phytochlorin and Breast-Neoplasms

We combined phosphoinositol-3-kinin inhibitor IPI-549 and photodynamic Chlorin e6 (Ce6) on carboxymethyl chitosan to develop a novel drug delivery nanoparticle (NP) system (Ce6/CMCS-DSP-IPI549) and evaluate its glutathione (GSH) sensitivity and targeting ability for breast cancer treatment. The NPs were spherical with a uniform size of 218.8 nm, a stable structure over 7 days. The maximum encapsulation efficiency was 64.42%, and NPs drug loading was 8.05%. The NPs released drugs within tumor cells due to their high GSH concentration, while they maintained structural integrity in normal cells, which have low GSH concentration. The cumulative release rates of IPI-549 and Ce6 at 108 h were 70.67% and 40.35% (at GSH 10 mM) and 8.11% and 2.71% (at GSH 2

Topics: Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Endothelial Cells; Female; Glutathione; Humans; Isoquinolines; Nanoparticles; Photochemotherapy; Photosensitizing Agents; Porphyrins; Pyrazoles; Pyrimidines

As a major therapeutic approach for cancer treatment, the effectiveness of chemotherapy is challenged by multidrug resistance (MDR). Herein, we fabricated novel redox-responsive, chondroitin sulfate-based nanoparticles that could simultaneously deliver quercetin (chemosensitizer), chlorin e6 (photosensitizer) and paclitaxel (chemotherapeutic agent) to exert enhanced chemo-photodynamic therapy for overcoming MDR and lung metastasis of breast cancer. In vitro cell study showed that nanoparticles down-regulated the expression of P-glycolprotein (P-gp) on MCF-7/ADR cells and thereby improved the anticancer efficacy of PTX against MCF-7/ADR cells. Moreover, NIR laser irradiation could induce nanoparticles to generate cellular reactive oxygen species (ROS), leading to mitochondrial membrane potential loss, and meanwhile facilitating lysosomal escape of drugs. Importantly, the novel nanoplatform exhibited effective in vivo MDR inhibition and anti-metastasis efficacy through enhanced chemo-photodynamic therapy. Thus, the study suggested that the multifunctional nanoplatform had good application prospect for effective breast cancer therapy.

Topics: Animals; Antineoplastic Agents; ATP Binding Cassette Transporter, Subfamily B, Member 1; Breast Neoplasms; Cell Proliferation; Chlorophyllides; Chondroitin Sulfates; Combined Modality Therapy; Doxorubicin; Drug Carriers; Drug Resistance, Neoplasm; Female; Gene Expression; Humans; Infrared Rays; Lasers; Lung Neoplasms; MCF-7 Cells; Nanoparticles; Paclitaxel; Photosensitizing Agents; Porphyrins; Quercetin; Reactive Oxygen Species; Xenograft Model Antitumor Assays

A promising direction in Biopharmaceuticals is the development of specific peptide-based systems to improve drug delivery. This approach may increase tumor specificity and drug penetration into the target cell. Similar systems have been designed for several antitumor drugs. However, for photodynamic therapy drugs, such studies are not yet enough. Previously, we have developed a method of inclusion of chlorin e6 (Ce6), a photosensitizer used in photodynamic therapy, in phospholipid nanoparticles with a diameter of up to 30 nm, and reported an increase in its effectiveness in the experiments in vivo. In this work, we propose to modify a previously developed delivery system for Ce6 by the addition of cell-penetrating (R7) and/or targeting NGR peptides. The interaction of the compositions developed with HepG2 and MCF-7 tumor cells is shown. The expression of CD13 protein with affinity to NGR on the surface of these cells has been studied using flow cytometry. The expression of this protein on the HepG2 cells and its absence on MCF-7 was demonstrated. After incubation of tumor cells with the resulting Ce6 compositions, we evaluated the cellular accumulation, photoinduced, and dark cytotoxicity of the drugs. After irradiation, the highest level of cytotoxicity was observed when R7 peptide was added to the system, either alone or in combination with NGR. In addition to R7, the NGR-motif peptide increased the internalization of Ce6 in HepG2 cells without affecting its photodynamic activity. In this work we also discuss possible mechanisms of action of the cell-penetrating peptide when attached to phospholipid nanoparticles.

Topics: Breast Neoplasms; Carcinoma, Hepatocellular; CD13 Antigens; Cell Survival; Cell-Penetrating Peptides; Chlorophyllides; Drug Carriers; Drug Compounding; Hep G2 Cells; Humans; Liver Neoplasms; MCF-7 Cells; Nanoparticles; Oligopeptides; Phospholipids; Photochemotherapy; Photosensitizing Agents; Porphyrins

Photodynamic therapy (PDT) holds a number of advantages for tumor therapy. However, its therapeutic efficiency is limited by non-sustainable reactive oxygen species (ROS) generation and heterogeneous distribution of photosensitizer (PS) in tumor. Herein, a "

Topics: Animals; Apoptosis; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Drug Carriers; Glutathione; Glutathione Disulfide; Humans; Hyaluronic Acid; Hydrogen Peroxide; Hydroxyl Radical; Low-Level Light Therapy; Manganese Compounds; Mice; Nanoparticles; Oxides; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Reactive Oxygen Species; Silicon Dioxide; Singlet Oxygen; Tissue Distribution

A two-pronged concept combining photodynamic therapy (PDT) and epithelial-mesenchymal transition (EMT) blockade in a minimalist nanoplatform was proposed to combat basal-like breast cancer (BLBC) metastasis. Based on PDT-mediated tumor killing and epalrestat (Epa)-mediated EMT blockade, as-prepared Ce6/Epa nanoparticles prevented BLBC metastasis effectively in vivo, providing a very promising two-pronged strategy against BLBC metastasis.

