REDV Peptide Conjugated Nanoparticles/pZNF580 Complexes for Actively Targeting Human Vascular Endothelial Cells

Herein, we demonstrate that the REDV peptide modified nanoparticles (NPs) can serve as a kind of active targeting gene carrier to condensate pZNF580 for specific promotion of the proliferation of endothelial cells (ECs). First, we synthesized a series of biodegradable amphiphilic copolymers by ring-opening polymerization reaction and graft modification with REDV peptide. Second, we prepared active targeting NPs via self-assembly of the amphiphilic copolymers using nanoprecipitation technology. After condensation with negatively charged pZNF580, the REDV peptide modified NPs/pZNF580 complexes were formed finally. Due to the binding affinity toward ECs of the specific peptide, these REDV peptide modified NPs/pZNF580 complexes could be recognized and adhered specifically by ECs in the coculture system of ECs and human artery smooth muscle cells (SMCs) <i>in vitro</i>. After expression of ZNF580, as the key protein to promote the proliferation of ECs, the relative ZNF580 protein level increased from 15.7% to 34.8%. The specificity in actively targeting ECs of the REDV peptide conjugated NPs/pZNF580 complexes was still retained in the coculture system. These findings in the present study could facilitate the development of actively targeting gene carriers for the endothelialization of artificial blood vessels

Multitargeting Peptide-Functionalized Star-Shaped Copolymers with Comblike Structure and a POSS-Core To Effectively Transfect Endothelial Cells

Gene therapy meets one serious bottleneck in clinical application, namely, lack of safe and efficient gene delivery systems. In order to solve this problem, we designed a lowly cytotoxic and highly efficient gene delivery system for the transfection of endothelial cells. Octa­(3-ammoniumpropyl)­octasilsesquioxane octachloride reacted with 2-bromoisobutyryl bromide under alkaline condition, and sequentially initiated 2-(dimethylamino)­ethyl methacrylate (DMAEMA) and poly­(ethylene glycol) monomethacrylate (PEGMA) via atom transfer radical polymerization (ATRP) to prepare the eight-arm copolymer with a biocompatible polyhedral oligomericsilsesquioxane (POSS). The side chain ends of the comblike PPEGMA were double-bonded to facilitate the attachment of CAGW or CAG-TAT-NLS functional peptide, thereby enabling the star-shaped copolymers with multifunction. The peptide-functionalized star-shaped copolymers were self-assembled into nanoparticles (NPs) and used to condense pEGFP-ZNF580 (pDNA) to prepare the NPs/pDNA complexes. These complexes had low toxicity as confirmed by MTT. The results of fluorescence microscopy and flow cytometry showed that these multitargeting functionalized gene complexes could effectively transfect endothelial cells. Their transfection efficiency is higher than the positive control PEI 25 kDa group. Moreover, the Western blot test, wound healing assay, and in vitro tube formation assay also demonstrated that the transfected cells showed high migration and enhanced angiogenesis. The pDNA was effective delivered and expressed in endothelial cells by these multitargeting functionalized gene complexes, and promoted cell migration and tube formation. These star-shaped copolymers with comblike structure and a POSS core are a potential gene carrier for gene therapy

MOESM1 of Oligohistidine and targeting peptide functionalized TAT-NLS for enhancing cellular uptake and promoting angiogenesis in vivo

Additional file 1: Figure S1. Hydrodynamic diameter distribution of REDV-TAT-NLS-Hn micelles characterized by DLS. Figure S2. Hydrodynamic diameter distribution of REDV-TAT-NLS-Hn/pZNF580 complexes (w/w = 1) characterized by DLS. Figure S3. Hydrodynamic diameter distribution of REDV-TAT-NLS-Hn/pZNF580 complexes (w/w = 2) characterized by DLS. Figure S4. Hydrodynamic diameter distribution of REDV-TAT-NLS-Hn/pZNF580 complexes (w/w = 3) characterized by DLS. Figure S5. Hydrodynamic diameter distribution of REDV-TAT-NLS-Hn/pZNF580 complexes (w/w = 4) characterized by DLS. Figure S6. Hydrodynamic diameter distribution of REDV-TAT-NLS-Hn/pZNF580 complexes (w/w = 5) characterized by DLS

CAGW Modified Polymeric Micelles with Different Hydrophobic Cores for Efficient Gene Delivery and Capillary-like Tube Formation

