PAMAM G4 Research Articles

Novel Hyperbranched Dialdehyde Carboxymethyl Cellulose-Grafted Gelatin Films as Potential Natural Antibacterial Materials

Carboxymethyl cellulose (CMC) has potential applications in food packaging given its good processibility and biological properties. In this research, dialdehyde carboxymethyl cellulose (DCMC) was first modified with polyamidoamine (PAMAM G0) to synthesize hyperbranched dialdehyde carboxymethyl cellulose (HPD). Then, a novel hyperbranched dialdehyde carboxymethyl cellulose-grafted gelatin (HPD/Gs) film was obtained by the “one-pot” method. The structural characteristics, degree of cross-linking, thickness, swelling behavior, water solubility, water vapor permeability, surface morphology, hydrophilicity, mechanical strength, antibacterial properties, and biocompatibility of HPD/Gs were characterized by using Fourier transform infrared (FTIR), scanning electron microscopy energy-dispersive X-ray spectroscopy (SEM-EDS), X-ray diffractometry (XRD), thermogravimetric analysis/derivative thermogravimetry (TGA/DTG), differential scanning calorimetry (DSC), and other analysis technologies. The results revealed that the prepared HPD/Gs film showed good thermal stability and mechanical properties. In particular, when the mass ratio of HPD to G was 1:1.5, the cross-linking degree, swelling behaviors, water vapor permeability (WVP), water contact angle, tensile strength, and elongation at break of the HPD/1.5G film reached 42.757%, 109.114%, 46.71 g·mm/m2·day·kPa (1 day), 115.4 ± 4.5°, 1.77 MPa, and 320.93%, respectively. Moreover, the HPD/Gs films had good cytocompatibility and antibacterial properties against Staphylococcus aureus and Escherichia coli. The research indicated that the designed HPD/Gs are a type of green antimicrobial material with potential value.

Efficient Removal of Uranyl Ions Using PAMAM Dendrimer: Simulation and Experiment

In this work, using atomistic molecular dynamics (MD) simulations and polymer-assisted ultrafiltration experiments, we explore the adsorption and removal of uranyl ions from aqueous solutions using poly(amidoamine) (PAMAM) dendrimers. The effects of uranyl ion concentration and the pH of the solution were examined for PAMAM dendrimers of generations 3, 4, and 5. Our simulation results show that PAMAM has a high adsorption capacity for the uranyl ions. The adsorption capacity increases with increasing concentration of uranyl ions for all 3 generations of PAMAM in agreement with experimental findings. We find that the number of uranyl ions bound to PAMAM is significantly higher in acidic solutions (pH < 3) as compared to neutral solutions (pH ∼ 7) for all uranyl ion concentrations. Additionally, we find an increase in the number of adsorbed uranyl ions to PAMAM with the increase in the dendrimer generation. This increase is due to the greater number of binding sites present for higher-generation PAMAM dendrimers. Our simulation study shows that nitrate ions form a solvation shell around uranyl ions, which allows them to bind to PAMAM binding sites, including the amide, amine, and carbonyl groups. In polymer-assisted ultrafiltration (PAUF) experiments, the removal percentage of uranyl ions by G3 PAMAM dendrimer increased from 36.3% to 42.6% as the metal ion concentration increased from 2.1 × 10-5 M to 10.5 × 10-5 M at a pH of 2. Our combined experiment and simulation study suggests that PAMAM is an effective adsorbent for removing uranyl ions from aqueous solutions.

High-Performance Multi-Dynamic Bond Cross-Linked Hydrogel with Spatiotemporal siRNA Delivery for Gene-Cell Combination Therapy of Intervertebral Disc Degeneration.

