Progress and challenges of lyotropic liquid crystalline nanoparticles for innovative therapies

https://doi.org/10.1016/j.ijpharm.2022.122299Get rights and content

Abstract

Since the late 20th century, we have witnessed a growing and substantial advance in nanomedicine, in part due to the development of multifunctional and multimodal nanoplatforms that have enabled improved efficacy, biocompatibility, and novel therapeutic applications. Non-lamellar liquid-crystalline nanoparticles, especially, reverse hexagonal and cubic bicontinuous mesophases, have gained the attention of the scientific-academic community due to their intriguing and functional characteristics, such as self-organization into two- and three-dimensional supramolecular structures, high symmetry, and ability to accommodate hydrophobic and hydrophilic small molecules, peptides, proteins, nucleic acids, and imaging agents. Furthermore, these particles can be easily modified with specific and/or bioresponsive molecules allowing targeting and improved therapeutic performance. In this contribution we provide an overview of advances in the design and architecture of LCNPs, strategies to overcome biological barriers and main findings about interactions with different types of interfaces. We highlight recent applications in topical, oral, pulmonary and intravenous drug delivery in preclinical in vivo studies. We discussed the current scenario and translational obstacles faced for clinical translation, as well as our perspectives.

Introduction

Nanoparticles as drug delivery systems and theranostic agents are now a reality due to their potential to increase the solubility of drugs and deliver them to a specific tissue (Shepherd et al., 2021). Among the various compositions of nanoparticles, those formed from an intermediate state between solid and liquid (Liquid Crystalline Nanoparticles - LCNPs) have attracted much attention in recent decades and are used in various fields, including nanomedicine (Waghule et al., 2021a). However, the translation of research findings into the clinic still faces limitations (Zhai et al., 2019) due to a combination of factors, particularly the search for an efficient design of nanoparticles and their complex biological interactions (Madheswaran et al., 2019). Understanding the transport mechanisms of nanoparticles across various biological barriers (Fig. 1) is critical for the development of safe and effective nanoscale delivery systems. Physicochemical parameters of nanoparticles such as particle size, dispersion, surface charge, functionalization, chemical composition, and others determine their destination and toxicity. Recently, Tejashree Waghule et al. (2021), investigated how the application of quality by design to liquid crystalline nanoparticles will provide a better understanding of products and biological processes and accelerate the transfer of this technology from bench to bedside for treatment of various diseases (Waghule et al., 2021a).

Nanoparticles applied topically to the skin and eye pass through the skin-, corneal- and retinal blood barriers, and nanoparticles given orally go through the intestinal barrier to enter the bloodstream (J. Jia et al., 2020a; Mitchell et al., 2021). However, the biological barrier function of the skin, eye, and mucosa has the disadvantage of limited drug uptake and impaired drug permeation and retention. The improved drug penetration/retention of LCNPs has been confirmed to overcome this disadvantage (Bessone et al., 2021, Nguyen et al., 2011, Ramirez et al., 2021, Silvestrini et al., 2020, Teba et al., 2021).

The skin is responsible for the barrier function of the body. It keeps homeostasis in balance and prevents microorganisms and foreign bodies from entering our body. Each layer of skin has different types of specialized cells that are in different stages of maturation. For example, the stratum corneum (SC), the outermost layer that contains dead, keratinized, and parallel-stacked cells, has a rigid and lipidic structure that prevents the penetration of hydrophilic and especially hydrophobic drugs with high molecular weight (Matsui and Amagai, 2015). In the epidermis, keratinocytes at different stages of keratinization form a uniform and parallel structure (Bouwstra et al., 2002). Mechanistic studies have shown that LCNPs are able to induce a temporary disorganization in the SC structure, creating small holes for the penetration of drug molecules allowing them to reach the deeper layers of the epidermis. In the epidermal layers, LCNPs have the property of fusing with cellular lipids and thus can better interact with the intercellular lipids of the skin matrix and minimize the skin barrier effect (Madheswaran et al., 2014, Uchino et al., 2019).

