|Year : 2016 | Volume
| Issue : 1 | Page : 1-6
Applications of nanomedicine in parasitic diseases
Sherif M Abaza PhD
Department of Medical Parasitology, Faculty of Medicine, Suez Canal University, Ismailia, Egypt
|Date of Submission||24-Mar-2016|
|Date of Acceptance||26-Apr-2016|
|Date of Web Publication||25-Oct-2016|
Sherif M Abaza
Department of Medical Parasitology, Faculty of Medicine, Suez Canal University, Ismailia
Source of Support: None, Conflict of Interest: None
Nanomedicine is defined as application of nanotechnology for treatment, monitoring, prevention, and control of biological diseases. To apply nanomedicine, the precise targets (cells and/or receptors) specific to the clinical disease should be identified and the suitable nanoparticles for delivery system to minimize the side effects of the original drug should be selected. One of these precise targets are macrophages, endothelial, dendritic as well as tumor cells. The main aim of the present review is to throw light on possible nanotechnology applications in parasitic diseases focusing on three main aspects: diagnosis, treatment, and vaccination.
Keywords: adjuvant, diagnosis, drug delivery system, gene therapy, macrophage, nanoparticles, parasitic diseases, treatment, vaccine
|How to cite this article:|
Abaza SM. Applications of nanomedicine in parasitic diseases. Parasitol United J 2016;9:1-6
Reduction of certain materials from macro to nano scale is associated with changes in some of their properties. For example, gold becomes liquid at room temperature, silicon turns into a conductor instead of an insulator, and aluminum becomes combustible. The synthesis and properties of semiconductor nanocrystals were studied, leading to the discovery of a number of metal and metal oxide nanoparticles (NPs) . This was followed by the use of NPs in nanomedicine for the diagnosis and monitoring as well as the treatment and control of biological diseases. The use of NPs is increasing as they have defined chemical, optical, and mechanical properties . In the new century, there has been a strong focus on applications in the diagnosis and treatment of cancer, e.g. formulation of therapeutic agents in polymeric, biocompatible, and submicron-sized composites. A recent review article discussed the principles of nanotechnology and applications of nanomaterials in medicine and techniques as well as the instruments used to investigate the physicochemical characteristics (size, shape, surface properties, composition, purity, and stability) of the commonly used nanomaterials .
Cytotoxicity associated with nanotechnology raised certain concerns in terms of the individual physicochemical properties (size, charge, concentration, and outer coating bioactivity) and environmental conditions (oxidative, photolytic, and mechanical stability). For example, some NPs were found to be cytotoxic only after oxidative and/or photolytic degradation of their core coatings. Few in vitro and in vivo studies have suggested that NPs can affect cell growth and viability in a dose-dependent manner. The most important factor of NPs toxicity is their stability, both in vivo and during synthesis and storage .
Nanomaterials include silver, gold, platinum, and magnetic NPs, quantum dots, liposomes, nanospheres, carbon nanorods, nanotubes, and others. Silver ,,,,,,,,, and gold ,,,,,,,,, NPs were the commonest elements used, and constituted the majority of publications in the last 10 years. However, other elements have been used such as calcium , iron oxide , and silica . Chitosan (CS) is another NP formula, formed of the natural polysaccharide Arabic gum, that attracted significant scientific interest during the last two decades because of several properties: 1) it is nontoxic, 2) it has various molecular weights, 3) it forms complexes with DNA that effectively protects it from degradation, 4) it enhances the penetration of large molecules across the mucosal surface, and 5) it is taken up by Payer’s patches. Therefore, it has been used in drug delivery, tissue engineering, and gene delivery ,,,,,,,,,,. Liposomes and microspheres are considered the most extensively studied carriers for drug delivery systems. Liposomes are nano-sized vesicles formed of layers of natural phospholipids and cholesterol surrounding an aqueous space. According to the size and number of layers, they are classified into three types: small unilamellar, large unilamellar, and multilamellar liposomes. According to the physicochemical characteristics of the drug, it is either entrapped in the aqueous space or inserted into the liposome layers ,,,,,. Nanospheres consist of synthetic or natural polymers (collagen or albumin), and the required drug is conjugated or encapsulated within the polymeric matrix ,,,. Other chemical components were also reported; e.g. self-assembling polypeptide (SAP), solid lipid (SL), polyamidoamine (PAMAM), polylactide-co-glycolide (PLGA), polyethylimine, γ polyglutamic acid (γ-PGA), and archaeosome vesicles (ARC) which are made from lipids extracted from Archaea. Their NPs are helpful to carry the antigen to dendritic cells (DCs) due to two important characteristics: its tiny size which allows it to cross the skin’s extracellular matrix, and its coating which mimics a bacteria cell wall causing the complement system to be activated ,,,,,.
