J. Microbiol. Infect. Dis., (2026), Vol. 16(2): 44–71

Review Article

10.5455/JMID.2026.v16.i2.1

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Nanoparticles as a novel approach to control oral multispecies biofilms: An update on progress

Sumaya Abusrewil^1,2, Rabia Daaeki¹, Om Alkhir Alshanta^2,3, James Alun Scott², William McLean^2,4, Gordon Ramage⁵ and Mohamed ElEmam^6*

¹Department of Operative Dentistry and Endodontics, School of Dentistry, University of Tripoli, Tripoli, Libya

²Glasgow Endodontology Group, Glasgow Dental School, School of Medicine, Dentistry and Nursing, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom

³National Health System, Ministry of Health, Tripoli, Libya

⁴Oral Sciences Research Group, Glasgow Dental School, School of Medicine, Dentistry and Nursing, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK

⁵Safeguarding Health Through Infection Prevention (SHIP) Research Group, Research Centre for Health, School of Health and Life Sciences, Glasgow Caledonian University, Glasgow, UK

⁶Department of Periodontology, School of Dentistry, University of Tripoli, Tripoli, Libya

*Corresponding Author: Mohamed ElEmam. Department of Periodontology, School of Dentistry, University of Tripoli, Tripoli, Libya. Email: m.elemam [at] uot.edu.ly

Submitted: 05/01/2026 Revised: 15/02/2026 Accepted: 23/02/2026 Published: 01/04/2026

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ABSTRACT

The human mouth is a complex environment that is heavily colonized by diverse communities of microorganisms, which establish dynamic, biofilm communities on various surfaces within the human oral cavity. Biofilm-related oral diseases are among the most common diseases linked to serious systemic diseases. The effective management of oral diseases remains a global challenge that demands the discovery of innovative therapeutic approaches for eradicating biofilm-associated infections. Nanoparticles (NPs) have emerged as a novel approach for targeting oral biofilms, and this paper aims to provide a comprehensive overview of recent in vitro, in vivo, ex vivo, and clinical studies, within the last four years (2022–2025). It intends to investigate the potential applications of different types of NPs in disrupting multispecies biofilms related to a range of oral diseases, including periodontitis, dental caries, endodontic diseases, peri-implantitis, and oral candidiasis. In general, nanoparticles, alone or conjugated with various substances, demonstrated effective antimicrobial properties against oral multispecies biofilms, making them a promising strategy for the management of biofilm-related oral diseases. Novel strategies are being highlighted with the use of NPs, such as the application of phototherapy and magnetic fields, aiming for the complete eradication of biofilms. While most of these approaches have not been implemented clinically, they hold promise for enhancing the management of oral diseases. Further research and more clinical trials are required to confirm their clinical efficacy and long-term safety.

Keywords: Antimicrobial, Multispecies biofilm, Nanoparticles, Oral biofilm, Oral diseases.

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Introduction

Oral health is essential to overall health, enabling communication, nutrition, and overall quality of life (WHO, 2022). However, oral diseases represent the most prevalent noncommunicable diseases, impacting nearly half of the global population (45% or 3.5 billion individuals) throughout their lives, from early life to old age (WHO, 2022). The oral cavity harbors one of the most diverse microbial communities within the human body (Project, 2012). The composition of these microbial populations can fluctuate with sugar-rich diets (Angarita-Díaz et al., 2022), oral hygiene, antimicrobials, and disease (Brown et al., 2025). The microorganisms found in the human oral cavity have been referred to as the oral microflora, oral microbiota, or oral microbiome (Dewhirst et al., 2010). The term microbiome was coined in 2001 by Joshua Lederberg as “the ecological community of commensal, symbiotic and pathogenic microorganisms that literally share our body space and have been all but ignored as determinants of health and disease” (Lederberg and McCray, 2001). Indeed, the human microbiome plays very important roles in the maintenance of health, yet when disrupted, these inherent microbes drive diseases (Jorth et al., 2014; Marsh and Zaura, 2017). There is mounting evidence demonstrating a clear correlation between dysbiosis of the oral microbiome and diseases such as cardiovascular diseases, diabetes, neurodegenerative diseases, and chronic inflammatory diseases (Pisano et al., 2023).

