and for each measurement parameter, to assess the influence of examiner expertise on measurement agreement. The database is available in Supporting information.

Results

Throughout the recruitment phase, 52 individuals indicated a willingness to participate in the study. All prospective participants met the inclusion criteria, and therefore none were excluded. Every image acquired during data collection was reviewed in terms of content and quality, accepted and subsequently analyzed, ensuring that no data were lost. The final sample comprised 35 women and 17 men, and 103 images of the PM muscle were captured from both sides. Given that each examiner acquired two images per muscle, a total of 416 SWE images were acquired and 412 images (n = 4 were excluded due to a data loss) were analyzed.

Table 1 provides an overview of the demographic and clinical profiles of the participants, including gender-based comparisons. Statistically significant differences in demographic variables were found between male and female participants: males were notably younger (p < 0.001), heavier (p < 0.001), and taller (p < 0.001). Differences were also observed in body composition; males exhibited a higher BMI (p = 0.009) and greater water volume (p < 0.001). Nonetheless, despite the BMI discrepancy, there was no significant difference in body fat percentage between sexes (p = 0.271). Clinically, both genders reported comparable levels of pain intensity, classified as moderate (p = 0.584), and similar degrees of disability (moderate disability, p = 0.209).

Table 1. Demographic and clinical characteristics of the sample analyzed.

Variables Subjects with Low Back Pain (n = 52) Difference (95% CI)
Females (n = 35) Males (n = 17)
Demographics
Age, years 32.5 ± 13.4 24.6 ± 7.7 7.9 (0.6;15.3) p < 0.001
Weight, kg 69.9 ± 13.6 92.8 ± 13.8 22.9 (14.3;31.4) p < 0.001
Height, m 1.65 ± 0.04 1.76 ± 0.08 0.11 (0.08;0.15) p < 0.001
BMI, kg/m2 25.7 ± 5.1 29.9 ± 4.9 4.2 (1.1;7.3) p = 0.009
Water volume, L 32.3 ± 3.17 48.5 ± 8.8 16.1 (10.6;21.6) p < 0.001
Body fat, % 33.0 ± 10.5 25.4 ± 12.2 7.6 (−6.5;21.7) p = 0.271
Clinical Characteristics
VAS, 0–10 4.9 ± 1.7 5.2 ± 1.9 0.3 (−0.8;1.4); p = 0.584
ODI, 0–100 24.8 ± 9.2 25.3 ± 9.0 0.5 (−0.2;1.3) p = 0.209

n: Number; CI: Confidence interval; BMI: Body Mass Index; VAS: Visual Analogue Scale; ODI: Oswestry Disability Index.

Reliability estimates for repeated measures of PM stiffness are shown in Table 2. For the experienced examiner, no significant differences emerged between the first and second trials in measurements of shear wave speed or Young’s modulus (both p > 0.05). Conversely, the novice examiner demonstrated significant trial-to-trial differences in both parameters (shear wave speed: p = 0.041; Young’s modulus: p = 0.048). Despite these discrepancies, both examiners achieved excellent intra-class correlation coefficients (ICCs > 0.9), although the experienced examiner exhibited superior consistency across trials (shear wave speed: ICC = 0.971 vs. ICC = 0.930; Young’s modulus: ICC = 0.977 vs. ICC = 0.904). Furthermore, the experienced examiner showed a lower minimal detectable change (MDC), suggesting greater sensitivity to true changes beyond measurement error (shear wave speed: MDC = 0.15 vs. 0.43; Young’s modulus: MDC = 2.06 vs. 7.24). Bland-Altman plots (Fig 2) visually represent these reliability findings, plotting the mean of both trials on the X-axis and their differences on the Y-axis.

Table 2. Test-retest reliability estimates to determine psoas major stiffness.

