Ultrasound-Assisted Immunoregulation: A Rising Star in Antibiofilm Therapeutics

Sonobactericide, an antibiotic-free alternative for antibacterial treatment, has proven extraordinary efficiency in preventing and managing biofilm-related bacterial infections. However, the pathophysiological processes underlying biofilm infections are complex. Considering the dysregulated immune-microenvironment associated with these infections, a dual-track strategy that combines ultrasound with immunoregulation presents a more compelling approach. Nanomaterials provide a golden bridge for ultrasound-assisted immunoregulation. The mechanical forces and reactive oxygen species induced by ultrasound-activated nanomaterials can not only inactivate bacteria enclosed in biofilm, but also disrupt dense biofilm architectures, endowing immunoregulators with superior biofilm penetration. Throughout the process of biofilm removal, immunoregulation plays a continuous role by primarily modulating the inflammatory response of macrophages and neutrophils. The activated immune cells are highly effective in eliminating residual bacteria and repairing damaged tissues. Such complementary action mechanism confers significant superiority over traditional antibiofilm therapeutics, particularly in the case of multidrug-resistant biofilm infections that do not respond to antibiotics. Article type: Perspective Ultrasound-Assisted Immunoregulation: A Rising Star in Antibiofilm Therapeutics Xuan Wu 1 | Can Wu 1 | Tong Ye 1 | Xiaotong Sun 1 | Lingfeng Xu 1 | Mengfei Pang 1 | Lulu Wang 2 | Xin Pang 1,* 1 School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China 2 Department of Pharmacy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China * Corresponding author. E-mail: [email protected] (X. P.)

Sonobactericide, an antibiotic-free alternative for antibacterial treatment, has proven extraordinary efficiency in preventing and managing biofilm-related bacterial infections. However, the pathophysiological processes underlying biofilm infections are complex. Considering the dysregulated immune-microenvironment associated with these infections, a dual-track strategy that combines ultrasound with immunoregulation presents a more compelling approach. Nanomaterials provide a golden bridge for ultrasound-assisted immunoregulation. The mechanical forces and reactive oxygen species induced by ultrasound-activated nanomaterials can not only inactivate bacteria enclosed in biofilm, but also disrupt dense biofilm architectures, endowing immunoregulators with superior biofilm penetration. Throughout the process of biofilm removal, immunoregulation plays a continuous role by primarily modulating the inflammatory response of macrophages and neutrophils. The activated immune cells are highly effective in eliminating residual bacteria and repairing damaged tissues. Such complementary action mechanism confers significant superiority over traditional antibiofilm therapeutics, particularly in the case of multidrug-resistant biofilm infections that do not respond to antibiotics.

u ltrasound; immunotherapy; bacterial infections; biofilm 1 | Introduction Biofilms, defined as surface-associated microbial communities, are ubiquitous across diverse ecosystems. Their formation initiates with bacterial adhesion to surfaces, either inert or biological, followed by encasement within a self-produced extracellular polymeric substance (EPS) matrix composed of lipids, proteins, nucleic acids, and polysaccharides. Clinically, biofilms are implicated in a wide spectrum of infections, ranging from exogenous device-associated infections (e.g., catheters, orthopedic implants, ventilator tubes) to chronic tissue-based infections such as periodontitis, osteomyelitis, endocarditis, and cystic fibrosis pneumonia [1]. The dense, complex architecture of biofilms, coupled with inherent spatial and functional heterogeneity within the microbial community, provides formidable protection against host immune clearance and impedes antibiotic penetration. Additionally, biofilm matrices can actively degrade antimicrobial agents through enzymatic activity or adsorption. Consequently, bacteria embedded within biofilms exhibit antibiotic tolerance levels up to 1000-fold higher than their planktonic counterparts [2]. This intrinsic resilience significantly complicates the clinical management of biofilm-associated infections, elevating risks of treatment failure and recurrent disease. As a category of recalcitrant infections, biofilm-associated diseases impose a substantial epidemiological and financial burden on global healthcare systems. If without effective interventions, it is projected that annual deaths attributable to antibiotic resistance could reach 10 million by 2050, surpassing cancer mortality rates [3]. Faced with a critically depleted antibiotic pipeline, the