In addition to aspects such as improved preocular retention and reduced ocular irritation, LCNPs may improve bioavailability by facilitating transcorneal/transconjunctival penetration. Barriers to ocular administration can be classified as: (i) static (corneal epithelium, endothelium, sclera, and conjunctiva), dynamic (blink reflex, lacrimal flow, and nasolacrimal drainage), (ii) metabolic (activation of phase I and phase II enzymes such as cytochrome p450, monoamine oxidase, lyosomal enzymes, and efflux protein), and (iii) intraocular microenvironment (blood-retinal barrier) (Rodrigues et al., 2020). Although developed nanoparticles (polymers, liposomes, and others) have been proposed as drug delivery systems for ocular use, rapid clearance from the precorneal region and rapid drainage from aqueous eye drops have been shown to limit most of these systems, resulting in low preocular retention and bioavailability (Lyu et al., 2021). On the other hand, several authors have consistently concluded that LCNPs adhere better to the corneal/conjunctival surface due to their mucoadhesive properties, small particle size, and increased surface area (Bessone et al., 2021, Ding et al., 2021, Kaul et al., 2021, Teba et al., 2021).

The mucoadhesive properties of LCNPs also favor oral administration, as they overcome the barrier of the oral mucosa. In addition, the use of lipids with low digestibility favors the retention and controlled release of the drug. The arrangement of lipid molecules gives LCNPs a rigid and fluid distinction that favors secondary transformations such as mixed micelles and liquid crystalline phase nanoparticles in the gastrointestinal environment (Rizwan et al., 2009). These secondary structures have properties such as good stability, sustained drug release and improved oral absorption, drug protection, and increased penetration. Therefore, incorporation of drugs into LCNPs is an efficient strategy to improve the oral bioavailability of insulin (Agrawal et al., 2017, Chung et al., 2002), antifungals (Xu et al., 2014) (Jain et al., 2018), antitumor (Swarnakar et al., 2014), antiplatelet (El-Laithy et al., 2019) and neuroprotective drugs (Elnaggar et al., 2015).

The major biological barriers that LCNPs must overcome after intravenous injection and during their transport to target cells are the andothelial barrier and the corona protein. However, how well they reach target cells or tissues depends on their behavior in the bloodstream. Nanoparticles circulate throughout the body after intravenous injection and accumulate passively in the target tissue through the permeation and retention effect (PRE), which prevents their elimination by the immune system (Tan et al., 2019a). Previous studies have shown that some properties of nanoparticles, such as size, shape, polydispersity, and surface chemistry, can be engineered to enhance their performance in biological systems (Fullstone et al., 2015, Shepherd et al., 2021). This affects cell interaction between the immune system and nanoparticles, modifies the blood clearance profile and target cell interaction, and thus supports effective drug delivery to cells or tissues (Mitchell et al., 2021). The prolonged circulation time and hemocompatibility of PEGylated LCNPs were confirmed almost a decade ago (Jain et al., 2015, Nisha et al., 2021, Zhai et al., 2019). PEGylated LCNPs were able to decrease the rate of LCNPs clearance by Kupffer and macrophage cells as well as protein adsorption, which are known to promote the corona effect and mask the ligands on the surface of the nanoparticles (Zhai et al., 2018). As a result, LCNPs were better delivered to the target tissue. These results are also encouraging for overcoming the blood–brain barrier (BBB), a semipermeable membrane composed of a monolayer of endothelial cells that separates the blood from the central nervous system (Elnaggar et al., 2015, Zhai et al., 2019). This semipermeable membrane often poses a major challenge for drug delivery to the brain because it acts as a physical barrier to the permeability of almost all drugs. In light of this and the confirmed ability of LCNPs to incorporate TweenTM 80, which is known to facilitate BBB crossing, the possibility of using LCNPs for brain therapies was envisioned (Abdelrahman et al., 2015, Azhari et al., 2021, Younus et al., 2018).