| Applications in parasitic diseases|| |
Regarding the diagnosis of malaria, recombinant heat shock protein 70 (HSP70) of P. falciparum conjugated with gold NPs and functionalized with anti-HSP70 monoclonal antibodies proved to be very sensitive in the detection of malaria antigen . Polystyrene NPs conjugated with polyclonal IgG antibodies specific to P. falciparum showed sensitive results . Specific DNA aptamers (oligonucleotide molecules selected from pools of random-sequence oligonucleotides in the P. falciparum DNA library to bind a wide range of relevant proteins) were conjugated to gold NPs to achieve rapid and early diagnosis of malignant malaria ,. The Wondfo rapid diagnostic test linked P. falciparum biomarkers (histidine-rich protein-2 and pan-Plasmodium lactic dehydrogenase, Pf-HRP2/PAN-pLDH) with gold NPs in an immunochromatographic (ICT) assay to achieve better diagnosis of malaria in clinical samples . Screening of blood films was successfully achieved by the detection of hemozoin or β-hematin (unique components of Plasmodium spp.-infected red blood cells) using surface-enhanced resonance Raman spectroscopy (SERRS) and atomic force microscopy (AFM). NPs were modified (tuning the core and shell with a silver shell) and used in SERRS allowing magnetic field. This tuning causes aggregation of β-hematin, and early diagnosis of malaria . While AFM was successfully used to generate transient vapor nanobubbles around the hemozoin in response to a short infrared picosecond laser pulse .
In the diagnosis of other parasitic diseases, the same concept of NPs conjugation with the parasite biomarkers was used. In toxoplasmosis, the use of specific agglutination of antigen-coated gold NPs in the detection of the corresponding antibody gave satisfactory agreement with ELISA results . An immunomagnetic bead-ELISA technique utilizing T. gondii IgG polyclonal antibodies coated with magnetic NPs to capture circulating surface antigen 1 gave better results than sandwich ELISA . In cryptosporidiosis, HSP70 was also conjugated with gold NPs to target HSP70 mRNA from C. parvum oocysts . An assay that used probes of oligonucleotide-functionalized gold NPs (complementary to the 18s rRNA sequence on C. parvum) proved its ability to detect the nucleic acids of C. parvum oocysts in stool samples . For amebiasis, fluorescent silica NPs were synthesized and conjugated with monoclonal anti-E. histolytica IgG1 for diagnosis of amebiasis. It showed high sensitivity results without cross reaction with other protozoa (E. dispar, E. moshkovskii, G. lamblia and Blastocystis spp.) . Synthesized iron oxide NPs were functionalized with CS and used to capture and remove Entamoeba spp. cysts after application of an external magnetic field. The investigators claimed that the synthesized NPs were well dispersed and suitable for water treatment. They suggested that cyst wall components (lectins and chitin) might interact with CS NPs, and recommended further studies to validate the use of NPs in water treatment, with a special emphasis on possible NPs toxicity . In leishmaniasis, gold NPs conjugated with four oligonucleotide probes, targeting DNA of Leishmania kinetoplastid, were used . Recently, gold NPs were also conjugated with labeled Leishmania spp. primers and magnetic beads for isothermal amplification of Leishmania DNA in blood samples of infected dogs. It was found that NPs have electrocatalytic activity for the rapid detection of the amplified DNA. This approach was found to be more sensitive and less expensive than the traditional PCR methods used in the diagnosis of visceral leishmaniasis (VL) .
When specific antibodies are coated with laser-irradiated gold NPs, they selectively attach to the parasite, and the heat produced from the irradiated laser will kill the parasite . Besides, the macrophage is a valid pharmaceutical target, as being a specialized host defense cell with well-known contribution to pathogenesis. Its surface contains receptors to recognize terminal galactose, mannose, fructose, or glucose residues of glycosides. Therefore, sugar-bearing liposomes are designed for improvement of new drugs to target the macrophage ,. At first, herbal medicine was not considered for development as novel formulations due to the lack of scientific justification and processing difficulties. However, NPs proved their use to deliver herbal medicine because of their unique physicochemical properties, their size and capacity to penetrate into the cell, as well as their ability to increase the therapeutic value by reducing toxicity and increasing bioavailability. That is besides its lower cost using natural herbs .