Experienced Examiner Novice Examiner
Trial 1 (n = 103) Trial 2 (n = 103) Trial 1 (n = 103) Trial 2 (n = 103)
Shear Wave Speed (m/s)
Mean 2.14 ± 0.32 2.15 ± 0.30 2.03 ± 0.27 2.09 ± 0.32
Error −0.01 ± 0.10 −0.06 ± 0.15
Absolute Error 0.08 ± 0.06 p > 0.05 0.13 ± 0.09 p = 0.041
ICC 0.971 (0.957;0.980) 0.930 (0.897;0.953)
SEM 0.05 0.15
MDC 0.15 0.43
CV (%) 3.7 6.3
Young’s Modulus (kPa)
Mean 15.79 ± 4.91 15.92 ± 4.54 15.02 ± 4.59 16.03 ± 5.67
Error −0.12 ± 1.40 −1.01 ± 3.05
Absolute Error 1.08 ± 0.89 p > 0.05 2.62 ± 1.85 p = 0.048
ICC 0.977 (0.966;0.985) 0.904 (0.858;0.935)
SEM 0.74 2.61
MDC 2.06 7.24
CV (%) 6.8 16.9

n: Number; CV: Coefficient of Variation; ICC: Intraclass Correlation Coefficient; SEM: Standard Error of Measurement; MDC: Minimal Detectable Change.

Fig 2. Bland-Altman plots comparing the difference between Trial 1 and Trial 2 measurements across different examiners and parameters.

Fig 2

(A) Shear wave speed: mean average for an experienced examiner, (B) Shear wave speed: mean average for a novice examiner, C) Young’s Modulus: mean average for an experienced examiner, (D) Young’s Modulus: mean average for a novice examiner. The solid red line represents the mean difference (bias) between trials, while the dashed lines indicate the 95% limits of agreement (mean difference ± 1.96 × standard deviation of the differences). Note: the dashed lines may appear slightly curved due to the smoothing method used by the plotting software, but they reflect constant limits of agreement across the x-axis.

Table 3 presents inter-examiner reliability metrics. Analysis of mean differences revealed that most of the measurements taken by the two examiners differed significantly (shear wave speed: both single and averaged measurements, p = 0.001; Young’s modulus: single measurement, p = 0.01). However, no statistically significant difference was found when comparing the average of two Young’s modulus measurements (p > 0.05). Reflecting these outcomes, inter-examiner ICC values were generally lower than intra-examiner values. Single measurements showed good reliability between examiners (shear wave speed: ICC = 0.854; Young’s modulus: ICC = 0.858). When the average of two measurements was used, reliability improved to good-to-excellent levels (shear wave speed: ICC = 0.857; Young’s modulus: ICC = 0.925). Correspondingly, MDC values were reduced when using averaged measurements, aligning with the improved ICCs.

Table 3. Inter-examiner reliability analysis: Single and mean average-measures scores to determine psoas major stiffness.

1 measurement
(n = 103 images per examiner)
Mean average of 2 measurements (n = 206 images per examiner)
Shear Wave Speed (m/s)
Mean 2.09 ± 0.20 2.10 ± 0.20
Error 0.11 ± 0.43 0.02 ± 0.24
Absolute Error 0.37 ± 0.25 p = 0.001 0.19 ± 0.13 p = 0.001
ICC 0.854 (0.785;0.901) 0.857 (0.788;0.903)
SEM 0.07 0.07
MDC 0.21 0.21
CV (%) 17.7 9.0
Young’s Modulus (kPa)
Mean 15.40 ± 3.14 15.69 ± 3.23
Error 0.76 ± 7.13 −0.12 ± 3.86
Absolute Error 5.96 ± 3.94 p = 0.01 3.15 ± 2.21 p > 0.05
ICC 0.858 (0.790;0.904) 0.925 (0.889;0.949)
SEM 1.18 0.88
MDC 3.28 2.45
CV (%) 38.7 20.1

n: Number; CV: Coefficient of Variation; ICC: Intraclass Correlation Coefficient; SEM: Standard Error of Measurement; MDC: Minimal Detectable Change.

Discussion

This investigation is, to our knowledge, the first to examine both intra- and inter-examiner reliability of SWE in evaluating PM muscle stiffness in individuals with LBP. Among the key findings, test-retest reliability demonstrated to be high by both novice and experienced examiners, with all reliability indices yielding near-perfect ICCs (>0.9). Despite these strong ICC values, analysis of mean differences revealed that the measurements taken by the experienced examiner were more consistently reproducible. In contrast, although the novice examiner also achieved excellent ICCs, significant trial-to-trial variability was evident, suggesting reduced repeatability relative to the experienced examiner.