Although knowledge of biological barriers and nanotechnology has opened the doors for the development of advanced nanoparticles, the nonspecific distribution of drugs and insufficient accumulation of therapeutics at disease sites still limit the widespread use of nanoparticles in clinical practice. In this issue, we point out that the physicochemical properties of engineered LCNPs are crucial for enhancing biological interactions, accumulating in the target tissue and thus leading to enhanced therapeutic effect. For that, advances in LCNPs architecture and design were deeply discussed. We also highlight recent applications in topical, oral, pulmonary and intravenous drug delivery in preclinical in vivo studies. Together with our perspectives and expert opinions, the current scenario, the progress, and prospects of LCNPs as well as its translational obstacles faced for clinical translation were presented.

Section snippets

Liquid crystalline mesophase and its nanoparticles

Liquid crystals (LC) are an intermediate state of matter formed by amphiphilic molecules that self-organize into one-, two-, and three-dimensional supramolecular structures called mesophases. This state exhibits properties that are intermediate between isotropic liquid and anisotropic solid crystal phases. In particular, the class of lyotropic liquid crystals (LLC) is formed by amphiphilic molecules and solvents (usually water) that assemble into molecular aggregates under certain conditions of

Delivery therapeutic molecules using LCNPs

The versatility and unique characteristics of LCNPs have allowed these platforms to be used in recent years to transport drugs of a different nature and administering them through different routes, wich oral, intravenous, and topical or transdermal. Table 2 summarizes the most recent results from the use of LCNPs for the in vivo delivery of therapeutic agents for a variety of diseases. These studies have proven that the rational design of LCNPs is extremely useful for the targeted and efficient

LCNPs in theranostic applications: Imaging and therapy

By trapping or decorating surfaces with imaging contrast agents, nanoparticles can be used for molecular imaging or be used in combination with a variety of therapies (e.g., chemotherapy, photodynamic therapy, neutron capture therapy, heat therapy, and magnet therapy) Kim et al., 2018a. The use of LCNPs as imaging or theranostic agents is still at an early stage, although important studies have been performed to clarify the uptake capacity of contrast agents and their destination in vivo (Bye

Challenges moving forward: Delivery of nucleic acids and vaccines

LCNPs are potential delivery systems for a variety of drugs, as demonstrated in this article. Advancement in the rational design of these platforms has led to the production of multifunctional LCNPs with highly controllable, bio-responsive, and targetable properties. The success of these LCNPs as drug delivery systems has led to their use for gene and vaccine delivery, particularly after the approval of Onpattro in 2018 and, more recently, COVID-19 vaccines from Moderna and Pfizer/BioNTech,

Future prospectives

LCNPs have evolved into systems with highly controlled geometry, surface charge, and physicochemical properties, and decoration of their surfaces with polymers and bioactive molecules have improved biocompatibility and active targeting. These nanoparticles are becoming multifunctional, multicomponent systems that can be used in a variety of delivery routes and are also biocompatible and biodegradable.

Currently, a reversed non-lamellar liquid crystalline technology platform has been the basis

Reviewer disclosures

Peer reviewers on this manuscript have no relevant financial or other relationships to disclose.

Funding

The research group carries out investigations on lyotropic liquid crystal systems in the framework of the National Institute of Science and Technology in Pharmaceutical Nanotechnology: a transdisciplinary approach INCT-NANOFARMA, which is supported by São Paulo Research Foundation (FAPESP, Brazil) Grant #2014/50928-2, and by ‘‘Conselho Nacional de Desenvolvimento Científico e Tecnológico’’ (CNPq, Brazil) Grant #465687/2014-8. A.V.P.S. and B.W.D are fellowship of “Coordenação de Aperfeiçoamento

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References (260)

  • L. Boge et al.