Chronic inflammatory diseases, such as intervertebral disc degeneration (IVDD), which affect the lives of hundreds of millions of people, still lack effective and precise treatments. In this study, a novel hydrogel system with many extraordinary properties is developed for gene-cell combination therapy of IVDD. Phenylboronic acid-modified G5 PAMAM (G5-PBA) is first synthesized, and therapeutic siRNA silencing the expression of P65 mixed with G5-PBA (siRNA@G5-PBA) is then embedded into the hydrogel (siRNA@G5-PBA@Gel) based on multi-dynamic bonds including acyl hydrazone bonds, imine linkage, π-π stacking, and hydrogen bonding interactions. Local and acidic inflammatory microenvironment-responsive gene-drug release can achieve spatiotemporal regulation of gene expression. In addition, gene-drug release from the hydrogel can be sustained for more than 28 days in vitro and in vivo, greatly inhibiting the secretion of inflammatory factors and the subsequent degeneration of nucleus pulposus (NP) cells induced by lipopolysaccharide (LPS). Through prolonged inhibition of the P65/NLRP3 signaling pathway, the siRNA@G5-PBA@Gel is verified to relieve inflammatory storms, which can significantly enhance the regeneration of IVD when combined with cell therapy. Overall, this study proposes an innovative system for gene-cell combination therapy and a precise and minimally invasive treatment method for IVD regeneration.

Bioaffinity Recovery of Linear CRGDS Peptides Grafted to Zwitterionic PAMAM Nanocarriers

Functional oligopeptides derived from natural proteins are often used as targeting groups or therapeutic drugs but always suffer from a dramatic bioaffinity decrease due to the loss of conformational restriction in proteins, which increases their entropy state. It is worth noting that the linear Cys-Arg-Gly-Asp-Ser (CRGDS) oligopeptide can spontaneously restore its natural conformation by anchoring one terminus and forming hydrogen bonds on another terminus, on protein-like zwitterionic fifth-generation polyamide-amine (G5 PAMAM) nanocarriers, thus exhibiting high bioaffinity. However, the different physicochemical properties of nanocarriers have an impact on the bioaffinity recovery of linear CRGDS. In this work, it is found that the bioaffinity decreases with the reduction of generation of PAMAM and zwitterionic G3 PAMAM is a turning point for bioaffinity recovery via a series of in vitro experiments. This indicates that the spontaneous restoration of the native conformation of the RGD segment requires proper surface group density and structural rigidity of the zwitterionic nanocarrier for reducing the entropic energy of CRGDS peptides before binding. This approach paves a way for functional reproduction of protein molecules using low-cost synthetic polymers for extensive biomedical applications.

Phenylboronic acid conjugated PAMAM G4 dendrimers augmented usnic acid delivery to gastric cancer cells

Here we report on the synthesis, characterization, and biological evaluation of Siglec’s targeted G4 PAMAM dendrimers loaded with usnic acid (UA). Siglec's targeted G4 dendrimers were obtained by conjugation of 4-carboxy phenylboronic acid (PBA) to the periphery of the G4 PAMAM dendrimer (PBA-PAMAM). UA-loaded PBA-PAMAM dendrimers (UA@PBA-PAMAM) showed average particle size (18 nm) without any sign of aggregation with amorphous nature, stability of the drug, and PBA-mediated anticancer efficacy. The UA@PBA-PAMAM dendrimer formulation showed a site-specific drug release profile. After 12 h, 17.6% and 95.7% of UA was released from UA@PBA-PAMAM dendrimer formulation in pH 7.4 and pH 5.0, respectively. In vitro anticancer results suggested the no cytotoxicity of G4 dendrimers after PBA conjugation. The cell viability of AGS cells was observed as 88.9 ± 0.7% (12 h) and 50.4 ± 1.9% (24 h) after pure UA treatment and 37.3 ± 0.6% (12 h) and 14.4% ± 1.2 (24 h) after treatment with UA@PBA-PAMAM. The pure UA had IC50 values of 44.9 ± 3.1 μg/mL (for 12 h) and 8.2 ± 1.1 μg/mL (for 24 h) while UA@PBA-PAMAM had an IC50 value of 8.4 ± 0.3 μg/mL (for 12 h) and 2.7 ± 0.4 μg/mL (for 24 h) in AGS cells. As a result of cytotoxicity studies, the dendrimer formulation UA@PBA-PAMAM maintained its effectiveness and was able to produce a long-term anticancer effect over time compared to the pure UA dose. The PBA-PAMAM dendrimer also significantly enhanced the cellular internalization in AGS gastric cancer cells. The new PBA-PAMAM design appears to be suitable nanocarriers for the UA to deliver gastric cancer cells.