Topics: Animals; Apoptosis; Breast Neoplasms; Cell Proliferation; Chlorophyllides; Enzyme Inhibitors; Epithelial-Mesenchymal Transition; Female; Humans; Lung Neoplasms; Mice; Mice, SCID; Nanoparticles; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Rhodanine; Thiazolidines; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

The combination therapy strategy based on both chemotherapy and photodynamic therapy (PDT) exhibits great potential for advanced cancer treatment. Multimodal nanodrug delivery systems based on both chemotherapeutic drug and photodynamic agent have been proven to possess excellent synergistic efficacy. In this study, 7-ethyl-10-hydroxycamptothecin (SN38) and chlorin e6 (Ce6) were co-assembled into novel carrier-free nanoparticles (SN38/Ce6 NPs) via simple antisolvent precipitation method. As expected, SN38/Ce6 NPs exhibited uniform morphology with a particle size of around 150 nm and a zeta potential of about -30 mV, good stability in aqueous solution/at lyophilized state and high cellular uptake efficiency against murine mammary carcinoma (4T1) cell lines. Besides, enhanced singlet oxygen generation capacity of the nanoparticles was both observed in test-tube and in 4T1 cell lines in contrast with Ce6 injection. Moreover, a ∼85 % inhibition rate of SN38/Ce6 NPs with laser was detected, which was significantly higher (P < 0.05) than those without laser (∼65 %) and injections (less than 20 %), verified the excellent synergistic antitumor efficacy of the nanoparticles due to combined chemo-photodynamic therapy, enhanced tumor accumulation and higher cellular internalization. Notably, chemical thermodynamic method and molecular dynamics (MD) simulations supplied solid data and visual images to estimate the driving forces for the self-assembly process of the carrier-free nanoparticles as primary hydrophobic interactions (π-π stacking) and subordinate hydrogen bonds. Conclusively, the above self-assembled carrier-free nanoparticles represented a promising synergistic anticancer strategy capable of maximal therapeutic efficacy and minimal systemic toxicity. Moreover, the application of thermodynamic method together with MD simulations in the investigation of NPs self-assembly process also provided new ideas for the assembly mechanism exploration of more complicated nanodrug delivery system.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Chlorophyllides; Drug Screening Assays, Antitumor; Irinotecan; Mice; Molecular Dynamics Simulation; Nanoparticles; Particle Size; Photochemotherapy; Photosensitizing Agents; Porphyrins; Surface Properties; Thermodynamics

Combining functional proteins with small molecular drugs into one entity may endow distinct synergistic advantages. However, on account of completely different physicochemical properties of such payloads, co-delivery through systemic administration for therapeutic purpose is challenging. Herein, we designed the protein-drug conjugate HSAP-DC-CAT (human serum albumin/Pt (IV)-dibenzocyclooctyne/chlorin e6-catalase) by modification of CAT and cisplatin pro-drug loaded HSA with pH-sensitive azide linker 3-(azidomethyl)-4-methyl-2,5-furandione (AzMMMan) followed by click chemistry assembly with DC. The dynamic covalent bonds between linker and proteins, on the one hand, can bridge proteins and small molecular drugs in the intermediate state for systemic delivery in the harsh in vivo environment; on the other hand, it can trigger traceless cleavage and release of drugs and proteins with full bioactivity in acidic microenvironment of tumor. The multifunctional HSAP-DC-CAT provides efficient cytosolic transduction in vitro, excellent blood half-lives after systemic administration, and significant antitumor outcome via integrated cisplatin-based chemotherapy and Ce6-based photodynamic therapy enhanced by catalase-induced manipulation of tumor hypoxia microenvironment. This study describes a universal formulation strategy for protein and small molecular drug by a bifunctional linker through amide reaction and click chemistry, with traceless in vivo release of therapeutic units.

Topics: Animals; Antioxidants; Breast Neoplasms; Catalase; Cell Line, Tumor; Chlorophyllides; Cisplatin; Click Chemistry; Delayed-Action Preparations; Drug Carriers; Drug Compounding; Female; Hydrogen-Ion Concentration; Mice, Nude; Photochemotherapy; Porphyrins; Prodrugs; Radiation-Sensitizing Agents; Serum Albumin, Human; Tumor Hypoxia; Tumor Microenvironment

Cancer treatment has always been a big problem for people. With the application of photodynamic therapy, the problem has been alleviated. However, the problem of tumor hypoxia affecting photodynamic therapy has been waiting to be resolved. Therefore, we report here that a redox nanocarrier (called RN) is prepared by hollow mesoporous silica sphere (HMSNs) and a redox-responsive polymer ligand. The nanocarrier is loaded with metformin and catalase, and the polymer is linked to the photosensitizer chlorin e6 (Ce6). Metformin inhibits the mitochondrial respiration of cancer cells, reducing the activity of cancer cells and increasing the oxygen concentration required for photodynamic therapy. Not only the effect of photodynamic therapy is enhanced, but also the effect of chemotherapy is increased to achieve super additive treatment. These RNs exhibit not only low cytotoxicity but also high biocompatibility in vitro experiments. In vitro Ce6 release studies have shown a higher release in the presence of glutathione (GSH). Confocal microscopy can further indicate that the nanoparticles are carried to the area around the nucleus of the cancer cells. In addition, treatment with a mouse tumor model demonstrated that RN has an effective therapeutic effect on tumors.

