{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:28:16Z","timestamp":1760059696787,"version":"build-2065373602"},"reference-count":49,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2025,6,30]],"date-time":"2025-06-30T00:00:00Z","timestamp":1751241600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Important Weak Subject Construction Project of Shanghai Pudong New Area Health Commission","award":["PWZbr2022-12","202140250"],"award-info":[{"award-number":["PWZbr2022-12","202140250"]}]},{"DOI":"10.13039\/100017950","name":"Shanghai Municipal Health Commission","doi-asserted-by":"publisher","award":["PWZbr2022-12","202140250"],"award-info":[{"award-number":["PWZbr2022-12","202140250"]}],"id":[{"id":"10.13039\/100017950","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>Background: Gallbladder hypomotility is a key pathogenic factor in cholelithiasis. Non-invasive interventions to enhance gallbladder contractility remain limited. Ultrasound therapy has shown promise in various muscular disorders, but its effects on gallbladder function are unexplored. Methods: This study employed low-intensity pulsed ultrasound (LIPUS) at a 3 MHz frequency and 0.8 W\/cm2 intensity with a 20% duty cycle to irradiate the gallbladder region of fasting guinea pigs. Gallbladder contractile function was evaluated through multiple complementary approaches: in vivo assessment via two-dimensional\/three-dimensional ultrasound imaging to monitor volumetric changes; quantitative functional evaluation using nuclear medicine scintigraphy (99mTc-HIDA); and ex vivo experiments including isolated gallbladder muscle strip tension measurements, histopathological analysis, \u03b1-smooth muscle actin (\u03b1-SMA) immunohistochemistry, and intracellular calcium fluorescence imaging. Results: Ultrasound significantly enhanced gallbladder emptying, evidenced by the volume reduction and increased ejection fraction. Scintigraphy confirmed accelerated bile transport in treated animals. Ex vivo analyses demonstrated augmented contractile force, amplitude, and frequency in ultrasound-treated smooth muscle. Histological examination revealed smooth muscle hypertrophy, \u03b1-SMA upregulation, and elevated intracellular calcium levels. Extended ultrasound exposure produced sustained functional improvements without tissue damage. Conclusions: Ultrasound effectively enhances gallbladder contractile function through mechanisms involving smooth muscle structural modification and calcium signaling modulation. These findings establish the experimental foundation for ultrasound as a promising non-invasive therapeutic approach to improve gallbladder motility and potentially prevent gallstone formation.<\/jats:p>","DOI":"10.3390\/bioengineering12070716","type":"journal-article","created":{"date-parts":[[2025,7,1]],"date-time":"2025-07-01T07:42:06Z","timestamp":1751355726000},"page":"716","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Ultrasound Improves Gallbladder Contraction Function: A Non-Invasive Experimental Validation Using Small Animals"],"prefix":"10.3390","volume":"12","author":[{"given":"Run","family":"Guo","sequence":"first","affiliation":[{"name":"Department of Ultrasound in Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Center of Gallbladder Disease, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tian","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Ultrasound in Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Center of Gallbladder Disease, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4624-3134","authenticated-orcid":false,"given":"Fan","family":"Ding","sequence":"additional","affiliation":[{"name":"Department of Ultrasound in Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Center of Gallbladder Disease, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li-Ping","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Ultrasound in Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Center of Gallbladder Disease, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1470-7942","authenticated-orcid":false,"given":"Fang","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Ultrasound in Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Center of Gallbladder Disease, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gang","family":"Zhao","sequence":"additional","affiliation":[{"name":"Center of Gallbladder Disease, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"},{"name":"Institute of Gallstone Disease, School of Medicine, Tongji University, Shanghai 200331, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Ultrasound in Medicine, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai 200120, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,6,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/0168-8278(92)90158-L","article-title":"Fasting gallbladder volume, postprandial emptying and cholecystokinin release in gallstone patients and normal subjects","volume":"14","author":"Stolk","year":"1992","journal-title":"J. Hepatol."