A microchip is a micro-fabricated device that includes a pump, valve, and flow channel to allow controlled release of single or multiple drugs on demand. It is implanted in patients who require pulsatile drug release. The drug reservoirs are covered by membranes formed of PLGA . Carbon nanotubes consist of graphite sheets rolled up into tubular forms. They can cross the cell membrane without membrane disruption to be localized into the cytosol and mitochondria. They are manufactured for drug delivery and gene therapy . Moreover, most of the new drug targets that emerged from high-throughput drug screening initiatives are insoluble or poorly soluble in water. Therefore, the therapeutic agents are conjugated or encapsulated or adsorbed on NPs or liposomes or nanospheres to overcome the drug solubility issue ,.
As sole treatment, targeting infected macrophages with NPs is a valuable and validated strategy for treatment of VL . Combined therapy by silver, CS, and curcumin NPs gave the highest effect and complete cure in giardiasis in experimentally infected animals . Silver and CS were evaluated singly or combined for in vivo treatment of toxoplasmosis in experimental animals. The combined treatment showed significant decrease in hepatic and splenic parasite burden. Microscopic examination revealed stoppage of movement and deformity in the shape of tachyzoites .
As a drug delivery system, quercetin conjugated with gold NPs was established for treatment of VL caused by wild-type resistant strains . On the other hand, CS proved to be a suitable drug delivery mean for several drugs used in VL treatment. Doxorubicin displayed significant reduction in Leishmania amastigotes (in vivo) and promastigotes (in vitro) , while amphotericin B  and rifampicin  gave significant results compared with control drugs without CS. Amphotericin B was also encapsulated in PLGA NPs and gave significantly effective results in comparison with the drug alone . A review article discussed all the studies that employed nanotechnology in drug delivery systems for amphotericin B in the treatment of VL. The reviewers concluded that CS and chondroitin sulfate NPs are the best ones nowadays due to their lower costs . In cutaneous leishmaniasis, glucantime formulated with liposomes was effective in the topical treatment of leishmanial ulcers caused by L. major in mice. It resulted in a significant decrease in lesion size and spleen parasite burden .
Isolated fungus from the soil (Trichoderma harzianum) conjugated with silver NPs increased the efficacy of triclabendazole in the treatment of fascioliasis . Binding of curcumin  and choloroquie  to CS increased chemical stability and enhanced bioavailability when evaluated in the treatment of malaria in infected mice. In cryptosporidiosis, polyvinyl alcohol conjugated with CS was proven to suppress the attachment of Cryptosporidium sporozoites to enterocytes in vitro . Albendazole bound to CS was effective in the treatment of alveolar echinococciosis caused by E. multilocularis  and visceral larva migrans caused by T. canis . In the first report, it was concluded that CS improved albendazole absorption, increased its bioavailability in vivo, is easily manufactured as capsules or tablets, and above all is nontoxic and of low cost. In the treatment of schistosomiasis mansoni, praziquantel (300 mg/kg) encapsulated in liposomes showed a significant reduction in worm burden and stool and intestinal egg counts as well as in the number of hepatic granulomas . Diminazene aceturate also encapsulated into liposomes, showed in vitro and in vivo significant results in treatment of Suda, caused by T. evans . Recently, miltefosine, an anticancer therapy, was enclosed in lipid nanocapsules and administered to S. mansoni-infected mice as a single oral dose (20 mg/kg), and its efficacy was compared with that of praziquantel. The results proved its potentiality in schistosomiasis mansoni, and the ability of nanomedicine to act as efficient drug delivery vehicle .