Topics: Animals; Breast Neoplasms; Catalase; Cell Line, Tumor; Cell Survival; Chlorophyllides; Combined Modality Therapy; Female; Glutathione; Humans; Metformin; Mice; Nanoparticles; Oxygen; Photochemotherapy; Porosity; Porphyrins; Radiation-Sensitizing Agents; Silicon Dioxide; Tumor Microenvironment

Currently, the development of polysaccharide, especially chitosan (CS), based drug delivery system to afford magnetic resonance imaging (MRI) guided theranostic cancer therapy remains largely unexplored. Herein, we successfully developed a CS derived polymer (Gd-CS-OA) through chemical conjugation of CS, octadecanoic acid (OA) and gadopentetic acid (GA). After self-assemble into glycolipid nanoparticles to loaded chlorin e6 (Ce6), the resulted Gd-CS-OA/Ce6 was able to realize MRI guided photodynamic therapy (PDT) of cancer. Our results revealed that Gd-CS-OA was able to increase the MRI sensitivity as compared to Gd-DTPA with decent residence time and preferable excretion behavior in vivo. Moreover, the Gd-CS-OA/Ce6 showed negligible hemolysis, satisfactory ROS generation and stability in physiological environments with preferable cellular uptake and enhanced in vitro cytotoxicity (through elevated ROS generation) on 4T1 cells. Most importantly, Gd-CS-OA/Ce6 demonstrated promising in vivo tumor targetability (enhanced penetration and retention effect) and powerful MRI guided tumor ablation through PDT on in situ 4T1 tumor model.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chitosan; Chlorophyllides; Disease Models, Animal; Drug Delivery Systems; Female; Gadolinium DTPA; Glycolipids; Magnetic Resonance Imaging; Mice; Mice, Inbred BALB C; Nanoparticles; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Stearic Acids; Tumor Burden

Aiming at developing a moderate and efficient sono-photodynamic therapy for breast cancer, tissue engineering scaffolds may provide an easy and efficient strategy to eliminate serious side effects in conventional surgery or chemotherapy, and thus, they are highly desired. However, the development of ideal sono-photodynamic therapeutic scaffolds is always hindered by the poor stability and incompatibility between the different biomaterial components. Herein, the Food and Drug Administration (FDA)-approved sono/photosensitizer Chlorin e6 (Ce6) was successfully and tightly incorporated into electrospun polycaprolactone/gelatin (PG) scaffolds via positively charged protonated g-C

Topics: Antineoplastic Agents; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell Survival; Chlorophyllides; Drug Screening Assays, Antitumor; Female; Gelatin; Graphite; Humans; Nitrogen Compounds; Particle Size; Photochemotherapy; Photosensitizing Agents; Polyesters; Porphyrins; Protons; Reactive Oxygen Species; Surface Properties

Photoactivity "on-off" switchable nano-agents could shield phototoxicity until reaching target region, which immensely promoted photodynamic therapy. However, the masking ratio of nano-agents in vivo was dynamic and positively correlated with the phototoxicity induced by laser irradiation, in which case the timing of laser irradiation was unpredictable to maximize antitumor efficacy.. Herein, low molecular weight chitosan and hydrophobic polymethylacrylamide derivatives were linked via GSH cleavable 3, 3'-dithiodipropionic acid to construct polymeric micelles (Ce6-CSPD). The doxorubicin loading nano-agent (Ce6-CSPD/DOX) could quench both photoactivity and fluorescence of photosensitizer chlorin e6 (Ce6) and doxorubicin (DOX) under physiological condition by homo-fluorescence resonance energy transfer (homoFRET).. Once internalized by tumor cells, the photoactivity as well as fluorescence of Ce6 was recovered rapidly when motivated by intracellular high GSH. Specifically, the fluorescence intensity and photoactivity of Ce6 were proven to be positive linear correlated, upon which appropriate timing of laser irradiation could be determined by referring to the dynamic fluorescence intensity in vivo. In addition, the theranostic nano-agents also possessed the capacity of monitoring the DOX release process. Accordingly, under the guidance of fluorescence intensity, the experimental group subjected to laser irradiation at 18 h postadministration acquired the highest antitumor inhibition efficacy compared to that at four hours and 48 h, which held great potential for maximizing chemo-photodynamic therapy and avoiding nonspecific phototoxicity precisely to normal organs.. In summary, we prepared homoFRET-based theranostic nano-agent (Ce6-CSPD/DOX) for monitoring PDT precisely and decreasing phototoxicity to normal organs before reaching target region. Under the guidance of dynamic fluorescence intensity, the appropriate laser irradiation timing could be monitored to maximize antitumor therapy efficacy, which offered opportunities for monitoring efficiency of chemo-photodynamic therapy in a timely and accurate manner.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Chitosan; Chlorophyllides; Doxorubicin; Drug Delivery Systems; Drug Liberation; Female; Humans; Lasers; Mice, Inbred BALB C; Micelles; Nanostructures; Photochemotherapy; Photosensitizing Agents; Polymers; Porphyrins; Precision Medicine; Rabbits; Spectrometry, Fluorescence

Metastasis is the main cause of treatment failure in breast cancer, and integrated phototheranostics is a promising strategy to achieve both precision theranostics and metastasis inhibition. In this work, a multifunctional phototheranostic nanoprobe composed of chlorin e6 (Ce6)-conjugated and polydopamine (PDA)-coated gold nanostars (AuNSs) was synthesized for simultaneous photoacoustic (PA) imaging, photothermal therapy (PTT) and photodynamic therapy (PDT). Under the irradiation of near infrared laser, AuNSs@PDA showed enhanced photothermal conversion and amplified PA imaging performance, compared with single AuNSs. By the covalent conjugation of Ce6, the AuNSs@PDA-Ce6 nanoprobe showed robust stability and excellent singlet oxygen (1O2) generation ability. Under the combination of PTT/PDT, the AuNSs@PDA-Ce6 nanoprobes significantly reduced the growth of 4T1 tumors and suppressed their lung metastasis. All the results demonstrated the considerable potential of AuNSs@PDA-Ce6 phototheranostic nanoprobes for precision theranostics and metastasis inhibition of breast cancer.