},{"key":"ref_2","first-page":"518","article-title":"Gallstone Disease: Common Questions and Answers","volume":"109","author":"Patel","year":"2024","journal-title":"Am. Fam. Physician"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e313","DOI":"10.1111\/j.1365-2982.2012.01935.x","article-title":"Disruption of gallbladder smooth muscle function is an early feature in the development of cholesterol gallstone disease","volume":"24","author":"Lavoie","year":"2012","journal-title":"Neurogastroenterol. Motil."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2576","DOI":"10.1111\/j.1572-0241.2006.00793.x","article-title":"Is it possible to predict the clinical course of gallstone disease? Usefulness of gallbladder motility evaluation in a clinical setting","volume":"101","author":"Colecchia","year":"2006","journal-title":"Am. J. Gastroenterol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Tong, K., Jing, C., Wang, T., Liu, K., Guo, W., and Zhang, Z. (2024). The roles of metal ions in gallstones formation. Asian J. Surg.","DOI":"10.1016\/j.asjsur.2024.05.243"},{"key":"ref_6","first-page":"148","article-title":"The Treatment of Gallstone Disease","volume":"117","author":"Gutt","year":"2020","journal-title":"Dtsch. Arztebl. Int."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"255","DOI":"10.21037\/tau.2016.02.04","article-title":"Clinical applications of low-intensity pulsed ultrasound and its potential role in urology","volume":"5","author":"Xin","year":"2016","journal-title":"Transl. Androl. Urol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"7964897","DOI":"10.1155\/2019\/7964897","article-title":"Efficacy and Safety of a Stimulator Using Low-Intensity Pulsed Ultrasound Combined with Transcutaneous Electrical Nerve Stimulation in Patients with Painful Knee Osteoarthritis","volume":"2019","author":"Kim","year":"2019","journal-title":"Pain Res. Manag."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1007\/s11255-016-1298-8","article-title":"Ultrasound induces contraction of the bladder smooth muscle","volume":"48","author":"Ren","year":"2016","journal-title":"Int. Urol. Nephrol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"119278","DOI":"10.1016\/j.carbpol.2022.119278","article-title":"Study on synergistic effects of carboxymethyl cellulose and LIPUS for bone tissue engineering","volume":"286","author":"Tang","year":"2022","journal-title":"Carbohydr. Polym."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Sun, L., An, S., Zhang, Z., Zhou, Y., Yu, Y., Ma, Z., Fan, X., Tang, L., and Guo, J. (2021). Molecular and Metabolic Mechanism of Low-Intensity Pulsed Ultrasound Improving Muscle Atrophy in Hindlimb Unloading Rats. Int. J. Mol. Sci., 22.","DOI":"10.21203\/rs.3.rs-389558\/v1"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Li, X., Zhong, Y., Zhou, W., Song, Y., Li, W., Jin, Q., Gao, T., Zhang, L., and Xie, M. (2023). Low-intensity pulsed ultrasound (LIPUS) enhances the anti-inflammatory effects of bone marrow mesenchymal stem cells (BMSCs)-derived extracellular vesicles. Cell. Mol. Biol. Lett., 28.","DOI":"10.1186\/s11658-023-00422-3"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"106739","DOI":"10.1016\/j.ultras.2022.106739","article-title":"Ca2+ signaling-mediated low-intensity pulsed ultrasound-induced proliferation and activation of motor neuron cells","volume":"124","author":"Truong","year":"2022","journal-title":"Ultrasonics"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1452","DOI":"10.1016\/j.ultrasmedbio.2017.03.003","article-title":"Optimal Ultrasound Exposure Conditions for Maximizing C2C12 Muscle Cell Proliferation and Differentiation","volume":"43","author":"Salgarella","year":"2017","journal-title":"Ultrasound Med. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1016\/j.neurom.2024.06.005","article-title":"Low-Intensity Ultrasound Tibial Nerve Stimulation Suppresses Bladder Activity in Rats","volume":"28","author":"He","year":"2025","journal-title":"Neuromodulation"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e14843","DOI":"10.1111\/nmo.14843","article-title":"Low-intensity pulsed ultrasound combined with ST36 modulate gastric smooth muscle contractile marker expression via RhoA\/Rock and MALAT1\/miR-449a\/DLL1 signaling in diabetic rats","volume":"36","author":"Han","year":"2024","journal-title":"Neurogastroenterol. Motil."