Abstract
Biosensors are of significant importance today in the rapid, sensitive, and reliable detection of biological molecules in many fields, including medicine; owing to this fact, the development of a strong and reliable diagnostic agent is a very interesting topic. Because of their unique features, among other nanomaterials, lipid-based vesicles such as liposomes, exosomes, and microvesicles represent a type of biocompatible and versatile biosensing membrane surface for rapid biomarker detection and diagnosis of diseases, enhancing the assay sensitivity and decreasing the detection limit. In this review, we have reviewed the recent diagnostic application of lipid-based vesicles as biosensing substances in both conventional and novel techniques for identifying targets, especially in medicine and biotechnology sciences. Eventually, we have highlighted several recent promising developments in a new generation of biosensors based on liposome–nanomaterial hybrids and exosomes for analyzing targets and possible further advances in the future.
Introduction
Lipid-based vesicles are biophospholipid bilayer vesicles such as liposomes and exosomes, which can be used in diagnosis and therapy. The term “vesicles” is employed particularly for liposomes, which are structures formed spontaneously by polar lipid particles or amphiphilic particles, each having a polar head bunch and a long hydrophobic tail. The interesting focal points of these lipid vesicles are to extend the effectiveness and helpfulness of drugs, increase the soundness of drugs through typifying, decrease the harmfulness of the drugs typified, make a difference to diminish the harmfulness of touchy tissues, and have the capacity to pair with committed ligands to realize focused treatments; liposomes are, moreover, non-toxic, adaptable, biocompatible, biodegradable, and non-immunogenic for systemic or non-systemic organizations (). These interesting properties have driven liposome applications in different areas of nanobiotechnology, chemical and biochemical analytics, nourishment and horticulture businesses, makeup, and quality or medicinal conveyance. In addition to clinical applications, liposomes can also be utilized for explanatory purposes (). The ease of functionalization of the inalienable utilitarian groups of lipids and their capacity to typify receptive atoms in their hydrophilic compartment make it possible to create a detecting substrate for effective bioaffinity detection. So the foremost critical necessity for biosensing is the arrangement of steady vesicles with a tall embodiment capacity and with dynamic destinations for atomic intuition (). Liposomes, combined with successful advances such as fluorescence, calorimetry, and optical spectroscopy, can display expansive flag speakers and accomplish ultrasensitive tests. Biosensors are one of the achievements of the 1960s that have been developed with the advancement of life sciences, chemistry, and engineering over the past few years. Lipid-based vesicle biosensors are analytical tools that can exploit biological intelligence to detect compounds and interact with them. The product of this reaction can be a chemical, optical, or electrical output. On the other hand, their widespread use in diagnostic processes has led to the development of a new generation of these vesicles along with other nanomaterials such as quantum dots, nanotubes, nanodiamonds, and graphene (; ). In this article, we will specifically discuss the application of lipid-based vesicles, such as liposomes, exosomes, and microvesicles, in a variety of biosensors used in medicine (Figure 1).
