A novel dual-targeting Pluronic/poly (lactic acid) polymersome containing transferrin and folic acid ligands (Tf/FA-F127-PLA) has been designed to study its application in the targeted drug delivery system. Both biotin and folic acid conjugated Biotin/FA-F127-PLA polymersomes (Ps) were prepared as the precursor.
The dual-targeting behaviors of Tf/FA-F127-PLA over C6 glioma cells were then fulfilled through connecting the precursor with biotinylated transferrin by using a three-step biotin-avidin technique.
Paclitaxel (PTX) was loaded successfully into Biotin/FA-F127-PLA and showed a burst release followed by a slow-release process in vitro.
It was also obtained that Tf/FA-F127-PLA had higher cytotoxicity and cellular uptake amount than non-targeted and single-targeted Ps do.
These results could be because more PTX-loaded Tf/FA-F127-PLA Ps entered C6 cells through both FA-Folate Receptor (FR) and Tf-Transferrin Receptor (TfR) specific affinity and thus possessed the better anti-tumor ability.
It was further proved that the uptake of Ps by C6 cells was through the endocytosis related to clathrin, caveolae, and lysosome, etc.
Furthermore, it was demonstrated that the uptake of dual-targeting Tf/FA-F127-PLA Ps by C6 cells was related to the endocytosis mediated by both FR and TfR.
These findings indicated that dual-targeting Tf/FA-F127-PLA Ps could be a potential carrier in targeted drug delivery systems.
Transferrin/Folate Dual-Targeting Pluronic F127/Poly(lactic acid) Polymersomes for Effective Anticancer Drug Delivery
A novel dual-targeting Pluronic/poly (lactic acid) polymersome containing transferrin and folic acid ligands (Tf/FA-F127-PLA) has been designed to study its application in the targeted drug delivery system. Both biotin and folic acid conjugated Biotin/FA-F127-PLA polymersomes (Ps) were prepared as the precursor.
The dual-targeting behaviors of Tf/FA-F127-PLA over C6 glioma cells were then fulfilled through connecting the precursor with biotinylated transferrin by using a three-step biotin-avidin technique.
Paclitaxel (PTX) was loaded successfully into Biotin/FA-F127-PLA and showed a burst release followed by a slow-release process in vitro.
It was also obtained that Tf/FA-F127-PLA had higher cytotoxicity and cellular uptake amount than non-targeted and single-targeted Ps do.
These results could be because more PTX-loaded Tf/FA-F127-PLA Ps entered C6 cells through both FA-Folate Receptor (FR) and Tf-Transferrin Receptor (TfR) specific affinity and thus possessed the better anti-tumor ability.
It was further proved that the uptake of Ps by C6 cells was through the endocytosis related to clathrin, caveolae, and lysosome, etc.
Furthermore, it was demonstrated that the uptake of dual-targeting Tf/FA-F127-PLA Ps by C6 cells was related to the endocytosis mediated by both FR and TfR.
These findings indicated that dual-targeting Tf/FA-F127-PLA Ps could be a potential carrier in targeted drug delivery systems.
Synthesis of Biotinylated PAMAM G3 Dendrimers Substituted with R-Glycidol and Celecoxib/Simvastatin as Repurposed Drugs and Evaluation of Their Increased Additive Cytotoxicity for Cancer Cell Lines
Recent achievement in anticancer therapy considers the application of repurposed drugs in optimal combinations with the use of specific carriers for their targeted delivery. As a result, new optimized medications with reduced side effects can be obtained.
In this study, two known anticancer drugs, celecoxib and/or simvastatin, were conjugated covalently with PAMAM G3 dendrimer and tested in vitro against human squamous carcinoma (SCC-15-15) and glioblastoma (U-118 MG) cells, as well as normal human fibroblasts (BJ).
The obtained conjugates were also substituted with biotin and R-glycidol to increase their affinity for cancer cells and were characterized with NMR spectroscopy and dynamic light scattering technique.
Conjugates furnished with two celecoxib and four simvastatin residues revealed the very high effectiveness and dramatically decreased the SCC-15 and U-118 MG cell viability at very low concentrations with IC50 equal to about 3 µM. Its action was 20-50-fold stronger than that of either drug alone or as a mixture.
Combined conjugate revealed also additive action since it was 2-8-fold more effective than conjugates with either single drug.
The combined conjugate revealed rather low specificity since it was also highly cytotoxic for BJ cells.
Despite this, it may be concluded that biotinylated and R-glycidylated PAMAM G3 dendrimers substituted with both celecoxib and simvastatin can be considered as a new perspective anticancer agent, effective in therapy of malignant, incurable glioblastomas.
