Table 3.
Delivery of anticancer therapeutic agents using silk-based nanoparticles.
| Type of Anticancer Therapeutic | Therapeutic Agent | Silk Source | Preparation Method | Particle Size | Functionalization/Surface Msodification | Target/ In Vitro/ In Vivo Model |
Outcome/Findings | Ref |
|---|---|---|---|---|---|---|---|---|
| Plant-derived therapeutic agents | Curcumin | B. mori silk fibroin | Precipitation with ionic liquids and high-power ultrasounds | 166–171 nm | Liver cancer/ Hep3B and Neuroblastoma/ KELLY/ ND |
Sustained drug release up to 3 days Drug bioavailability Cytotoxic activity towards cancer cells No toxic effect in healthy cells |
[196] | |
| Suspension-enhanced dispersion by supercritical CO2 (SEDS) |
<100 nm | Colon cancer/ HCT-116/ ND |
Time-dependent intracellular uptake ability Improved inhibition effects on colon cancer cells No toxic effect in healthy cells |
[197] | ||||
| Desolvation and cross-linking with genipin | 217 nm | Murine breast cancer/ 4T1/ Tumor in mice |
Binary drug loading (5-FU and curcumin) High loading efficacy Improvement in the cytotoxic activity and bioavailability compared with free drugs Toxic effect toward cancer cells in vitro and in vivo |
[191] | ||||
| B. mori silk fibroin-chitosan blend | Microdot capillary method | <100 nm | Breast cancer/ MCF-7 and MDA-MB-453/ ND |
Sustained drug release over 9 days Efficacy against Her2-overexpressing cancer cells |
[198] | |||
| Resveratrol | B. mori silk sericin | Desolvation with DMSO and pluronic F-68 | 200–400 nm | Colon cancer/ Caco-2/ ND |
High drug encapsulation levels and stable drug release profile over 72 h High intra-cellular internalization efficiency The anticancer effect, but no toxicity towards healthy cells |
[199] | ||
| Triptolide/ celastrol |
B. mori silk fibroin | Desolvation with acetone and ethanol | 166 nm/ 170 nm | Pancreatic cancer/ MIA PaCA-2 and PANC-1/ ND |
Improved bioavailability and pharmacokinetic properties compared to free drugs The pH-dependent sustained drug release over 192 h Increased therapeutic efficiency compared to free drugs |
[200] | ||
| Emodin | B. mori silk fibroin | Lyophilisation of silk fibroin with emodin-loaded liposomes | 316 nm | Breast cancer/ MCF-7, BT-474 and MDA-MB-453/ ND |
Silk coating of liposomes decreased drug release rate compared to uncoated liposomes Longer intracellular retention of silk coated liposomes than liposomes w/o coating lead to the longer availability of emodin for down-modulation of various Her2/neu pathways |
[201,202] |
||
| α-mangostin | B. mori silk fibroin | Desolvation and crosslinking with EDC or PEI | 300 nm | Colon cancer/ Caco-2 and Breast cancer/ MCF-7/ ND |
Increase in water solubility of the drug Maintained α-mangostin’s apoptotic effect Increased cytotoxic effect on cancer cells than the free drug Reduction of hematoxicity compared to free drug |
[203] | ||
| Nucleic acid-based therapeutic agents | siRNA (anti-LUC) |
B. mori silk fibroin-oligochitosan blend | Desolvation with acetone | 250–450 nm | Lung cancer/ H1299/ ND |
Enhanced particle loading capacity than oligochitosan polyplexes Enhanced serum stability of siRNA than naked nucleic acid Increased gene silencing effect compared with oligochitosan polyplexes |
[27] | |
| pDNA encoding GFP |
A. pernyi silk fibroin (ASF) | Self-assembly with PEI/DNA complexes | 230–360 nm | Colon cancer/ HCT-116/ ND |