Topics: Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Gold; Humans; Indoles; Lung Neoplasms; Photoacoustic Techniques; Photochemotherapy; Photosensitizing Agents; Polymers; Porphyrins

Hyaluronic acid (HA)-based nanocarriers are of great interest in the drug delivery field due to the tumor targetability via CD44-mediated recognition and endocytosis. However, sufficient tumor-specific release of encapsulated cargoes with steady controllability is necessary to optimize their outcome for cancer therapy. In this study, we constructed a light-activated nanocarrier TKHCENPDOX to enable on-demand drug release at the desired site (tumor). Particularly, TKHCENPDOX encapsulating doxorubicin (DOX) was self-assembled from a HA-photosensitizer conjugate (HA-TK-Ce6) containing reactive oxygen species (ROS)-sensitive thioketal (TK) linkers. Following i.v. injection, TKHCENPDOX was accumulated in the MDA-MB-231 breast tumor xenograft more efficiently through preventing drug leakage in the bloodstream and the HA-mediated targeting effect. Upon internalization into tumoral cells, 660 nm laser irradiation generated ROS during a photodynamic (PDT) process to cleave the TK linker next to Ce6, resulting in light-induced TKHCENPDOX dissociation and selective DOX release in the tumor area. Consequently, TKHCENPDOX showed a remarkable therapeutic effect and minimized toxicity in vivo. This strategy might provide new insight for designing cancer-selective nanoplatforms with active targeting and locoregional drug release simultaneously.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Doxorubicin; Drug Carriers; Drug Liberation; Female; Humans; Hyaluronic Acid; Light; Mice, Inbred BALB C; Mice, Inbred ICR; Nanoconjugates; Nanoparticles; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species; Xenograft Model Antitumor Assays

Nanomedicines have been demonstrated as promising strategies for cancer therapy due to the advantages in pharmacokinetics and drug targeting delivery to tumor tissues. However, creation of delivery platforms able to intrinsically and spatially optimize drug cellular uptake during the entire delivering process remain challenging. To address this challenge, here we report on tumor microenvironment-adaptable self-assembly (TMAS) of pentapeptides regulated by the pH-sensitive cis/trans isomerization of 4-amino-proline (Amp) amide bonds for enhanced drug delivery and photodynamic therapeutic (PDT) efficacy. We found that decreasing solution pH led to the cis → trans isomerization of Amp amide bonds, thus promoting reversible self-assembly of pentapeptide FF-Amp-FF (AmpF) into superhelices and nanoparticles upon alternating exposure to neutral and mild acidic conditions. Co-assembly of peptide AmpF with its derivative containing a photosensitizer Chlorin e6 (AmpF-C) allows for creation of TMAS systems undergoing a morphological transition adaptable to the pH gradient present in cellular uptake pathway. Ex vivo studies revealed that TMAS nanomedicines prolonged circulation in the animal body and improved accumulation at tumor sites compared to morphology-persistent nanomedicines. In addition to the optimized cellular uptake, the morphological transition of TMAS into nanofibers in cytoplasm caused an enhanced intracellular ROS level compared to nanoparticle counterparts, thus leading to a lowered half lethal dose value for cancer cells. The combined advantages of TMAS eventually allowed in vivo PDT therapy for significant inhibition of tumor growth, thus demonstrating the improved drug delivery efficiency and therapeutic efficacy of TMAS systems toward new-generation nanomedicines.

Topics: Animals; Breast Neoplasms; Cell Survival; Chlorophyllides; Delayed-Action Preparations; Female; Isomerism; Mice; Nanostructures; Oligopeptides; Photochemotherapy; Photosensitizing Agents; Porphyrins; Proline; Tumor Microenvironment

Photodynamic therapy (PDT) is a clinically approved cancer treatment approach that relies on the generation of excess reactive oxygen species (ROS) to eradicate tumor cells by inducing oxidative stress. Unfortunately, if the tumor's endogenous glutathione (GSH) is overexpressed, it will eliminate the ROS and restrict the therapeutic efficacy of PDT. Herein, we report a H

Topics: Acrolein; Administration, Intravenous; Animals; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chlorophyllides; Combined Modality Therapy; Female; Glutathione; Humans; Hydrogen Peroxide; Mice; Oxidative Stress; Photochemotherapy; Porphyrins; Xenograft Model Antitumor Assays