Regarding inter-examiner agreement, significant differences in mean scores were identified between examiners, despite good ICC values for both shear wave speed and Young’s modulus. However, when the mean of two measurements was calculated, no significant differences were observed between examiners for Young’s modulus. This approach improved agreement and accuracy, as evidenced by lower absolute error, SEM, and MDC values. Therefore, conducting a second measurement and calculating the mean average is justified for novel examiners for improving intra-examiner reliability. While experienced examiners obtained almost perfect ICCs obtained for a single measurement (>0.97) with no significant differences between trials, novice examiners have a wider improvement margin (as significant differences between trials were found). Therefore, even if the small improvement margin and the extra time required for multiple measurements is not worthy for experienced examiners, novice examiners may benefit from calculating a mean average of multiple trials.

The variability observed in both intra- and inter-examiner reliability when novice examiners were involved may be attributed to inconsistencies in transducer manipulation, including fluctuations in applied pressure, angulation or alignment. Additionally, differences in the interpretation of SWE images could have contributed to this variability, specially regarding precise delineation of the muscle boundaries [32], which may be hindered by potential histological alterations commonly associated with chronic pain [21,3337].

Future studies should explore these hypotheses by involving novice and experienced examiners assessing healthy subjects. If reliability differences persist in healthy individuals, this would support the transducer handling hypothesis. Conversely, reduced variability in healthy subjects would point toward the impact of muscle histological changes in patients with LBP.

A comparison with Zhou et al. [23] (who reported ICCs of 0.89–0.92 using an examiner with 16 years of clinical experience and 3 years of SWE experience) suggests that experienced examiners are less affected by potential histological changes, supporting the importance of extensive training and experience in probe handling. The reliability observed in Zhou’s study aligns with the results obtained here, regardless of whether participants had pain or were asymptomatic.

The comparison of stiffness scores from asymptomatic individuals reported by Zhou et al. [23] to those in this study should be interpreted with caution as different devices were used and there are no established reliability values for deep muscles. Nevertheless, this contrast revealed higher Young’s modulus values in LBP patients (13.8 ± 3.7 kPa in asymptomatic individuals versus 15.79 ± 4.91 kPa in LBP patients). This difference may indicate that LBP, particularly through its chronic nature, contributes to increased muscle stiffness, possibly linked to the high prevalence of myofascial trigger points in the PM. These differences in stiffness between healthy and LBP populations should be highlighted considering that the mean BMI of the population in our study exceeded 25 points, indicating overweight or obesity, which could imply a greater presence of fatty infiltration in the analyzed muscles [38]. The presence of fatty infiltrates may reduce tissue stiffness and therefore be associated with a lower Young’s modulus, indicating that, in subjects with normal and comparable BMIs, this contrast could be even more notable [39]. Further research is needed to explore whether SWE can effectively differentiate between LBP patients and asymptomatic individuals, the impact of BMI on SWE errors, and its correlation with clinical severity indicators of LBP.

To enhance measurement accuracy, for longitudinal or follow-up assessments, the participation of an experienced examiner is particularly important. Their lower MDC values allow for more accurate identification of true physiological changes in muscle stiffness, thereby decreasing the risk of mistaking measurement variability for actual clinical change. When posible, a single examiner is recommended to be responsible for both acquiring and interpreting SWE images in both clinical practice and research contexts. Variability in technique across different examiners can introduce inconsistencies, potentially leading to misinterpretation of changes in muscle stiffness. When the involvement of multiple examiners is unavoidable, averaging at least two measurements is advised, as this approach minimizes error and enhances measurement reliability, making it appropriate for both clinical and investigative use.

All these findings reinforce the critical role of examiner expertise and methodological rigor in obtaining reliable SWE measurements, particularly in clinical populations.

Despite the valuable results, this study has some limitations that should be acknowledged. The SWE-obtained parameters have not been compared with other clinical variables like the Thomas test, the range of motion in hip extension or the previous physical activity levels, factors that could enrich the clinical contribution of our research. In addition, the sample size is small and the design does not include a group of healthy subjects, which would allow to contrast the results of individuals with similar characteristics except for the presence or absence of LBP. Additionally, the fact that we only evaluated one portion of the PM at rest can restrict the generalizability of our results. Future studies could improve these aspects in their methodology to complement our findings and facilitate their extrapolation and integration in clinical settings.