PEI/DNA complexes coated with RGD-rich ASF Increased target specificity in comparison with PEI/DNA complexes alone Higher uptake of silk coated complexes in cancer cells due to the affinity of the RGD peptides from ASF for integrins, Lower post-transfection cell toxicity than uncoated complexes |
[204] | ||
| siRNA (anti-CK2, anti-ASH2L, anti-Cyclin D1) |
B. mori silk sericin-albumin | Desolvation with ethanol | 127–142 nm | poly-L-lysine (PLL)-conjugated and hyaluronic acid (HA)-conjugated | Laryngeal cancer/ Hep-2/ ND |
Particles modified with PLL for siRNA binding and decorated HA to target cancer cells High siRNA entrapment Downregulation of target CK2, ASH2L and Cyclin D1 genes Higher silencing effect comparing with naked siRNA |
[205] | |
| siRNA (anti-STAT3) |
Bioengineered N. clavipes spider silk (MS2KN) | Salting out with potassium phosphate | 202 nm | poly-L-lysine (KN) | Macrophages/ J774/ ND |
Approach for cancer immunotherapy Protection of CpG-siRNA therapeutics from degradation by serum nucleases CpG-STAT3-siRNA targeted delivery to TLR9-positive macrophages Prolonged siRNA presence in macrophages than naked siRNA Prolonged silencing effect on STAT3 expression than naked siRNA |
[80] | |
| pDNA encoding LUC |
Bioengineered N. clavipes spider silk (15mer) | Self-assembly with pDNA | 186 nm | poly-L-lysine and RGD-conjugated | Cervical cancer/ HeLa/ ND |
High pDNA delivery efficiency Increased integrin-mediated transfection with RGD sequences than non-conjugated constructs |
[82] | |
| 99 nm | poly-L-lysine and ppTG1-conjugated | Melanoma/ MDA-MB-435/ ND |
High transfection rates Controlled enzymatic degradation rate of the silk-based pDNA complexes enables the regulation of the release profile of genes from the complexes |
[83] | ||||
| 90 nm | poly-L-lysine and Lyp1 or F3 peptide-conjugated | Melanoma/ MDA-MB-435 and Breast cancer/ MDA-MB-231/ Tumors in mice |
Enhanced pDNA delivery than non-functionalized complexes Low cytotoxicity Functionalization with F3 tumor homing peptide was the most effective in pDNA delivery to cancer cells |
[84] | ||||
| Protein-based therapeutic agents | Lactoferrin | S. cynthia ricini Eri silk | Milling | 200–300 nm | Breast cancer/ MCF-7 and MDA-MB-231/ ND |
Sustained release of therapeutic agents Higher stability in presence of proteolytic enzymes than bovine lactoferrin alone EGFR or TfR2 receptors-mediated endocytosis of nanoparticles Cytotoxic properties towards cancer cells |
[206] | |
| Peptides from ovoalbumin (C16-OVA) | Recombinant A. diadematus spider silk |
Salting-out with potassium phosphate using micromixing device | 369–386 nm | Bone marrow derived cells (BMDC)/ in vivo mouse model |
Potential approach for cancer vaccine immunotherapy Preferential uptake by immunological cells Localization in lysosomes Particles cleaved by lysosomal cathepsins to release transported peptide Antigen-specific proliferation of T-cells and cytotoxicity of released peptides in vivo |
[207] | ||
| Inorganic agents | IONPs/ Dox |
Bioengineered N. clavipes spider silk (MS1, MS2, EMS2) | Salting-out with potassium phosphate |
500 nm | ND | The addition of silk did not influence magnetic properties of IONPs Efficient incorporation and sustained release of incorporated drug (Dox) Not cytotoxic in vitro |
[110] | |
| ND | H2.1 peptide-conjugated (anti-Her2) | Breast cancer/ SK-BR-3/ ND |
Specific affinity of functionalized magnetic silk particles towards Her2-overexpressing cancer cells, Efficient binding of doxorubicin Ability to generate heat upon application of magnetic field (MF) Induction of hyperthermia in targeted cancer cells |