Development of 177Lu-Cetuximab-PAMAM dendrimeric nanosystem: a novel theranostic radioimmunoconjugate.

Epidermal growth factor receptors (EGFRs) are overexpressed in a wide range of tumors and are attractive candidates to target in targeted therapies. This study aimed to introduce a novel radiolabeled compound, 177Lu-cetuximab-PAMAM G4, for the treatment of EGFR-expressing tumors. In this study, the cetuximab mAb was bound to PAMAM G4 and labeled with 177Lu via DTPA-CHX chelator. The synthesized nanosystem was confirmed by different analyses such as DLS, FT-IR, TEM, and RT-LC. Cell viability of the radioimmunoconjugate was assessed over the EGFR-expressing cell line of SW480. The biodistribution of 177Lu-Cetuximab-PAMAMG4 was determined in different intervals after injection of the radiolabeled compound in normal and tumoral nude mice via scarification and SPECT images. The average size of PAMAM G4 and PAMAM-Cetuximab-DTPA-CHX nanoparticles were 2 and 70nm, respectively. 177Lu-Cetuximab-PAMAMG4 was prepared with radiochemical purity of more than 98%. The survival rates of SW480 cells at 24, 48, and 72h post-treatment with177Lu-Cetuximab-PAMAMG4 (500nM) were 18%, 15%, and 14%, respectively. The biodistribution studies showed a significant accumulation of 177Lu-Cetuximab-PAMAM in the EGFR-expressing tumor. According to the results, this new agent can be considered as an efficient therapeutic complex for tumors expressing EGFR receptors.

Conjugation of Short Oligopeptides to a Second-Generation Polyamidoamine Dendrimer Shows Antibacterial Activity

The growing evolution of bacterial resistance to antibiotics represents a global issue that not only impacts healthcare systems but also political and economic processes. This necessitates the development of novel antibacterial agents. Antimicrobial peptides have shown promise in this regard. Thus, in this study, a new functional polymer was synthesized by joining a short oligopeptide sequence (Phe-Lys-Phe-Leu, FKFL) to the surface of a second-generation polyamidoamine (G2 PAMAM) dendrimer as an antibacterial component. This method of synthesis proved simple and resulted in a high conjugation yield of the product FKFL-G2. To determine its antibacterial potential, FKFL-G2 was subsequently analyzed via mass spectrometry, a cytotoxicity assay, bacterial growth assay, colony-forming unit assay, membrane permeabilization assay, transmission electron microscopy, and biofilm formation assay. FKFL-G2 was found to exhibit low toxicity to noncancerous NIH3T3 cells. Additionally, FKFL-G2 had an antibacterial effect on Escherichia coli and Staphylococcus aureus strains by interacting with and disrupting the bacterial cell membrane. Based on these findings, FKFL-G2 shows promise as a potential antibacterial agent.

Carbon Nanotube-Incorporated Nanofibers for Immunosensor Preparation against CD36.