Environmental stimuli, including pH, light, and temperature, have been utilized for activating controlled drug delivery to achieve efficient antitumor therapeutics while minimizing undesirable side effects. In this study, a multifunctional nanoplatform based on hollow mesoporous copper sulfide nanoparticles (H-CuS NPs) was developed by loading the interior cavity of the NPs with a drug-loaded phase-change material (PCM, 1-tetradecanol). Doxorubicin (DOX) and chlorin e6 (Ce6) were selected as the model chemotherapeutic drug and photosensitizer, respectively, which were encapsulated in H-CuS NPs via the PCM to form H-CuS@PCM/DOX/Ce6 (HPDC) NPs. When exposed to near infrared laser irradiation, this nanocomplex could produce a strong photothermic effect and thus induce the controlled release of DOX and Ce6 from the melting PCM. Subsequently, the DOX-mediated chemotherapeutic effect and Ce6-mediated photodynamic effect further contributed to enhanced tumor eradication. The efficacy of this multimodal cancer treatment combining chemo-, photothermal, and photodynamic therapies was systematically evaluated both in vitro and in vivo using a 4T1 mouse mammary tumor cell line and a mouse model bearing breast cancer. Moreover, this nanoplatform exhibited minimal systemic toxicity and good hemocompatibility and may provide an effective strategy for the delivery of multiple therapeutic agents and application of multimodal cancer treatments.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Copper; Doxorubicin; Drug Delivery Systems; Female; Hyperthermia, Induced; Infrared Rays; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Mice, Nude; Nanoparticles; Photochemotherapy; Photosensitizing Agents; Porphyrins; Sulfides; Xenograft Model Antitumor Assays

Topics: Animals; Apoptosis; Breast Neoplasms; Cell Proliferation; Chlorophyllides; DNA, Catalytic; Drug Carriers; Early Growth Response Protein 1; Female; Gene Silencing; Genetic Therapy; Humans; Metal-Organic Frameworks; Mice; Mice, Inbred BALB C; Mice, Nude; Nanoparticles; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Reactive Oxygen Species; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Photodynamic therapy (PDT) as a treatment method has many advantages such as minimal invasiveness, repeatable dosage, and low systemic toxicity. Issues with conventional PDT agents include the limited availability of endogenous oxygen and difficulty in accumulation at the tumor site, which has hindered the successful treatment of tumors. Herein, we developed catalase-encapsulated hyaluronic-acid-based nanoparticles loaded with adamantane-modified photosensitizer for enhanced PDT of solid tumors. Chlorin e6 (Ce6) as the photosensitizer was modified with adamantane to yield adamantane-modified Ce6 (aCe6). The obtained nanosystem (HA-CAT@aCe6) could target overly expressed CD44 receptors on cancer cells, supplying oxygen by converting endogenous hydrogen peroxide (H

Topics: Adamantane; Animals; Breast Neoplasms; Catalase; Cell Line, Tumor; Chlorophyllides; Female; Humans; Hyaluronic Acid; Mice, Nude; Nanocapsules; Photochemotherapy; Photosensitizing Agents; Porphyrins

Hypoxia is a potent tumor microenvironmental (TME) factor promoting immunosuppression and metastatic progression. For current anticancer therapeutic strategies, the combination of hypoxia alleviation and photodynamic therapy (PDT) might be a useful approach to further improve anticancer efficacy. In this study, we alleviated tumor hypoxia using a prolonged oxygen-generating phototherapy hydrogel (POP-Gel), which effectively elevated the oxygen level and shrank the hypoxic regions of tumors for up to 5 days evaluated by photoacoustic (PA) imaging and immunofluorescence staining, meeting the requirement of the "once injection, sustained treatment" strategy and significantly increasing PDT efficacy. The long-period improvement of the tumor hostile environment downregulated the expression of hypoxia inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF), further preventing tumor growth and metastasis. More importantly, the enhanced PDT triggered a more intense immune response, improving the inhibition of triple negative breast cancer growth even tumor elimination. The POP-Gel may contribute useful insights into the combination of hypoxia alleviation and PDT.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Chlorophyllides; Female; Humans; Hydrogels; Mice; Mice, Inbred BALB C; Mice, Nude; Oxygen; Photochemotherapy; Photosensitizing Agents; Porphyrins; Tumor Hypoxia

Conformation of protein is vital to its function, but may get affected when processing to manufacture products. It is therefore important to understand structural changes during each step of production. In this study, we investigate secondary structure changes in the targeting protein Epidermal Growth Factor (EGF) during synthesis of theranostic bifunctional nanoparticle, devised for Photodynamic therapy of breast cancer. We acquired FTIR spectra of EGF; unconjugated, post treatment with α-lipoic acid, attached to gold nanoparticle, and bound to the bifunctional nanoprobe. We observed decreasing disordered structures and turns, and increasing loops, as the synthesis process progressed. There was an overall increase in β-sheets in final product compared to pure EGF, but this increase was not linear and fluctuated. Previous crystal structure studies on EGF-EGFR complex have shown loops and β-sheets to be important in the binding interaction. Since our study found increase in these structures in the final product, no adverse effect on binding function of EGF was expected. This was confirmed by functional assays. Such studies may help modify synthesis procedures, and thus secondary structures of proteins, enabling increased functionality and optimum results.

