Smart nanoparticle vaccines integrate nanotechnology artificial intelligence and immunoengineering for precision immunization

Abstract

The rise of multidrug-resistant microbes, rapidly evolving viruses, and recurring pandemics underscores the urgent need for advanced vaccine technologies. Nanoparticle-based vaccines have emerged as a transformative approach capable of overcoming the major shortcomings of traditional immunization methods. Their nanoscale architecture allows precise antigen targeting, enhanced stability, and controlled release, leading to more potent and durable immune protection. These smart systems can carry multiple antigens or adjuvants, mimic natural pathogens, and efficiently activate immune cells to elicit strong humoral and cellular responses. Various nanoparticle types, lipid-based, polymeric, inorganic, and biomimetic, demonstrate broad potential against infectious, inflammatory, and neoplastic diseases in both humans and animals. However, critical barriers remain in mass production, regulatory harmonization, and long-term safety assurance. The integration of nanotechnology with artificial intelligence (AI) and bioengineering now enables rational vaccine design, predictive modeling, and personalized immunization strategies. AI-driven optimization of nanoparticle formulations and immune response prediction are accelerating translational progress. The convergence of these disciplines is shaping a new generation of vaccines that are safer, more effective, and adaptable to global health challenges, paving the way toward precision vaccination for the modern era.

Graphical Abstract

Introduction

Humanity is currently facing an array of pressing public health threats, ranging from the emergence of multidrug-resistant bacteria and rapidly evolving viruses to the growing prevalence of superbugs, drug-resistant parasites, zoonotic infections, and secondary malignancies [1]. These challenges, intensified by the recent global pandemic, continue to endanger both human and animal health [2]. The repeated occurrence of epidemic and pandemic events has profound consequences, contributing to increased mortality, disrupting economies, and hindering sustainable development. Such recurring crises highlight the urgent need for comprehensive biodefense and preparedness strategies capable of mitigating future health emergencies [3]. The misuse and overreliance on antibiotics and other therapeutic agents have fueled the widespread escalation of antimicrobial resistance, now recognized as one of the most serious health concerns of the twenty-first century [4]. Simultaneously, environmental degradation and climate change have accelerated the emergence and transmission of infectious diseases, further burdening healthcare systems worldwide [5]. This multifaceted crisis underscores the importance of coordinated global responses. Among the most effective preventive approaches, vaccination remains a cornerstone strategy to curb antimicrobial resistance, limit the spread of infectious agents, and reduce the overall disease burden [6].

The immune system, an intricate and dynamic defense network, serves as the body’s primary shield against external infectious and non-infectious stimuli. The innate immune system comprises a diverse population of circulating cells, including natural killer cells, granulocytes such as neutrophils, basophils, eosinophils, and mast cells, as well as antigen-presenting cells like macrophages and dendritic cells [7]. These components are responsible for detecting and eliminating invading particles through phagocytosis and subsequent degradation. The recognition of pathogens relies on several classes of pattern-recognition receptors, each playing a critical role in initiating and modulating immune responses [8]. The ultimate goal of vaccination is to activate both innate and adaptive immunity, prompting B- and T-lymphocyte responses that ensure long-term protection. When an antigen is introduced into the body, B cells generate antibodies to neutralize pathogens, whereas T cells identify and destroy infected or abnormal host cells, thereby reducing infection spread and maintaining immune homeostasis [9]. Recent progress in nanotechnology has revolutionized modern medicine, leading to the emergence of nanovaccines, nanoscale therapeutics, advanced diagnostic systems, and innovative platforms for drug screening and gene delivery. Nanovaccine technology, in particular, has opened new horizons in immunization by addressing the limitations of traditional vaccine design [10]. These novel platforms employ nanoparticles composed of lipids, polymers, proteins, or other biocompatible materials to optimize antigen delivery to target cells, thereby enhancing immune activation and protection against a broad range of diseases, including cancer, infectious, autoimmune, and inflammatory conditions [11].

Nanoparticles can be engineered to improve antigen stability and immunogenic potential. They may serve not only as vehicles for antigen delivery but also as immunogenic agents themselves. Due to their tunable size, composition, and surface properties, nanoparticles enable precise targeting of immune cells while minimizing off-target effects [12]. They also protect antigens and adjuvants from enzymatic degradation and ensure efficient uptake by antigen-presenting cells. By eliciting both humoral and cellular immune responses, nanovaccines offer durable protection. Their sustained stability allows extended antigen exposure, promoting dendritic cell maturation and efficient T-cell activation, which in turn supports the balanced release of cytokines essential for robust immune regulation [13]. Moreover, nanoparticle-based formulations can enhance B-cell activation and antibody production while facilitating targeted delivery to lymph nodes through lymphatic drainage, resulting in strong, long-lasting, and antigen-specific immunity [14].

This review explores the pivotal role of nanoparticles in reshaping vaccine development and delivery. It emphasizes their capacity to strengthen antigen presentation, amplify immune activation, and overcome conventional vaccine constraints. Furthermore, it discusses the mechanisms, types, and applications of nanoparticle systems in vaccine innovation, as well as the opportunities and challenges that define their path toward more effective, stable, and globally accessible immunization strategies.

From traditional vaccines to nanoparticle-based platforms

Traditional vaccines

Traditional vaccines represent one of the most remarkable milestones in the history of medicine. Their origins trace back to the late eighteenth century, when Edward Jenner demonstrated that exposure to a mild cowpox infection could protect individuals against smallpox [15]. This groundbreaking discovery led to the first successful vaccine against a contagious disease and marked the beginning of preventive immunization [16]. Over time, scientists continued to refine these early methods, conducting numerous experimental efforts to develop vaccines targeting specific infectious agents[17]. The classical vaccine development process follows a structured pathway that includes exploratory research, preclinical testing, and three phases of clinical trials before proceeding to regulatory review and approval. Traditional vaccines are categorized according to their mechanisms of action and the nature of the immune response they elicit [18].

Inactivated vaccines

Inactivated vaccines are produced from pathogens that have been killed by physical or chemical means. They include vaccines against influenza, typhoid, cholera, hepatitis A, rabies, and several bacterial infections such as Brucella and Mycoplasma [19]. Although these formulations are generally safe and stable, they tend to generate a weaker immune response compared with live vaccines. Consequently, multiple booster doses are often required to maintain sufficient and long-term immunity [20].

Live attenuated vaccines

Live attenuated vaccines contain weakened forms of pathogens that maintain their ability to replicate without causing disease. These vaccines elicit robust and long-lasting immune protection due to their close resemblance to natural infections [21]. The following subsections will address both viral and bacterial live attenuated vaccines, detailing their mechanisms, clinical relevance, and associated challenges [22].

Viral live attenuated vaccines

Examples of viral live attenuated vaccines include those against measles, rotavirus, Newcastle disease, and bovine viral diarrhea. These vaccines work by introducing a weakened version of the virus into the host, promoting an immune response similar to that produced by a natural infection [21]. The immune system responds by generating antibodies and memory cells, therefore providing long-term protection against future infections [23].

While viral live attenuated vaccines are effective, they do present certain safety risks, particularly for immunocompromised individuals. There is a small risk that the attenuated virus could revert to a virulent form, potentially causing disease in vulnerable populations [24]. Additionally, these vaccines can interact with maternal antibodies, potentially lowering vaccine efficacy or leading to adverse reactions. Furthermore, because of tissue-specific replication, there is a risk of secondary bacterial infections following vaccination [25].

Bacterial live attenuated vaccines

Bacillus Calmette-Guérin (BCG) vaccine stands out as a prominent instance of a bacterial live attenuated vaccine. It is primarily used to prevent tuberculosis (TB) and is derived from a strain of Mycobacterium bovis that has been attenuated [26]. The BCG vaccine’s mechanism of action contains stimulating a robust immune response against the tuberculosis bacillus. It induces both humoral and cellular immunity, enhancing the body's ability to fight TB infections [27].

Despite its clinical significance, BCG has restrictions. Its effectiveness varies considerably based on geographical location, the epidemiological context, and the specific population exposed [28]. This variability may be attributed to factors such as age, genetic background, and environmental conditions [29].

Recent advancements in nanoparticle-based platforms hold promise for addressing some of these challenges. These platforms can enhance vaccine stability, optimize delivery mechanisms, and improve the immune response [30]. For instance, nanoparticles can be engineered to target specific cells or tissues, potentially increasing vaccine efficacy while minimizing adverse reactions. This tailored approach offers the possibility of creating more effective live attenuated vaccines that can overcome the limitations associated with traditional formulations [31].

Subunit and conjugate vaccines

Subunit, recombinant, polysaccharide, and conjugate vaccines are composed of specific antigenic components of a pathogen, such as proteins, polysaccharides, or surface capsules. Vaccines for hepatitis B, human papillomavirus, and shingles are among the most notable examples [32]. These formulations are well-tolerated and suitable for individuals with weakened immunity, as they do not contain live organisms. Nonetheless, because they lack certain molecular structures that strongly stimulate the immune system, they often require adjuvants or repeated booster doses to achieve durable protection [33, 34].

Toxoid vaccines

Toxoid vaccines are designed to induce immunity against bacterial toxins rather than the bacteria themselves. They are produced by chemically or thermally inactivating bacterial toxins while preserving their antigenic properties [35]. Classic examples include vaccines against diphtheria and tetanus. These vaccines stimulate the production of neutralizing antibodies that block the effects of the toxins. Depending on the formulation, they can be administered via various routes, including intramuscular, subcutaneous, oral, or nasal delivery [36].