FIGURE 1
Optical biosensors based on liposomes
In concept, optical biosensors serve as an elective alternative to customary explanatory measures; they are nano-biomolecular devices containing a bio-recognition-detecting component that coordinates with an optical transducer system to deliver a signal relative to the concentration of a measured analyte. Optical biosensing methodologies are basically preferred because of their high sensitivity, robustness, and potential to be coordinated on a single chip without complexity in their pretreatment and probable impact on target atoms (). For a long time, many analysts have connected the optical or fiber-optical tests within the inside fluid environment of liposomes or in screens of conjugates with the liposome layers to make strides in the concentration and sensitivity of the biosensors (). In an interesting study, Zhang et al. illustrated a liposome-based chemosensory by embedding the fluorescent sensors (calcein, spiropyran, and monochlorobimane) into the layer of the liposomes for the specific detection of extracellular analytes such as Zn2+, Ca2+, and GSH for biomedical applications, especially in the IVF setting (). In order to screen particle channel capacities, Szekacs et al. proposed a novel sensor stage for optical waveguide lightmode spectroscopy (OWLS) assays by depositing liposomes containing the particle channels onto the hydrophilic polytetrafluoroethylene (PTFE) layer. The pharmacological performance of the sensor was assessed by observing the movement of Na+ and Cl- particles through gramicidin and GABAA receptor channels individually (). Huang et al. proposed a test chemotherapy agent delivery stage based on the fiber-optic activated topical release of photo-thermally responsive liposomes embedded with Au nanoparticles, employing a 200-µm optical fiber and a 65-mW continuous wave diode laser. The measured pattern of the activated fluorescein discharge from the liposomes illustrated that the AuNP-embedded liposomes appeared to have a more effective activated release than conventional liposomes without AuNPs (). Clear et al. planned a liposome-based nanosensor containing lipid-conjugated optical tests for observing pH in biomedical tests (Figure 2A). In order to fine-tune the sensor pKa, they applied two lipid-anchored pH-sensitive optical tests conjugated within the liposome layer, each with enhanced deep-red cyanine dyes containing indoline nitrogen atoms. They also accomplished the next sensing dynamic extension by changing the liposome layer composition from zwitterionic POPC to anionic POPC: POPS (1:1), which raised the pKa value of the two optical probes by 0.6 and 0.9, respectively (). By taking advantage of PDA properties, Park et al. created coordinate PDA liposome-based multiplex biosensor arrays employing a novel immobilization method for the simultaneous and quantitative detection of six species of pathogens. They chose ethylenediamine as an ideal interlinker between individual PDA liposomes, with a final concentration of 1 mM. Their novel technique improved the signal intensity and sensitivity of PDA chips for bioanalysis (). Liposomes have been connected to SPR biosensors for the characterization of cell membrane-related natural components and drug-induced layer impacts. The interaction between a few drugs and the immobilized liposomes has been checked in real time using the liposomes directly attached to the SPR biosensor surface (; ). The ProteOn GLC Lipid Kit and the Biacore L1 sensor chip are two examples of sensor chips on the market that use covalently connecting alkyl groups to the chip surface to capture liposomes through lipophilic intelligence (Larsson et al., 2020). The ProteOn liposome capturing kit, including the LCP sensor chip and the LCP capturing reagent kit, advertised a novel strategy for liposome capture. The ProteOn XPR36 framework may be a label-free SPR biosensor, with the array 6 × 6 arrangement providing an ideal platform to directly analyze liposomes and small-molecule drugs intelligently. In order to show the effective performance of liposomes on localized surface plasmon resonance (LSPR) detection, Kawasaki et al. examined the biosensing capabilities using the immobilization of liposome or phospholipid single-layer structures on Au nanostructures of LSPR sensor chips. They concluded that the DMPC single layer was more effective in enhancing the sensitivity than the liposomes, probably since its measure was closer to the range of the near-field effect. Their proposed biosensor clearly recognized the 100-nM target protein of carbonic anhydrase from bovine (CAB) with an evaluated detection range of 10 nM from wavelength determination by interaction with the phospholipid single layer in Figure 2B(a, b) (). Liposome-based fluorescence biosensors have become a popular instrument for studying lipid–protein interactions by exploiting the signal amplification method (). Scaglione and Rintoul created a sensitive liposome-based fluorometric biosensor to assess the function of crevice intersections using the liposome containing the fluorescent probe