development of novel therapeutic strategies is urgently needed to combat biofilm-related infections, particularly to address existing resistance and prevent the emergence of new mechanisms [4]. Recent advancements in interfacial engineering have facilitated the development of materials exhibiting unique photothermal, photodynamic, sonothermal, sonodynamic, wavethermal, and wavedynamic properties, which are activated by exogenous energy sources such as light, ultrasound, and microwaves [5-9]. These materials hold significant promise for treating bacterial infections and cancers, primarily by generating cytotoxic reactive oxygen species (ROS) or localized hyperthermia. Among these stimulations, ultrasound wave stands out for its superior tissue penetration depth, non-invasiveness, and minimal collateral damage, making it particularly suitable for targeting deep-seated infections [10-12]. Specifically, such ultrasound-activated bactericidal approach, termed sonobactericide, has emerged as a promising alternative to conventional antibiotics for combating biofilm-associated infections. By utilizing ultrasound pressure waves to activate sonosensitizers, sonobactericide generates heat, mechanical forces, or ROS to disrupt bacterial biofilms [13]. A key benefit of this approach is its ability to precisely deliver ultrasound energy to deep-seated biofilm foci while sparing surrounding healthy tissues. Crucially, unlike traditional antibiotics that target specific bacterial pathways, sonobactericide employs a multifaceted mechanism of action. It can simultaneously prevent bacterial adhesion, degrade the biofilm extracellular polymeric substance (EPS), and disrupt intracellular pathogenic processes. This non-specific targeting, coupled with a low mutagenesis potential, renders sonobactericide broadly effective against diverse bacterial biofilms without inducing significant resistance. Despite its considerable efficacy in preventing and managing biofilm-associated infections, sonobactericide monotherapy often falls short of achieving complete biofilm eradication, because this treatment primarily focuses on eliminating bacteria, whereas biofilm infections involve inherently complex pathophysiological processes that extend beyond mere bacterial presence. The host immune system plays a vital role in bacterial clearance and rarely develops resistance. As part of the innate defense response, immune cells secrete bactericidal agents, including ROS and nitric oxide. Furthermore, phagocytic immune cells effectively ingest and induce intracellular killing of invading bacteria [14]. However, bacterial biofilms establish a formidable defense by creating both chemical and physical barriers at the infection site, concurrently fostering an immunosuppressive microenvironment. This protective architecture significantly impedes direct contact between immune cells and the entrapped bacteria, leading to “frustrated phagocytosis” [15]. Compounding this challenge, biofilms actively subvert host immunity through the release of virulence factors. These factors suppress phagocyte activation and diminish inflammatory cytokine secretion, thereby severely hampering the recruitment of innate immune cells essential for pathogen eradication. Moreover, pathogenic bacteria within biofilms can strategically conceal antigenic epitopes to evade immune surveillance and may even eliminate antigen-presenting cells, facilitating persistent immune evasion. This cascade of events ultimately shifts the adaptive immune response from homeostasis towards a state of suppression, rendering conventional antibiofilm approaches highly challenging. Given the profound dysregulation of the immune microenvironment by biofilm infections, an integrated strategy combining immunomodulation with sonobactericide presents a compelling therapeutic paradigm. This coordinated action leverages ultrasound to eliminate bacteria shielded within biofilm barriers while simultaneously restoring the local immune microenvironment, enabling effective pathogen clearance and tissue repair (Figure 1). Consequently, this dual-pronged approach achieves potent and safe disinfection without inducing cross-resistance. FIGURE 1 | Schematic illustration of ultrasound-assisted immunoregulation (UAI) for combating bacterial biofilm infections. Ultrasound-activated nanomaterials generate mechanical forces and reactive oxygen species (ROS), which synergistically disrupt dense biofilm architectures and kill enclosed bacteria. This disruption enhances the penetration and efficacy of co-delivered immunomodulators. Subsequently, the reactivated immune cells efficiently clear residual pathogens and facilitate the repair of infection-associated damaged tissue. 