While conventional vaccines have played an essential role in eradicating or controlling many infectious diseases and have a well-established safety profile, they also present notable drawbacks. Their limitations include reduced genetic stability, risk of mutation in live vaccines, and relatively weak or short-lived immunity in inactivated formulations [37]. These challenges have motivated the search for next-generation vaccine platforms capable of inducing stronger and more precise immune responses, leading to the emergence of nanoparticle-based vaccines [38].

Nanoparticle-based vaccines

NPs are ultrafine structures with unique physicochemical properties that make them suitable for a wide range of biomedical applications, particularly in vaccine development. NPs play a crucial role in vaccine formulation by acting as both delivery systems and immune-stimulating adjuvants [39]. Nanovaccines offer several advantages over traditional vaccines, including reduced antigen degradation, enhanced stability, improved immunogenicity, and more efficient uptake and processing by antigen-presenting cells (APCs) [40]. Nanocarriers such as liposomes, dendrimers, and virosomes can promote cytokine production and enhance antibody responses, making them effective platforms for vaccine delivery [30]. These nanomaterials can also improve bioavailability, stabilize sensitive cargo, minimize off-target effects, and enable active targeting. In addition, peptide-drug conjugation represents a promising strategy for further investigation [41]. Figure 1 illustrates common vaccine carriers (Fig. 1).

Fig. 1.

Representative schematic illustration of major nanoparticle-based delivery systems, including A virus-like particles, B liposomes, C immune-stimulating complexes (ISCOMs), D polymeric nanoparticles, E inorganic nanoparticles, F emulsion-based systems, and G exosomes (40)

The properties of NPs including size, surface charge, composition, and hydrophilic or hydrophobic nature decisively influence their interactions with biological systems and immune responses. Small nanoparticles can more efficiently cross biological barriers and epithelial layers compared to larger particles [42]. Their size, which can be comparable to intracellular components, enables efficient cellular uptake and the induction of robust immune responses against intracellular pathogens. Among different geometries, spherical NPs have confirmed superior immunogenic potential [43].

Surface characteristics, such as charge and hydrophobicity, also play serious roles in targeting strategies. Hydrophobic nanoparticles tend to be more readily taken up by antigen-presenting cells (APCs), eliciting stronger immune activation [44]. Equally, hydrophilic nanoparticles often generate weaker immune responses. Moreover, negatively charged nanoparticles have been found to interact more effectively with macrophages, while positively charged variants show a stronger affinity for dendritic cells [45].

Nanoparticle formulations composed of polymers, lipids, proteins, emulsions, inorganic materials, and virus-like particles can be engineered to carry antigens and adjuvants. Many of these formulations are already approved for medical or veterinary applications [46]. Importantly, nanoparticles can protect encapsulated antigens from degradation, enhancing vaccine stability and ensuring effective delivery to target immune cells [47].

Beyond serving as delivery vehicles, certain NPs possess inherent adjuvant properties, stimulating immune responses without demanding additional adjuvant components. This is a significant advantage, as it simplifies the formulation and can lead to improved immune activation [48].

One of the most important benefits of nanoparticle-based vaccines is their tunable antigen release profiles, which can prolong immune stimulation and support the development of long-term immunity. By engineering NPs for targeted delivery such as to draining lymph nodes scientists can amplify antigen presentation and antibody production [49].

Surface modifications, such as attaching ligands or antibodies specific to cell surface receptors, enhance the precision of targeting plans. This allows NPs to be directed toward specific immune subsets or tissues, substantially improving the effectiveness of the vaccine [50]. For instance, modifications can be made to enhance the uptake by specialized APCs, such as dendritic cells, thereby facilitating better activation of T cells [51]. NPs vaccines offer flexibility in administration routes, including ocular, oral, nasal, intradermal, intramuscular, and subcutaneous methods. Among these, subcutaneous injection is frequently measured the most immunogenic method, as it delivers the formulation directly into tissues rich in immune cells, promoting high antibody titers [51, 52]. As vaccine science advances, nanoparticle-based systems are expected to play a defining role in the future of global immunization (Table 1) [53]

Table 1.

Comparative overview of traditional and nanoparticle-based vaccines

Vaccine type	Composition and mechanism	Advantages	Limitations	Representative formulations / applications	Refs
Inactivated vaccines	Pathogens killed by heat or chemicals; stimulate immune response without replication	Safe, stable, and easy to store	Weak immune response; need booster doses	Inactivated influenza vaccine, inactivated rabies vaccine, hepatitis A vaccine, typhoid vaccines	[54, 55]
Live attenuated vaccines	Contain weakened pathogens that replicate without causing disease	Strong, long-lasting immunity mimicking natural infection	Risk in immunocompromised individuals; possible reversion to virulence	MMR vaccine (measles component), Rotarix/Rotateq, live attenuated bovine viral diarrhea vaccine	[56, 57]
Subunit/conjugate vaccines	Contain purified antigens (proteins/polysaccharides) conjugated to carriers	Highly specific; safe for all populations	Require adjuvants and booster doses	Engerix-B/Recombivax HB (hepatitis B), Gardasil/Cervarix (HPV), Shingrix	[58, 59]
Nanoparticle-based vaccines	Antigens encapsulated or displayed on nanoscale carriers such as lipid, polymeric, or inorganic systems	Targeted delivery, sustained release, enhanced antigen stability	Manufacturing complexity; regulatory and safety concerns	BNT162b2 and mRNA-1273 (mRNA-LNP formulations) for SARS-CoV-2; NVX-CoV2373 (recombinant spike protein nanoparticle vaccine); nanoparticle-based platforms under investigation for cancer, tuberculosis, and HIV	[60, 61]

As the field continues to evolve, the diverse applications of nanoparticle-based vaccines are driving research toward optimizing formulation designs, delivery strategies, and enhancing both the magnitude and durability of immune responses [62]. Their versatility and adaptability position nanoparticle platforms not only to combat infectious diseases but also to address non-communicable conditions, including cancer and autoimmune disorders. By focusing on platform-specific targeting strategies instead of general properties, we can provide a clearer understanding of how to harness the potential of nanoparticles to improve vaccine efficacy and targeted immune responses [63].

Emerging challenges and nanoparticle vaccine platforms

Although the advancement of nanovaccine technologies has brought remarkable progress in preventing infectious, inflammatory, allergic, and malignant diseases, several obstacles continue to impede their full clinical translation. Achieving uniformity in nanoparticle size, morphology, and composition remains a major technical hurdle, as these attributes are critical to their pharmacological performance, biological interactions, and physicochemical stability [64]. Another key challenge involves managing potential toxicity and unintended immune responses that may result from nanoparticle accumulation in specific tissues or organs. Ensuring long-term stability and shelf life under variable environmental conditions is also demanding, particularly for lipid-based formulations that are highly sensitive to temperature changes [65]. Furthermore, large-scale manufacturing, regulatory approval processes, and the safe disposal of nanomaterial waste add to the complexity and cost of production. Messenger RNA-based nanovaccines, despite their success in recent years, face additional issues such as selecting optimal antigen targets, achieving high-quality mRNA synthesis and purification, and ensuring efficient encapsulation within lipid nanocarriers for stable and targeted delivery [66].

Categories of nanoparticles used in vaccine design

Nanoparticles possess unique properties that make them valuable tools for enhancing immune responses and achieving precise antigen delivery. Their tunable structure allows for targeted, sustained, and controlled release of vaccine components [67]. Several nanoparticle platforms have been investigated for vaccine development, including lipid-based systems, polymeric carriers, self-assembled protein or peptide structures, biomimetic particles, and inorganic nanomaterials. Among these, lipid-based nanoparticles have attracted particular attention due to their biocompatibility, biodegradability, and adaptability for different vaccine formulations [68, 69].

Lipid nanoparticles

Lipid-based nanoparticles, commonly known as lipid nanocarriers, represent an important advance in vaccine delivery systems. These spherical particles are primarily composed of ionizable lipids, which not only provide structural integrity but also enhance the efficiency of drug and vaccine delivery [70]. Their biodegradability, high compatibility with biological systems, and strong safety profile make them particularly suitable for vaccine applications [71].

Lipid NPs have proven capable of encapsulating messenger RNA (mRNA), which is critical for modern vaccines. By protecting mRNA from enzymatic degradation, these nanoparticles facilitate the entry of mRNA into cells, where it can stimulate robust immune responses [72]. This ability has positioned lipid NPs as foundational components in the development of contemporary vaccines and therapeutic nanomedicines [73].

Since the introduction of lipid nanoparticle-mRNA formulations, they have emerged as a potent alternative to conventional vaccine platforms. This is largely due to their unique attributes. First, mRNA is a non-infectious and non-integrating agent that can encode a wide variety of antigens, making it highly flexible for vaccine development [74, 75]. For instance, in a cytomegalovirus vaccine, six separate mRNAs were combined, where five encoded a pentameric antigen and one encoded a glycoprotein antigen. This flexibility allows innovative vaccine designs tailored to specific pathogens [76].

Another important advantage of mRNA vaccines using lipid nanoparticles is the speed at which they can be manufactured. GMP-grade lipid nanoparticle-mRNA vaccines can be produced rapidly for specific antigens compared with traditional vaccine approaches, such as recombinant proteins and inactivated vaccines. This rapid development is crucial for responding effectively to emerging infectious diseases [77].