Conclusion

SWE test-retest reliability for evaluating PM muscle stiffness in individuals with chronic LBP was excellent for both novice and experienced examiners, with ICCs exceeding 0.9, providing highly consistent measurements when performed by the same assessor. In contrast, inter-examiner reliability was notably lower, reflecting considerable variability in SWE outcomes between examiners with differing levels of experience. The accuracy of SWE largely depends on consistency in the evaluator conducting the assessment. When multiple assessors must be involved, averaging multiple measurements is recommended to improve reliability and reduce the potential for measurement error.

Supporting information

S1 File. Raw data.

(PDF)

pone.0330076.s001.pdf (98.3KB, pdf)

Data Availability

All relevant data are within the paper and its Supporting information files.

Funding Statement

This study was financially supported by Banco Santander (https://www.bancosantander.es) in the form of a researcher training scholarship award (CT25/24) received by GR-R. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding was received for this study.

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PLoS One. 2025 Oct 9;20(10):e0330076. doi: 10.1371/journal.pone.0330076.r001

Author response to Decision Letter 0


Transfer Alert

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25 May 2025

Decision Letter 0

Masatoshi Nakamura

23 Jun 2025

Dear Dr. Buffet-García,

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Additional Editor Comments (if provided):

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Partly

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2. Has the statistical analysis been performed appropriately and rigorously? -->?>

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available??>

The PLOS Data policy

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English??>

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Thank you for the opportunity to review this study. The present manuscript investigates the intra- and inter-rater reliability of shear wave elastography (SWE) measurements of the psoas major (PM) muscle. The study is well designed and clearly written, and the results suggest that SWE may be a reliable method for assessing PM stiffness in patients with low back pain (LBP). I appreciate the authors’ contribution to this area of musculoskeletal research. Nevertheless, I have several comments that I believe would strengthen the manuscript, particularly regarding the methodology.

Introduction:

-The Introduction is somewhat lengthy. The detailed description of the PM functional anatomy may be reduced or moved to the Discussion if necessary.

Methods:

-Was participants’ physical activity level controlled prior to the SWE assessment? For example, performing physical exercise or manual labor immediately before measurement could influence the mechanical properties of the PM. This potential confounder should be discussed

-Given that the PM is located relatively deep, was the depth of the region of interest (ROI) recorded during SWE acquisition? Depth can influence modulus values. If the depth varied across measurements or between participants, this should be considered in the interpretation.

-In each SWE measurement session, was only one SWE image acquired? If so, this should be stated clearly. Since SWE values tend to be quite sensitive to how transducer is manipulated, acquiring and averaging multiple scans would reduce measurement error and improve reliability. Please clarify.

-The classification thresholds for intraclass correlation coefficient (ICC) (e.g., moderate, good, excellent) should be explicitly defined, preferably with references.

-It would enhance the clinical impact of the study if SWE values were compared to physical examination findings associated with PM stiffness, such as the Thomas test or hip extension range of motion. Furthermore, including healthy volunteers as a comparison group would have helped interpret whether elevated stiffness values are indeed associated with LBP.

Results:

-There seems to be a discrepancy in participant numbers. Table 1 reports 16 men and 31 women, which sums to 47, yet the total number of participants is stated as 52. Please confirm and correct this inconsistency.

Discussion

-Lines 338–344: The average BMI of both male and female participants exceeds 25, indicating that some may be classified as overweight or obese. Obesity can lead to intramuscular fatty infiltration, including in the PM. Could the relatively higher Young’s modulus observed in this study be attributed, at least in part, to fatty infiltration rather than LBP per se?

-Please add the limitations of this study.

Table

-As mentioned in the main text, please include p-values to support the statistical significance of the comparisons.

Reviewer #2: Thank you for the opportunity to review this manuscript aimed to evaluate the intra and inter-examiner reliability of shear wave elastography for calculating the PM stiffness in patients with LBP. While the approach is rigor and the findings are important, I would like to point out some methodological and interpretation concerns.