Recently, polymeric micelles with different biodegradable hydrophobic cores, such as poly­(lactide-co-glycolide) (PLGA) and poly­(lactide-co-3­(S)-methyl-morpholine-2,5-dione) (PLMD), have been used for gene delivery. The biodegradable hydrophobic cores should play an important role in gene delivery. However, little research has focused on selectively promoting proliferation and migration of endothelial cells (ECs) as well as vascularization by altering hydrophobic cores of polymeric micelles. Herein, we prepared two kinds of CAGW peptide (selective adhesion for ECs) modified micelles with PLGA and PLMD as hydrophobic cores, respectively, and poly­(ethylene glycol) (PEG) and polyethylenimine (PEI) as mixed hydrophilic shell. Their ability of condensing pEGFP-ZNF580 (pZNF580) to form gene complexes was proved by agarose gel electrophoresis assay. MTT results showed that the relative cell viability of the micelles with PLMD cores was higher than control groups and the micelles with PLGA cores. The cellular uptake ability of these CAGW modified gene complexes was higher than the complexes without CAGW target function. A similar trend was also found in transfection tests in vitro, which further demonstrated the effect of CAGW peptide and different hydrophobic cores on gene delivery. The number of migrated cells treated by the gene complexes with PLGA cores was 82 (nontarget group) and 115 (target group), whereas the complexes with PLMD cores was 88 (nontarget group) and 120 (target group). Capillary-like tube formation of CAGW peptide modified complexes with PLMD core group was much higher (about 6 times) than the PEI­(10 kDa)/pZNF580 group. These results demonstrated that transfection efficiency, cell proliferation, migration, and vascularization could be promoted by altering hydrophobic cores and CAGW modification

Synthesis of In-Chain-Functionalized Polystyrene-<i>block</i>-poly(dimethylsiloxane) Diblock Copolymers by Anionic Polymerization and Hydrosilylation Using Dimethyl-[4-(1-phenylvinyl)phenyl]silane

Synthesis of In-Chain-Functionalized Polystyrene-block-poly(dimethylsiloxane) Diblock Copolymers by Anionic Polymerization and Hydrosilylation Using Dimethyl-[4-(1-phenylvinyl)phenyl]silan

Synthesis, Self-assembly, and Crystal Structure of a Shape-Persistent Polyhedral-Oligosilsesquioxane-Nanoparticle-Tethered Perylene Diimide

A novel organic−inorganic hybrid with two polyhedral oligosilsesquioxane (POSS) nanoparticles covalently attached to perylene diimide (PDI) via a rigid 1,4-phenylene linkage (POSS-PDI-POSS) was designed and synthesized to examine the effect of bulky and well-defined nanoparticle side chains on the self-assembly behavior of PDI derivatives. The molecules were self-assembled directly by slow evaporation of a cast drop from solution in tetrahydrofuran to give rise to uniform crystalline nanobelts with dimensions typically of 0.2 mm × 1 μm × 50 nm. The phase behavior and crystal structure of the sample were then elucidated via a combination of different experimental techniques such as differential scanning calorimetry (DSC), wide-angle X-ray diffraction (WAXD), selected area electron diffraction (SAED) in transmission electron microscopy (TEM), polarized light microscopy, and atomic force microscopy. One-dimensional (1D) WAXD and DSC revealed that only one crystalline phase exists. Based on the 2D WAXD fiber pattern obtained from the oriented POSS-PDI-POSS samples, the crystalline structure was determined to be a triclinic unit cell with dimensions of a = 6.577 nm, b = 5.213 nm, c = 1.107 nm, α = 93.26°, β = 94.85°, and γ = 92.73°, which was confirmed by SAED experiments on the single crystals with different crystal zone orientations. The detailed molecular conformational analysis indicated that the steric hindrance of the POSS nanoparticles covalently attached to PDI via a rigid 1,4-phenylene linkage makes it difficult to achieve a continuous stacking of PDIs. Instead, the molecules dimerized to maximize the π−π interaction. The dimers then became the building blocks and packed themselves into the unit cell. This strong tendency for dimerization was supported by concentration-dependent ultraviolet/visible absorption spectra, florescence spectra, and tandem mass spectroscopy with traveling wave ion mobility separation. The combined SAED and TEM results showed that the c*-axis of the crystal is along the elongated direction of the single-crystal nanobelt and the normal direction of the π−π stacking is along the a*-axis. A crystal structure with six dimers as one supramolecular motif in one unit cell was proposed to account for the unusually large unit cell dimensions. The complex structure could be attributed to the longitudinal, transverse, and slightly rotational offsets between the PDIs in the dimers and interdigitated neighboring dimers due probably to both electrostatic interactions and steric demands. The molecular packing scheme in the crystal was simulated using Cerius2 software, and the resulting diffraction data agreed well with the experimental results. The rationale for such 1D nanostructured morphology formation is also discussed