    Cubosomes post-loaded with antimicrobial peptides: Characterization, bactericidal effect and proteolytic stability

    Int. J. Pharm.

    (2017)
  • L. Boge et al.

    Freeze-dried and re-hydrated liquid crystalline nanoparticles stabilized with disaccharides for drug-delivery of the plectasin derivative AP114 antimicrobial peptide

    J. Colloid Interface Sci.

    (2018)
  • L. Boge et al.

    Cubosomes for topical delivery of the antimicrobial peptide LL-37

    Eur. J. Pharm. Biopharm.

    (2019)
  • R. Brusini et al.

    Advanced nanomedicines for the treatment of inflammatory diseases

    Adv. Drug Deliv. Rev.

    (2020)
  • M. Caillaud et al.

    Small interfering RNA from the lab discovery to patients’ recovery

    J. Control. Release

    (2020)
  • M. Chountoulesi et al.

    Liquid crystalline nanoparticles for drug delivery: The role of gradient and block copolymers on the morphology, internal organisation and release profile

    Eur. J. Pharm. Biopharm.

    (2021)
  • R. Delshadi et al.

    Development of nanoparticle-delivery systems for antiviral agents: A review

    J. Control. Release

    (2021)
  • L.V. Depieri et al.

    RNAi mediated IL-6 in vitro knockdown in psoriasis skin model with topical siRNA delivery system based on liquid crystalline phase

    Eur. J. Pharm. Biopharm.

    (2016)
  • Y. Diebold et al.

    Applications of nanoparticles in ophthalmology

    Prog. Retin. Eye Res.

    (2010)
  • Y. Ding et al.

    Targeted delivery of LM22A-4 by cubosomes protects retinal ganglion cells in an experimental glaucoma model

    Acta Biomater.

    (2021)
  • Y.D. Dong et al.

    Applications of X-ray scattering in pharmaceutical science

    Int. J. Pharm.

    (2011)
  • B.P. Dyett et al.

    Delivery of antimicrobial peptides to model membranes by cubosome nanocarriers

    J. Colloid Interface Sci.

    (2021)
  • Y.E. Elakkad et al.

    Tenoxicam loaded hyalcubosomes for osteoarthritis

    Int. J. Pharm.

    (2021)
  • A.E. Eldeeb et al.

    Formulation and evaluation of cubosomes drug delivery system for treatment of glaucoma: Ex-vivo permeation and in-vivo pharmacodynamic study

    J. Drug Deliv. Sci. Technol.

    (2019)
  • M.A. Elfaky et al.

    Development, Optimization, and Antifungal Assessment of Ocular Gel Loaded With Ketoconazole Cubic Liquid Crystalline Nanoparticles

    J. Pharm. Sci.

    (2021)
  • H.M. El-Laithy et al.

    Stabilizing excipients for engineered clopidogrel bisulfate procubosome derived in situ cubosomes for enhanced intestinal dissolution: Stability and bioavailability considerations

    Eur. J. Pharm. Sci.

    (2019)
  • D.A. Ferreira et al.

    Cryo-TEM investigation of phase behaviour and aggregate structure in dilute dispersions of monoolein and oleic acid

    Int. J. Pharm.

    (2006)
  • C. Fong et al.

    Micellar Fd3m cubosomes from monoolein – long chain unsaturated fatty acid mixtures: Stability on temperature and pH response

    J. Colloid Interface Sci.

    (2020)
  • M. Fornasier et al.

    Cubosomes stabilized by a polyphosphoester-analog of Pluronic F127 with reduced cytotoxicity

    J. Colloid Interface Sci.

    (2020)
  • M.M. Gabr et al.

    Hexagonal Liquid Crystalline Nanodispersions Proven Superiority for Enhanced Oral Delivery of Rosuvastatin. In Vitro Characterization and In Vivo Pharmacokinetic Study

    J. Pharm. Sci.

    (2017)
  • L. Gan et al.