Major comments

1. While psoas major CSA changes and fatty infiltration were clearly described in L98-112, these changes did not necessarily align with the main aim, which was to evaluate the reliability of stiffness measurements. The introduction should reduce or eliminate these descriptions to focus on the need for a validated method to quantify muscle stiffness in LBP patients.

2. To further strengthen the rationale for this study, the authors should mention the prior reliability study in healthy subjects by Zhou et al. in the introduction. Following this, the authors should hypothesize why these findings of high reliability may not be generalizable to patients with LBP.

3. L181. The study analyzes muscles from both sides of the body. It is possible that data from the left and right sides of the same participant are not independent. I wonder if this lack of independence can cause the intraclass correlation coefficient (ICC) to increase. The potential influence of this non-independence on the statistical results should be discussed or addressed within the statistical model.

4. L254. While the authors assessed consistency within and between examiners using ICC(3,1) and ICC(3,2), they may overlook systematic error, which would be an issue in clinical settings. To provide a more comprehensive evaluation of reliability, I suggest adding an ICC(2,1), which incorporates both random and systematic error. Also, please calculate the SEM and MDC using the ICC(2,1) value.

Specific comments

Abstract

5. L66. The conclusion states, "Excellent test-retest reliability was observed for the measurement of PM muscle stiffness... by both experienced and novice evaluators." However, this statement should be revised to acknowledge the statistically significant difference between the two trials for the novice examiner.

Introduction

6. L95�”including” is a typo.

Methods

7. L191. What is the rationale behind including 52 participants, despite having confirmed that the minimum sample size was 29? Could using a larger sample size lead to an overestimation of reliability?

8. L232. Please add the region of interest size.

9. The manuscript does not explain how Young's modulus was calculated from the shear wave speed. To ensure reproducibility, the authors must provide the formula used and any assumptions made (e.g., tissue density).

10. Please add the information on imaging processing, such as smoothing an so on.

11. The authors have two values of each muscle in each examiner. Which data (trial 1 or 2?) do the author use to calculate ICC(3,1)?

Results

12. L264: The statement "Every image acquired during data collection was deemed valid" is concerning. Were there pre-defined criteria for image acceptability? The absence of any excluded images in 416 acquisitions seems unlikely and requires explanation.

13. L265. The manuscript has stated that the total number of participants is 52. However, 31 females and 16 males add up to 47. Please correct.

14. L267. Did you acquire two consecutive images without removing the probe, or did you reposition the probe before the second measurement?

15. L272. This section includes data for "Water volume" and “body composition”, but the method to measure these variables is not described in the Methods section.

16. Please consider adding the coefficient of variation alongside ICC to provide a more complete assessment of measurement variability, especially given the differing scales of shear wave velocity and Young’s modulus.

17. Bland-Altman plots are presented in the results, but this analysis is not described in the Methods section. Furthermore, the authors should analyze the presence or absence of systematic and proportional error using the plots and statistical tests.

Discussion

18. L338. In the Discussion, the authors compared their Young's modulus values with data from the study by Zhou et al. However, the two studies used different ultrasound systems (Canon vs. Aixplorer). Without established inter-machine reliability, particularly for deep muscles, this direct comparison is not valid. Therefore, I believe this comparison should be avoided.

19. The authors should acknowledge that evaluating only one portion of the PM at rest is a limitation that restricts the generalizability of the study.

Figure/Caption

20. Figure 1: The figure, particularly the left-hand schematic, appears very similar to a figure in the publication with PMID: 40150065. If this figure has been adapted or inspired by another source, that source must be appropriately cited. Also, in Figure1’s caption, “mayor” is a typo.

21. Figure 2: The caption for the Bland-Altman plots should explain what the solid red lines and the dashed lines represent. Additionally, the dashed lines appear curved in the figure; please confirm if this is an error or an intended feature of the plotting method.

22. Table2. In trial 2, the novice examiner measured a mean shear wave speed of 2.09 m/s and a Young’s modulus of 16.03 kPa, while the experienced examiner measured a higher shear wave speed of 2.15 m/s but a lower Young’s modulus of 15.9 kPa. Is this discrepancy due to different calculation methods, or is it a reporting error?