NBD-PE. Quenching of the probe with cobalt chloride permitted the measurement of the permeability of the liposomes in the presence and absence of major intrinsic protein 26 (MIP26) (). Imamura and colleagues created a label-free fluorescently displayed biosensor manufactured with different phospholipid liposomes, encapsulating calcein molecules to separate different target proteins and their concentrations. They used three methodologies in their work: 1) considering a proper concentration of a fluorescent particle for encapsulation within the liposome; 2) incorporating cholesterol into the liposome membrane to improve the fluidity of the membrane and the resultant fluorescence; and 3) assessing the interaction qualities of different phospholipid liposome species with target molecules to recognize the most effective phospholipids. They also found that the protein discrimination by the exhibited sensor with a single shot was way better than that with many shots of a microwell (). For identifying the intramural heterogeneity of EGFR activity, Weitsman and his colleagues designed a genetically encoded Crkll-based biosensor focused on lipopolyplexes bearing EGFR-binding peptides that experience conformational changes in phosphorylation by EGFR. They found a critical correlation between high EGFR activity in tumor cells and macrophage-tumor cell proximity, which partially accounts for the intramural heterogeneity in EGFR activity. ) displayed a real-time in vivo fluorescence imaging strategy for observing and evaluating tumor drug uptake during warming by exploiting the inherent fluorescence of Dox and in vivo microscopy probes. Their result illustrated that time lapse in vivo fluorescence imaging can screen medication uptake in real time during hyperthermia-interceded medication delivery, and in vivo fluorescence intensity can predict tumor Dox accumulation (). Nanosensor detection systems have provided an attractive alternative to focused imaging of cancer cells. Wang et al. have also developed a fluorescence “turn-on” nanosensor for the location of MUC1 in living cells by designing a polydiacetylene (PDA) liposome-based sensing framework functionalized with a Cy3-labeled MUC1 binding aptamer with a limit of discovery around 0.8 nM. In the presence of MUC1, the highly specific interaction with the Cy3-aptamer leads to the dye dissociation from the liposome interface, recovering the red fluorescence ().
FIGURE 2
Colorimetric biosensor based on liposomes
Several liposome-based colorimetric biosensors have been outlined to screen for very low levels of pesticides by joining a dynamic protein inside the nanoenvironment of the liposomes. A plasmonic colorimetric sensor for highly sensitive visual detection of telomerase activity in cancer cells has been created by integrating an etching substrate, Au nanobipyramids, with a liposome-based signal enhancement procedure. In reaction to high expression of telomerase, HRP was discharged from the liposomes, following the expansion of the TMB substrate, and then, the formation of TMB2+ led to the morphological advancement of Au NBPs, which gave rise to tremendous LSPR responses with location limits comparable to one HeLa cell for localized surface plasmon resonance spectrometer estimation and a visual detection limit of 20 cells (
Electrochemical biosensors based on liposomes
The liposomes have been integrated into two forms of electrochemical biosensor stages: strip-organized immunoassays and microfluidic chips. Within the final decade, many analysts created an arrangement of strip-organized based immunoassay based lateral stream guideline (
Bhuvana et al. outlined a liposome-based nanocomposite by covalently immobilizing a 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine (DOPE) liposome on the MPA layer on the gold surface for electrochemical sensing of name-free DNA. The single-stranded DNA (ssDNA) probes were immobilized on the DOPE-AuNP nano-composite through a basic gold–thiol linkage to separate between the hybridized and single base mismatch targets in the presence of [Fe (CN) 6] 3-/4- and [Ru (NH3) 6] 3+ redox probes without signal amplification with a detection limit of 0.1 × 10–12 M. Their results collectively affirmed the non-fused round nature of the DOPE on the MPA layer and the high steadiness of the DOPE-AuNP nanocomposite on the gold surface (Figure 3A) (
FIGURE 3

(A) Formation of a spherical DOPE–AuNP nano-composite and DNA detection. (B) Dropping the Molt-4 cells also caused an increase in EIS. High sensitivity of Molt-4 cells was observed with lectin liposome. (C) Dropping the boronic acid liposome caused a decrease in EIS. In this stage, higher detection occurred due to the target screening of sialic acid in Molt-4 cells.
New generation of liposome-nanomaterial hybrids and exosomes for analyzing targets
In order to improve the diagnostic efficiency of liposomes, studies have been conducted on the use of other nanomaterials along with liposomes to form hybrids or supermolecules. A new generation of natural lipid vesicles called exosomes has been developed to take a new step.