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Chen, F., Guo, R., Chen, T., Liu, L., Ding, F., Zhao, G., and Zhang, B. (2025). The Therapeutic Potential of Low-Intensity Pulsed Ultrasound in Enhancing Gallbladder Function and Reducing Inflammation in Cholesterol Gallstone Disease. Bioengineering, 12.","DOI":"10.3390\/bioengineering12010034"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"448","DOI":"10.3390\/livers3030032","article-title":"Perspective on Quantitative Structure\u2013Toxicity Relationship (QSTR) Models to Predict Hepatic Biotransformation of Xenobiotics","volume":"3","author":"Rai","year":"2023","journal-title":"Livers"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1111\/j.1651-2227.2006.tb02350.x","article-title":"Ultrasonographic evaluation of fatty meal stimulated gallbladder contraction in the diagnosis of biliary dyskinesia in children","volume":"95","author":"Cay","year":"2006","journal-title":"Acta Paediatr."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"967","DOI":"10.2967\/jnumed.113.131490","article-title":"Hepatobiliary scintigraphy in 2014","volume":"55","author":"Ziessman","year":"2014","journal-title":"J. Nucl. Med."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1016\/j.suc.2024.04.007","article-title":"Biliary Dyskinesia and Hyperkinesis","volume":"104","author":"Tseng","year":"2024","journal-title":"Surg. Clin. N. Am."},{"key":"ref_22","first-page":"385","article-title":"Gallbladder motility in children with celiac disease before and after gluten-free diet","volume":"34","author":"Das","year":"2021","journal-title":"Ann. Gastroenterol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"114283","DOI":"10.1016\/j.yexcr.2024.114283","article-title":"Mechanism of denervation muscle atrophy mediated by Ach\/p38\/MAPK pathway in rats with erectile dysfunction caused by nerve injury","volume":"442","author":"Jie","year":"2024","journal-title":"Exp. Cell Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"G501","DOI":"10.1152\/ajpgi.00140.2005","article-title":"Identification of a spontaneously active, Na+-permeable channel in guinea pig gallbladder smooth muscle","volume":"289","author":"Petkov","year":"2005","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Zhang, L., Liu, X., Gao, L., Ji, Y., Wang, L., Zhang, C., Dai, L., Liu, J., and Ji, Z. (2020). Activation of Piezo1 by ultrasonic stimulation and its effect on the permeability of human umbilical vein endothelial cells. Biomed. Pharmacother., 131.","DOI":"10.1016\/j.biopha.2020.110796"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"546","DOI":"10.1302\/0301-620X.80B3.0800546","article-title":"Dose-related effects of shock waves on rabbit tendo Achillis. A sonographic and histological study","volume":"80","author":"Rompe","year":"1998","journal-title":"J. Bone Jt. Surg. Br."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Wang, J., Ren, L., Liu, X., Xu, W., Liu, M., Hu, P., Wang, T., Liu, J., and Ling, Q. (2023). Transcriptomics Reveals Molecular Features of the Bilateral Pelvic Nerve Injury Rat Model of Detrusor Underactivity. Biomolecules, 13.","DOI":"10.3390\/biom13081260"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5517","DOI":"10.7150\/thno.33876","article-title":"Focused ultrasound activates voltage-gated calcium channels through depolarizing TRPC1 sodium currents in kidney and skeletal muscle","volume":"9","author":"Burks","year":"2019","journal-title":"Theranostics"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1007\/s11427-015-4958-5","article-title":"The role of TRPP2 in agonist-induced gallbladder smooth muscle contraction","volume":"59","author":"Zhong","year":"2016","journal-title":"Sci. China Life Sci."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Przystupski, D., and Ussowicz, M. (2022). Landscape of Cellular Bioeffects Triggered by Ultrasound-Induced Sonoporation. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231911222"},{"key":"ref_31","first-page":"401","article-title":"Effects of Extracellular Matrix Changes Induced by a High-Fat Diet on Gallbladder Smooth Muscle Dysfunction","volume":"29","author":"Zhong","year":"2024","journal-title":"FBL"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Keshavarz, M., Ruppert, A.L., Meiners, M., Poharkar, K., Liu, S., Mahmoud, W., Winterberg, S., Hartmann, P., Mermer, P., and Perniss, A. (2024). Bitter tastants relax the mouse gallbladder smooth muscle independent of signaling through tuft cells and bitter taste receptors. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-69287-6"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1002\/j.2040-4603.2016.tb00717.x","article-title":"Functions of the Gallbladder","volume":"6","author":"Housset","year":"2016","journal-title":"Compr. Physiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1046\/j.1365-2362.2000.00639.x","article-title":"Gallbladder motility and cholesterol crystallization in bile from patients with pigment and cholesterol gallstones","volume":"30","author":"Portincasa","year":"2000","journal-title":"Eur. J. Clin. Investig."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2066","DOI":"10.3748\/wjg.v13.i14.2066","article-title":"Surgical anatomy of innervation of the gallbladder in humans and Suncus murinus with special reference to morphological understanding of gallstone formation after gastrectomy","volume":"13","author":"Yi","year":"2007","journal-title":"World J. Gastroenterol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"7487","DOI":"10.1021\/acs.jcim.3c00954","article-title":"Molecular Insights into the MLCK Activation by CaM","volume":"63","author":"Fang","year":"2023","journal-title":"J. Chem. Inf. Model."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1007\/s10620-005-2852-x","article-title":"Controversies concerning pathophysiology and management of acalculous biliary-type abdominal pain","volume":"50","author":"Rastogi","year":"2005","journal-title":"Dig. Dis. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1007\/s11605-010-1342-4","article-title":"The cholecystokin provocation HIDA test: Recreation of symptoms is superior to ejection fraction in predicting medium-term outcomes","volume":"15","author":"Falk","year":"2011","journal-title":"J. Gastrointest. Surg."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.jss.2014.01.004","article-title":"Abdominal ultrasound versus hepato-imino diacetic acid scan in diagnosing acute cholecystitis\u2014What is the real benefit?","volume":"188","author":"Kaoutzanis","year":"2014","journal-title":"J. Surg. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"443","DOI":"10.21037\/qims-20-680","article-title":"Review on experimental study and clinical application of low-intensity pulsed ultrasound in inflammation","volume":"11","author":"Xu","year":"2021","journal-title":"Quant. Imaging Med. Surg."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1038\/nrm2592","article-title":"Mechanotransduction in development: A growing role for contractility","volume":"10","author":"Wozniak","year":"2009","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1152\/japplphysiol.00980.2014","article-title":"Ultrasound-induced modulation of cardiac rhythm in neonatal rat ventricular cardiomyocytes","volume":"118","author":"Fleischman","year":"2015","journal-title":"J. Appl. Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1016\/j.biomaterials.2007.10.050","article-title":"The effect of cyclic strain on embryonic stem cell-derived cardiomyocytes","volume":"29","author":"Gwak","year":"2008","journal-title":"Biomaterials"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1006\/jmcc.1998.0865","article-title":"Orientation change of cardiocytes induced by cyclic stretch stimulation: Time dependency and involvement of protein kinases","volume":"31","author":"Kada","year":"1999","journal-title":"J. Mol. Cell. Cardiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1569","DOI":"10.12669\/pjms.37.6-WIT.4839","article-title":"Low intensity pulsed ultrasound information technology intervention in diagnosis and prediction of Muscle Atrophy","volume":"37","author":"Sun","year":"2021","journal-title":"Pak. J. Med. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e1211","DOI":"10.1016\/j.urology.2015.07.026","article-title":"Low-intensity Pulsed Ultrasound Improves Erectile Function in Streptozotocin-induced Type I Diabetic Rats","volume":"86","author":"Lei","year":"2015","journal-title":"Urology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.7150\/ijbs.79685","article-title":"Low-intensity pulsed ultrasound promotes skeletal muscle regeneration via modulating the inflammatory immune microenvironment","volume":"19","author":"Qin","year":"2023","journal-title":"Int. J. Biol. Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"111097","DOI":"10.1016\/j.cellsig.2024.111097","article-title":"Low-intensity pulsed ultrasound (LIPUS) promotes skeletal muscle regeneration by regulating PGC-1\u03b1\/AMPK\/GLUT4 pathways in satellite cells\/myoblasts","volume":"117","author":"Duan","year":"2024","journal-title":"Cell. Signal."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1973","DOI":"10.1016\/j.ultrasmedbio.2024.08.016","article-title":"Jingle Cell Rock: Steering Cellular Activity With Low-Intensity Pulsed Ultrasound (LIPUS) to Engineer Functional Tissues in Regenerative Medicine","volume":"50","author":"Marcotulli","year":"2024","journal-title":"Ultrasound Med. Biol."}],"container-title":["Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-5354\/12\/7\/716\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:02:14Z","timestamp":1760032934000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-5354\/12\/7\/716"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,30]]},"references-count":49,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2025,7]]}},"alternative-id":["bioengineering12070716"],"URL":"https:\/\/doi.org\/10.3390\/bioengineering12070716","relation":{},"ISSN":["2306-5354"],"issn-type":[{"type":"electronic","value":"2306-5354"}],"subject":[],"published":{"date-parts":[[2025,6,30]]}}}