The increased serum concentration of CD36 is significantly associated with atherosclerosis, insulin resistance, and diabetes mellitus. Currently, there is no sensor system used for the detection of CD36 in the clinical field. Therefore, there is a need to develop a sensor system for the detection of CD36. The large surface area/volume ratio and controllable surface conformation of electrospun nanofibers (ENs) make them highly attractive for immunosensor applications. In the present study, PS/MWCNT-PAMAM ENs were produced and used as an immobilization matrix of Anti-CD36. Thus, the electrochemical behavior of the developed nanocomposite-based ENs and their usage potential were investigated for immunosensor applications. First, an oxidized multiwall carbon nanotube (MWCNT-OH) was synthesized and modified with a polyamidoamine generation 3 (PAMAM G3) dendrimer. The synthesized MWCNT-PAMAM nanocomposite was mixed with polystyrene (PS) solutions at different ratios to produce bead-free, smooth, and uniform PS/MWCNT-PAMAM ENs. PS/MWCNT-PAMAM ENs were accumulated on a screen-printed carbon electrode (SPCE) using the electrospinning technique. A biofunctional surface on the PS/MWCNT-PAMAM EN-coated SPCE was created using carbodiimide chemistry by covalent immobilization of Anti-CD36. The analytic performance characteristics of the developed PS/MWCNT-PAMAM/Anti-CD36 immunosensor were determined by performing electrochemical measurements in the presence of the CD36 protein. The linear detection range was found to be from 5 to 40 ng/mL, and the limit of detection was calculated as 3.94 ng/mL for CD36. The developed PS/MWCNT-PAMAM/Anti-CD36 immunosensor also displayed high tolerance to interference substances, good repeatability, and high recovery percent (recovery%) for artificial blood serum analysis.

Effect of PEGylation on drug uptake, biodistribution, and tissue toxicity of efavirenz–ritonavir loaded PAMAM G4 dendrimers

The present investigations aimed to compare the efficiency of PAMAM G4 (PG4) and PEGylated PAMAM G4 (PPG4) dendrimers as novel nanocarriers for the treatment of HIV-1. Synthesized PG4 and PPG4 dendrimers were confirmed by electrospray ionization and particle size with its morphology. The anti-human immunodeficiency virus (HIV) drug efavirenz (EFV) with a booster dose of ritonavir (RTV) was encapsulated into PG4 and PPG4 formerly noted as PG4ER and PPG4ER, respectively. Further, evaluated for dendrimers mediated solubilization, drug release, cytotoxicity, drug uptake, plasma, and tissue pharmacokinetics, and histopathology. PG4ER and PPG4ER both promoted a prolonged release of EFV in weakly acidic pH 4 up to 84 h and 132 h, respectively. The results of the cytotoxicity assay and drug uptake study showed that PPG4ER was safe and biocompatible up to 12.5 µg/ml. The plasma pharmacokinetic profile of EFV and RTV was significantly increased by PPG4ER with prolonged t 1/2 up to three times as compared to free EFV–RTV and PG4ER. Histopathological analysis showed remarkably lower tissue toxicity in PPG4ER as compared to free EFV–RTV. Therefore, overall data suggested that PPG4 has a great potential for prolonged release of EFV and RTV with enhanced bioavailability and lower toxicity.

Electrochemical sensing of methotrexate in the presence of folic acid using PAMAM dendrimer-functionalized multiwalled carbon nanotube-modified electrode.

This works presents a novel electrochemical sensor based on the third-generation poly(amidoamine) dendrimer (G3 PAMAM)-functionalized multiwalled carbon nanotube (MWCNT)-modified screen-printed graphite electrode (SPGE) for the simple and sensitive detection of methotrexate (MTX). The carboxylated MWCNTs were covalently functionalized with amino groups of G3 PAMAM and characterized using different techniques. The sensing ability of the designed nanosensor (MWCNTs-PAMAM/SPGE) was tested using differential pulse voltammetry (DPV), chronoamperometry (CHA), linear sweep voltammetry (LSV), and cyclic voltammetry (CV). To investigate the electrocatalytic activity of PAMAM-functionalized MWCNTs, a comparative electrochemical analysis was carried out and it was determined that PAMAM-functionalized MWCNT-modified SPGE showed good electrocatalytic performance for MTX oxidation compared to the unmodified SPGE. The MWCNT-PAMAM/SPGE lead to a reduced overpotential of MTX oxidation of about 300 mV and enhanced current of about 9 μA of the unmodified SPGE. Experiments were performed for the quantitative determination of MTX using the DPV technique. The response peak current linearly related against MTX concentration in the ranges from 0.01 to 110.0 μM and a limit of detection (LOD) equal to 0.003 μM. Also, MWCNT-PAMAM/SPGE exhibits good catalytic ability toward MTX determination in the presence of folic acid (FA), and the separation of their oxidation peaks (peak potential difference = 320 mV) simultaneously detected the above compounds. To prove the applicability of the MWCNT-PAMAM/SPGE sensor, the concentrations of MTX and FA in pharmaceutical products and biological samples were determined. The calculated recoveries were close to 100%, which indicates that the method might be assumed to be accurate.