[126] | ||||
| IONPs/ Dox |
B. mori silk fibroin | Salting-out with potassium phosphate | 171–206 nm | Breast cancer/ MCF-7 and MCF-7-ADR/ tumor xenograft in mice |
High drug loading efficiency pH-dependent drug release up to 4 days Efficient magnetic targeting and intracellular delivery into both MCF-7 and MCF-7/ADR Ability to overcome multidrug resistance (MDR) The magnetic targeting to tumor in vivo |
[208] | ||
| IONPs/ Mtx |
Suspension-enhanced dispersion by supercritical CO2 (SEDS) |
112 nm | Skin from guinea pig (ex vivo studies) | High drug loading efficiency Magnetic nanoparticles for transdermal drug delivery Improved penetration of drugs across the skin upon application of MF |
[195] | |||
| IONPs/ Cur |
Salting-out with potassium phosphate | 90–350 nm | Breast cancer/ MDA-MB-231/ ND |
The magnetic targeting to cancer cells in vitro Higher uptake of drug-loaded nanoparticles than free drug Lower viability of cancer cells than control cells |
[209] | |||
| IONPs/ ODN (anti-c-myc) |
B. mori silk fibroin mixed with PEI | Salting-out with sodium phosphate | <200 nm | Breast cancer/ MDA-MB-231/ ND |
Magnetic-silk/PEI core-shell nanoparticles Lower surface charge and reduced cytotoxicity than magnetic-PEI-coated particles High cellular uptake, efficient magnetofection level |
[210] | ||
| MnO2/ Dox/ICG |
B. mori silk fibroin | Self-assembly induced by organic solvent | 140 nm | Breast cancer/ 4T1/ tumor-bearing mice |
Strong and stable photothermal effect upon NIR irradiation Effective tumor-specific accumulation via EPR effect Combination chemotherapy, PDT and PTT under the guidance of NIR/MR imaging Reduced systemic toxicity |
[211] | ||
| Photo-sensitive or photo-dynamic agents | ICG | B. mori silk fibroin | Desolvation with acetone | 210 nm | Glioblastoma/ C6/ Tumor xenograft in mice |
A therapeutic nano-platform for imaging and PTT of glioblastoma High encapsulation efficiency of photosensitive agent and slow drug release profile in vitro Increased stability of PTT effect under NIR irradiation than free drug Internalization of particles by cancer cells in vitro Accumulation of particles in site of tumor and tumor growth suppression in vivo |
[212] | |
| Ce6/ 5-FU |
B. mori silk fibroin | Desolvation with acetone | 278.2–364.9 nm | cRGDfk and Ce6-conjugated | Gastric cancer/ MGC-803/ Tumor xenograft in mice |
Combination of targeted drug delivery and (PDT) Active tumor targeting of integrin receptor-overexpressing cells Together with laser irradiation, the drug-loaded particles reduced the tumor burden Biocompatibility and safety in vivo |
[127] | |
| IR780 | B. mori silk sericin-cholesterol | Self-assembly induced by DMSO | 105 nm | FA-conjugated | Gastric cancer/ BGC-823/ ND |
Efficient incorporation of photosensitive substance IR780 Improved photo-stability and water solubility of IR780 Efficient absorption by FA-positive cancer cells Excellent PDT and PTT cytotoxicity towards cancer cells under NIR irradiation |
[47] |
EDC, ethylcarbodiimide; PEI, polyethylenimine; FA, folic acid; Dox, doxorubicin; 5-FU, 5′-fluorouracil; Ce6, chlorin e6; DMSO, dimethyl sulfoxide; CK2, casein kinase II; ASH2L, ASH2 like, histone lysine methyltransferase complex; GFP, green fluorescent protein; LUC, firefly luciferase; PLL, poly-L-lysine; HA, hyaluronic acid; IONPs, iron oxide nanoparticles; ICG, indocyanine green; Mtx, methotrexat; Cur, curcumin; MR, magnetic resonance; NIR, near-infrared, PDT, photodynamic therapy; PTT, photothermal therapy.