Topics: Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Epidermal Growth Factor; ErbB Receptors; Female; Gold; Humans; Metal Nanoparticles; Neoplasm Proteins; Photochemotherapy; Photosensitizing Agents; Porphyrins; Protein Binding; Protein Structure, Secondary; Spectroscopy, Fourier Transform Infrared; Thioctic Acid

The development of imaging-guided smart drug delivery systems for combinational photodynamic/chemotherapy of the tumor has become highly demanded in oncology. Herein, redox-responsive theranostic polymeric nanoparticles (NPs) were fabricated innovatively using low molecular weight heparin (LWMH) as the backbone. Chlorin e6 (Ce6) and alpha-tocopherol succinate (TOS) were conjugated to LMWH via cystamine as the redox-sensitive linker, forming amphiphilic Ce6-LMWH-TOS (CHT) polymer, which could self-assemble into NPs in water and encapsulate paclitaxel (PTX) inside the inner core (PTX/CHT NPs). The enhanced near-infrared (NIR) fluorescence intensity and reactive oxygen species (ROS) generation of Ce6 were observed in a reductive environment, suggesting the cystamine-switched "ON/OFF" of Ce6. Also, the in vitro release of PTX exhibited a redox-triggered profile. MCF-7 cells showed a dramatically higher uptake of Ce6 delivered by CHT NPs compared with free Ce6. The improved therapeutic effect of PTX/CHT NPs compared with mono-photodynamic or mono-chemotherapy was observed in vitro via MTT and apoptosis assays. Also, the PTX/CHT NPs exhibited a significantly better in anti-tumor efficiency upon NIR irradiation according to the results of in vivo combination therapy conducted on 4T1-tumor-bearing mice. The in vivo NIR fluorescence capacity of CHT NPs was also evaluated in tumor-bearing nude mice, implying that the CHT NPs could enhance the accumulation and retention of Ce6 in tumor foci compared with free Ce6. Interestingly, the anti-metastasis activity of CHT NPs was observed against MCF-7 cells by a wound healing assay, which was comparable to LMWH, suggesting LMWH was promising for construction of nanocarriers for cancer management.

Topics: alpha-Tocopherol; Animals; Antineoplastic Agents, Phytogenic; Apoptosis; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chlorophyllides; Drug Compounding; Drug Delivery Systems; Female; Heparin, Low-Molecular-Weight; Humans; Infrared Rays; Mice, Inbred BALB C; Mice, Nude; Microscopy, Electron, Transmission; Nanoparticles; Neoplasm Transplantation; Optical Imaging; Paclitaxel; Photochemotherapy; Porphyrins; Random Allocation; Reactive Oxygen Species; Theranostic Nanomedicine; Tumor Burden; Whole Body Imaging

The combination of chemotherapy with photodynamic therapy (PDT) has attracted broad attention as it can overcome limitations of conventional chemo-treatment by using different modes of action. However, the efficacy of PDT to treat solid tumors is severely affected by hypoxia in tumors.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Combined Modality Therapy; Doxorubicin; Drug Carriers; Drug Compounding; Female; Humans; Magnetic Resonance Imaging; Manganese Compounds; MCF-7 Cells; Mice; Mice, Nude; Nanoparticles; Oxides; Oxygen; Photoacoustic Techniques; Photochemotherapy; Photosensitizing Agents; Polyesters; Polyethylene Glycols; Porphyrins; Theranostic Nanomedicine; Xenograft Model Antitumor Assays

Incorporation of a desired stimuli-responsive unit in a stereospecific manner at the specific location within a nonlinear block copolymer architecture is a challenging task in synthetic polymer chemistry. Herein, we report a facile and versatile method to synthesize AB

Topics: Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Click Chemistry; Delayed-Action Preparations; Doxorubicin; Female; Humans; Photochemotherapy; Polyesters; Polyethylene Glycols; Polymethacrylic Acids; Porphyrins; Singlet Oxygen

An ideal cancer therapeutic strategy is expected to possess potent ability to not only ablate primary tumors but also prevent distance metastasis and relapse. In this study, human serum albumin was hybridized with hemoglobin by intermolecular disulfide bonds to develop a hybrid protein oxygen nanocarrier with chlorine e6 encapsulated (C@HPOC) for oxygen self-sufficient photodynamic therapy (PDT). C@HPOC realized the tumor-targeted co-delivery of photosensitizer and oxygen, which remarkably relieved tumor hypoxia. C@HPOC was favorable for more efficient PDT and enhanced infiltration of CD8

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Cell Proliferation; Cell Survival; Chlorophyllides; Drug Screening Assays, Antitumor; Female; Humans; Mice; Mice, Inbred BALB C; Nanoparticles; Oxygen; Photochemotherapy; Photosensitizing Agents; Porphyrins; Serum Albumin, Human; Tumor Cells, Cultured

Multiple therapeutic modalities, such as photodynamic (PDT) and photothermal (PTT) therapies, have been jointly applied to produce a synergistic effect for tumor eradication based on the hyperthermia and generation of reactive oxygen species (ROS) mediated by photoactive agents. Effective delivery of highly efficient photosensitizers and photothermal agents is the key for combination of PDT/PTT. Herein, we propose a strategy to functionalize Prussian blue (PB) nanoparticles (NPs) with Chlorin e6 (Ce6)-imbedded erythrocyte membrane vesicles. This nanoplatform can address the major issues of these two capable photoactive agents, such as limited biocompatibility, lack of functional chemical groups, and poor bioavailability due to rapid blood clearance or self-aggregation. Specifically, PB NPs were packaged within Ce6-imbedded erythrocyte membrane vesicles, named as PB@RBC/Ce6 NPs, to take advantage of both biological functions of natural erythrocyte membranes and the unique physicochemical properties of synthetic nanoagents. Compared to bare PB NPs or free Ce6, PB@RBC/Ce6 NPs exhibited considerably enhanced cellular uptake and accumulation in tumoral tissues. Moreover, the PB@RBC/Ce6 NP-mediated PDT/PTT combination therapies produced a notable effect in boosting the necrosis and late apoptosis of tumor cells in vitro, and further showed a synergistic therapeutic effect against an orthotopic tumor model in vivo.