Mixed-surface polyamidoamine polymer variants retain nucleic acid-scavenger ability with reduced toxicity.

Nucleic acid-binding polymers can have anti-inflammatory properties and beneficial effects in animal models of infection, trauma, cancer, and autoimmunity. PAMAM G3, a polyamidoamine dendrimer, is fully cationic bearing 32 protonable surface amines. However, while PAMAM G3 treatment leads to improved outcomes for mice infected with influenza, at risk of cancer metastasis, or genetically prone to lupus, its administration can lead to serosal inflammation and elevation of biomarkers of liver and kidney damage. Variants with reduced density of cationic charge through the interspersal of hydroxyl groups were evaluated as potentially better-tolerated alternatives. Notably, the variant PAMAM G3 50:50, similar in size as PAMAM G3 but with half the charge, was not toxic in cell culture, less associated with weight loss or serosal inflammation after parenteral administration, and remained effective in reducing glomerulonephritis in lupus-prone mice. Identification of such modified scavengers should facilitate their development as safe and effective anti-inflammatory agents.

Some things old, new and borrowed: Delivery of dabrafenib and vemurafenib to melanoma cells via self-assembled nanomicelles based on an amphiphilic dendrimer

Two clinically approved anticancer drugs targeting BRAF in melanoma patients – dabrafenib (DAB) and vemurafenib (VEM) – have been successfully encapsulated into nanomicelles formed upon self-assembly of an amphiphilic dendrimer AD based on two C18 aliphatic chains and a G2 PAMAM head. The process resulted in the formation of well-defined (∼10 nm) core-shell nanomicelles (NMs) with excellent encapsulation efficiency (∼70% for DAB and ∼60% for VEM) and good drug loading capacity (∼27% and ∼24% for DAB and VEM, respectively). Dynamic light scattering (DLS), transmission electron microscopy (TEM), small-angle x-ray scattering (SAXS), nuclear magnetic resonance (NMR), isothermal titration calorimetry (ITC), and molecular simulation (MS) experiments were used, respectively, to determine the size and structure of the empty and drug-loaded nanomicelles (DLNMs), along with the interactions between the NMs and their cargoes. The in vitro release data revealed profiles governed by Fickian diffusion; moreover, for both anticancer molecules, an acidic environment (pH = 5.0) facilitated drug release with respect to physiological pH conditions (pH = 7.4). Finally, both DAB- and VEM-loaded NMs elicited enhanced response with respect to free drug treatments in 4 different melanoma cell lines.

PEGylated G4 dendrimers as a promising nanocarrier for piperlongumine delivery: Synthesis, characterization, and anticancer activity