The human mouth is a complex environment, heavily colonized by diverse communities of microorganisms, including viruses, protozoa, archaea, bacteria, and fungi (Wade, 2013) (Fig. 1). In a recent update, it has been established by Human Oral Microbiome Database (HOMD) that there are 834 bacterial taxa in the human mouth and aerodigestive tract, and only 49% of the oral taxa are officially named (homd.org). Additionally, 101 fungal species have been reported to colonize the oral cavity of healthy individuals (Ghannoum et al., 2010). Looking at the ecology of our own body, the complexity of the human oral microbiome represents a challenge for microbiologists, with diverse niches where highly complex microbial communities of hundreds of microbial species communicate and interact. This, of course, raises the inevitable question: “What are all these species doing there?” (Janus et al., 2016). This question does not have a simple answer, and it may be appropriate to begin by explaining the concept of biofilms within an individual’s oral microbiome. In fact, the microbial communities colonizing the oral cavity, often in the form of spatially organized complex structures, are known as biofilms (Pisano et al., 2023). The biofilms have been described as highly organized microbial communities encased in a protective extracellular polymeric substances that adhere to a biotic or abiotic substrate (Hall-Stoodley et al., 2004; Pinto et al., 2020). These communities exist within enclosed biofilms, constituting both synergistic and antagonistic interactions (Diaz and Valm, 2020; Hwang, 2022).

Biofilms are inherently tolerant to both antimicrobials and host immune defenses, making them the underlying cause of many persistent microbial infections (Hall-Stoodley et al., 2004). Although biofilm control measures are implemented to protect health and prevent oral diseases, they may lead to unintended consequences, such as antimicrobial resistance. For example, amoxicillin and metronidazole are commonly prescribed antibiotics in oral care. Although they effectively treat severe bacterial infections, their use can promote antibiotic resistance, which is often associated with an overgrowth of Candida albicans (C. albicans), manifesting as oral thrush (Brown et al., 2025). When resistant microbes persist, they can alter oral and systemic environments through ecological changes and genetic adaptation, which may drive collateral damage to human health (Brown et al., 2025).

Beyond conventional antibiotic and antimicrobial therapies, the potential application of nanoparticles in controlling oral biofilm diseases has substantially progressed and attracted increasing attention. These tiny particles are thought to possess great potential to improve diagnosis, treatment and prevention of numerous oral diseases because of their unique physicochemical and biological characteristics (Bapat et al., 2019; Moraes et al., 2021), including nanoscale size, large surface area, surface reactivity and their electronic and optical properties, making them suitable candidates for many uses (Khan et al., 2019).

Until recently, the field of nanotechnology has been swamped with studies investigating mono-species biofilm models, which are very limited in scope and complexity. However, there has been a shift toward the construction of multispecies consortia in scientific publications. Multispecies biofilms have been observed to display higher levels of tolerance to antimicrobial therapies than the corresponding mono-species biofilms (Dong et al., 2023). Understanding interkingdom/interspecies interactions and the polymicrobial nature of oral biofilms provides important insights for developing novel therapeutic approaches. Therefore, considering multispecies biofilm models is important when testing the antimicrobial efficacy of nanoparticles. This paper aims to provide a recent update on nanoparticle-based therapies for disrupting and eradicating oral multispecies biofilms in clinical trials, in vitro, in vivo, and ex vivo studies. To the best of our knowledge, this review is the first to provide an update on the antibiofilm efficacy of nanoparticles against oral multispecies biofilm models associated with oral infections.

Biofilm-Related Oral Diseases

The oral biofilm is the origin of various oral diseases, including dental caries, endodontic infections, periodontal diseases (Maddi and Scannapieco, 2013), peri-implantitis (Lafaurie et al., 2017), and oral candidiasis (candidosis) (Rautemaa and Ramage, 2011) (Fig. 1). Dental caries or tooth decay, as it is commonly referred to, is caused by dental plaque, which is a community of microorganisms in the form of biofilms (Mosaddad et al., 2019). Pulpal and periapical tissue infections, driven by biofilms (Ricucci and Siqueira, 2010; Ricucci et al., 2017), have long been seen as an extension of the caries process due to the dominance of dental caries as a gateway for dentin infection (Ørstavik, 2019). However, it has been said that tissue infections and responses are likely an older and more general biological manifestation than tooth decay. Apical periodontitis may be perceived as a tissue response to pulp infection, as a way of “taming” and “coping” with virulence expressions from infectious microorganisms (Ørstavik, 2019), or it may be viewed as a protective mechanism against the spread of infection, where the process of self-induced destruction of apical periodontitis provides a space for the infiltration of specialized immune cells and creates a barrier thereby preventing the spread of infection (Nair, 2004).