Despite these advantages, several limitations remain. Naked or poorly protected mRNA is inherently unstable and can be rapidly degraded by extracellular and intracellular ribonucleases. Under physiological conditions, its half-life is generally short and may range from minutes to a few hours depending on sequence design, chemical modification, and cellular context. This instability creates important challenges for storage, transport, and in vivo delivery [53]. In addition, lipid nanoparticles themselves may be sensitive to temperature fluctuations, which can compromise encapsulation efficiency, particle integrity, and delivery performance [78]. Safety considerations also require careful attention, including reactogenicity, inflammatory responses, and the need to optimize biodistribution while minimizing unintended tissue exposure [79]. Therefore, both mRNA engineering and lipid nanoparticle design must be carefully optimized before these formulations can be broadly and reliably applied in clinical settings [80].

To address these challenges, the engineering of mRNA molecules and the design of lipid nanoparticles have been optimized significantly. These advancements contributed to the rapid development and clinical assessment of COVID-19 mRNA vaccines, such as mRNA-1273 and BNT162b2 [79]. Both vaccines utilize ionizable lipid NPs to deliver nucleoside-modified mRNA encoding the full-length spike protein of SARS-CoV-2. Using a prime-boost vaccination strategy, these vaccines elicit high levels of antigen-specific antibodies and generate robust T helper 1 cell responses [81](Fig. 2).

Fig. 2.

Mechanism of mRNA delivery and antigen presentation mediated by ionizable lipid nanoparticles in COVID-19 vaccines (82)

Significantly, both vaccines demonstrated comparable efficacy of approximately 95% in phase III clinical trials. Furthermore, additional COVID-19 vaccines utilizing lipid nanoparticle-mRNA formulations are currently undergoing assessment in various clinical phases [83]. Preliminary studies have shown that certain vaccine candidates can confer protective effects by employing innovative strategies, such as delivering self-amplifying RNA or formulating cocktails of mRNAs that encode different viral proteins [84]. This flexibility and rapid manufacturability make lipid nanoparticle-mRNA formulations an important frontier in vaccine development, providing a platform that can be mobilized quickly against emerging infectious pathogens [82].

Interbilayer crosslinked multilamellar vesicles

Interbilayer crosslinked multilamellar vesicles consist of an aqueous interior enclosed by multiple lipid bilayers. Their dense lipid structure shields the internal contents from serum degradation, allowing for the stable delivery of vaccine antigens to immune tissues such as lymph nodes, where they promote strong antibody responses [85]. These vesicles can encapsulate both hydrophilic and hydrophobic molecules, making them suitable for the co-delivery of antigens and adjuvants. However, their complex production process, potential to provoke unintended immune reactions, and difficulties in scaling up for industrial manufacturing present practical limitations that restrict their widespread use [86].

Lipoprotein nanoparticles

Lipoproteins are natural complexes composed of lipids and proteins that function in the transport of various biomolecules, including hormones, vitamins, and genetic material. Their amphiphilic nature, characterized by a hydrophobic core and a hydrophilic surface, enables them to carry a wide range of therapeutic agents, making them effective vaccine delivery systems [87]. Engineered recombinant lipoproteins with built-in immunostimulatory domains have shown promise in the design of subunit vaccines against pathogens such as meningococcus, dengue virus, and Clostridium difficile [88]. Nevertheless, their tendency toward instability in the bloodstream, the complexity of their production, and the potential for inducing undesired immune responses or toxicity remain significant concerns in their clinical development [89].

Liposomes

Liposomes are vesicular structures composed of phospholipid bilayers that encapsulate an aqueous core. Their amphiphilic nature allows them to carry both hydrophilic and hydrophobic molecules, making them ideal for vaccine and drug delivery [90]. Liposomes have been widely used to deliver immunostimulatory molecules, nucleic acids, and antigens, thereby facilitating antibody production and immune cell activation [91]. They can serve as both carriers and adjuvants, enhancing antigen stability and prolonging immune stimulation. The effectiveness of liposome-based vaccines depends on parameters such as particle size, surface charge, and lipid composition [92]. Despite their versatility, liposomes may experience instability during storage, including leakage or fusion of encapsulated materials. In addition, their production often involves complex and expensive processes, and maintaining temperature stability during storage remains a key challenge [93].

Solid lipid nanoparticles

Solid lipid nanoparticles consist of a solid lipid core stabilized by surfactants or cationic lipids. They can bind to DNA or other antigens to form stable complexes, facilitating their application in vaccine formulations [94]. These systems protect encapsulated biomolecules from enzymatic degradation and can carry both hydrophilic and hydrophobic agents, making them suitable for a wide range of therapeutic applications [95]. Despite these benefits, solid lipid nanoparticles may experience issues with stability during storage, as some bioactive molecules can be expelled from the core over time. Their high water content and limited ability to penetrate certain biological barriers, such as the skin, may also restrict their application [96].

Exosomes

Exosomes are nanosized vesicles naturally secreted by cells, typically ranging from 50 to 100 nm in diameter [97]. They are composed of lipid bilayers and carry various biomolecules, including proteins and nucleic acids. Because of their natural origin and biocompatibility, exosomes represent safe and efficient carriers for vaccine delivery [98]. They can traverse biological barriers such as the blood–brain barrier and deliver therapeutic antigens to specific target cells. Exosome-based vaccines have demonstrated potential in enhancing immune responses against cancer and viral infections, and current research continues to explore their role in immunization against diseases such as influenza, Ebola, and COVID-19 [99].

Virosomes

Virosomes are hybrid structures that combine viral envelope proteins with liposomal membranes. They typically range between 20 and 50 nm in size and possess surface glycoproteins derived from viruses such as influenza or herpes [100]. These viral components enhance antigen presentation and immune system activation. Virosomes have been successfully used in approved influenza vaccines to strengthen immune responses by mimicking natural viral entry pathways without the risks associated with live virus exposure [101].

Emulsion systems

Emulsion-based formulations, generally composed of oil-in-water mixtures, have become valuable tools for vaccine delivery. They allow the incorporation of antigens, adjuvants, and surfactants, improving the magnitude and quality of immune responses [102]. Such emulsions have been applied in vaccines targeting infections caused by influenza viruses, human papillomavirus, and hepatitis B virus. Notably, certain emulsion adjuvants have been approved for clinical use due to their demonstrated ability to enhance antigen presentation and induce both humoral and cellular immune responses [103].

lyotropic liquid crystalline lipid nanoparticles (LLCNP)

Lyotropic liquid crystalline lipid nanoparticles (LLCNPs) have recently emerged as an additional class of lipid-based nanocarriers with relevance to drug and gene delivery, including RNA-based delivery. Unlike conventional liposomes, LLCNPs possess ordered non-lamellar internal architectures, particularly inverse cubic and inverse hexagonal mesophases, commonly described as cubosomes and hexosomes. These structures provide interconnected aqueous channels, high lipid-water interfacial area, and tunable pore size, allowing the loading of hydrophilic and hydrophobic drugs, peptides, proteins, nucleic acids, and imaging agents. Their polymer-based outer corona can also be modified to improve colloidal stability, prolong circulation, or introduce targeting ligands. Recent evidence further suggests that pH-dependent cubic and hexagonal mesophase behavior in ionizable lipid nanoparticles may influence mRNA transfection efficiency, indicating that internal lipid architecture can affect endosomal escape and nucleic acid delivery. In Alzheimer’s disease nanomedicine, LLCNPs may therefore be useful for delivering poorly soluble neuroprotective agents, anti-inflammatory compounds, peptides, or gene-modulating cargos. However, their application in neurodegenerative disease remains at an early stage, and further studies are needed to clarify their brain biodistribution, blood–brain barrier transport, structural stability, neurotoxicity, and long-term safety [104–106].

Polymeric nanoparticles

Polymeric nanoparticles have emerged as adaptable delivery vehicles capable of carrying therapeutic or immunogenic compounds that can either be encapsulated within the polymeric matrix or adsorbed onto its surface. These nanoparticles may be synthesized chemically or derived from natural sources such as polysaccharides, proteins, or lipids [107]. Naturally derived polymeric nanoparticles generally possess superior characteristics in terms of biodegradability, compatibility with biological tissues, and minimal toxicity compared with synthetic forms. Their structural designs vary and include nanospheres, micelles, and polymeric assemblies [108]. Commonly utilized polymers in vaccine and adjuvant formulations include poly(lactic-co-glycolic acid), chitosan, hyaluronic acid, silk fibroin, alginate, polyethyleneimine, polypropylene sulfide, and polystyrene[109]. The fabrication of polymeric nanoparticles typically follows one of two routes, either through the polymerization of monomers or the dispersion of preformed polymers [110].

These nanoparticles can incorporate multiple adjuvants or antigens, thereby protecting them from premature degradation and improving their structural stability. The controlled release of encapsulated molecules can be achieved via diffusion or through gradual polymer degradation, allowing prolonged antigen exposure and sustained immune activation [111]. Furthermore, polymeric nanoparticles can be functionalized with ligands, antibodies, or targeting molecules to direct their cargo toward specific cells or tissues, thereby reducing off-target effects and improving vaccine efficacy [112]. Some are designed to release their payload in response to environmental triggers such as changes in pH, enzymatic activity, or temperature. Cellular uptake generally occurs through endocytosis, after which the nanoparticles release their content intracellularly. This feature makes polymeric nanoparticle vaccines particularly attractive for cancer immunotherapy, as they can deliver antigens to tumor or inflamed tissues [113].

These systems also protect fragile antigenic materials from enzymatic breakdown within the body, ensuring that the immune system encounters them in their active form. Other biodegradable polymers, including polylactic acid, polyethylene glycol, and polycaprolactone, have been widely used as adjuvants to enhance immune responses [114]. Polymeric micelles, which are self-assembled amphiphilic structures, have shown potential for the efficient presentation of viral antigens in their native conformation, although their instability in biological environments remains a key limitation [115]. Chitosan nanoparticles have been successfully employed to strengthen mucosal immunity, particularly in the respiratory tract, improving protection against infections such as tuberculosis [116].