Topics: Animals; Apoptosis; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chlorophyllides; Combined Modality Therapy; Erythrocyte Membrane; Female; Ferrocyanides; Humans; Hypothermia, Induced; Light; Materials Testing; Mice, Inbred BALB C; Mice, Nude; Nanoparticles; Neoplasm Transplantation; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species; Tissue Distribution

Mesoporous silica nanoparticles (MSNs), with their large surface area and tunable pore sizes, have been widely applied for anticancer therapeutic cargos delivery with a high loading capacity. However, easy aggregation in saline buffers and limited blood circulation lifetime hinder their delivery efficiency and the anticancer efficacy. Here, new multifunctional MSNs-supported red-blood-cell (RBC)-mimetic theranostic nanoparticles with long blood circulation, deep-red light-activated tumor imaging and drug release were reported. High loading capacities were achieved by camouflaging MSNs with RBC membrane to co-load an anticancer drug doxorubicin (Dox) (39.1 wt%) and a near-infrared photosensitizer chlorin e6 (Ce6) (21.1 wt%). The RBC membrane-coating protected drugs from leakage, and greatly improved the colloidal stability of MSNs, with negligible particle size change over two weeks. Upon an external laser stimuli, the RBC membrane could be destroyed, resulting in 10 times enhancement of Dox release. In a 4T1 breast cancer mouse model, the RBC-mimetic MSNs could realize

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Chlorophyllides; Disease Models, Animal; Doxorubicin; Drug Carriers; Drug Liberation; Erythrocyte Membrane; Lasers; Mice; Nanoparticles; Photosensitizing Agents; Porphyrins; Silicon Dioxide

Topics: Albumins; Animals; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chlorophyllides; Gadolinium; Hyperthermia, Induced; Magnetic Resonance Imaging; Mice; Nanoparticles; Photochemotherapy; Photosensitizing Agents; Porphyrins; Theranostic Nanomedicine; Treatment Outcome

Photodynamic therapy (PDT) has been applied in cancer treatment by utilizing reactive oxygen species to kill cancer cells. However, a high concentration of glutathione (GSH) is present in cancer cells and can consume reactive oxygen species. To address this problem, we report the development of a photosensitizer-MnO2 nanosystem for highly efficient PDT. In our design, MnO2 nanosheets adsorb photosensitizer chlorin e6 (Ce6), protect it from self-destruction upon light irradiation, and efficiently deliver it into cells. The nanosystem also inhibits extracellular singlet oxygen generation by Ce6, leading to fewer side effects. Once endocytosed, the MnO2 nanosheets are reduced by intracellular GSH. As a result, the nanosystem is disintegrated, simultaneously releasing Ce6 and decreasing the level of GSH for highly efficient PDT. Moreover, fluorescence recovery, accompanied by the dissolution of MnO2 nanosheets, can provide a fluorescence signal for monitoring the efficacy of delivery.

Topics: Animals; Breast Neoplasms; Chlorophyllides; Delayed-Action Preparations; Female; Glutathione; Humans; Manganese Compounds; MCF-7 Cells; Mice, Nude; Nanostructures; Oxidation-Reduction; Oxides; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species

To avoid the overheating effect of excitation light and improve the efficacy of photodynamic therapy (PDT) of upconversion nanoplatform, a novel nanoprobe based on 808 nm-excited upconversion nanocomposites (T-UCNPs@Ce6@mSiO2) with low heating effect and deep penetration has been successfully constructed for targeted upconversion luminescence, magnetic resonance imaging (MRI) and high-efficacy PDT in HER2-overexpressed breast cancer. In this nanocomposite, photosensitizers (Ce6) were covalently conjugated inside of mesoporous silica to enhance the PDT efficacy by shortening the distance of fluorescence resonance energy transfer and to decrease the cytotoxicity by preventing the undesired leakage of Ce6. Compared with UCNPs@mSiO2@Ce6, UCNPs@Ce6@mSiO2 greatly promoted the singlet oxygen generation and amplified the PDT efficacy under the excitation of 808 nm laser. Importantly, the designed nanoprobe can greatly improve the uptake of HER2-positive cells and tumors by modifying the site-specific peptide, and the in vivo experiments showed excellent MRI and PDT via intravenous injection by modeling MDA-MB-435 tumor-bearing nude mice. Our strategy may provide an effective solution for overcoming the heating effect and improving the PDT efficacy of upconversion nanoprobes, and has potential application in visualized theranostics of HER2-overexpressed breast cancer.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Contrast Media; Female; Hot Temperature; Humans; Magnetic Resonance Imaging; Mice; Mice, Inbred BALB C; Mice, Nude; Nanocapsules; Nanocomposites; Photochemotherapy; Photosensitizing Agents; Porphyrins; Receptor, ErbB-2; Theranostic Nanomedicine; Treatment Outcome