Periodontitis is a chronic multifactorial inflammatory disease associated with dysbiotic plaque biofilms and characterized by progressive destruction of tooth-supporting tissues, including loss of clinical attachment and alveolar bone (Papapanou et al., 2018). The global burden of severe periodontitis has been significant and has increased over the past thirty years. In 2019, approximately 1.1 billion cases of severe periodontitis were reported worldwide (Chen et al., 2021). Periodontitis is thought to be an inflammatory disease initiated by commensal bacteria (Van Dyke, 2017). It was hypothesized that P. gingivalis may function as a keystone pathogen in periodontal disease by employing sophisticated mechanisms to disturb the normally homeostatic host-microbiome, thereby causing dysbiosis of the periodontal microbiome leading to inflammation (Hajishengallis et al., 2011; Hajishengallis et al., 2012). The term “keystone” has been used in the ecological literature to characterize species that were thought to have a disproportionately significant impact on the structure of their community (Paine, 1969), which has been expanded in the human microbiome. However, the suggestion that specific pathogens promote dysbiosis and disease is questioned due to the absence of a definitive link between any assumed keystone pathogen and the initiation of disease in humans (Van Dyke et al., 2020). Indeed, biofilm dysbiosis has long been considered the starting point of periodontal disease, followed by inflammation and ultimately bone loss (Yost et al., 2015; Lee et al., 2016). Recently, it has been proposed that gingival inflammation may actually precede and subsequently induce biofilm dysbiosis, thereby triggering further inflammation (Van Dyke et al., 2020). Although the management of periodontitis primarily consists of non-surgical periodontal debridement (Kwon et al., 2021), it is not fully effective and fails in high-risk individuals. This is likely due to the complex interplay between host response, genetics, and environmental factors (Van Dyke et al., 2020). Peri-implantitis is a pathological condition affecting the tissues surrounding dental implants, characterized by inflammation and bone loss. Patients with a history of chronic periodontitis, poor plaque control, and lack of regular maintenance following implant therapy are at a higher risk for peri-implantitis (Schwarz et al., 2018).

Fig. 1. Oral microbiome in health and disease. The oral microbiome is a complex ecosystem, consisting of bacteria, fungi, viruses, archaea, and protozoa that live in a dynamic equilibrium. Disease occurs when this balance is disrupted (dysbiosis), often due to poor oral hygiene, high sugar intake, or systemic changes.

Oral candidiasis is the most common opportunistic fungal infection of the oral cavity (Akpan and Morgan, 2002), predominantly caused by the overgrowth of C. albicans. Other non-albicans species also contribute to oral candidiasis, although to a lesser extent (Rajendra Santosh et al., 2021). Chronic atrophic candidiasis “denture stomatitis” is a type of candidiasis that occurs in denture wearers, manifested by localized chronic erythema of tissues in contact with the denture base (Akpan and Morgan, 2002). In partially dentate patients, it has been noted that poor denture hygiene and traumatic denture fit can lead to complications beyond denture stomatitis, such as dental caries and periodontal diseases (McReynolds et al., 2023). In fact, a complex polymicrobial biofilm containing fungi and bacteria can grow on the surface of denture materials and form denture plaque, which induces a localized inflammatory response (Hannah et al., 2017; Brown et al., 2022). Fungi are eukaryotes that possess distinct biological traits and occupy a greater biovolume than bacteria. Therefore, we cannot apply our essential perspectives on bacterial biofilms when seeking a comprehensive understanding of fungal biofilms (Ramage et al., 2025).

To this end, when evaluating antibiofilm properties, it is important to consider a comprehensive perspective on oral infections. This is relevant in clinical contexts, as the interactions between yeast and bacteria may have the potential to complicate the infections (Du et al., 2021). One of the most thoroughly researched mechanisms of Candida-bacteria interactions is physical attachment. A strong interkingdom synergy was shown to exist through the physical scaffold of hyphae, which offers a potential niche for bacteria to colonize and form biofilms on the existing Candida biofilm. This phenomenon is referred to as “mycofilms” (Kean et al., 2017; Kean et al., 2018). It has been demonstrated that C. albicans conferred protection upon a number of bacterial species against different antimicrobials (Young et al., 2020; Abusrewil et al., 2021). C. albicans was found to enhance the tolerance of Pseudomonas aeruginosa (P. aeruginosa) biofilm to an antibiotic (meropenem) (Alam et al., 2020). In contrast, Streptococcus mutans (S. mutans) was shown to enhance C. albicans biofilm tolerance to fluconazole via secreted bacterial extracellular polysaccharides, which provided protection against the antifungal drug (Kim et al., 2018).