2 | Nanotechnology in UAI Current immunomodulators face significant limitations, including poor bioavailability, inadequate target specificity, and restricted penetration through the dense biofilm matrix. Consequently, a facile combination of ultrasound with antibiofilm immunomodulation often proves insufficient for effectively overcoming these barriers and achieving complete infection eradication. The rapid development of nanotechnology offers a promising solution to bridge this gap. Diverse nanocarriers, such as liposomes, metal-organic frameworks (MOFs), and cell membrane-derived nanoparticles, have been engineered to co-deliver sonosensitizers and immunoregulatory agents (Table 1). Following functionalization with targeting ligands, these nanosystems can selectively accumulate at infected sites, improving the biodistribution and therapeutic efficacy of their payload. Furthermore, nanotechnology enables the flexible design of ultrasound-responsive platforms. Exogenous ultrasound triggers the controlled release of encapsulated sonosensitizers and immunomodulators, minimizes premature drug leakage, maximizes local drug concentrations within the biofilm microenvironment, and enhances immunotherapy response rates. Notably, some nanomaterials, like piezoelectric nanosheets, exhibit intrinsic sonosensitivity [16]. By leveraging nanoscale effects, these systems can significantly amplify sonobactericidal efficacy, synergizing with immunotherapy even without exogenous sensitizers. Nanomaterials possessing inherent sonosensitivity or loaded with small-molecule sonosensitizers are collectively termed nano-sonosensitizers [13]. Compared to conventional sonosensitizers, nano-sonosensitizers offer superior targeting precision to infection foci [17], enhanced water solubility, and amplified cavitation effects, thereby boosting sonobactericide outcomes [18]. TABLE 1 | Representative types of nanomaterials as carriers for sonosensitizers and immune agents. | Liposome | Indocyanine green | Anti-H. pylori antibody | H. pylori Infections | [19] | | Iron-based covalent organic framework | Curcumin | The released double-stranded DNA | Implant-related infections | [20] | | Macrophage membrane-decorated organosilica framework | Tetra (4-carboxyphenyl) porphyrin | Cytosine-phosphate-guanine oligodeoxynucleotide or microRNA-21-5p | Holistic Biofilm-Related Infections | [21] | | Neutrophil/macrophage membranes decorated MnO2-hydrangea nanoparticle | Porphyrinic | Mn2+ | Orthopaedic biofilm infections | [22] | | Cell membrane nanovesicles | Meso-tetrakis (4-sulfonatophenyl) porphyrin | MEDI4893 | MRSA myositis | [23] | However, the biocompatibility and safety of nanomaterials warrant careful consideration. Strategies such as polyethylene glycol (PEG) surface modification or the use of biodegradable materials can enhance biocompatibility [24]. Rigorous evaluation of cytotoxicity, histocompatibility, biological effects, and biodegradability is essential for clinical translation [25]. Beyond augmenting ultrasound bioeffects and facilitating ultrasound-assisted immunomodulation, nanotechnology holds potential to remodel the biofilm-associated immunosuppressive microenvironment and restore immune surveillance. Through sophisticated engineering, certain nanoplatforms can even enable sequential and adaptive immune modulation tailored to different stages of biofilm infection. In brief, the rapid progress in nanotechnology unlocked novel avenues and significantly strengthened the potential of ultrasound-assisted immunomodulation for combating biofilm-related bacterial infections (Figure 2). FIGURE 2 | Schematic illustration of nanotechnology to augment UAI. 2.1 | Ultrasound-Induced Mechanical Forces Regulate Macrophage Macrophages are central to research in ultrasound-assisted immunomodulation. As pivotal innate immune cells, they are broadly classified into pro-inflammatory (M1) and anti-inflammatory (M2) phenotypes [26, 27]. M1 macrophages exhibit potent defensive capabilities, including high phagocytic activity for bacterial clearance and debris removal, as well as the induction of inflammatory responses [28]. Conversely, M2 macrophages are essential for inflammation resolution, angiogenesis, and wound healing [29, 30]. During early biofilm infection, M2 macrophage predominance impedes eradication efforts, potentially leading to persistent/recurrent infections and tissue necrosis. Consequently, strategies leveraging nanomaterials to reprogram macrophage polarization and modulate innate immune interactions are crucial for antibiofilm design. However, the dense biofilm matrix severely restricts the penetration of externally administered nanomaterials. Ultrasound waves, harnessing significant mechanical energy, can generate physical forces capable of disrupting biofilm architectures, thereby enhancing the penetration of therapeutic agents. This mechanical disruption often relies on microbubbles (MBs) as key sonosensitizers. These microscale