Drug delivery system in herbal medicine: For arthropods, an aqueous extract of neem leaves conjugated with silver NPs proved to be a potent insecticide against mosquitoes larvae, pupas, and adults for control of vector-transmitted diseases . Recently, several publications utilized synthesized silver NPs with different herbs (Barleriacristata spp., Aristolochiaindica spp., and Pteridiumaquilinum spp.) for testing against larvae of different mosquitoes (species of Anopheles, Aedes and Culex), and they showed significant larvicidal effects compared with using herbs alone ,,,. For parasitic diseases, essential oil of Achyrocline satureioides was given in an ananoencapsulated formula to experimentally T. evansi-infected mice and its efficacy was investigated in comparison with diminazeneaceturate and free essential oil. The encapsulated form showed better anti-trypanosomal activities without hepatic or renal toxicity, i.e. the use of nanotechnology reduced the side effects of the therapeutic agents . Synthesized palladium NPs using Eclipta prostrata leaf aqueous extract  and synthesized silver NPs using Euphorbia prostrata leaf extracts  or Acacia auriculiformis extracts  were tested against P. berghei, L. donovani, and the bovine filarial parasite Setaria cervi, respectively. The results showed the efficacy of NPs as potent drug delivery systems in herbal medicine manufacturing.
Immunization and vaccination
With variable effects, several NPs were formulated to optimize vaccine development as they protect DNA vaccines from degradation, and the use of chemical components (SAP, SL, PAMAM, PLGA, γ-PGA, and ARC) is considered a hopeful approach in vaccine development .
As prophylactic candidate: Since SAP NPs have the ability to induce CD8+ and CD4+T cells, they were used to stimulate long-lasting immune responses to specific epitope of P. falciparum circumsporozoite protein , and to vaccinate mice against toxoplasmosis .
As an adjuvant: To improve the immune response in schistosomiasis, calcium NPs were used with anti-idiotypic antibody (NP30) to enhance cellular and humoral immune response , while CS was given with gene encoding S. japonicum ferritin . In VL, cationic SL NPs were used with a tri-fusion gene of L. tarentolae (lizard species nonpathogenic to humans) in a DNA vaccine. Genes encoding A2 antigen and cysteine proteinases (CPA and CPB) were used and yielded promising results . Immunoliposomes when given with soluble Leishmania antigen (SLA) improved immunization . In Chagas disease, ARC act as strong adjuvants when given with soluble T. cruzi antigens .
As delivery system: Human P. falciparum Pfs25 specific antibodies block parasite infectivity to mosquitoes, but the extent of blocking was insufficient for an effective transmission blocking vaccine. Recombinant Pfs25H was then conjugated with Pseudomonas aeruginosa exoprotein A and formulated in gel NPs to improve blockage of infectivity to mosquitoes. This blocking vaccine is in phase 1 human trial in USA . In schistosomiasis, S. mansoni antigen (Rho) is involved in a number of cell signaling pathways with effects on actin cytoskeleton, gene transcription, and membrane trafficking. It was conjugated with CS NPs containing Rho and CpG (as an adjuvant to induce interleukin-12) and produced moderate protection, with a reduction in hepatic granulomas . Lysine-modified PAMAM was used as a novel delivery system for S. japonicum DNA vaccine . In leishmaniasis, PLGA NPs ,, alginate microspheres , SL NPs , and cationic liposomes , were used as delivery systems with several antigens such as leishmanial cysteine proteinase I, L. major antigen, soluble Leishmania antigen, and Leishmania kinetoplasmid membrane protein-11 to induce protection against VL. In malaria, several trials were reported; PLGA NPs were used as a delivery system for P. vivax sporozoites antigen (VMP001) , whereas merozoite surface protein-1 was conjugated with iron oxide NPs  or combined NPs of PE and γ-PGA  or anionic NPs ,, or quantum dots NPs .
| Concluding remarks|| |
- In the last decade, the use of NPs received considerable interest because of their defined properties, and they were used in the development of diagnostic methods, therapeutic targets, and in protection and vaccination of tropical parasitic diseases.
- AFM and scanning tunneling are two main instruments developed to manipulate nanostructures and to use NPs in the early diagnosis of fatal parasitic diseases such as malaria and VL.
- Targeting infected macrophages with NPs is a valuable and validated strategy for treatment.
- NPs are novel drug delivery systems for herbal medicine to be used as potent insecticides against mosquitos’ different stages, i.e. new strategy in the control of vector-transmitted diseases.
- Silver and CS NPs are considered potent therapies in the treatment of toxoplasmosis and giardiasis.
- Metal NPs, CS, and liposomes are conjugated with several drugs, e.g. praziquantel, choloroquie, amphotericin B, rifampicin, and albendazole in the treatment of schistosomiasis, malaria, VL, and visceral larva migrans.
- NPs act as vaccine candidates against toxoplasmosis and malaria, as an adjuvant to improve immune response against schistosomiasis, VL and Chagas disease, and as a vaccine delivery system against malaria, schistosomiasis, and VL.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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