In fact, the escalating issue of antibiotic resistance poses a significant threat, making infectious diseases harder, or even impossible to treat with current antibiotics (English and Gaur, 2010). However, physical removal, including toothbrushing and non-surgical therapies, remains one of the most significant methods for plaque control in contemporary practice. These practices aim to maintain a predominance of immature plaque species while preventing overgrowth of pathogenic species (Brown et al., 2025). Oral mouthwash is widely considered to be an adjunct to mechanical oral hygiene procedures, although increasing evidence suggests that over-the-counter mouthwashes, such as chlorhexidine (CHX), appear to promote bacterial communities linked to caries and the emergence of antimicrobial resistance genes (Bartsch et al., 2024). In fact, bacteria and fungi have adopted numerous mechanisms of resistance that enable them to evade antimicrobials (McKeegan et al., 2002). As a result, the control of the oral biofilms that are resistant to the available antibiotics and antimicrobials demands the discovery of innovative therapeutic approaches for eradicating biofilm-associated infections.

Nanotechnology

Advancements in nanotechnology have greatly impacted the development of nanomodified biomaterials. According to the Oxford English Dictionary, the prefix “nano” is derived from a Greek word meaning “dwarf” or something extremely small. A nanometer (nm) is a unit of length in the International System of Units, which is equal to one billionth of a meter (10−9 m) (oed.com). Nanotechnology refers to the application of nanoscience to practical uses by controlling the shape and size of various devices and systems at the nanometer scale (Szczyglewska et al., 2023). Although nanoscience appears to be a relatively recent field of research, metallic nanoparticles have been synthesized for centuries. Their use began during the dawn of glass-making in ancient Egypt and Mesopotamia as early as the 14^th and 13^th centuries BCE (Schaming and Remita, 2015).

Nanomaterials (NMs) have attracted considerable attention in various disciplines owing to their unique physical and chemical properties (Sajid, 2022). The European Commission Recommendation defines a nanomaterial as “a natural, incidental or manufactured material consisting of solid particles that are present, either on their own or as identifiable constituent particles in aggregates or agglomerates, and where 50% or more of these particles in the number-based size distribution fulfill at least one of the following conditions: one or more external dimensions of the particles are in the size range from 1 to 100 nm; the particle has an elongated shape, where two external dimensions are smaller than 1 nm and the other dimension is larger than 100 nm; the particle has a plate-like shape, where one external dimension is smaller than 1 nm and the other dimensions are larger than 100 nm” (European Commission, 2022). Nanomaterials can be categorized into three groups based on their origin: naturally occurring, incidental, and synthetic (engineered). Naturally formed NMs are found in various parts of the Earth’s spheres, including the atmosphere, hydrosphere (oceans, rivers, and lakes), lithosphere (soils, rocks, and lava), and biosphere (bodies of organisms, insects, plants, animals, and humans) (Jeevanandam et al., 2018). Incidental NMs are byproducts generated incidentally during industrial or natural processes that lead to the production of NPs, whereas synthetic NMs are created either through grinding or by using physical, chemical, biological, or hybrid methods (Jeevanandam et al., 2018). Nanoparticles are the most synthetically produced NMs (Nentwich, 2011).

A nanoparticle was defined as a particle of any shape with dimensions of 1 to 100 nm. In contrast, a microparticle has dimensions ranging from 0.1 to 100 μm, though the boundary between micro-sized and nano-sized particles is still debatable (Vert et al., 2012), as definitions vary across disciplines. Numerous types of NPs have been reported with a diverse array of applications across multiple domains of medicine and biology. These applications include drug delivery, cancer therapy, tissue engineering, regenerative medicine, biomolecule detection, and antimicrobial activity (Rudramurthy and Swamy, 2018). Nanoparticles can generally be classified into organic NPs (including dendrimers, micelles, liposomes, and ferritin), inorganic NPs (comprising metal and metal oxides), and carbon-based NPs (such as fullerenes, graphene, and carbon nanotubes) (Ealia and Saravanakumar, 2017) (Fig. 2). Nanoparticles can exist in either amorphous or crystalline forms. Nanocrystals are a type of nanoparticles referring specifically to the crystalline form (Bai et al., 2019). Nanospheres, in contrast, are a class of NPs characterized by a matrix structure. However, despite the name, nanospheres are not required to have a spherical shape (Kaeokhamloed et al., 2022).

Fig. 2. Classification of nanoparticles and their applications. Nanoparticles are generally classified into three categories: organic, inorganic, and carbon-based nanomaterials. Numerous types of NPs have been reported with diverse applications in various fields of biology and medicine, including drug delivery, cancer therapy, tissue engineering, biomolecule detection, and antimicrobial activity.