The advantages of polymeric nanoparticles lie in their structural tunability, capacity for precise control of particle size and surface chemistry, and ability to accommodate a wide range of therapeutic molecules [117]. However, challenges persist, including their potential for aggregation, cytotoxicity under certain conditions, and the complexity of large-scale production. Their physical and chemical stability during storage can also be problematic [118]. Polymeric nanoparticle platforms have been explored for a wide range of medical applications, including targeted cancer therapy, gene delivery, and immunization against infectious diseases such as malaria, HIV, COVID-19, and tuberculosis [119]. Several notable examples have advanced through preclinical and clinical stages.

The NVX-CoV2373 vaccine, developed by Novavax, utilizes polymeric nanoparticles to deliver recombinant spike proteins of SARS-CoV-2 arranged in trimeric structures, combined with an immune-stimulatory adjuvant. Clinical trials demonstrated high efficacy following a two-dose regimen, with substantial protection observed after the second administration [120].

Another example is the GBP510 vaccine, which employs nanoparticle bioengineering to elicit robust immune responses against COVID-19. This candidate has successfully completed early-phase human trials, with a design aimed at optimizing immune recognition and inducing a strong humoral response [121].

The TAK-101 vaccine, designed for celiac disease and simian immunodeficiency virus infection, incorporates gliadin encapsulated within poly(lactic-co-glycolic acid) nanoparticles alongside Toll-like receptor agonists. Studies in primate models have indicated that these formulations can induce durable and potent immune protection [49].

While polymeric nanoparticles have confirmed significant promise, it is essential to compare their performance with other nanoparticle platforms, such as lipid-based, inorganic, and virus-like particles [122]. For example, lipid-based nanoparticles excel at delivering mRNA and exhibit excellent cell membrane fusion properties. In contrast, polymeric nanoparticles offer safer storage solutions and greater design versatility [123]. The controlled release of cargo from polymeric nanoparticles stands in stark contrast to the often fast release seen in lipid-based systems. Understanding these differences is crucial for selecting the optimal platform based on specific vaccine requirements or therapeutic goals [124].

Inorganic nanoparticles

Inorganic NPs represent a distinct class of nanomaterials distinguished by their high biocompatibility, hydrophilicity, and remarkable structural stability. These nanoparticles can be broadly classified into two main categories: solid forms, such as calcium phosphate and hydroxyapatite, and porous variants, including mesoporous silica and porous silicon [125]. Their unique chemical and physical properties render them highly favorable for various biomedical applications, including drug and vaccine delivery, tumor imaging, and infectious disease prevention. Inorganic nanoparticles offer several advantages, as well as controlled antigen release, long-term stability, and targeted delivery, which cooperatively contribute to improved vaccine performance and therapeutic precision [125, 126].

Among the most extensively studied inorganic NPs are gold, silica, carbon-based materials, and quantum dots. Gold nanoparticles are particularly noteworthy due to their excellent optical, chemical, and electronic properties, which differ significantly from those of bulk gold [127]. They can form colloidal dispersions in aqueous solutions that exhibit intense light absorption and scattering at specific wavelengths due to surface plasmon resonance [128]. This optical property facilitates various diagnostic and photothermal applications while also establishing gold nanoparticles as effective immune enhancers in vaccine formulations. These nanoparticles can be conjugated with antigens or drugs to improve targeted delivery, minimize systemic side effects, and enhance vaccine stability during storage [129]. Research demonstrates that gold nanoparticle-conjugated vaccines can elicit strong antibody responses without intrinsic toxicity, leading to increased efficacy across diverse infectious models, including influenza and coronavirus vaccines [130].

In addition to their role in immunization, gold nanoparticles also exhibit antimicrobial and anticancer properties. They can disrupt bacterial membranes, produce reactive oxygen species that damage cellular macromolecules, and induce apoptosis through oxidative stress trails [131]. When engineered with additional elements like platinum or copper, they can promote glucose oxidation within tumor microenvironments, presenting potential applications in cancer therapy [132]. While gold nanoparticles hold great promise, certain restrictions exist, including cytotoxicity at high concentrations, synthesis complexities, a tendency to aggregate during storage, and the potential for inflammatory or immunogenic side effects [133].

Another significant category is silica nanoparticles, commonly referred to as nanosilica. Typically amorphous and spherical, they can be finely tailored in size, porosity, and surface chemistry for various biomedical applications. Their high surface area and modifiable structure make silica nanoparticles excellent candidates for drug and antigen delivery [134]. These materials are cost-effective, thermally stable, and generally regarded as safe for biomedical use. Mesoporous silica nanoparticles, characterized by tunable pore sizes and surface functionalities, are particularly advantageous for controlled release systems [135]. Recent innovations have produced large-pore mesoporous silica nanoparticles capable of encapsulating macromolecules such as proteins and nucleic acids, facilitating their use in cancer vaccines and immunotherapy [136]. These particles can efficiently deliver tumor antigens to dendritic cells within lymphoid tissues, triggering effective cytotoxic T-cell activation and antitumor immunity.

Furthermore, silica-based nanoparticles are biodegradable and biocompatible, allowing for informal functionalization with adjuvants or targeting ligands, enhancing their immunostimulatory potential in subunit vaccines [137]. Continuing research is exploring their clinical applications in various cancers, including ovarian, prostate, and brain malignancies. Conversely, challenges persist, particularly regarding the complexity of their synthesis, environmental safety, and potential long-term toxicity [138]. Prolonged exposure to silica nanoparticles may induce oxidative stress, inflammation, and cellular damage, underscoring the importance of careful design and evaluation prior to large-scale biomedical application [139].

Developing research focuses on hybrid inorganic nanoparticles that combine properties from different materials to enhance performance. For instance, gold–silica core–shell nanoparticles offer improved stability and versatility [140]. These hybrids can optimize drug release profiles and provide multifunctional capabilities, such as imaging, drug delivery, and photothermal therapy in a single construct. Additional area of interest is magnetic nanoparticles, which can be manipulated with external magnetic fields for targeted delivery and localized treatment, enhancing drug concentrations at disease sites while reducing systemic exposure [125].

The expansion of quantum dots nanoscale semiconductor particles also demonstrations promise in bioimaging and diagnostics. Their unique electronic properties allow for fluorescent tagging of cells, enabling real-time tracking of cellular processes [141]. Quantum dots can be functionalized to ensure targeted delivery of therapeutic agents, making them versatile tools in both research and clinical settings [142]. Moreover, the biocompatibility of inorganic nanoparticles can be enhanced through various surface modification techniques, including polymer coatings and biofunctionalization with peptides or antibodies [143]. These alterations not only improve biocompatibility but also allow for selective targeting of specific tissues or cell types, thereby maximizing therapeutic efficacy while minimizing adverse effects [144].

Finally, the field of inorganic nanoparticles remains to evolve, revealing their multifaceted roles in vaccine expansion and therapeutic interventions [145]. By comparing the numerous classes of inorganic nanoparticles and discussing their complementary benefits and limitations, this section can effectively broaden the understanding of their potential in advancing medical science and addressing pressing health challenges [146]. Incorporating discussions on emerging research, clinical trials, and comparative analyses with other NPs platforms will provide a more broad view of the contributions and challenges of inorganic nanoparticles in the present and future landscape of vaccine technology [32].

Virus like particle platforms

Virus-like particles (VLPs) are assemblies of viral structural proteins devoid of any genetic material. They mimic the outer architecture of true viruses, making them an excellent platform for vaccine expansion, as they can replicate the native structure and appearance of viral antigens without posing any risk of disease [147]. Because VLPs present highly ordered and repetitive antigen displays, they are powerful immune stimulators that provoke both cellular and antibody-mediated responses [148]. This typical makes them attractive candidates for vaccines against numerous infectious agents, including hepatitis B, malaria, and the recent coronavirus [149].

The effectiveness of VLPs lies in their ability to induce robust immune responses. By recapitulating the geometry of native virions, these particles reliably engage antigen-presenting cells, which are crucial for activating both B and T cells. This arrangement is critical for generating strong, durable immunity, which explains their growing role in vaccine research [150]. Products built on this concept are broadly regarded as effective, safe, and stable, capable of being produced in diverse formats. In many licensed preparations, classical aluminum salt adjuvants are comprised to intensify antibody formation through local retention of antigens at the injection site and continued stimulation of innate pathways [151].

Even though their advantages, several important restrictions exist. First, VLPs may be less stable than some other nanosystems and can lose structural integrity during processing or storage. Maintaining stability is crucial for ensuring that the vaccine retains its potency and efficacy throughout its shelf life [152]. Second, widespread purification is often required to remove process-related impurities, making production labor-intensive. This purification step can significantly increase manufacturing costs and complicate the scale-up process [153]. Third, sensitivity to temperature and acidity narrows storage options, necessitating cold-chain logistics that can be a barrier to distribution, particularly in low-resource settings. Fourth, particles produced in bacterial systems can carry trace endotoxins, which may lead to annoying inflammatory reactions unless rigorously removed [154].

Recent progresses have aimed to overcome some of these limitations. Genetic engineering techniques are being employed to enhance the stability of VLPs. By modifying the structural proteins or introducing stabilizing components, researchers are developing VLPs that can withstand harsher conditions without compromising their effectiveness [155]. Additionally, new purification methods, such as affinity chromatography and other high-throughput techniques, are being explored to streamline the production process and reduce the time and cost associated with purification [156].