    Self-assembled liquid crystalline nanoparticles as a novel ophthalmic delivery system for dexamethasone: Improving preocular retention and ocular bioavailability

    Int. J. Pharm.

    (2010)
  • Y.A. Garbovskiy et al.

    Liquid Crystalline Colloids of Nanoparticles

    Solid State Physics - Advances in Research and Applications.

    (2010)
  • M. Godlewska et al.

    Voltammetric and biological studies of folate-targeted non-lamellar lipid mesophases

    Electrochim. Acta

    (2019)
  • S. Guillot et al.

    Nanostructured monolinolein miniemulsions as delivery systems: Role of the internal mesophase on cytotoxicity and cell internalization

    Int. J. Pharm.

    (2017)
  • J.E. Harris et al.

    Rapid skin repigmentation on oral ruxolitinib in a patient with coexistent vitiligo and alopecia areata (AA)

    J. Am. Acad. Dermatol.

    (2016)
  • S.Y. Helvig et al.

    Materialia Hexosome engineering for targeting of regional lymph nodes

    Materialia

    (2020)
  • E. Jabłonowska et al.

    Lipid membranes exposed to dispersions of phytantriol and monoolein cubosomes: Langmuir monolayer and HeLa cell membrane studies

    Biochim. Biophys. Acta - Gen. Subj.

    (2021)
  • V. Jain et al.

    Paclitaxel loaded PEGylated gleceryl monooleate based nanoparticulate carriers in chemotherapy

    Biomaterials

    (2012)
  • P. Jelinkova et al.

    Nanoparticle-drug conjugates treating bacterial infections

    J. Control. Release

    (2019)
  • S. Jia et al.

    Visible light-triggered cargo release from donor acceptor Stenhouse adduct (DASA)-doped lyotropic liquid crystalline nanoparticles

    J. Colloid Interface Sci.

    (2019)
  • S. Jia et al.

    Hexaarylbiimidazoles(HABI)-functionalized lyotropic liquid crystalline systems as visible light-responsive materials

    J. Colloid Interface Sci.

    (2020)
  • H.M. Abdelaziz et al.

    Liquid crystalline assembly for potential combinatorial chemo–herbal drug delivery to lung cancer cells

    Int. J. Nanomed.

    (2019)
  • H.M. Aboud et al.

    Novel in situ gelling vaginal sponges of sildenafil citrate-based cubosomes for uterine targeting

    Drug Deliv.

    (2018)
  • M.A.S. Abourehab et al.

    Cubosomes as an emerging platform for drug delivery: a review of the state of the art

    J. Mater. Chem. B

    (2022)
  • A.K. Agrawal et al.

    “Liquid Crystalline Nanoparticles”: Rationally Designed Vehicle To Improve Stability and Therapeutic Efficacy of Insulin Following Oral Administration

    Mol. Pharm.

    (2017)
  • M.Z. Ahmed et al.

    Liquid crystalline nanoparticles for nasal delivery of rosuvastatin: Implications on therapeutic efficacy in management of epilepsy

    Pharmaceuticals

    (2020)
  • N. Alcaraz et al.

    Clickable Cubosomes for Antibody-Free Drug Targeting and Imaging Applications

    Bioconjug. Chem.

    (2018)
  • B. Angelov et al.

    DNA/Fusogenic Lipid Nanocarrier Assembly: Millisecond Structural Dynamics

    J. Phys. Chem. Lett.

    (2013)
  • A. Angelova et al.

    Liquid Crystalline Nanostructures as PEGylated Reservoirs of Omega-3 Polyunsaturated Fatty Acids: Structural Insights toward Delivery Formulations against Neurodegenerative Disorders

    ACS Omega

    (2018)
  • S. Assenza et al.

    Curvature and bottlenecks control molecular transport in inverse bicontinuous cubic phases

    J. Chem. Phys.

    (2018)
  • Cited by (6)

    View full text