Researchers recognized the significance of nanoparticles when they discovered their unique physicochemical properties, attributed to their ultrasmall sizes (Khan et al., 2019). It has been suggested that the nanoscale size of these materials results in a large surface area, which could potentially promote interactions between the surface of living organisms and nanomaterials (Navya and Daima, 2016). Metallic nanoparticles and nitric-oxide-releasing NPs possess intrinsic antimicrobial properties, killing microbes by emulating the mechanisms employed by phagocytic cells through the generation of substantial amounts of reactive oxygen species (ROS) and reactive nitrogen species (RNS) (Jamil et al., 2017). It has been thought that NPs exert their antimicrobial effects by simultaneously targeting multiple vital biological processes or metabolic pathways of microbes, thereby making it unlikely for genetic mutations conferring resistance to these agents to occur (Jamil et al., 2017; Wang et al., 2017). Recently, the emergence of bacterial resistance against metallic NPs, by employing multiple mechanisms, has become a growing concern (Kamat and Kumari, 2023), which requires deep investigations.

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Literature Search

Databases

An electronic search was conducted using the Web of Science – Core Collection (WOS) and PubMed databases, and the search engine of Google Scholar, covering the timeline between January 2022 and September 2025. All records identified through databases were imported into EndNote. Duplicates were removed, and original research articles were initially screened by scanning the titles and abstracts to exclude non-relevant publications.

Inclusion and exclusion criteria

The inclusion criteria were as follows: (1) original research articles, (2) studies assessing the antimicrobial properties of nanoparticles against oral biofilms, (3) in vitro, in vivo and ex vivo studies involving multispecies (≥ 3 species) and saliva-derived biofilm models related to oral diseases, (4) clinical trials involving oral biofilms, (5) articles published in English. The exclusion criteria were as follows: (1) literature and systematic reviews, (2) studies involving planktonic microorganisms, mono- and dual-species biofilm models, (3) studies not assessing the antibiofilm efficacy of NPs against multispecies or saliva-derived biofilms (4) studies involving non-oral biofilms, (5) articles not published in English.

Search results

Following the exclusion of duplicates and non-relevant articles, a total of 48 original articles have been considered eligible for this review, as shown in the Prisma Diagram Figure 4. Table 1 summarizes the antimicrobial properties of various types of nanoparticles, with key findings illustrated in Figure 3.

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Discussion

In recent years, nanoparticles have been investigated as a promising tool for disrupting and removing microbial biofilms, which play a key role in the development and progression of oral infectious diseases. There is a burgeoning field of nanoparticle science that opens potential applications in the management of oral biofilms. It has been established that the mechanisms of action of NPs are influenced by their type, shape, size, dose, and concentration. Despite extensive research, the comprehensive mechanism underlying the action of NPs remains unclear (Rawat et al., 2021).

Metal and metal-oxide NPs have received much attention in the literature because of their potent antifungal and antibacterial activities (Tanweer et al., 2022; Wang et al., 2023; Jin et al., 2023; Teixeira et al., 2024; Bankar et al., 2024). One of the most extensively used nanosystems in dentistry is silver NPs (AgNPs) (Elizabeth et al., 2019), well known for their potent antimicrobial properties (Fernandez et al., 2021). Researchers have used AgNPs as a root canal irrigant (Pérez-Sáenz et al., 2025) and incorporated them into mouthwashes (Tomiyama et al., 2024; Raghav et al., 2025; Nemoda et al., 2025), resins used in denture fabrication (Teixeira et al., 2023; De Castro et al., 2024; Teixeira et al., 2024), titanium implants (Pérez-Tanoira et al., 2022), and glass ionomer cement (GIC) (Guo et al., 2025), which exhibited remarkable antibiofilm activity. It has been thought that AgNPs demonstrate their antimicrobial effects by penetrating microbial cells, generating ROS and free radicals, and additionally modulating microbial signal transduction pathways (Dakal et al., 2016).