Synthesis of Spherical Nanoparticle Hybrids via Aerosol Thiol-Ene Photopolymerization and Their Bioconjugation
Hybrid nanomaterials possess the properties of both organic and inorganic components and find applications in various fields of research and technology.
In this study, aerosol photopolymerization is used in combination with thiol-ene chemistry to produce silver poly(thio-ether) hybrid nanospheres.
In aerosol photopolymerization, a spray solution of monomers is atomized, forming a droplet aerosol, which then polymerizes, producing spherical polymer nanoparticles.
To produce silver poly(thio-ether) hybrids, silver nanoparticles were introduced to the spray solution.
Diverse methods of stabilization were used to produce stable dispersions of silver nanoparticles to prevent their agglomeration before the photopolymerization process.
Successfully stabilized silver nanoparticle dispersion in the spray solution subsequently formed nanocomposites with non-agglomerated silver nanoparticles inside the polymer matrix.
Nanocomposite particles were analyzed via scanning and transmission electron microscopy to study the degree of agglomeration of silver nanoparticles and their location inside the polymer spheres.
The nanoparticle hybrids were then introduced onto various biofunctionalization reactions.
A two-step bioconjugation process was developed involving the hybrid nanoparticles: (1) conjugation of (biotin)-maleimide to thiol-groups on the polymer network of the hybrids, and (2) biotin-streptavidin binding.
The biofunctionalization with gold-nanoparticle-conjugates was carried out to confirm the reactivity of -SH groups on each conjugation step.
Fluorescence-labeled biomolecules were conjugated to the spherical nanoparticle hybrids (applying the two-step bioconjugation process) verified by Fluorescence Spectroscopy and Fluorescence Microscopy.
The presented research offers an effective method of synthesis of smart systems that can further be used in biosensors and various other biomedical applications.
The Influence of Genes on the “Killer Plasmid” of Dinoroseobacter shibae on Its Symbiosis With the Dinoflagellate Prorocentrum minimum
The marine bacterium Dinoroseobacter shibae shows a Jekyll-and-Hyde behavior in co-culture with the dinoflagellate Prorocentrum minimum:
In the initial symbiotic phase,
it provides the essential vitamins B12 (cobalamin) and B1 (thiamine) to the algae.
In the later pathogenic phase it kills the dinoflagellate.
The killing phenotype is determined by the 191 kb plasmid and can be conjugated into other Roseobacters.
From a transposon-library of D. shibae we retrieved 28 mutants whose insertion sites were located on the 191 kb plasmid.
We co-cultivated each of them with P. minimum in L1 medium lacking vitamin B12.
V5, Biotin conjugated |
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| AS15-2965 | Agrisera AB | 100 µg | 443 EUR |
S, Biotin conjugated |
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| AS15-2975 | Agrisera AB | 100 µg | 443 EUR |
T7, Biotin conjugated |
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| AS15-2977 | Agrisera AB | 100 µg | 443 EUR |
Urease, Biotin conjugated |
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| AS09-560 | Agrisera AB | 10 mg | 401 EUR |
Lysozyme Biotin conjugated |
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| AR-6637-11 | ImmunoBioscience | 5mg | 167.3 EUR |
Bromelain, Biotin conjugated |
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| AS09-553 | Agrisera AB | 10 mg | 401 EUR |
Fibrinogen, Biotin conjugated |
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| IMS06-038-312 | Agrisera AB | 100 µl (1mg/ml) | 348 EUR |
PPI | papain, Biotin conjugated |
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| AS09-546 | Agrisera AB | 10 mg | 401 EUR |
Biotin Conjugated Protein A |
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| BA1025-0.5 | BosterBio | 0.5ml | 157.2 EUR |
Biotin Conjugated Protein A |
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| BA1025-1 | BosterBio | 1ml | 242.4 EUR |
Biotin Conjugated Protein G |
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| BA1026-0.5 | BosterBio | 0.5ml | 279.6 EUR |
Biotin Conjugated Protein G |
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| BA1026-1 | BosterBio | 1ml | 486 EUR |
Biotin Conjugated Protein G |
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| MBS176618-05mL | MyBiosource | 0.5mL | 260 EUR |
Biotin Conjugated Protein G |
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| MBS176618-1mL | MyBiosource | 1mL | 430 EUR |
With 20 mutant strains no algal growth beyond the axenic control lacking B12 occurred.
Several of these genes were predicted to encode proteins from the type IV secretion system (T4SS).
They are apparently essential for establishing the symbiosis.
With five transposon mutant strains, the initial symbiotic phase was intact but the later pathogenic phase was lost in co-culture. In three of them the insertion sites were located in an operon predicted to encode genes for biotin (B7) uptake. Both P. minimum and D. shibae are auxotrophic for biotin.
We hypothesize that the bacterium depletes the medium from biotin resulting in apoptosis of the dinoflagellate.