spheres feature a gas core stabilized by a polymer, protein, or phospholipid shell. Upon ultrasound exposure, the gas-filled nuclei undergo oscillatory expansion and contraction, while phase-change droplets can generate localized microstreams through inertial cavitation. The resulting shear stresses disrupt biofilm integrity, creating pores and channels that facilitate the entry of antimicrobial agents and immunomodulators [31-33]. Notably, the magnitude and effect of these ultrasound-induced mechanical forces are governed by parameters such as frequency, amplitude, and duration, necessitating optimization based on experimental objectives. For instance, a biomimetic nano-sonosensitizer (EMB-Hu) was engineered by coating erythrocyte membranes onto MBs loaded with Fe 3 O 4 nanoparticles and hydroxyurea [15]. Exploiting the unique acoustic properties of MBs, EMB-Hu exhibited stable oscillation under ultrasound, disrupting biofilms via an ”exocytosis”-like mechanism. This enhanced the release and penetration of Fe 3 O 4 nanoparticles and hydroxyurea, increasing their diffusion within the biofilm and boosting antibacterial efficacy. Importantly, the released Fe 3 O 4 nanoparticles served as signaling molecules, polarizing macrophages towards the pro-inflammatory M1 phenotype. This significantly promoted macrophage infiltration into infected tissues and enhanced phagocytic clearance of disrupted biofilms. Similarly, Xiu et al. developed ultrasound-responsive catalytic MBs with a piperacillin and Fe 3 O 4 nanoparticles-loaded shell surrounding an air core for treating biofilm-induced pulmonary infections [31]. Ultrasound stimulation triggered biofilm disruption via MBs and enhanced penetration of piperacillin and Fe 3 O 4 nanoparticles. Subsequently, the Fe 3 O 4 nanoparticles, aided by piperacillin, degraded EPS and eradicated bacteria. Furthermore, the Fe 3 O 4 nanoparticles polarized macrophages towards the M1 phenotype, contributing to biofilm elimination. Thus, by integrating physical biofilm disruption with macrophage immunomodulation, ultrasound-responsive MBs demonstrate potent efficacy against bacterial biofilm infections. In summary, ultrasound-induced mechanical forces hold great potential for immunotherapy, particularly by facilitating immunomodulator release and overcoming biofilm delivery barriers (Figure 3). When coupled with MBs, the mechanical effects are amplified, effectively disrupting the biofilm matrix, increasing porosity, enabling deep penetration of therapeutics, and ultimately reshaping the immunosuppressive microenvironment. FIGURE 3 | Schematic illustration of ultrasound-induced mechanical forces to regualte macrophage. 2.2 | Ultrasound-Induced ROS Regulate Macrophage Ultrasound, functioning as a remote trigger, can not only induce mechanical forces on MBs but alos activate sonosensitizers, typically organic small molecule compounds, to initiate electron transfer processes. These electrons subsequently catalyze surrounding oxygen-containing substrates (O 2 and H 2 O) to produce ROS via sonoluminescence or pyrolysis. Some inorganic piezoelectric nanosheets also contribute to ROS generation via electron polarization under ultrasound stimulation. I Critically, ROS serve dual functions: exerting direct antibacterial effects and acting as signaling molecules to stimulate the secretion of inflammatory cytokines [34]. Within the biofilm microenvironment, elevated ROS levels significantly enhance M1 macrophage polarization [35]. This occurs through the induction of oxidative stress, damaging vital bacterial biomolecules such as proteins, nucleic acids, and lipid membranes. Consequently, a synergistic therapeutic strategy has emerged, utilizing nano-sonosensitizers as dual-functional agents for both ROS generation and immunomodulation, thereby combining direct bacterial inhibition with macrophage immunomodulation [36]. Under ultrasound assistance, these nano-sonosensitizers efficiently generate ROS to kill bacteria and downregulate biofilm-associated genes. Simultaneously, they stimulate macrophage polarization towards the pro-inflammatory M1 phenotype, characterized by potent phagocytosis and bactericidal activity, primarily through activation of the PI3K-AKT and MAPK signaling pathways. Furthermore, the transient hypoxia resulting from rapid oxygen consumption during ROS production upregulates hypoxia-inducible factor 1α (HIF-1α) expression. This disrupts macrophage metabolic homeostasis, promoting glycolysis and enhancing M1 macrophage infiltration, which collectively amplifies the inflammatory immune response against biofilms [37]. Apart from their essential roles in pathogen recognition, phagocytosis, and digestion, macrophages can paradoxically reservoirs for bacterial persistence, shielding pathogens from external antimicrobial agents. B Intracellular bacteria