Silver NPs have also been conjugated with CHX to make their antimicrobial properties more durable. This conjugation exhibits effective synergistic antimicrobial properties (Ivanova et al., 2023; Abbaszadegan et al., 2024). For instance, the incorporation of CHX and AgNPs into mesoporous silica NPs demonstrated significant antimicrobial effects against periodontitis- and caries-related biofilms, and modulating biofilm toward a healthy microbial profile (Fang et al., 2024; Wang et al., 2024). While AgNPs have not been universally recognized as safe by all authorities, no systemic toxicity from ingestion has been reported, and the clinical significance of their potential cytotoxicity remains unknown (Yin et al., 2020). However, in laboratory studies, CHX-AgNPs demonstrated cytotoxicity on various cell lines (Ivanova et al., 2023), and there have been ongoing concerns regarding the potential cytotoxicity of AgNPs on human cells (Khan et al., 2021), gut dysbiosis, and neurobehavioral alterations (Lyu et al., 2021). Accordingly, AgNPs have been conjugated with different substances to reduce their potential cytotoxic effects. Silver NPs have been conjugated with ebselen, a selenorganic compound with antioxidant and biological properties (Santi et al., 2021). Silver NPs and ebselen demonstrated synergistic antibiofilm effects against periodontal pathogens, thereby lowering the required concentrations of AgNPs. This combination showed high cytocompatibility and anti-inflammatory properties (Liang et al., 2023). Furthermore, biocompatible nanostructured gels containing AgNPs and CuNPs exhibited excellent antibiofilm ability (Berrio et al., 2024). In another study by Choi and others, it has been demonstrated that nano silver fluoride, composed primarily of AgNPs, chitosan and fluoride, can effectively inhibit biofilm formation and tooth demineralization through the sustained release of silver fluoride (Choi et al., 2025).

Table 1. Summary of nanoparticles used against oral multispecies biofilms and their main findings.

The increase in antibiotic resistance is an ongoing concern for public health. It has been established that the matrix of extracellular polymeric substances is a protective barrier that plays a significant role in microbial resistance and prevents drugs from penetrating biofilms (Pinto et al., 2020). When antibiotics are warranted, one approach that has been thought to minimize antimicrobial resistance is to use antibiotic-loaded NPs, such as AgNPs, for targeted delivery to enhance the actions of antibiotics (Gurunathan et al., 2014). Kang et al. (2024) synthesized novel magnetic NPs (MNPs) loaded with rhamnolipid (a glycolipid surfactant) and vancomycin. The antimicrobial efficacy of the MNPs was intensified in the presence of a magnetic field. This enabled the efficient penetration of positively charged NPs into the deeper layer of subgingival biofilms, along with the diffusion of rhamnolipid, vancomycin, and Ag, which together exhibited synergistic antimicrobial effects resulting in substantial biofilm eradication (Kang et al., 2024). Magnetic NPs also effectively inhibit cariogenic biofilms (Chen et al., 2025a), enhance the penetration of an endodontic sealer, and exert significant antimicrobial effects under a magnetic field (Guo et al., 2022). In other studies, another strategy was implicated, wherein positively charged agents were used with NPs to intensify their antimicrobial properties (He et al., 2023; Li et al., 2024). For instance, polyethyleneimine was used as a capping agent for AgNPs to impart a positive charge, thereby facilitating electrostatic interactions with the negatively charged bacterial surface in a cariogenic biofilm model (He et al., 2023). Additionally, silver ion-doped Prussian blue (a photothermal therapy agent) encased within cationic guar gum was shown to have great effectiveness in eradicating cariogenic biofilms following the photothermal therapy (PTT) (Li et al., 2024).

Given the escalating challenges associated with effective antimicrobial therapies, phototherapy has attracted considerable attention as a potential alternative to conventional antibiotic therapy (Abelha and Caires, 2021). Photothermal therapy uses photothermal agents that convert absorbed light into heat, resulting in heat generation that could eradicate microbial cells (Huo et al., 2021). In contrast, photodynamic therapy (PDT) is a highly promising strategy that uses light-activated non-toxic agents to trigger the production of ROS without inducing microbial resistance (Ribeiro et al., 2022) or signs of adverse effects on host tissues in vivo (Liu et al., 2022). The combination of PDT, PTT, and chemodynamic therapy with a nanocomposite (Bi₂S₃/Cu-TCP) exhibits potent antibacterial effects against periodontal pathogens, especially in animal models of periodontitis. This approach led to notable therapeutic outcomes, such as reduction of inflammation and bone preservation (Kong et al., 2023).

Fig. 3. Key terms in the literature. Most studies were conducted in vitro, often using human saliva-derived biofilms, inoculated anaerobically on a biological substrate (enamel specimens), thereby simulating complex in vivo conditions. Silver nanoparticles have been used extensively in research for the management of biofilm-related oral diseases (e.g., dental caries and periodontitis). Biofilm inhibition and eradication are typically evaluated via colony counting, CLSM, SEM, and PCR. The figure was created using Word Cloud.