Clinical and preclinical programs span a broad range, with notable success stories emphasizing the effectiveness of VLPs. Hepatitis B and human papillomavirus particle vaccines are already in widespread use worldwide, demonstrating the real-world applicability of this platform [157]. Moreover, platforms displaying coronavirus spike proteins have advanced into human testing, showcasing their versatility in addressing emerging infectious diseases [158]. Current development continues for vaccines targeting influenza, human immunodeficiency virus, and malaria. By offering features such as multivalent displays, where multiple antigens can be existing simultaneously, VLPs can enhance the breadth of immunity against diverse viral strains [159].

Inventions are ongoing to emerge in the use of VLPs for not only infectious diseases but also therapeutic applications. For instance, VLPs are being explored for their potential in cancer immunotherapy, where they can be designed to present tumor-associated antigens, so stimulating the body's immune response to recognize and attack cancer cells [160]. This dual application highlights the adaptability of VLP platforms in both preventive and therapeutic settings. In summary, virus-like particles represent a promising and dynamic platform for vaccine development [161]. While they offer unique advantages in stimulating robust immune responses, addressing their limitations, such as stability, purification challenges, and storage requirements, is crucial for maximizing their potential in global vaccination strategies [162]. Continuous research and technological advancements will likely enhance the utility and effectiveness of VLP platforms, paving the way for their broader application in combating various diseases [161].

Self assembling peptide and protein vaccines

Self-assembling peptide and protein vaccines represent an exciting frontier in vaccine expansion, harnessing the natural ability of short peptides and engineered proteins to impulsively organize into nanoscale architectures such as fibers, particles, and hydrogels [163]. These ordered structures offer significant advantages in terms of controlled antigen presentation, improved immunogenicity, and the capability to direct cargo to key immune cells [163].

Recent progresses in protein design have enabled the development of multivalent displays, allowing multiple antigens to be presented simultaneously. This multivalency enhances the likelihood of causing a robust immune response by ensuring that different regions of the immune system can recognize and react to various pathogen components [164]. For instance, ferritin-based particles can form symmetric cages that have been explored for presenting antigens from complex pathogens, such as the Epstein-Barr virus, coronavirus, and various oncological targets [164]. The inherent structure of ferritin not only stabilizes the encapsulated antigens but also optimizes their presentation to the immune system. Moreover, self-assembling peptides can be tailored for specific immune responses. By regulating the peptide sequence or altering the physical properties of the resulting nanostructures, scientists can elicit precise T-cell or antibody-dominated responses [165]. This modulability offers a notable advantage in responding swiftly to emerging pathogens, as modifications can be made rapidly to adapt to new viral variants or bacterial strains [166].

The modular nature of self-assembling peptides permits for rapid revision to altering pathogenic landscapes, making them mainly useful in scenarios where time is of the essence, such as during an outbreak [167]. Their biocompatibility ensures minimal adverse reactions in therapeutic requests, while their ability to be engineered for specific pathways supports their role in personalized vaccines [168]. Despite their impressive potential, self-assembling peptide and protein vaccines do harbor some limitations. Long-term stability can be a concern, as environmental conditions may impact the integrity of the peptides and their assembled structures [164]. This instability can lead to challenges in maintaining efficacy over time, particularly in varying storage conditions.

Moreover, there is a risk of aggregation during production, which can compromise the consistency and effectiveness of the vaccine [169]. Maintaining a consistent structure is critical, as variations in size or shape could significantly alter the immune response. Robust quality control processes are necessary to ensure that these vaccines maintain their intended physical and functional features throughout the manufacturing procedure [170]. Research in this field is rapidly developing, with many studies focusing on improving the stability and manufacturing methods associated with self-assembling peptide and protein vaccines [171]. Innovations such as stabilizing agents or alternative assembly strategies are being explored to enhance the resilience of these nanostructures [53]. Also, constant clinical trials are assessing the performance of these vaccine platforms against various infectious diseases, with promising results. Table 2 summarizes the main categories of NPs used in vaccine design, including self-assembling peptide and protein vaccines, emphasizing their structural composition, mechanisms, advantages, and limitations (Table 2).

Table 2.

Main categories of nanoparticles used in vaccine design

Nanoparticle category	Structural composition / examples	Mechanism and application in vaccines	Major advantages	Key limitations / challenges	Refs
Lipid-based nanoparticles (LNPs)	Ionizable lipids forming spherical carriers; includes multilamellar vesicles, lipoproteins, liposomes, solid lipid NPs, exosomes, virosomes, and emulsions	Encapsulate mRNA or protein antigens for delivery to immune cells; protect cargo from enzymatic degradation; enhance antigen presentation and immune activation	Biocompatible, biodegradable, adaptable, and highly efficient for mRNA delivery; proven success in COVID-19 vaccines	Require cold-chain storage; potential instability and aggregation; complex large-scale manufacturing	[172–174]
Polymeric nanoparticles	Natural or synthetic polymers such as PLGA, chitosan, alginate, PEG, or PCL; structures include nanospheres, micelles, and polymeric assemblies	Carry or adsorb antigens/adjuvants; release them via diffusion or polymer degradation; can be functionalized for targeted delivery to immune tissues	Tunable structure, controlled release, prolonged antigen exposure, strong immune activation, suitable for mucosal delivery	Aggregation tendency, cytotoxicity at high doses, stability and scalability issues	[175, 176]
Inorganic nanoparticles	Gold, silica, calcium phosphate, hydroxyapatite, carbon-based materials, quantum dots, mesoporous silica	Serve as carriers or immune enhancers; deliver antigens to dendritic cells; can provide imaging or photothermal capabilities	High stability, controlled antigen release, strong adjuvant potential, dual therapeutic–diagnostic (“theranostic”) use	Possible cytotoxicity, complex synthesis, aggregation risk, oxidative stress, and long-term safety concerns	[177, 178]
Virus-like particle (VLP) platforms	Self-assembled viral structural proteins lacking genetic material; e.g., Hepatitis B, HPV, or coronavirus spike-based VLPs	Mimic native viruses to trigger robust humoral and cellular immune responses without infection risk	High immunogenicity, safety, and established clinical use; repetitive antigen display enhances immune recognition	Structural instability, need for stringent purification, temperature sensitivity, potential endotoxin contamination	[179, 180]
Self-assembling peptide and protein nanostructures	Peptides or engineered proteins that self-organize into fibers, cages, or hydrogels (e.g., ferritin-based nanocages)	Enable multivalent antigen display and controlled presentation; target antigen-presenting cells to elicit specific immune pathways	Modular, biocompatible, and rapidly adaptable to emerging pathogens; capable of eliciting tailored T-cell or antibody responses	Limited long-term stability, potential aggregation, and challenges in maintaining uniform structure during production	[181, 182]

Nanovaccines properties

Nanoparticles can carry antigens inside the particle or attach them on the surface. Encapsulation shields fragile molecules from degradation and allows timed release, while surface display promotes direct interaction with immune receptors [182]. The overall immune effect depends on size, shape, surface charge, composition, dose, and the route by which cells take up the particles. Small dimensions improve tissue penetration and foster contact with dendritic cells and macrophages at sites of infection or vaccination [183]. By protecting antigen from early breakdown and by extending exposure, nanoparticles enhance priming of both B cells and T cells [184].

Targeting motifs can be added to bind receptors on antigen presenting cells, which increases uptake and speeds delivery to processing pathways. Once inside these cells, the particles disassemble and antigens are loaded onto major histocompatibility complexes for presentation to lymphocytes [185]. In some settings the immune system may also recognize the carrier itself, producing antibodies to the nanoparticle as well as to the intended antigen [186].

Materials used as carriers often act as adjuvants in their own right. They sustain antigen release, concentrate vaccine components in lymphoid tissues, and activate innate sensors, which together permit protective immunity with smaller antigen quantities [187]. Lipid based carriers are especially effective for messenger RNA delivery. They protect the nucleic acid, promote cytosolic entry, and drive robust activation of cytotoxic T cells by enabling efficient expression of encoded proteins in antigen presenting cells [188]. Nanoplatforms can also package multiple antigens in a single construct, for example several coronavirus spike variants, with the goal of broad cross protection [189, 190]

Technological and emerging innovations in nanoparticle-based vaccine research

Technological innovation and preclinical evaluation

Ongoing advances in nanoparticle-based vaccine research continue to highlight the transformative potential of these systems across preventive and therapeutic medicine. Nanoparticles are increasingly incorporated into modern vaccine platforms because of their capacity to improve antigen delivery, enhance immune activation, and increase formulation precision [32]. Recent progress has also strengthened vaccine stability, immunogenicity, and safety profiles. In this context, advanced microfluidic technologies are gaining importance in preclinical evaluation, as they allow the simulation of physiological conditions in vitro and support the prediction of nanoparticle behavior, including biodistribution, degradation, and immune activation, before progression to animal or human studies [191]. In addition to platform design, analytical characterization has become increasingly important for translational development. Techniques such as dynamic light scattering are widely used to assess particle size, size distribution, and colloidal stability, all of which directly influence vaccine performance, reproducibility, and regulatory readiness [192].

In parallel, membrane-coated nanovaccines have emerged as promising candidates, particularly in cancer immunotherapy. These systems incorporate cellular membranes or tumor-associated components onto nanoscale carriers and are being investigated for their ability to suppress tumor progression and metastasis while simultaneously stimulating potent immune responses [193]. Notable examples include exosome-based vaccine systems, which are under early clinical evaluation in several malignancies and may improve both tumor targeting and immune activation [194].