may evade clearance for extended periods, and infected macrophages can act as “Trojan horses”, disseminating bacteria to distant sites and contributing to metastatic infections. Targeting this vulnerability, copper overload-induced dysregulation of macrophage copper metabolism has proven effective for eliminating intracellular bacteria. By integrating copper and porphyrin sonosensitizer into a metal-organic framework (CTMM), the ROS generated by ultrasound activation successfully disrupt biofilm structure, interfere with bacterial growth and metabolism, and promote macrophage chemotaxis and M1 polarization for enhanced biofilm eradication [38]. Concurrently, the ultrasound-induced ROS enable CTMM to induce cuproptosis-like and ferroptosis-like stress within macrophages, leading to the complete elimination of intracellular bacteria. This integrated approach, combining ultrasound-triggered biofilm destruction with targeted macrophage immunomodulation, represents a promising “Trojan War” strategy against implant-associated infections. Collectively, nano-sonosensitizers harness ultrasound energy to generate ROS, facilitating macrophage immunomodulation that enhances phagocytic activity and pro-inflammatory responses against biofilm infections. Three Key mechanisms underpinning the promotion of M1 macrophage polarization include the activation of PI3K-AKT and MAPK signaling pathways, the induction of hypoxia-mediated metabolic reprogramming towards glycolysis, and triggering of cupferroptosis-like stress within macrophages (Figure 4). FIGURE 4 Schematic illustration of ultrasound-induced ROS to regulate macrophage. 2.3 | Ultrasound Assists Multi-Immunoregulation Biofilm formation is a complex, multi-stage process involving attachment, maturation, and detachment. However, current ultrasound-assisted immunomodulation approaches often target specific infection stages, lacking adaptability to the distinct immunological requirements at each phase. During the initial stage (Stage I) of biofilm infection, neutrophils play a critical role in recognizing and ingesting bacteria, producing ROS and antimicrobial peptides, and constructing neutrophil extracellular traps (NETs) to capture pathogens. Notably, NETs can also trigger M1 macrophage polarization, influencing the progression to Stage II. In Stage II, M1 macrophages become the primary effectors, specializing in phagocytosis and bacterial clearance. Enhancing their antibacterial activity is thus a key therapeutic goal at this stage. However, excessive or uncontrolled M1 polarization coupled with persistent NET release can amplify neutrophil infiltration, perpetuate NET formation, and exacerbate tissue damage [39]. Subsequently, in Stage III, following biofilm elimination, M2 macrophages contribute significantly to tissue repair by secreting anti-inflammatory cytokines and promoting regeneration. Given the complexity of cytokine storms and dysregulated innate immunity across biofilm infection stages, a comprehensive ultrasound-assisted multi-immunomodulation strategy holds significant appeal. Such an approach aims to target all infection phases and normalize the function of key immune cells (e.g., neutrophils and macrophages). Accordingly, macrophage cell membrane-coated sonosensitive nanoadjuvants have been engineered for sequential delivery of immunomodulators like cytosine-phosphate-guanine oligodeoxynucleotide (CpG-ODN) or microRNA-21-5p (miR-21-5p) [23]. When exposed to ultrasound irradiation, the embedded sonosensitizer generates singlet oxygen for bacterial eradication. Specifically, in Stage I, CpG-ODN-loaded nanoadjuvants promote NET formation to capture unattached pathogens. During Stage II, these nanoadjuvants drive the repolarization of M2 macrophages towards the M1 phenotype, countering biofilm-induced immune evasion and enabling clearance of residual bacteria. Finally, in the healing phase (Stage III), administration of miR-21-5p-loaded nanoadjuvants suppresses excessive inflammation and enhances tissue repair by boosting M2 macrophage polarization. This sequential strategy, integrating infection elimination, tissue restoration, and stage-specific immune modulation, demonstrates considerable clinical promise compared to conventional antibiofilm therapies. While effective, this chronological approach necessitates administering multiple immunomodulators, introducing complexity in formulation design and treatment regimens. In contrast, nano-sonosensitizers capable of self-adaptive targeting and immunomodulation in response to the distinct microenvironments associated with different infection stages offer a more streamlined solution. Biofilm infections create unique microenvironments characterized by hypoxia, low pH, and elevated hydrogen peroxide levels, which normalize upon biofilm clearance. Leveraging these microenvironmental