Chitosan is another compound that has attracted significant attention among researchers. Chitosan, a natural polysaccharide derived from the deacetylation of chitin, has been considered as a valid treatment alternative for antimicrobials against oral biofilms (Costa et al., 2017; Sales et al., 2025). In the study by Rajan et al. (2025), although chitosan NPs showed statistically significant antibiofilm effects compared with the control, the encapsulation of an antimicrobial peptide (nisin) within chitosan NPs showed a synergistic effect, where a concentration as low as 6.25 mg/ml resulted in nearly complete inhibition of biofilm formation. This study even reported that this combination showed no cytotoxic effects on human gingival fibroblasts (HGFs), which exhibited gap closure that was comparable to the control group. In another study, cervical cavities prepared on extracted molars, inoculated with 28-day multispecies biofilms, were restored with GIC conjugated with either chitosan or fluoride-loaded chitosan NPs. Although the antibiofilm efficacy was not assessed, the assessment of teeth restored with GIC incorporated with chitosan, particularly chitosan NPs, demonstrated significant decreases in the depth of carious lesions and increases in calcium, phosphorus, and fluoride levels at the restoration-root junction. This modified material offers a potential promising material for restorative applications owing to its controlled release properties (Altınışık et al., 2024).

It is worth mentioning that antimicrobial peptides, combined with NPs, are small peptides that are widely distributed in nature and derived from a wide range of organisms. These peptides are known for their antimicrobial activities against fungi, bacteria, eukaryotic parasites, and viruses (Brown and Hancock, 2006). Nisin is a type of bacterial antimicrobial peptide and a part of the family of lantibiotics (lanthionine-containing antibiotics), produced by strains of Lactococcus lactis, a lactic acid bacterium (Breukink and De Kruijff, 1999; Da Silva Oliveira et al., 2024). Antimicrobial peptides are increasingly being investigated, particularly as an alternative to traditional antibiotics (Rima et al., 2021). The study by Radaic and the group reported that solid lipid NPs loaded with nisin were more effective as oral antibiofilm and anticancer agents than free nisin (Radaic et al., 2022).

Fig. 4. Prisma flow diagram.

Researchers have developed novel nanocomposites against cariogenic biofilms by integrating resin-modified glass ionomer (RMGIC) (Su et al., 2025) and calcium phosphate (AlSahafi et al., 2022) cements with dimethylaminohexadecyl methacrylate; a type of quaternary ammonium methacrylate (QAM). The antimicrobial mechanism of action is assumed to be electrostatic (Li et al., 2014), when the positively-charged quaternary amine N+ binds to the negatively-charged microbial cell membrane, leading to membrane disruption. Researchers have also incorporated cerium oxide NPs into poly methyl methacrylate, used in denture fabrication (Mangal et al., 2023; Jin et al., 2023). The positively charged cerium oxide NPs (nanoceria) can be adsorbed onto bacterial membranes via electrostatic attraction. Upon adsorption, the nanoparticles may affect bacterial viability through two principle mechanisms: the induction of oxidative stress and disruption of nutrient transport (Zhang et al., 2019).

However, a few randomized controlled clinical trials have been undertaken to evaluate the efficacy of different nanoparticles on oral biofilms. The addition of zirconium oxide NPs to 3D printable acrylic resin enhanced peri-implant tissue health, reduced surface roughness, and minimized biofilm formation (Albadwi et al., 2024). Another clinical study compared the antibacterial efficacy of AgNPs, Ca(OH)₂ NPs (nano-paste), and Ca(OH)₂ when used as intracanal medicaments during endodontic retreatment (Fahim et al., 2022). The results indicated that the three medications reduced total bacterial counts, but the difference between their efficacy was not statistically significant. However, post-operative pain was significantly reduced following the application of nano-Ag and nano-Ca(OH)₂ at the 48- and 72-hours intervals. In contrast, in an in vitro and in vivo study, potent antimicrobial effects for nano-Ca(OH)₂ (aqueous and viscous formulas) were demonstrated when compared to the conventional Ca(OH)₂, particularly in the viscous form (Rattanakijkamol et al., 2025).

Two clinical studies investigated the antimicrobial property of AgNPs among orthodontic and gingivitis patients, comparing their efficacy with that of CHX, the gold standard for chemical plaque control (Soundarajan and Rajasekar, 2023; Raghav et al., 2025). Although the results indicated improved plaque control and gingival health, as well as reduced microbial colony-forming unit counts, their efficacy was less than that of CHX. In contrast, another study found that both AgNPs and CHX mouthrinses exhibited comparable efficacy as adjuncts to scaling and root planing in patients with chronic periodontitis (Sachdeva et al., 2023). However, it is worth noting that the use of antimicrobials such as CHX mouthwash was found to alter the composition of the oral microbiome (Bescos et al., 2020; Brookes et al., 2021). This major shift was correlated with a marked reduction in saliva pH and buffering capacity, with elevated levels of lactate and glucose in saliva—conditions that promote dental caries (Bescos et al., 2020).

