Multifunctional Lactobionic Acid-Modified Dendrimers for Targeted Drug Delivery to Liver Cancer Cells: Investigating the Role Played by PEG Spacer

We report the development of a lactobionic acid (LA)-modified multifunctional dendrimer-based carrier system for targeted therapy of liver cancer cells overexpressing asialoglycoprotein receptors. In this study, generation 5 (G5) poly­(amidoamine) (PAMAM) dendrimers were sequentially modified with fluorescein isothiocyanate (FI) and LA (or polyethylene glycol (PEG)-linked LA, PEG-LA), followed by acetylation of the remaining dendrimer terminal amines. The synthesized G5.NHAc-FI-LA or G5.NHAc-FI-PEG-LA conjugates (NHAc denotes acetamide groups) were used to encapsulate a model anticancer drug doxorubicin (DOX). We show that both conjugates are able to encapsulate approximately 5.0 DOX molecules within each dendrimer and the formed dendrimer/DOX complexes are stable under different pH conditions and different aqueous media. The G5.NHAc-FI-PEG-LA conjugate appears to have a better cytocompatibility, enables a slightly faster DOX release rate, and displays better liver cancer cell targeting ability than the G5.NHAc-FI-LA conjugate without PEG under similar experimental conditions. Importantly, the developed G5.NHAc-FI-PEG-LA/DOX complexes are able to specifically inhibit the growth of the target cells with a better efficiency than the G5.NHAc-FI-LA/DOX complexes at a relatively high DOX concentration. Our results suggest a key role played by the PEG spacer that affords the dendrimer platform with enhanced targeting and therapeutic efficacy of cancer cells. The developed LA-modified multifunctional dendrimer conjugate with a PEG spacer may be used as a delivery system for targeted liver cancer therapy and offers new opportunities in the design of multifunctional drug carriers for targeted cancer therapy applications

Acetylated Polyethylenimine-Entrapped Gold Nanoparticles Enable Negative Computed Tomography Imaging of Orthotopic Hepatic Carcinoma

Developing an effective computed tomography (CT) contrast agent is still a challenging task for precise diagnosis of hepatic carcinoma (HCC). Here, we present the use of acetylated polyethylenimine (PEI)-entrapped gold nanoparticles (Ac-PE-AuNPs) without antifouling modification for negative CT imaging of HCC. PEI was first linked to fluorescein isothiocyanate (FI) and then utilized as a vehicle for the entrapment of AuNPs. The particles were then acetylated to reduce its positive surface potential. The designed Ac-PE-AuNPs were characterized by various techniques. We find that the Ac-PE-AuNPs with a uniform size distribution (mean diameter = 2.3 nm) are colloidally stable and possess low toxicity in the studied range of concentration. Owing to the fact that the particles without additional antifouling modification were mainly gathered in liver, the Ac-PE-AuNPs could greatly improve the CT contrast enhancement of normal liver, whereas poor CT contrast enhancement appeared in liver necrosis region caused by HCC. As a result, HCC could be easily and precisely diagnosed. The designed Ac-PE-AuNPs were demonstrated to have biocompatibility through in vivo biodistribution and histological studies, hence holding an enormous potential to be adopted as an effective negative CT contrast agent for diagnosis of hepatoma carcinoma

Manganese Dioxide-Entrapping Dendrimers Co-Deliver Protein and Nucleotide for Magnetic Resonance Imaging-Guided Chemodynamic/Starvation/Immune Therapy of Tumors

Development of a nanoscale drug delivery system that can simultaneously exert efficient tumor therapeutic efficacy while creating the desired antitumor immune responses is still challenging. Herein, we report the use of a manganese dioxide (MnO2)-entrapping dendrimer nanocarrier to codeliver glucose oxidase (GOx) and cyclic GMP-AMP (cGAMP), an agonist of the stimulator of interferon genes (STING) for improved tumor chemodynamic/starvation/immune therapy. Methoxy poly(ethylene glycol) (mPEG)- and phenylboronic acid (PBA)-modified generation 5 (G5) poly(amidoamine) dendrimers were first synthesized and then entrapped with MnO2 nanoparticles (NPs) to generate the hybrid MnO2@G5-mPEG–PBA (MGPP) NPs. The created MGPP NPs with an MnO2 core size of 2.8 nm display efficient glutathione depletion ability, and a favorable Mn2+ release profile under a tumor microenvironment mimetic condition to enable Fenton-like reaction and T1-weighted magnetic resonance (MR) imaging. We show that the MGPP-mediated GOx delivery facilitates enhanced chemodynamic/starvation therapy of cancer cells in vitro, and further codelivery of cGAMP can effectively trigger immunogenic cell death (ICD) to strongly promote the maturation of dendritic cells. In a bilateral mouse colorectal tumor model, the dendrimer delivery nanosystem elicits a potent antitumor performance with a strong abscopal effect, greatly improving the overall mouse survival rate. Importantly, the dendrimer-mediated codelivery not only allows the coordination of Mn2+ with GOx and cGAMP for respective chemodynamic/starvation-triggered ICD and augmented STING activation to boost systemic antitumor immune responses, but also enables T1-weighted tumor MR imaging, potentially serving as a promising nanoplatform for enhanced antitumor therapy with desired immune responses

^99mTc-Labeled Multifunctional Low-Generation Dendrimer-Entrapped Gold Nanoparticles for Targeted SPECT/CT Dual-Mode Imaging of Tumors

Development of cost-effective and highly efficient nanoprobes for targeted tumor single-photon emission computed tomography (SPECT)/computed tomography (CT) dual-mode imaging remains a challenging task. Here, multifunctional dendrimer-entrapped gold nanoparticles (Au DENPs) modified with folic acid (FA) and labeled with ^99mTc were synthesized for targeted dual-mode SPECT/CT imaging of tumors. Generation 2 (G2) poly­(amidoamine) (PAMAM) dendrimers (G2-NH₂) conjugated with cyclic diethylenetriamine pentaacetic anhydride (cDTPAA) via an amide linkage and FA via a spacer of polyethylene glycol (PEG) were used for templated synthesis of Au core NPs, followed by labeling of ^99mTc via chelation. The thus created multifunctional Au DENPs were well-characterized. It is shown that particles with an average Au core diameter of 1.6 nm can be dispersed in water, display stability under different conditions, and are cytocompatible in the studied concentration range. Further results demonstrate that the multifunctional nanoprobe is able to be utilized for targeted SPECT/CT dual-mode imaging of cancer cells having FA receptor (FAR)-overexpression in vitro and the established subcutaneous tumor model in vivo within a time frame up to 4 h. The formed multifunctional Au DENPs synthesized using dendrimers of low-generation may be employed as an effective and economic nanoprobe for SPECT/CT imaging of different types of FAR-expressing tumors

Haematoxylin and eosin staining of endometrium (original magnification, 200×).

<p>(A) eutopic endometrium. (B) ectopic endometrium. The glandular tissue (arrow) was obviously observed in eutopic endometrium (A). In ectopic endometrium(B), the neovascularization (arrow) was observed under the columnar epithelial cells (arrowhead).</p

Fe content in the surgically induced endometriotic lesions at different time points after intravenous injection of HA-Fe₃O₄ NPs (2 mg Fe per mouse, in 1 mL PBS).

<p>Fe content in the surgically induced endometriotic lesions at different time points after intravenous injection of HA-Fe₃O₄ NPs (2 mg Fe per mouse, in 1 mL PBS).</p

T₂ signal intensity value of the wall of ectopic endometriotic lesions at different time points.

<p>T₂ signal intensity value of the wall of ectopic endometriotic lesions at different time points.</p

The surgically induced ectopic endometriotic lesions at four weeks post operation.

<p>The tubal cystic structure with a size of 23.8×4.7×3.9 mm was noticed at the fixed site (long arrow). The ectopic lesions were full of liquids and the small dendritic vessels on the surface of the wall were also clearly observed. Note that a strip of the adhesion tissues (arrowhead) was also observed between the ectopic lesions and abdominal wall.</p

Construction of Hybrid Alginate Nanogels Loaded with Manganese Oxide Nanoparticles for Enhanced Tumor Magnetic Resonance Imaging

Development of sensitive contrast agents for positive magnetic resonance (MR) imaging of biosystems still remains a great challenge. Herein, we report a facile process to construct hybrid alginate (AG) nanogels (NGs) loaded with manganese oxide (Mn₃O₄) nanoparticles (NPs) for enhanced tumor MR imaging. The obtained AG/PEI-Mn₃O₄ NGs with a mean size of 141.6 nm display excellent colloidal stability in aqueous solution and good cytocompatibility in the studied concentration range. Moreover, the hybrid NGs have a high <i>r</i>₁ relaxivity of 26.12 mM^–1 s^–1, which is about 19.5 times higher than that of PEI-Mn₃O₄ NPs with PEI surface amine acetylated (PEI.Ac–Mn₃O₄ NPs). Furthermore, the AG/PEI-Mn₃O₄ NGs presented longer blood circulation time and better tumor MR imaging performances <i>in vivo</i> than PEI.Ac–Mn₃O₄ NPs. With the good biosafety confirmed by histological examinations, the developed AG/PEI-Mn₃O₄ NGs may be potentially used as an efficient contrast agent for enhanced MR imaging of different biosystems

MR images of the ectopic endometriotic lesions at different time points.

<p>On T₁WI, the EUTs (arrow) appeared as ill defined cystic mass with low signal (A). On axial FS-T₂WI before injection (B), the EUTs appeared to have slightly high signal intensity surrounding with intermediate signals of fibrous walls. At 15 min (C), 30 min (D), 60 min (E), and 120 min (F) post injection, the wall of lesions were more clearly outlined and the lesion to background contrast was obviously improved compared with (A).</p