In oncology, nanoparticle-based delivery systems are also attracting considerable attention for both immunization and targeted drug delivery. In lung cancer models, for example, liposomes and polymeric nanoparticles have been used to transport chemotherapeutic or immunomodulatory agents directly to tumor tissue, thereby improving treatment precision and reducing systemic toxicity [195]. Gold nanoparticles and quantum dots are likewise being explored because of their structural and functional properties, which facilitate the delivery of drugs or antigens to tumor cells and their surrounding microenvironment. Liposomal formulations combined with immune checkpoint inhibitors have also entered clinical investigation, reflecting the growing translational relevance of these nanosystems [196].

Animal models remain essential for validating the efficacy, safety, and immunogenicity of nanoparticle-based vaccines. Commonly used species include rodents, rabbits, sheep, pigs, dogs, and non-human primates such as macaques and African green monkeys [197]. Small animals such as mice and guinea pigs remain particularly valuable because of their relatively low maintenance costs and well-characterized immune systems. However, model selection must be aligned with disease biology to preserve translational relevance [198]. Tuberculosis is a clear example, since standard laboratory mice do not fully recapitulate classical TB pathology. In properly selected animal models, encapsulation of vaccine antigens into nanoparticles or microparticles has repeatedly been associated with stronger immune responses and improved protective efficacy compared with conventional formulations [199]. PLGA nanoparticles, for instance, have shown promise in enhancing the immunogenicity of tuberculosis vaccines in multiple animal systems [200].

Self-assembling peptide nanovaccines and virus-like particles continue to represent important innovations within the preclinical pipeline. Self-assembling peptide systems organize into nanoscale architectures that improve dendritic cell uptake, support B-cell activation, and promote antigen transport to lymphoid tissues [201]. These properties can generate robust and durable immune responses against diverse infectious agents in animal models. Recent studies have demonstrated strong immune responses against pathogens such as HIV and influenza. Similarly, virus-like particles remain attractive as non-replicating vaccine platforms, with preclinical evidence supporting their effectiveness against SARS-CoV-2 and other infectious agents [202]. Clinical studies are also continuing to evaluate VLP-based approaches for hepatitis B and human papillomavirus, underscoring their broad utility in vaccine development [203].

Innovative cargoes and platforms

Cargo types

Recent developments in nanotechnology have introduced a diverse set of nanoscale architectures that enhance both vaccine delivery and immune modulation [204]. Within these systems, the nature of the cargo plays a central role in determining biological performance. Broadly, nanoparticle cargoes may include antigens, nucleic acids, and adjuvants [205].

Antigens can be encapsulated within nanocarriers to improve their stability, preserve structural integrity, and facilitate targeted delivery to immune cells [206]. Platforms such as nanocages and dendrimers are particularly suitable for this purpose because they permit controlled release and surface customization. Nucleic acid cargoes, including RNA-based constructs, have also gained substantial attention. Dendrimer-RNA systems, for example, have shown promise in inducing durable immune responses against viral and parasitic pathogens [207]. Adjuvant incorporation represents another major application. By co-delivering or conjugating immunostimulatory molecules, nanocarriers can amplify immune activation and improve vaccine efficacy. Dendrimer-based systems, in particular, have been explored as scaffolds for adjuvant presentation and controlled immunomodulation [208]. This translational relevance is already evident in lipid nanoparticle formulations used for mRNA delivery, where efficient encapsulation, protection of nucleic acid cargo, and intracellular release are central determinants of vaccine performance. Likewise, adjuvant selection remains critical for shaping both the magnitude and durability of immune responses, particularly in formulations designed for rapid adaptation to emerging pathogens [208, 209].

Emerging platforms

Among the emerging platforms, nanocages have attracted considerable interest. These hollow, porous nanoscale structures can encapsulate and release therapeutic or biological molecules in a controlled manner [210]. Their large surface area, tunable pore size, and modifiable surface chemistry allow careful control over drug loading, stability, and release kinetics. Nanocages may be organic or inorganic in composition. Organic nanocages, including protein-based or DNA-based structures, are generally biocompatible and interact minimally with living cells, thereby limiting toxicity [211]. Inorganic nanocages, such as those based on gold, silica, or carbon, often provide greater structural stability and can incorporate imaging or magnetic reporter components, which expands their theranostic potential [212, 213].

Dendrimers represent another important class of nanocarriers. These highly branched nanosized polymers possess structural uniformity, polyvalent surface groups, high solubility, and relatively low intrinsic toxicity, making them useful scaffolds for conjugating antigens, nucleic acids, and adjuvants [214]. Dendrimer-based carriers have demonstrated improved delivery efficiency and, in some experimental settings, reduced dose requirements. Dendrimer-RNA formulations have shown durable immune protection against influenza, Ebola, and Toxoplasma gondii in animal studies [215]. More broadly, surface engineering strategies such as PEGylation remain important for improving nanoparticle circulation time, steric stabilization, and resistance to premature immune clearance. These design principles continue to inform vaccine-oriented nanocarrier development (Fig. 3). More generally, modular platform design has become a major translational advantage in nanovaccine development, because it allows rapid substitution of antigen cargo while preserving a validated carrier architecture. This flexibility is especially important in outbreak settings, where speed of reformulation can strongly influence public health responsiveness [216].

Fig. 3.

Summary of nanoparticle design strategies for the effective delivery of small molecules and biologics. A Structural engineering enables modulation of release kinetics through stimulus-responsive, porous, and core–shell designs. B Amphiphilic nanomaterials support combination therapy by enabling the co-delivery of hydrophobic and hydrophilic agents. C Nanoengineered biomaterials can overcome biological barriers, including extracellular and intracellular constraints, thereby facilitating cellular uptake and intracellular release. D Nanoparticle systems may also help overcome biological resistance mechanisms, including P-glycoprotein-mediated efflux, while supporting gene silencing, organelle targeting, and apoptosis induction in cancer cells. E Cell membrane coating promotes immune evasion, prolongs circulation, and enhances homotypic targeting. F Encapsulation and nanoparticle-biomolecule interactions protect biologics against environmental stress and denaturation. G In addition, nanoparticles can support in situ vaccination by promoting tumor antigen release, immune priming, T-cell recruitment, and accumulation at the target site [217]

Spherical nucleic acids are another notable example of structural nanomedicine. These systems consist of a nanoparticle core surrounded by a dense shell of radially oriented oligonucleotides, including DNA, RNA, or related analogs [218]. Their modular design allows precise control over size, composition, conjugation strategy, and oligonucleotide arrangement, which makes them versatile platforms for diagnostics and therapeutics. They have already shown potential in DNA- and mRNA-based diagnostics as well as in antisense and RNA interference applications [218, 219].

Outer membrane vesicles are naturally released spherical vesicles derived from the outer membrane of Gram-negative bacteria. They reproduce many antigenic features of bacterial surfaces while lacking replicative capacity, which makes them attractive as immunostimulatory but non-pathogenic platforms [220]. OMVs can be engineered to display antigens of bacterial, viral, or tumor origin and are capable of activating both innate and adaptive immunity. Their pathogen-like surface architecture enhances immune recognition, although issues such as proteolytic degradation, antigen misfolding, and suboptimal presentation remain under active investigation [221, 222].

Metal–organic frameworks have also emerged as highly tunable carriers for vaccine antigens. These porous crystalline materials are formed through coordination between metal ions and organic linkers, creating structures capable of controlled encapsulation and release of biomolecules [223]. In vaccine applications, MOFs have been used to deliver viral antigens such as the SARS-CoV-2 spike protein, where they appear to enhance innate immune activation and cytokine production through Toll-like receptor-associated pathways [224].

Nanogels provide another flexible platform. These soft, water-swollen polymer networks can encapsulate antigens or adjuvants while maintaining high biocompatibility. Their cross-linked architecture supports sustained release and can be engineered for selective immune-cell targeting [225]. Because nanogels can also be loaded with multiple antigens, they offer potential for multivalent vaccine designs that provide protection against several pathogens simultaneously [226].

Bacterial and probiotic spore-based carriers also deserve inclusion among emerging platforms because of their promise in mucosal immunization. Spores, especially those derived from Bacillus species, are naturally resistant to harsh environmental conditions and therefore provide durable vehicles for antigen presentation [227]. Through recombinant or non-recombinant approaches, multiple antigens have been displayed on spore surfaces, resulting in strong mucosal and systemic immune responses [227]. Probiotic-based spores further combine carrier stability with the immunomodulatory properties of beneficial microbes, which makes them especially attractive for oral vaccination strategies that aim to reduce cold-chain dependence and manufacturing costs [228].

Finally, combinational nanovaccines are being developed to integrate multiple antigens and adjuvants within a single nanoparticle formulation. These systems are intended to broaden immune coverage and increase protective efficacy [229]. They have been explored in contexts such as influenza, tuberculosis, cancer, and HIV, where cross-reactive or multipronged immune responses may be desirable. At the same time, such formulations also introduce added challenges, including formulation instability, antigen interference, and more complex safety assessment requirements [230].

Challenges and future directions in nanoparticle-based vaccines

Regulatory and quality assurance considerations

The regulation of NPs-based vaccines presents unique challenges due to their complex physicochemical nature and wide-ranging applications across both medical and non-medical sectors [231]. Regulatory authorities such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), and the National Medical Products Administration (NMPA) oversee their approval within existing legal frameworks for biologics and medical devices [30]. For nanomedicine, these agencies require comprehensive evaluations of safety, quality, and efficacy, including preclinical studies, pharmacokinetic analyses, and large-scale clinical trials. Post-marketing surveillance also plays a critical role in confirming continued safety after product approval [232, 233].

For vaccines incorporating nanoparticles, additional assessments focus on antigen selection, formulation stability, release kinetics, and biodistribution [231]. However, the challenges encountered can vary significantly across different types of nanoparticles, which necessitates tailored regulatory approaches [164].

Lipid-based nanoparticles, for instance, are known for their role in mRNA vaccine delivery. They face specific challenges concerning cold-chain storage requirements and potential instability [231]. Regulatory bodies require rigorous testing to assess the integrity of lipid compositions and their ability to protect mRNA from degradation [234]. Additionally, the potential for lipotoxicity and immunogenicity must be evaluated, particularly since lipid nanoparticles interact closely with biological membranes, which can lead to unintended side effects [235].

Polymeric nanoparticles pose their own set of regulatory hurdles. Their synthesis often includes various polymers, which can differ significantly in biocompatibility and toxicity profiles. Regulatory agencies need for thorough valuations of the polymers used, particularly when they are derived from synthetic sources [236]. The variability in degradation rates and release profiles of encapsulated antigens necessitates wide pharmacokinetic studies to ensure consistent therapeutic outcomes. Also, the complex manufacturing processes involved in polymeric nanoparticles can complicate scalability and product consistency, which are critical attentions for regulatory approval [237].

Inorganic nanoparticles, such as silica or AuNPs, bring another dimension to regulatory challenges. Their unique optical and electronic properties may present risks that differ from those of organic nanoparticles [238]. For instance, concerns about heavy metal toxicity or the formation of reactive oxygen species during degradation require comprehensive toxicological evaluations [239]. Additional studies focusing on the nanoparticles’ behavior in biological systems including their biodistribution, long-term stability, and potential for accumulation in non-target organs, are needed to meet safety standards [240].

VLPs and self-assembling peptide nanostructures further complicate the regulatory landscape. VLPs, while presenting lower risks of infection, demand thorough examination of their structural integrity and stability since they are often derived from complex viral proteins [241]. Regulatory authorities must ensure that VLPs maintain their immunogenicity through production, storing, and administration. In the case of peptide nanostructures, challenges arise around confirming consistent assembly and uniformity during production, which are critical for effectiveness [242].

For all NPs types, standardized methods for evaluating immune responses and long-term safety remain essential. Each stage ranging from Phase I safety trials to Phase III efficacy studies must address the specific behavior of nanoparticles within biological systems to meet regulatory standards [243]. Addressing these risks through thorough toxicological evaluations is essential before clinical deployment. Emerging safety frameworks, as outlined by major regulatory organizations, aim to ensure that these vaccines adhere to rigorous safety benchmarks [244].

Also, the maintenance of cold-chain logistics is crucial for the effective distribution of these vaccines, particularly in lower-resource environments. Addressing these logistical constraints will be key to ensuring global access to these advanced vaccine technologies [245].

Global regulatory harmonization and social considerations

An important challenge in the field of nanoparticle-based vaccines is achieving constant global regulatory standards. Different regions apply varied regulatory frameworks, which can lead to inconsistencies in testing protocols, safety thresholds, and manufacturing requirements [246]. Efforts by international organizations, including the WHO, are crucial in striving for unified approaches to ensure equitable access and reliable quality across diverse regions [247].

In addition to regulatory challenges, public perception and vaccine hesitancy remain substantial barriers to widespread acceptance of nanoparticle-based vaccines. Factors contributing to these issues include misinformation, cultural beliefs, and unequal access to healthcare services [248]. Enhancing public education and fostering transparency in scientific communication, combined with community engagement, are vital to building trust and encouraging acceptance of advanced vaccine technologies [249].

Manufacturing and scalability challenges

One of the serious challenges in developing nanoparticle-based vaccines is the intricacy of nanoparticle design, which complicates large-scale production [250]. Effective manufacturing requires precise control over critical parameters like size, shape, surface charge, and purity to maintain reproducibility [251]. Top-down approaches, such as milling and lithography, can lead to contamination and uneven particle size distribution. Meanwhile, bottom-up techniques, counting sol–gel synthesis and self-assembly, usually yield better uniformity but require sophisticated manufacturing infrastructure [251].

Scalability also presents potential financial and technical challenges. Production often demands high-grade raw materials, sterile conditions, and specialized equipment, with even trace impurities able to alter nanoparticle behavior. Recent research has focused on streamlining manufacturing processes to enhance yield, consistency, and cost efficiency [232].

Advances in scalable nanoparticle production

Recent novelties are underway to optimize the synthesis of nanoparticles, utilizing automation, high-throughput screening, and precision engineering [250]. New engineering methods, such as the Silicon Scalable Lipid Nanoparticle Generation platform, have appeared to allow large-scale, reproducible lipid nanoparticle production. These advancements became especially significant during the COVID-19 pandemic, facilitating rapid vaccine deployment [233].

Analytical technologies such as electron microscopy, atomic force microscopy, and dynamic light scattering are employed to characterize nanoparticle morphology and distribution. Establishing standardized synthesis and testing protocols is essential to ensure consistent performance across the board [252]. The implementation of automated production systems, stringent process controls, and real-time quality monitoring will minimize human error, maintaining product reliability [233].

Quality control and preclinical assessment

Quality assurance is a fundamental concern in the development of NP-based vaccines. National and international pharmacopeias, including the U.S. Pharmacopeia, have recognized analytical guidelines to verify nanoparticle identity, purity, and composition [253]. For mRNA vaccine formulations, further standards regarding encapsulation efficiency, impurity profiles, and content uniformity have been developed [254].

Preclinical evaluations encompass assessments of physicochemical parameters, stability, immunogenicity, and toxicity in both in vitro and in vivo contexts. These studies are vital for understanding how nanoparticles interact with biological systems and lay the groundwork for regulatory approval [255]. Through adherence to rigorous safety and quality frameworks, researchers and manufacturers can foster regulatory confidence and facilitate the transition of nanovaccines from laboratory to clinical application [256] (Table 3).

Table 3.

Challenges and key strategies in nanoparticle vaccine development

Challenge category	Underlying issue	Impact on vaccine development	Proposed or emerging solutions	Expected outcome	Refs
Manufacturing and scalability	Maintaining uniform nanoparticle size, charge, and purity during mass production	Batch variability; reduced reproducibility	Automation, microfluidic synthesis, scalable lipid NP generation	Consistent, cost-effective vaccine production	[257–259]
Safety and toxicity	Potential accumulation in organs, oxidative stress, or DNA damage	Inflammatory or genotoxic responses	Rigorous toxicological testing; biocompatible materials (PLGA, chitosan)	Improved biosafety and reduced systemic toxicity	[260, 261]
Cold chain and stability	Sensitivity of lipid NPs to temperature fluctuations	Limited global distribution, especially in low-resource settings	Thermostable formulations, lyophilization, AI-predicted stability optimization	Expanded accessibility and longer shelf-life	[262, 263]
Regulatory approval	Lack of standardized evaluation for nanoparticle-based products	Delayed clinical translation and commercialization	Harmonized global guidelines (FDA, EMA, WHO frameworks)	Faster, transparent approval processes	[253, 264]
Public acceptance	Vaccine hesitancy, misinformation, and cultural barriers	Lower vaccination coverage, inequity	Community education, ethical communication, equitable distribution	Increased public trust and acceptance	[265, 266]

Artificial intelligence and computational design in nanovaccine development

The expanding role of AI in vaccine innovation

Artificial intelligence is reforming the landscape of nanoparticle-based vaccine design by integrating computational intelligence into every stage of expansion, including theoretical design and formulation, testing, and clinical translation [266]. Over machine learning and deep learning algorithms, AI systems can process vast datasets of immunological, genomic, and structural information, allowing researchers to identify optimal antigen targets, predict immune responses, and accelerate candidate selection with unprecedented precision [267, 268]. These advancements are not limited to lipid-based platforms; AI techniques are being applied across a variety of NPstypes, including polymeric nanoparticles, AuNPs, and protein-based nanocages, enhancing the effectiveness of each class in its application for immunization [269].

Optimization of nanoparticle properties using AI

AI technologies are also being engaged to optimize the physicochemical characteristics of various nanoparticle platforms such as size, surface charge, shape, and composition which are critical factors influencing vaccine delivery and immunogenicity [267]. For instance, while lipid nanoparticles have shown great promise in delivering mRNA vaccines, polymeric nanoparticles offer advantages in stability and controlled release [270]. AI-driven showing can predict how these parameters interact with biological systems, enabling the development of nanoparticles that achieve efficient antigen delivery, minimal toxicity, and controlled release profiles across multiple platforms. This flexibility improves the ability to tailor vaccines for specific diseases or patient demographics [271].

In vaccine formulation, machine learning models have confirmed exceptional capacity in mapping complex relationships between formulation mechanisms and performance outcomes across different types of NPs [272]. For instance, predictive algorithms can analyze how variations in polymer ratios or different coating materials affect the stability and encapsulation efficiency of polymeric or gold nanoparticles [273]. As a outcome, researchers can design formulations that are both thermally stable and suitable for large-scale manufacturing across various NP types, effectively addressing one of the key bottlenecks in nanovaccine production [32].

Predicting immune responses and safety profiles

Additional vital function of AI in vaccine science is the prediction of immune outcomes and potential adverse events across different nanoparticle platforms. AI models trained on extensive immunological datasets can anticipate how specific formulations whether lipid-based, polymeric, or metallic will behave in vivo. This capability guides the selection of the safest and most effective candidates before human testing [274]. Furthermore, AI-assisted tools used to model cytokine release patterns, antibody titers, and cell-mediated immune responses allow researchers to fine-tune formulations for desired immunological results regardless of the fundamental nanoparticle technology [275].