PAMAM dendrimers are well-known delivery vehicles for various therapeutic agents like synthetic drugs, natural phytoconstituents, and genetic materials. The tunable physicochemical properties and variety of surface interactions make them attractive nano delivery system. However, plain dendrimers particularly higher generation dendrimers are reported to cause dose-dependent toxicities. Therefore, in this study, PEGylated G4 PAMAM dendrimers were synthesized via a click chemistry approach and used for the delivery piperlongumine (PL) for the treatment of colon cancer. PL is an amide alkaloid and potential anticancer agent, but it has poor aqueous solubility. PL was loaded into the dendrimer cavity through both physical loading and electrostatic chemical interactions. The prepared PL-loaded PEGylated G4 PAMAM dendrimers (PL@PEGylated G4) possessed a nanoscale size about 20 nm, monodispersity with a PDI value of 0.098 ± 0.002 and high drug solubility 226 ± 9.4 μg/mL. The drug release profile of PL@PEGylated G4 indicated a sustained release of encapsulated PL. After 24 h, about 43.2 ± 4.2 % and 64.9 ± 4.5 % of PL was released in PBS and SAB, respectively. The PEGylated dendrimers showed good biocompatibility during hemolytic toxicity study. In cytotoxicity studies, PL@PEGylated G4 showed potential cytotoxicity (IC50 = 5.8 μg/mL, p < 0.05) and greater induction of apoptosis in HCT 116 human colon cancer cells in comparison to pure PL, as demonstrated by the MTT and AO/EB assay, respectively. Overall, this biocompatible PEGylated G4 dendrimer-based formulation of PL could be a potential nanomedicine for cancer therapy.

Long-circulation zwitterionic dendrimer nanodrugs for phototherapy of tumors

The development of stealth and effective antitumor nanodrugs has been drawing great attention. Herein, generation five poly(amide amine) dendrimer (G5 PAMAM) was modified by zwitterionic material carboxybetaine methacrylamide (CBMAA) on its surface to prepare zwitterionic dendrimer (G5-CBMAAn). The results showed that G5-CBMAA30 had the longest blood circulation time due to its thickest zwitterionic layer, and its residual rate after injection into mice at 2 and 12 h was as high as 47.22 % and 14.37 %, respectively. Nanodrug G5-CBMAA30-ICG was prepared by containing indocyanine green (ICG) in the cavity of G5-CBMAA30. G5-CBMAA30-ICG had better tumor targeting ability and antitumor effect than free ICG in mice after laser irradiation, and the tumor inhibition rate was 96.6 % after 14 days’ treatment. The prepared G5-CBMAA30-ICG has great potential applications in the field of antitumor by phototherapy.

One-pot synthesis of PAMAM-grafted hyperbranched cellulose towards enhanced thermal stability and antibacterial activity

Cellulose is widely used as a biomaterial because of its good biodegradability and biocompatibility. In this study, the periodate oxidation method was used to prepare dialdehyde cellulose (DAC), which was then modified with polyamidoamine dendrimer (PAMAM G0) by the hyperbranched crosslinking synthesis to obtain novel dialdehyde cellulose-based polymers (DPs) with high thermal stability and antibacterial activity. The characterization found that PAMAM was successfully grafted onto the cellulose backbone. Moreover, the thermal weight-loss temperature of DPs decreased, the char yields increased, and the crystallinity decreased significantly as the amount of PAMAM increased. The crystallinity was about 46.50%. Specifically, when the ratio of the aldehyde group in DAC to the amine group in PAMAM was 1:1, the maximum thermal-denaturation temperature and weight-loss temperature (Tmax) of the DPs reached 136 ℃ and 365.8 ℃, respectively. In addition, DPs showed a uniform long-strip structure with a good particle size distribution, preventing the self-assembly and aggregation of cellulose and DAC. Disk diffusion tests showed that the DPs had significant antibacterial activity against Escherichia coli and Staphylococcus aureus, with the inhibition zone diameters reaching 13–15 mm. Thus, DPs with good thermal stability and antibacterial activity could be used as a new type of cellulose-based functional material.

Aqueous Heat Method for the Preparation of Hybrid Lipid-Polymer Structures: From Preformulation Studies to Protein Delivery.