23. Table 3. I was not sure how the mean, error, and absolute error were calculated.

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Reviewer #2: No

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PLoS One. 2025 Oct 9;20(10):e0330076. doi: 10.1371/journal.pone.0330076.r003

Author response to Decision Letter 1


9 Jul 2025

Response to Reviewers

Shear Wave Elastography for measuring the elastic properties of the Psoas Major muscle: an intra- and inter-examiner reliability study

We would like to thank the Editor and Reviewers for their comments and suggestions for the improvement of our manuscript. We have carried out an in-depth revision to ensure that the document is adapted to the style requirements of the journal and to respond to the Reviewers' proposals. Changes are detailed below:

Academic Editor

Journal requirements:

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Response: We have revised the formal aspects of the text and files to adapt them to the journal's requirements.

2. We note that your Data Availability Statement is currently as follows: [All relevant data are within the manuscript and its Supporting Information files.]

Please confirm at this time whether or not your submission contains all raw data required to replicate the results of your study. Authors must share the “minimal data set” for their submission. PLOS defines the minimal data set to consist of the data required to replicate all study findings reported in the article, as well as related metadata and methods (https://journals.plos.org/plosone/s/data-availability#loc-minimal-data-set-definition).

For example, authors should submit the following data:

- The values behind the means, standard deviations and other measures reported;

- The values used to build graphs;

- The points extracted from images for analysis.

Authors do not need to submit their entire data set if only a portion of the data was used in the reported study.

If your submission does not contain these data, please either upload them as Supporting Information files or deposit them to a stable, public repository and provide us with the relevant URLs, DOIs, or accession numbers. For a list of recommended repositories, please see https://journals.plos.org/plosone/s/recommended-repositories.

If there are ethical or legal restrictions on sharing a de-identified data set, please explain them in detail (e.g., data contain potentially sensitive information, data are owned by a third-party organization, etc.) and who has imposed them (e.g., an ethics committee). Please also provide contact information for a data access committee, ethics committee, or other institutional body to which data requests may be sent. If data are owned by a third party, please indicate how others may request data access.

Response: Thank you. We confirm that our submission contains all required data.

3. When completing the data availability statement of the submission form, you indicated that you will make your data available on acceptance. We strongly recommend all authors decide on a data sharing plan before acceptance, as the process can be lengthy and hold up publication timelines. Please note that, though access restrictions are acceptable now, your entire data will need to be made freely accessible if your manuscript is accepted for publication. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If you are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process.

Response: Thank you for your recommendation. We confirm that all data is reported in the manuscript. If any additional information is required, we will be pleased to provide it.

4. Your ethics statement should only appear in the Methods section of your manuscript. If your ethics statement is written in any section besides the Methods, please move it to the Methods section and delete it from any other section. Please ensure that your ethics statement is included in your manuscript, as the ethics statement entered into the online submission form will not be published alongside your manuscript.

Response: We have deleted the ethics statement from the Declarations section and have left it only in the Methods section following your recommendations.

Reviewer #1

Thank you for the opportunity to review this study. The present manuscript investigates the intra- and inter-rater reliability of shear wave elastography (SWE) measurements of the psoas major (PM) muscle. The study is well designed and clearly written, and the results suggest that SWE may be a reliable method for assessing PM stiffness in patients with low back pain (LBP). I appreciate the authors’ contribution to this area of musculoskeletal research. Nevertheless, I have several comments that I believe would strengthen the manuscript, particularly regarding the methodology.

Response: Thank you for this positive feedback.

Introduction:

-The Introduction is somewhat lengthy. The detailed description of the PM functional anatomy may be reduced or moved to the Discussion if necessary.

Response: We appreciate your suggestion. We have reduced the detailed description of the PM anatomy to focus on its influence on LBP.

Methods:

-Was participants’ physical activity level controlled prior to the SWE assessment? For example, performing physical exercise or manual labor immediately before measurement could influence the mechanical properties of the PM. This potential confounder should be discussed.

Response: Despite being factors that could affect muscle properties, our study design did not take them into account as these factors are hard to control in the clinical practice. However, even if these factors were not controlled, the results we obtained further reinforce the real reliability of the procedure. Nevertheless, we have included this aspect in our limitations.