shifts, self-adaptive nano-sonosensitizers can be designed with responsive activities. For example, a hollow Cu 2 MoS 4 nano-sonosensitizer nano-sonosensitizer exhibits pH-dependent enzyme-mimicking properties [40]. In the acidic microenvironment typical of active infection, it shows enhanced peroxidase-like activity, significantly boosting ultrasound-triggered ROS generation to kill bacteria and promote M1 macrophage polarization. Following biofilm removal and the return to a neutral pH, the same nano-sonosensitizer spontaneously exhibits catalase-like activity, scavenging ROS to promote anti-inflammatory M2 macrophage polarization and facilitate tissue repair. This intrinsic self-adaptive immunomodulation capability, combined with potent antibiofilm effects, provides a sophisticated and efficient method for managing complex infections. In summary, the integration of ultrasound with advanced nano-sonosensitizers enables multi-immunomodulation across biofilm infection stages primarily through two complementary strategies. The first involves the concurrent use of nano-sonosensitizers and nano-immunomodulators to deliver precise, stage-specific immunomodulation chronologically, effectively targeting diverse immune cells like neutrophils and macrophages throughout the infection process, as illustrated in Figure 5a. The second strategy employs a single, intelligent nano-sonosensitizer designed to autonomously reprogram macrophage-mediated inflammatory responses based on the prevailing microenvironmental cues, adapting its immunomodulatory function as the infection progresses and resolves, as shown in Figure 5b. FIGURE 5 | Schematic illustration of multi-immunoregulation assisted by ultrasound. 3 | Conclusion and Outlook Ultrasound-assisted immunomodulation represents a promising therapeutic strategy against biofilm-associated bacterial infections. Ultrasound-activated nanomaterials generate mechanical forces and reactive oxygen species (ROS) that synergistically disrupt dense biofilm structures and inactivate embedded bacteria, while concurrently enhancing the penetration of co-delivered immunomodulators. Throughout this process, targeted immunomodulation continuously regulates inflammatory responses in key immune cells—notably macrophages and neutrophils. The reactivated immune system effectively clears residual pathogens and promotes repair of infection-damaged tissues. This dual-action synergy offers superior efficacy against persistent biofilm infections, particularly those caused by multidrug-resistant strains refractory to conventional antibiotics. Owing to its exceptional therapeutic outcomes and deep-tissue penetration capabilities, ultrasound-assisted immunomodulation holds significant potential for clinical translation in precision medicine. Further optimization of ultrasound parameters, rational selection of sonosensitizers, and refinement of treatment protocols may unlock new avenues for managing challenging conditions such as implant-associated infections, osteomyelitis, and orthopedic biofilms. The next generation of combinatorial therapies integrating diverse sonobactericidal modalities (e.g., sonothermal therapy) is poised to broaden the clinical scope of ultrasound-assisted immunomodulation. Future efforts should expand beyond overcoming frustrated macrophage activation to encompass regulation of adaptive immune cells—including T cells, B cells, and dendritic cells—thereby establishing comprehensive immunomodulatory frameworks. Advanced sonosensitive platforms featuring multifunctional, stimuli-responsive designs will further potentiate targeted antibiofilm efficacy. These innovations could ultimately redefine standards for managing complex biofilm infections through spatially and temporally controlled immune reprogramming. Acknowledgments This work was supported by the National Natural Science Foundation of China (82274114 and 82001955), Outstanding Youth Foundation Youth Fund of Henan Natural Science Foundation (242300421091), the Joint Fund of Science and Technology Development Program of Henan Province (232301420087), and Young Elite Scientists Sponsorship Program by CACM (2022-QNRC2-A04). Author Contributions X.W. and C.W. conceived the idea. X.T.S., T.Y., L.F.X., and M.F.P. contributed to the manuscript writing. L.L.W. and X.P revised the manuscript. Competing Interests The authors declare no competing interests.

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Authors Metrics & Citations Metrics Article Usage 258views 205downloads Citations Download citation XUAN WU, Can Wu, Tong Ye, et al. Ultrasound-Assisted Immunoregulation: A Rising Star in Antibiofilm Therapeutics. Authorea. 01 July 2025. DOI: https://doi.org/10.22541/au.175139666.69190967/v1 DOI: https://doi.org/10.22541/au.175139666.69190967/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu.