Table 3.
Features and findings of research on microneedle system for nucleic acid drug delivery
| Type of MN | Combined delivery system | Applications | Target nucleic acid | Features and findings | References | |
|---|---|---|---|---|---|---|
| Solid | Non (naked) |
Gene expression, Cancer vaccine |
HBsAg pDNA Luciferase pDNA |
Adjuvant (Flt3L, Fms-like tyrosine kinase 3 ligand) was added Immune response: MN + adjuvant + pDNA > IM + adjuvant pDNA ~ MN + pDNA Protective efficacy against tumor cell challenge was observed in mice |
Zhou et al. (2010) | |
| Solid | Non (naked) | Gene knockdown | GAPDH siRNA |
5' cholesterol modification was introduced into siRNA to enhance cellular uptake Evaluation of in vivo biodistribution of Cy5-labelled siRNA was included (Major distribution in MN-applied tissues compared to major organs) |
Deng et al. (2016) | |
| Solid hollow |
Non (Naked), Electroporation, Cationic polymer/lipid |
Immunization study | Ovalbumin pDNA | Permeability of DNA have been compared between EP, SC, solid MN, EP + solid MN, and hollow MN (33-gauge hypodermic needle). Characterization and optimization of complexes using commercial transfection reagents (Lipofectamine, Superfect) and cationic polymer (PEI). Hollow MN showed a much higher immune response than SC injection. The complex elicited a stronger immune response than the naked, with the strongest response in PEI complex | Pamornpathomkul et al. (2017) | |
| Solid |
Non (naked), Cationic polymer/lipid |
Gene expression |
Luciferase pDNA EGFP pDNA |
Longer MN and longer duration time showed stronger expression than shorter one. And naked DNA showed better expression than complexed one DNA → MN showed 87-times higher expression than MN → DNA |
Yan et al. (2014) | |
| Solid |
Cationic polymer (PEI) |
Gene expression, Immunization study |
Luciferase pDNA Hepatitis B HBsAg pDNA |
IM (naked) vs. MN (naked) vs. IM (complex) vs. MN (complex) PEI-pDNA loaded MN showed a 4.7-fold higher immune response than IM injection and a 2.6-fold higher immune response than naked DNA loaded MN |
Yin et al. (2013) | |
| Solid |
Cationic polymer- cell penetrating peptide |
Gene expression |
Luciferase pDNA EGFP pDNA |
Mannosylated PEI-CPP-pDNA complexes targeting mannose receptor positive dendritic cells. The complex exhibited 150-fold higher expression than naked pDNA in vitro expression study | Hu et al. (2014) | |
| Solid |
Cationic liposome (DOTAP) |
Gene knockdown | GAPDH siRNA |
Utilization of sponge Haliclona sp. Spicules as solid MN. Liposome-siRNA complex %Knockdown: MN + complex > SC + complex > MN + siRNA |
Liang et al. (2020) | |
| Solid |
Lipid nanoparticle (DOTAP) |
Gene expression |
EGFP pDNA β-galactosidase pDNA |
In vitro transfection efficiency and in vitro permeation study were involved DOTAP based lipid nanoparticle were tested with different charge condition (positive, neutral, negative). Positive charged nanoparticle only showed expression |
Chabri et al. (2004) and Coulman et al. (2005) | |
| Solid | Electroporation |
Gene expression, Fluorescently labelled RNA transfection |
RFP pDNA, Cy5 labelled siRNA |
Gold coated solid MN used for electroporation. Transfection efficiency and tissue damage were evaluated at various voltages (range 0—50 V) for the combination of MN and electroporation. MN alleviated the EP conditions, allowing it to decrease to 35 V with maximum transfection efficiency. MN combined with electroporation enhanced transfection of nucleic acids. Roller type MN was utilized to enhance transfection efficiency. Longer MN showed better expression | Wei et al. (2014) and Huang et al. (2018) | |
| Coated | Non (naked) |
Gene expression, Immunization study |