-Given that the PM is located relatively deep, was the depth of the region of interest (ROI) recorded during SWE acquisition? Depth can influence modulus values. If the depth varied across measurements or between participants, this should be considered in the interpretation.

Response: The depth of the region of interest varied between participants depending on their anthropometric characteristics, without setting a fixed reference value, as we have clarified in the Methods section. In our opinion, as discussed in the previous section and far from being a negative aspect, these possible variations reinforce the good reliability results that we obtained.

-In each SWE measurement session, was only one SWE image acquired? If so, this should be stated clearly. Since SWE values tend to be quite sensitive to how transducer is manipulated, acquiring and averaging multiple scans would reduce measurement error and improve reliability. Please clarify.

Response: Thank you. To avoid doubts, we have clarified that a single SWE image was acquired per muscle in each measurement session. Participants were asked to attend at four different times. The first two visits were in day one (in the morning with examiner A and in the afternoon with examiner B) and the second two visits were in day two (in the morning with examiner B and in the afternoon with examiner A).

Regarding the impact of averaging multiple scans, this is already discussed (declaring that a mean average calculation improved inter-examiner concordance compared to a single measurement, reaching no significant differences in Young’s modulus).

If you ask to calculate a mean average for improving intra-examiner reliability, we believe that investing time in conducting a second measurement and calculating the mean average is justified for novel examiners. Experienced examiners obtained almost perfect ICCs obtained for a single measurement (>0.97), with no significant differences between trials. Therefore, the small improvement margin and the extra time required for multiple measurements is not worthy. In contrast, novice examiners have more margin (as significant differences between trials were found) and they may benefit from calculating a mean average of multiple trials. We included this reflection in Discussion.

-The classification thresholds for intraclass correlation coefficient (ICC) (e.g., moderate, good, excellent) should be explicitly defined, preferably with references.

Response: Thank you for your appreciation, we have included the interpretation of the ICCs following the recommendations of Koo et al. (https://doi.org/10.1016/j.jcm.2016.02.012) in the Statistical analysis section.

-It would enhance the clinical impact of the study if SWE values were compared to physical examination findings associated with PM stiffness, such as the Thomas test or hip extension range of motion. Furthermore, including healthy volunteers as a comparison group would have helped interpret whether elevated stiffness values are indeed associated with LBP.

Response: Thank you very much for your suggestion. We totally agree and we are actually working on it, but this would require a totally different design (case-control study) and including both designs in a single article would increase significantly the length of the manuscript. While we recognize the clinical relevance of these studies, we believe that the first step should be confirm the reliability of the procedure to focus posteriorly on observational studies (correlation between muscle stiffness with clinical severity indicators, sensitivity and specificity to classify asymptomatic individuals or clinical populations, differences between cases and controls…)

We have added these aspects to the limitations and will take them into account for future research.

Results:

-There seems to be a discrepancy in participant numbers. Table 1 reports 16 men and 31 women, which sums to 47, yet the total number of participants is stated as 52. Please confirm and correct this inconsistency.

Response: Thank you for your comment, we apologize for the inconsistency. We have corrected the corresponding data on the total number of 47 subjects.

Discussion

-Lines 338–344: The average BMI of both male and female participants exceeds 25, indicating that some may be classified as overweight or obese. Obesity can lead to intramuscular fatty infiltration, including in the PM. Could the relatively higher Young’s modulus observed in this study be attributed, at least in part, to fatty infiltration rather than LBP per se?

Response: Thank you for your comment. We cannot confirm or discard this hypothesis as we did not conduct a correlation analysis. These analyses are planned after publishing the reliability results of the procedure (not only the association between SWE scores and BMI, but also the influence of BMI on SWE errors). For this purpose, a larger sample size and a wider range of BMIs would be needed to support the conclusions on enough statistical power.

We believe that this comment is a good opportunity to introduce the possible implication of BMI and fatty infiltration on SWE results. The possible increase of fatty infiltration in our sample could have reduced tissue stiffness, thus associating lower values of Young's modulus and making the differences between subjects with comparable BMIs even more significant.

-Please add the limitations of this study.

Response: Thank you. We have added this section at the end of the Discussion.

Table

-As mentioned in the main text, please include p-values to support the statistical significance of the comparisons.