Liposome nanomaterial (NM)
Two-dimensional and three-dimensional nanomaterials have a wide capacity to hybridize with liposomes. Imran et al. used self-assembled DOTAP liposomes on graphene oxide/reduced graphene oxide (GO/rGO) composites supported on a gold electrode for the ultra-sensitive and selective label-free detection of target DNA in the presence of gold nanoparticles, with the lowest detection limit of 1.29 × 10−17 M and a broad linear range of 10–17-10–8 M. The liposomes can be introduced as tethered, intact vesicles for improved drug and DNA delivery applications in therapeutics and diagnosis (Figure 4A). The bilayered liposome/AuNP structures on AuE, GO, and rGO do not appear to have any selectivity for DNA detection in graphene materials on a gold surface by physisorption without utilizing thiol self-assembled monolayers. They used this vesicle structure for label-free sensing of DNA in the energetic range from 10 to 15 to 10–9 nM [Figure 4B(a, b)] (
FIGURE 4

(A). Schematic image of graphene oxide-supported liposomes for liposomal sensors clearly in point-of-care test label-free electrochemical DNA biosensing. (B). Ultra-sensitive and selective label-free electrochemical DNA detection at layer-by-layer self-assembled graphene oxide and vesicle liposome nanoarchitecture. (C).(a) Quantum dot incorporation models of hybrid delivery systems. A, hydrophobic quantum dot-encapsulated shell model; B, hydrophilic quantum dot-linked shell model; C, hydrophilic quantum dot-encapsulated core model. (b) Temporal internalization of L-QD by confocal microscopy and representative NIR images. These images represented the denuded mouse after lipid-CdHgTe was injected via the tail vein for 10 seconds, and 5, 10, and 20 min.
Exosomes and MV
Exosomes are liposome-like characteristic vesicles with many parts in typical physiological work (
FIGURE 5

(A) Schematic image of echogenic exosomes as ultrasound contrast agents. (B) Ultrasound images before and after the injection of echogenic exosomes into the synovial space of a Sprague–Dawley rat. Images were taken with a Vevo 3,100 Imaging System and a 40 MHz transducer. (B) Schematic schema for the stages of the exosomal electrochemical sensor model for the detection of 21 miRNAs: (a) immobilization of the MCF-10A exosome with different biomarkers on the SCPE-GNP; (b) adding p19 to the MCF-10A exosome/SCPE-GNP; (c) adding a hybrid of the 21miR-probe to the p19/MCF-10A exosome/SCPE-GNP.
In another curiously novel study, Ghazizizedeh et al. suggested a double stage to identify any spiked infections and specific SARS-CoV-2 antigens consecutively. They presented a common bed-receptor surface as microparticle vesicle-galactins1 (MV-gal1) with the capacity of glycan binding to screen spiked viruses. The sensor has the capacity to screen for any spiked viruses and detect COVID-19 (LOD: 4.57 102 copies/mL) by utilizing the natural bed receptor and a specific antibody in the point-of-care test (
Conclusion
In order to diagnose, treat, and examine diseases, it is necessary to have an image with high quality and resolution (
Statements
Author contributions
All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Publisher’s note
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Summary
Keywords
biosensor, lipid-based vesicle, exosome, liposome, microvesicle
Citation
Ghazizadeh E and Nasery Z (2023) The viewpoint of nanolipid vesicles (liposomes, exosomes, and microvesicles) as biosensors in medical health advances. Front. Nanotechnol. 5:1230407. doi: 10.3389/fnano.2023.1230407
Received
08 June 2023
Accepted
25 July 2023
Published
15 September 2023
Volume
5 - 2023
Edited by
Arindam Pramanik, Amity University, India
Reviewed by
Ahmad Mobed, Tabriz University of Medical Sciences, Iran
Wei Shao, Westlake University, China
Krishna Kant, University of Vigo, Spain
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Copyright
© 2023 Ghazizadeh and Nasery.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Elham Ghazizadeh, elgh@tf.uni-kiel.de
† These authors have contributed equally to this work
Disclaimer
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.