Luciferase pDNA, Gag encoding pDNA (HIV model antigen SIV-gag) |
Multilayer coated MN enabling rapid release of pDNA in pH-responsive manner. PolyI:C was used for adjuvant. 140-fold higher level of expression with MN compared to ID injection (reporter gene). The dried formulation maintained the bioactivity of pDNA for 28 days at room temperature. tenfold higher antibody titers with MN compared to other treatments (ID, IM, IM + EP) | Demuth et al. (2013) | |
| Coated | Non (naked) | Cancer vaccine |
Hepatitis C virus 3/4A pDNA |
MN had an immune response similar to that of the gene gun at similar doses. MN (3.2 μg) showed protective efficacy in tumor cell challenge and equivalent immune response to a 30-fold higher dose of IM (100 μg) | Gill et al. (2010) | |
| Coated | Non (naked) |
Gene expression, Immunization study |
Luciferase pDNA Influenza HA pDNA |
Coated MN showed protective efficacy on viral challenge and improved immune response compared to the same dose of IM. Immunization study using clodronate-liposomes suggest that dendritic and macrophages may not provide a major contribution to protective efficacy of MN | Song et al. (2012) and Kim et al. (2013) | |
| Coated | Non (naked) | Immunization study |
Nucleosomal histones of Leishmania spp pDNA |
Comparison of immune responses by administration route of pDNA cocktails of plasmids encoding four different antigens. Immune response: SC < ID < MN | Moreno et al. (2017) | |
| Coated | Non (naked) |
Gene expression, Immunization study |
EGFP pDNA Ovalbumin pDNA |
MN showed a tenfold higher immune response compared to IM injection. Gene expression and immune response: IM < MN | Zhang et al. (2018) | |
| Coated |
Cationic polymer (PEI) |
Cancer vaccine | p53 pDNA | PEI-pDNA complex coated on polycaprolactone MN by layer-by-layer assembly. pH-responsive pDNA release promoted in the acidic environment (pH 5.5). pH responsive MNs showed better tumor suppression than non-multilayered MNs or IV injections | Li et al. (2019b) | |
| Coated |
Cationic liposome (DOTAP) |
Gene knockdown | CXCL1 gene siRNA | The nano-patch was coated with DOTAP-PEG-cholesterol-siRNA complex and methylcellulose for viscosity improving agent. Fluvax (one of the vaccines) was added to induce CXCL1 expression. The integrity of nucleic acid was maintained in the dry state | Haigh et al. (2014) | |
| Coated |
Nanoparticle (PEI-deoxycholate) |
Gene expression, Immunization study |
Alzheimer beta- amyloid pDNA |
Multilayer MN enhanced rapid release at physiological pH. Mannosylated PEI-deoxycholic acid-pDNA complexes were treated with SC or MN. MN showed a higher immune response compared to the SC group. Trehalose was used as a stabilizing agent | Kim et al. (2014) and Duong et al. (2018a) | |
| Coated | Nanoparticle (PEI-deoxycholic acid) |
Cancer vaccine (melanoma) |
Ovalbumin pDNA | Rapid release at physiological pH was induced with a multilayer MN (pH-responsive layer). Poly I:C was co-administered as an adjuvant | Duong et al. (2018b) | |
| Dissolving | Non (naked) |
Gene expression, Cancer vaccine |
Luciferase mRNA Ovalbumin mRNA |
Naked mRNA was loaded into PVP-based dissolving MN. The concentrated PVP solution lowered the in vivo transfection efficiency of mRNA in PVP solution Longer MNs showed higher mRNA expression in vivo |
Koh et al. (2018) | |
| Dissolving | Nanoparticle (PLGA-based) |
Gene expression, Immunization study |
GFP pDNA Ebolavirus protein pDNA |
The PLGA-Poly(L-lysine)-poly(γ-glutamate)-DNA complex was loaded onto PVA-based dissolving MN. Sucrose stabilized pDNA structure of MN at room temperature Immune response: MN + NP + pDNA > IM + NP + pDNA > IM + pDNA > MN + pDNA |
Yang et al. (2017) | |
| Dissolving |
Cationic polymer (PEI) |