Liposomes with adjuvant properties are utilized to carry biomolecules, such as proteins, that are often sensitive to the stressful conditions of liposomal preparation processes. The aim of the present study is to use the aqueous heat method for the preparation of polymer-grafted hybrid liposomes without any additional technique for size reduction. Towards this scope, liposomes were prepared through the combination of two different lipids with adjuvant properties, namely dimethyldioctadecylammonium (DDA) and D-(+)-trehalose 6,6′-dibehenate (TDB) and the amphiphilic block copolymer poly(2-(dimethylamino)ethyl methacrylate)-b-poly(lauryl methacrylate) (PLMA-b-PDMAEMA). For comparison purposes, PAMAM dendrimer generation 4 (PAMAM G4) was also used. Preformulation studies were carried out by differential scanning calorimetry (DSC). The physicochemical characteristics of the prepared hybrid liposomes were evaluated by light scattering and their morphology was evaluated by cryo-TEM. Subsequently, in vitro nanotoxicity studies were performed. Protein-loading studies with bovine serum albumin were carried out to evaluate their encapsulation efficiency. According to the results, PDMAEMA-b-PLMA was successfully incorporated in the lipid bilayer, providing improved physicochemical and morphological characteristics and the ability to carry higher cargos of protein, compared to pure DDA:TDB liposomes, without affecting the biocompatibility profile. In conclusion, the aqueous heat method can be applied in polymer-grafted hybrid liposomes for protein delivery without further size-reduction processes.

Comparative Evaluation of Lipofectamine and Dendrimer for Transfection of Short RNA Into Human T47D and MCF-10A Cell Lines.

Purpose: Non-viral transfection approaches are extensively used in cancer therapy. The future of cancer therapy lies on targeted and efficient drug/gene delivery. The aim of this study was to determine the transfection yields of two commercially available transfection reagents (i.e. Lipofectamine 2000, as a cationic lipid and PAMAM G5, as a cationic dendrimer) in two breast cell lines: cancerous cells (T47D) and non-cancerous ones (MCF-10A). Methods: We investigated the efficiencies of Lipofectamine 2000 and PAMAM G5 for transfection/delivery of a labeled short RNA into T47D and MCF-10A. In addition to microscopic assessments, the cellular uptakes of the complexes (fluorescein tagged-scrambled RNA with Lipofectamine or PAMAM dendrimer) were quantified by flow cytometry. Furthermore, the safety of the mentioned reagents was assessed by measuring cell necrosis through the cellular PI uptake. Results: Our results showed significantly better efficiencies of Lipofectamine compared to PAMAM dendrimer for short RNA transfection in both cell types. On the other hand, MCF-10A resisted more than T47D to the toxicity of higher concentrations of the transfection reagents. Conclusion: Altogether, our research demonstrated a route for comprehensive epigenetic modification of cancer cells and depicted an approach to efficient drug delivery, which eventually improves both short RNA-based biopharmaceutical industry and non-viral strategies in epigenetic therapy.

Development of an Ultrasmall and Biocompatible Platinum Nanozyme Encapsulated by Zwitterionic Dendrimer for Highly Sensitive Detection of Glucose.

Many kinds of noble metal nanoparticles can mimic the peroxidase-like function of horseradish peroxidase, which results in their wide applications in bio-related detection and drug delivery. However, those metal nanoparticles usually have low stability and reduced catalytic activity in biological complex medium. Herein, a zwitterionic peroxidase-like enzyme has been developed, which has high stability in fibrinogen solutions and high sensitivity for glucose detection. Maleic anhydride, cysteamine, and zwitterionic peptide EKEKC (EK-5) were used to modify generation 5 poly(amido amine) dendrimers (G5 PAMAM) to prepare zwitterionic dendrimer G5MEKnC with nonfouling properties. Finally, the G5MEKnC-encapsulated platinum nanoparticles (Ptn-G5MEK50C) were prepared by entrapping the platinum nanoparticles (1.40 nm) in the catalytic centers in the interior of G5MEK50C. Pt55-G5MEK50C showed high stability in the buffer solution and the fibrinogen solution within 4 days. They also displayed high biocompatibility toward HeLa cells based on cytotoxicity results and morphological observations. Furthermore, the catalytic oxidation of 3,3',5,5'-tetramethylbenzidine with H2O2 by Pt55-G5MEK50C followed the Michaelis-Menten equation, which confirmed their peroxidase-like properties. The catalytic mechanism was due to the generation of •OH from H2O2. More importantly, the peroxidase-like ability of Pt55-G5MEK50C was successfully used to establish a method for the determination of glucose concentration with a broad linear range of 1-2000 μM and a low detection limit of 0.1 μM. This method was highly accurate for the determination of glucose concentration in plasma. The zwitterionic dendrimer template enhanced the properties of Pt55-G5MEK50C. Taken together, a new kind of biocompatible nanozyme has been developed and successfully used for the sensitive detection of glucose in bio-related medium.