Integration of big data and AI for personalized nanovaccines

The convergence of artificial intelligence with big data analytics is paving the way for highly individualized vaccine strategies. By analyzing patient-specific data, including genomic sequences, immune profiles, and environmental factors, AI can support the design of truly personalized nanovaccines tailored to each individual’s immunological makeup [276]. This approach has the potential to maximize vaccine efficacy across diverse populations and minimize adverse effects associated with generalized immunization strategies [277].

However, achieving this vision requires overcoming several challenges, such as ensuring the availability of large, diverse, and high-quality datasets, validating AI-generated predictions in biological systems, and establishing transparent regulatory frameworks for AI-driven biomedical research [278]. Continued collaboration between computational scientists, immunologists, and clinicians will be critical to advancing the safe and ethical use of AI in vaccine development [279].

The future of AI-driven nanovaccine design

Looking ahead, AI is expected to become an indispensable component of vaccine innovation. The integration of predictive modeling, automation, and high-throughput screening could allow the real-time optimization of nanoparticle synthesis and vaccine performance [280]. These systems will enable rapid response to emerging infectious threats by streamlining the design of antigen constructs, delivery systems, and adjuvant combinations within days rather than months [281].

Ultimately, the fusion of artificial intelligence and nanotechnology promises to revolutionize global immunization strategies. As these technologies mature, they will contribute to developing safer, more effective, and widely accessible vaccines, marking a new era in preventive medicine driven by precision, personalization, and predictive science [282] (Table 4).

Table 4.

Integration of artificial intelligence and nanotechnology in vaccine innovation

AI application area	Function in nanovaccine development	Advantages of AI integration	Examples of application	Future outlook / expected impact	Refs
Antigen and epitope prediction	Identify optimal immune targets from genomic/proteomic databases	Reduces time and cost of antigen discovery	Prediction of SARS-CoV-2 spike epitopes using deep learning	Rapid vaccine design adaptable to emerging variants	[283–285]
Nanoparticle optimization	Model relationships between particle properties and biological outcomes	Enhances delivery efficiency and minimizes toxicity	AI-tuned lipid compositions for stable mRNA vaccines	More precise and stable nanocarrier formulations	[286, 287]
Immune response forecasting	Predict cytokine profiles, antibody titers, and T-cell activation	Minimizes adverse reactions, optimizes dosage	AI simulation of immune response dynamics in preclinical trials	Personalized immune modeling for tailored vaccination	[288, 289]
Manufacturing automation	Monitor and control production variables in real time	Improves reproducibility and reduces human error	Automated microfluidic NP fabrication platforms	Scalable and consistent large-scale vaccine output	[290, 291]
Personalized nanovaccine design	Integrate genomic, clinical, and environmental data for individualized vaccines	Maximizes efficacy and minimizes side effects	mRNA-4157 and fluoropolymer-based cancer nanovaccines	Precision immunization and personalized global health	[291, 292]

Conclusion

Nanoparticle-based vaccines represent a major advancement in immunology, offering a powerful alternative to conventional vaccination strategies. Their unique physicochemical properties enable precise antigen delivery, enhanced immunogenicity, and sustained immune activation, while their versatility allows for the incorporation of multiple antigens and adjuvants into a single formulation. These features not only improve vaccine efficacy but also simplify administration and broaden protective coverage against complex or rapidly evolving pathogens. Moreover, the ability to design nanoparticles that mimic natural infection processes supports balanced humoral and cellular immune responses. Despite these advantages, translating nanovaccine technologies into clinical use still faces several challenges, including large-scale production, long-term stability, regulatory approval, and the management of potential toxicity or immunogenic side effects. The future of nanovaccine development lies in the integration of nanotechnology with emerging tools in artificial intelligence, bioengineering, and computational modeling. These interdisciplinary approaches will accelerate antigen discovery, optimize nanoparticle design, and enable the creation of personalized vaccines tailored to individual immune profiles. As research advances, nanoparticle-based vaccines are expected to become a cornerstone of preventive medicine, offering new solutions for infectious diseases, cancers, and chronic conditions. Their continued evolution promises not only to strengthen global immunization programs but also to promote equitable, effective, and sustainable healthcare outcomes worldwide.

Abbreviations

ABC

ATP-binding cassette (transporter superfamily)

ABCB6/7/8/10

ATP-binding cassette subfamily B member 6/7/8/10 (mitochondrial/inner-membrane transporters)

AEC-II

Type II alveolar epithelial cell

AI

Artificial intelligence

APP

Amyloid precursor protein

ARE

Antioxidant response element

ATP

Adenosine triphosphate

BAX

Bcl-2–associated X protein

BBB

Blood–brain barrier

Bcl-2

B-cell lymphoma 2

BDNF

Brain-derived neurotrophic factor

BPD

Bronchopulmonary dysplasia

CKD

Chronic kidney disease

CRISPR

Clustered regularly interspaced short palindromic repeats (genome editing system)

CSF

Cerebrospinal fluid

DANCR

Differentiation antagonizing non-protein coding RNA

DC

Dendritic cell

DIC

Disseminated intravascular coagulation

DLCO

Diffusing capacity of the lung for carbon monoxide

DNA

Deoxyribonucleic acid

DOX

Doxorubicin

EBV

Epstein–Barr virus

EMA

European Medicines Agency

EV / EVs

Extracellular vesicle(s)

FDA

(U.S.) Food and Drug Administration

GDNF

Glial cell line–derived neurotrophic factor

GFR

Glomerular filtration rate

GPX

Glutathione peroxidase

GPC1

Glypican-1

H2O2

Hydrogen peroxide

hAEC-EVs

Human amniotic epithelial cell–derived extracellular vesicles

HIF-1α

Hypoxia-inducible factor-1 alpha

HIV

Human immunodeficiency virus

HIE

Hypoxic–ischemic encephalopathy

HPV

Human papillomavirus

HSFs

Heat shock factors

HSP / HSP70/HSP72

Heat shock protein(s) / 70-kDa family / inducible 72-kDa form

IKK

IκB kinase

IL

Interleukin

IL-1β

Interleukin-1 beta

IL-1RA

Interleukin-1 receptor antagonist

IL-6

Interleukin-6

IL-10

Interleukin-10

ISEV

International Society for Extracellular Vesicles

iPSC(s)

Induced pluripotent stem cell(s)

LNP(s)

Lipid nanoparticle(s)

MALAT1

Metastasis-associated lung adenocarcinoma transcript 1

MAPK

Mitogen-activated protein kinase

MEVs

Milk/breast-milk–derived extracellular vesicles

MET

Mesenchymal–epithelial transition factor (c-MET)

miRNA(s)

MicroRNA(s)

MISEV

Minimal Information for Studies of Extracellular Vesicles (ISEV guidelines)

MOF(s)

Metal–organic framework(s)

mRNA

Messenger RNA

MPTP

Mitochondrial permeability transition pore

MRP1/2/7

Multidrug resistance-associated protein 1/2/7 (ABC transporters)

MSC(s)

Mesenchymal stem cell(s)

MSC-EVs

Mesenchymal stem cell–derived extracellular vesicles

NAC

N-acetylcysteine

NEC

Necrotizing enterocolitis

NF-κB

Nuclear factor kappa-light-chain-enhancer of activated B cells

NMPA

National Medical Products Administration (China)

NRF1/NRF2

Nuclear respiratory factor 1/2 (mitochondrial biogenesis regulators)

Nrf2

NF-E2–related factor 2 (antioxidant master regulator)

OMV(s)

Outer membrane vesicle(s)

PCL

Polycaprolactone

PGC-1α

Peroxisome proliferator-activated receptor-γ coactivator-1 alpha

PEG

Polyethylene glycol

PI3K/Akt

Phosphoinositide 3-kinase / protein kinase B pathway

piRNA(s)

PIWI-interacting RNA(s)

PLGA

Poly(lactic-co-glycolic acid)

PPARγ

Peroxisome proliferator-activated receptor gamma

PTEN

Phosphatase and tensin homolog

RDS

Respiratory distress syndrome

RNA

Ribonucleic acid

ROS

Reactive oxygen species

SAPN(s)

Self-assembling peptide nanovaccine(s)

SARS-CoV-2

Severe acute respiratory syndrome coronavirus 2

siRNA(s)

Small interfering RNA(s)

SOD

Superoxide dismutase

STAT3

Signal transducer and activator of transcription 3

STING

Stimulator of interferon genes

TFAM

Mitochondrial transcription factor A

Th1/Th2/Th17

T-helper cell subsets 1/2/17

TGF-β

Transforming growth factor beta

TLR-4

Toll-like receptor 4

TNF-α

Tumor necrosis factor alpha

Treg

Regulatory T cell

USP14

Ubiquitin-specific peptidase 14

VEGF

Vascular endothelial growth factor

VLP(s)

Virus-like particle(s)

WHO

World Health Organization

Species shorthand

A. muciniphila

Akkermansia muciniphila

L. plantarum

Lactiplantibacillus plantarum

Author contributions

Iman morshedi 1, Zahra roodaki 2, Pejman gheibi 3, Ghazal.zamani 4, Amirhossein.jaberi 5, JaafarAlsadeq Hasan 6, Shams Chaloob 7, Mohammad Teymourzadeh 8*, Sepideh KarkonShayan 9*: Conceptualization, Writing—original draft, Writing—review & editing and Supervision.

Funding

No funding.

Data availability

Data sharing not applicable to this article as no datasets were generated or analyzed during the current study.

Declarations

Ethical approval and consent to participate

Not applicable.

Consent for Publication

Not applicable.

Competing Interest

The authors declare no competing interests.