In the study by Su and colleagues (Su et al., 2025), the nano composite integrated into RMGIC was shown to decrease plaque buildup by selectively inhibiting cariogenic bacteria through the suppression of sucrose synthesis and carbohydrate metabolism. The nanocomposite effectively prevented enamel demineralization during orthodontic treatment. In additional clinical studies, while the antibiofilm activity was not evaluated, the remineralization efficacy of various nanoparticle-based materials on white spot lesions (initial caries) has been demonstrated. These materials include nanohydroxyapatite, nanosilver fluoride (Annadurai et al., 2024), and nanosilver, which was more effective than CHX and fluoride mouthwashes in reducing white spot lesions (Ali et al., 2022). Furthermore, combining nanosilver with fluoride varnish serves as a mineralizing agent and offers a non-invasive alternative treatment option for cavitated caries lesions in pediatric patients (Juárez-López et al., 2024).

In the articles reviewed above, numerous studies have used in vitro oral biofilm models grown into microtiter plates, seeking to assess the antibiofilm efficacy of NPs. In fact, in vitro model systems hold many advantages; they are cost-effective to establish and can be consistently reproduced and controlled. Additionally, various factors, including pH, temperature, fluid flow rates, and microbial composition, can be monitored and adjusted to reflect the dynamic environmental conditions present within the oral cavity (Brown et al., 2019; Brown et al., 2025). In contrast, in vivo animal models could capture the complexity inherent in biological events (Boraschi and Italiani, 2016; Boraschi et al., 2021). A number of animal models have documented promising outcomes for nanoparticle-based dental materials in terms of reducing caries severity (Liu et al., 2022; Chen et al., 2025b), demineralization (Li et al., 2024), and periapical lesions (Guo et al., 2022; Cheng et al., 2024). Crucially, the use of valid in vitro and in vivo models is essential for evaluating both the beneficial and harmful effects of nanomaterials, thereby predicting potential risks to human health (Boraschi and Italiani, 2016). In this collection, several studies used undefined (saliva-derived) biofilm models. Others used engineered biofilm models developed from a number of known species (≥3 species) that are usually well-characterized laboratory strains. This type of model is commonly used because it is easy to manipulate, simple to analyze and interpret, and cost-effective (Tan et al., 2017). However, the interspecies and interkingdom interactions that may modify tolerance against NPs have not been investigated.

Despite the ongoing development of valid models, a caveat to this is that the number of clinical trials is currently limited to a few, with the absence of long-term clinical follow-up, making it difficult to draw definitive conclusions regarding the impact of nanoparticle-based materials on oral biofilms and dysbiosis. Notably, nanomedical studies account for only 0.8% of all registered clinical trials, underscoring key translational challenges, including high costs, regulatory complexities, and clinical design limitations (Gultepe et al., 2026). Such limitations in clinical studies highlight the need for more research, especially regarding the long-term effects of NPs on human tissues, such as cytotoxicity, immune responses, and systemic distribution, which have not been thoroughly investigated (Govindarajan et al., 2025). In addition, comprehensive regulation of nano-waste usage and disposal is essential to mitigate the risk of developing microbial resistance to NPs. Although this issue has only recently emerged, adherence to regulatory standards and vigilant monitoring of NP interactions with the environment are critical measures to keep this problem from reaching the scale of multidrug resistance (Kamat and Kumari, 2023).

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Concluding Remarks

The aforementioned experimental and clinical studies highlight the potential application of nanoparticles as an effective strategy in controlling oral biofilms, enhancing oral health, and improving the antimicrobial and antiadhesion efficacy of dental materials, including dental implants, oral hygiene products, restorative materials, and orthodontic adhesives. Furthermore, some nanoparticles reduced the volume of periapical lesions and inhibited enamel demineralization, thereby reducing the severity of caries and promoting remineralization and tissue regeneration. The paper underscores the importance of choosing suitable biofilm models, where interspecies and interkingdom interactions that may modify microbial tolerance against NPs are not overlooked. It has been thought that the multitargeted mechanism of action exhibited by numerous nanoparticles, makes it unlikely for microorganisms to develop resistance, thereby presenting a valuable approach to addressing the worldwide issue of antimicrobial resistance. Contrary to early assumptions, emerging evidence indicates that bacteria can develop resistance to NPs. As the field of nanotechnology continues to evolve, more clinical trials are required to examine their antibiofilm effectiveness and address the challenges related to their potential cytotoxicity and long-term safety.

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Acknowledgment

The authors would like to thank Mohamed Salah Altabibi for the preparation of the figures.

Data availability

No datasets were generated during the current study.

Conflicts of interest

The authors declare no conflict of interest.

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