Margetuximab conjugated-PEG-PAMAM G4 nano-complex: a smart nano-device for suppression of breast cancer.

Breast cancer due to its high incidence and mortality is the second leading cause of death among females. On the other hand, nanoparticle-based drug delivery is one of the most promising approaches in cancer therapy, nowadays. Hence, margetuximab- and polyethylene glycol-conjugated PAMAM G4 dendrimers were efficiently synthesized for targeted delivery of quercetin (therapeutic agent) to MDA-MB-231 breast cancer cells. Synthesized nano-complexes were characterized using analytical devices such as FT-IR, TGA, DLS, Zeta potential analyzer, and TEM. The size less than 40nm, - 18.8 mV surface charge, efficient drug loading capacity (21.48%), and controlled drug release (about 45% of drug release normal pH after 8h) were determined for the nano-complex. In the biomedical test, the cell viability was obtained 14.67% at 24h of post-treatment for 800 nM concentration, and IC50 was ascertained at 100 nM for the nano-complex. The expression of apoptotic Bax and Caspase9 genes was increased by more than eightfolds and more than fivefolds after treatment with an optimal concentration of nanocarrier. Also, more than threefolds of cell cycle arrest was observed at the optimal concentration synthetics, and 27.5% breast cancer cell apoptosis was detected after treatment with 100 nM nano-complex. These outputs have been indicating the potential capacity of synthesized nano-complex in inhibiting the growth of breast cancer cells.

Synthesis and Properties of α-Mangostin and Vadimezan Conjugates with Glucoheptoamidated and Biotinylated 3rd Generation Poly(amidoamine) Dendrimer, and Conjugation Effect on Their Anticancer and Anti-Nematode Activities.

α-Mangostin and vadimezan are widely studied potential anticancer agents. Their biological activities may be improved by covalent bonding by amide or ester bonds with the third generation poly(amidoamine) (PAMAM) dendrimer, substituted with α-D-glucoheptono-1,4-lactone and biotin. Thus, conjugates of either ester- (G3gh4B5V) or amide-linked (G32B12gh5V) vadimezan, and equivalents of α-mangostin (G3gh2B5M and G32B12gh5M, respectively), were synthesized, characterized and tested in vitro against cancer cells: U-118 MG glioma, SCC-15 squamous carcinoma, and BJ normal human fibroblasts growth, as well as against C. elegans development. α-Mangostin cytotoxicity, stronger than that of Vadimezan, was increased (by 2.5–9-fold) by conjugation with the PAMAM dendrimer (with the amide-linking being slightly more effective), and the strongest effect was observed with SCC-15 cells. Similar enhancement of toxicity resulting from the drug conjugation was observed with C. elegans. Vadimezan (up to 200 µM), as well as both its dendrimer conjugates, was not toxic against both the studied cells and nematodes. It showed an antiproliferative effect against cancer cells at concentrations ≥100 µM. This effect was significantly enhanced after conjugation of the drug with the dendrimer via the amide, but not the ester bond, with G32B12gh5V inhibiting the proliferation of SCC-15 and U-118 MG cells at concentrations ≥4 and ≥12 μM, respectively, without a visible effect in normal BJ cells. Thus, the drug delivery system based on the PAMAM G3 dendrimer containing amide bonds, partially-blocked amino groups on the surface, larger particle diameter and higher zeta potential can be a useful tool to improve the biological properties of transported drug molecules.