Cancer vaccine (melanoma) |
STAT3 siRNA | PEI-siRNA complex loaded into dissolving MN (matrix composition: dextran40: PVP17: HA = 4:1:1). Naked siRNA showed lower transfection than complex siRNA (in vitro). Dose dependence was confirmed with four dose levels (33, 66, 132, and 264 μg) | Pan et al. (2018) | |
| Dissolving | Peptide nanoparticle |
Gene expression, Cancer vaccine |
Luciferase pDNA HPV-16 E6 and E7 pDNA |
RALA (one of cell penetrating peptide)-pDNA complex loaded into PVA-based dissolving MN. Lyophilization was used to increase the loading capacity, and trehalose was used as a stabilizer for the process. 57 µg of pDNA was loaded into the array. pDNA loaded in MN was stable for 28 days at room temperature. Complex-MN group showed higher immune response than complex-IM | Cole et al. (2018) | |
| Dissolving | Electroporation |
Gene expression, Cancer vaccine (melanoma) |
Luciferase pDNA 2CMVmIL-12 pDNA (IL-12 subunits) |
Maltose-based dissolved MNs mounted on metal electrodes array. EP enhanced the expression of IL-12. pDNA delivery via dissolving MN combined with electroporation inhibited tumor growth and extended survival | Lee et al. (2011) | |
| Dissolving | Photothermal therapy | Cancer vaccine (melanoma) | p53 pDNA | Photothermal dye (named IR 820) and pDNA were loaded into hyaluronic acid-based dissolving MN. Near-infrared rays increase the temperature of the tissues where IR820 is distributed and have anticancer effects. Combination with photothermal therapy (PTT) effectively inhibited the growth of subcutaneous tumors | Xu et al. (2020) | |
| Hollow | Cationic lipid (Lipofectamine) | Gene knockdown | β-galactosidase pDNA EGFP pDNA | A commercial hollow MN device (MicronJet) was used, enabling reproducible amounts of siRNA. Transfection studies using ex vivo human skin | Dul et al. (2017) | |
| Hollow | Cationic lipid (Lipofectamine) | Gene expression | Luciferase mRNA | A commercial hollow MN device (MicronJet) was used. Transfection studies using ex vivo porcine skin. There were no significant differences in luciferase expression between naked mRNA and liposome complexed mRNA | Golombek et al. (2018) | |
| Hollow | Cationic niosome | Gene expression, Immunization study |
EGFP pDNA Ovalbumin pDNA |
Cationic niosome-pDNA complex (composition: span20/cholesterol/cationic lipids = 2.5/2.5/0.5 in molar ratio) was delivered in vivo by hollow MNs. MN injection of niosome complexes showed a higher immune response than SC injection In vivo immune response: Niosome complex > Lipofectamine complex > Naked pDNA |
Pamornpathomkul et al. (2018) | |
MN microneedle, HBsAg hepatitis B surface antigen, GAPDH glyceraldehyde-3-phosphate dehydrogenase, EP electroporation, SC subcutaneous, PEI polyethyleneimine, EGFP enhanced green fluorescent protein, IM intramuscular, CPP cell penetrating peptide, DOTAP 2, 3-dioleoyloxy-propyl-trimethylammoniumchlorid, RFP red fluorescent protein
MN microneedle, HIV human immunodeficiency virus, SIV simian immunodeficiency virus, ID intradermal, IM intramuscular, EP electroporation, HA hemagglutinin, SC subcutaneous, EGFP enhanced green fluorescent protein, PEI polyethyleneimine, IV intravenous, DOTAP 2, 3-dioleoyloxy-propyl-trimethylammoniumchlorid, CXCL1, C-X-C motif chemokine ligand 1, PEG polyethylene glycol
MN microneedle, PVP polyvinylpyrrolidone, PLGA poly(lactide-co-glycolide), PVA polyvinyl alcohol, GFP green fluorescent protein, NP nanoparticle, IM intramuscular, PEI polyethyleneimine, STAT3 signal transducer and activity of transcription 3, HA hyaluronic acid, HPV human papillomavirus, EP, CMV, cytomegalovirus, IL interleukin, EP electroporation, EGFP enhanced green fluorescent protein, SC subcutaneous