Response: Thank you for your suggestions. We have added the p-values of the comparisons to the tables.

Reviewer #2

Thank you for the opportunity to review this manuscript aimed to evaluate the intra and inter-examiner reliability of shear wave elastography for calculating the PM stiffness in patients with LBP. While the approach is rigor and the findings are important, I would like to point out some methodological and interpretation concerns.

Response: Thank you for this positive feedback.

Major comments

1. While psoas major CSA changes and fatty infiltration were clearly described in L98-112, these changes did not necessarily align with the main aim, which was to evaluate the reliability of stiffness measurements. The introduction should reduce or eliminate these descriptions to focus on the need for a validated method to quantify muscle stiffness in LBP patients.

Response: Thank you for your recommendation. We have eliminated these descriptions to avoid information overload and to focus on the objective of the study.

2. To further strengthen the rationale for this study, the authors should mention the prior reliability study in healthy subjects by Zhou et al. in the introduction. Following this, the authors should hypothesize why these findings of high reliability may not be generalizable to patients with LBP.

Response: Thank you for your appreciation. We have mentioned the previous study by Zhou et al. in our introduction and hypothesized that their results may not be extrapolated to individuals with LBP due to their specific muscular characteristics.

3. L181. The study analyzes muscles from both sides of the body. It is possible that data from the left and right sides of the same participant are not independent. I wonder if this lack of independence can cause the intraclass correlation coefficient (ICC) to increase. The potential influence of this non-independence on the statistical results should be discussed or addressed within the statistical model.

Response: We agree with the reviewer that measurements taken from both sides of the same participant cannot be considered fully independent data points, and this is an important methodological consideration that has been now discussed in the manuscript. However, we believe that this decision does not artificially inflate the ICC values reported. In fact, the inclusion of bilateral data could theoretically lead to underestimation or overestimation of ICCs.

In our study, we took several steps to mitigate the potential impact of non-independence: 1) repositioning protocols for both the patient and the probe were strictly standardized across all trials; (2) the order of side evaluation was randomized; and (3) previous research (e.g., doi: 10.3390/bioengineering10080904) has shown that anthropometric and sociodemographic factors are not significantly associated with SWE measurement errors. Moreover, our sample displayed considerable heterogeneity in these variables, which reduces the likelihood that demographic similarity introduced any systematic bias. In fact, in populations with musculoskeletal pain, asymmetries are often more pronounced, which further supports the relevance of analyzing each side independently for reliability purposes. Based on these considerations, we believe that the reported ICCs are robust and not substantially influenced by any lack of independence between sides. Please check the section “Examiners” where we addressed this comment.

4. L254. While the authors assessed consistency within and between examiners using ICC(3,1) and ICC(3,2), they may overlook systematic error, which would be an issue in clinical settings. To provide a more comprehensive evaluation of reliability, I suggest adding an ICC(2,1), which incorporates both random and systematic error. Also, please calculate the SEM and MDC using the ICC(2,1) value.

Response: After carefully reviewing the guidelines provided by Koo and Li (2016), we chose to report ICC(3,1) and ICC(3,2) based on a two-way mixed-effects model for consistency, which is more appropriate when the same raters are fixed and of primary interest, as is the case in our study. ICC(2,1), based on absolute agreement in a random-effects model, is generally applied when the goal is to generalize the results to a larger population of raters, which is not the case in our design.

Moreover, absolute agreement is more suitable for categorical or rater-dependent decisions, where any dis

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Decision Letter 1

Masatoshi Nakamura

27 Jul 2025

Shear Wave Elastography for measuring the elastic properties of the Psoas Major muscle: an intra- and inter-examiner reliability study

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Reviewer #1: The author has sufficiently addressed all of my comments. The revised manuscript has been improved accordingly and is now suitable for publication.

Reviewer #2: Thank you for addressing all my comments. I feel you have responded carefully to the concerns I raised. I have just one point, which may be a difference in perspective: while I agree with the interpretation of ICC(3,1) and ICC(2,1), I believe that since the SEM and MDC are also derived from ICC(3,1), they fail to account for the influence of systematic error.

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Acceptance letter

Masatoshi Nakamura

PONE-D-25-25182R1

PLOS ONE

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