Multidrug resistance (MDR) of breast cancer is a major obstacle in chemotherapy of cancer treatments. Recently the anti-tumor effects of Chlorin e6 (Ce6) mediated photodynamic therapy (Ce6-PDT) were reported in skin cancer and hepatoma in vitro. However, its therapeutic potential in killing human breast cancer especially those with MDR and the differences between MCF-7 and MCF-7/ADR after PDT treatment has not been fully investigated.. MTT assay was used to measure cell survival rate of MCF-7 cells and MCF-7/ADR cells. Intracellular reactive oxygen species (ROS) generation was measured by monitoring the fluorescence intensity of dichlorofluorescein (DCF) by flow cytometry. Nuclear morphology changes and DNA damage in both MCF-7 and MCF-7/ADR after Ce6-PDT were analyzed by hochest33342 staining and comet assay. Western blot and monodansylcadaverine (MDC) staining were used to monitor autophagic response in MCF-7/ADR.. Ce6-PDT induced cell viability decrease, intracellular ROS generation, and DNA damage in concentration-dependent and cell-specific manner, and MCF-7 was more sensitive to Ce6-PDT than MCF-7/ADR cells at the same PDT condition. PDT treatment could trigger cell death via apoptosis in MCF-7 cells but autophagic cell death in MCF-7/ADR cells.. These results suggested that MCF-7 was more sensitive to Ce6-PDT than MCF-7/ADR, and PDT treatment could trigger apoptotic response in MCF-7 cells, but stimulate autophagic response in MCF-7/ADR cells.

Topics: Breast Neoplasms; Cell Survival; Chlorophyllides; Dose-Response Relationship, Drug; Dose-Response Relationship, Radiation; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Humans; MCF-7 Cells; Photochemotherapy; Photosensitizing Agents; Porphyrins; Treatment Outcome

A programmed supramolecular nanocarrier was developed for multistage targeted photodynamic therapy. This smart nanocarrier exhibited enhanced cellular uptake and controlled mitochondria targeting, as well as an excellent photodynamic therapeutic effect after light irradiation.

Topics: Amino Acid Sequence; Apoptosis; beta-Cyclodextrins; Breast Neoplasms; Cell Proliferation; Chlorophyllides; Drug Carriers; Female; Humans; Light; MCF-7 Cells; Microscopy, Confocal; Mitochondria; Nanoparticles; Peptides; Photochemotherapy; Photosensitizing Agents; Porphyrins

Sono-Photodynamic therapy (SPDT), a new modality for cancer treatment, is aimed at enhancing anticancer effects by the combination of sonodynamic therapy (SDT) and photodynamic therapy (PDT). In this study, we investigated the antitumor effect and possible mechanisms of Chlorin e6 (Ce6) mediated SPDT (Ce6-SPDT) on breast cancer both in vitro and in vivo. MTT assay revealed that the combined therapy markedly enhanced cell viability loss of breast cancer cell lines (MDA-MB-231, MCF-7 and 4T1) compared with SDT and PDT alone. Propidium iodide/hoechst33342 double staining reflected that 4T1 cells with apoptotic morphological characteristics were significantly increased in groups given combined therapy. Besides, the combined therapy caused obvious mitochondrial membrane potential (MMP) loss at early 1 h post SPDT treatment. The generation of intracellular reactive oxygen species (ROS) detected by flow cytometry was greatly increased in 4T1 cells treated with the combination therapy, and the loss of cell viability and MMP could be effectively rescued by pre-treatment with the ROS scavenger N-acetylcysteine (NAC). Further, Ce6-SPDT markedly inhibited the tumor growth (volume and weight) and lung metastasis in 4T1 tumor-bearing mice, but had no effect on the body weight. Hematoxylin and eosin staining revealed obvious tissue destruction with large spaces in the Ce6-SPDT groups, and TUNEL staining indicated tumor cell apoptosis after treatment. Immunohistochemistry analysis showed that the expression level of VEGF and MMP were significantly decreased in the combined groups. These results indicated that Ce6-mediated SPDT enhanced the antitumor efficacy on 4T1 cells compared with SDT and PDT alone, loss of MMP and generation of ROS might be involved. In addition, Ce6-mediated SPDT significantly inhibited tumor growth and metastasis in mouse breast cancer 4T1 xenograft model, in which MMP-9 and VEGF may play a crucial role.

Topics: Animals; Apoptosis; Biological Transport; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Combined Modality Therapy; Female; Humans; Intracellular Space; Lung Neoplasms; Matrix Metalloproteinase 9; Mice; Mitochondria; Neoplasm Metastasis; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species; Ultrasonic Therapy; Vascular Endothelial Growth Factor A; Xenograft Model Antitumor Assays

Red blood cells are attached to iron oxide nanoparticles pre-coated with chlorine e6, a photosensitizer, and then loaded with a chemotherapeutic drug, doxorubicin, to enable imaging-guided combined photodynamic and chemotherapy of cancer, achieving excellent synergistic therapeutic effects in an animal tumor model. This work highlights the great promise of integrating cell-based drug-delivery systems with nanotechnology as a biocompatible multifunctional platform for applications in cancer theranostics.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Cell Tracking; Chlorophyllides; Combined Modality Therapy; Doxorubicin; Erythrocyte Transfusion; Erythrocytes; Magnetic Resonance Imaging, Interventional; Magnetite Nanoparticles; Mice; Mice, Inbred BALB C; Phototherapy; Porphyrins

Sono-photodynamic therapy (SPDT) is a new modality for cancer treatment. Some studies have reported enhanced tumor cytotoxicity when sonodynamic therapy (SDT) is combined with photodynamic therapy (PDT). In this study, we investigated the cytotoxic effect of SPDT-activated chlorin e6 (Ce6) on MDA-MB-231 cells. Ce6 was found to localize mainly in mitochondria, with maximal uptake within 4 h. Cell survival was estimated by MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltertrazolium bromide tetrazolium) assay 24 h after irradiation; the combined therapy enhanced cytotoxicity to a greater extent. Apoptosis was analyzed using annexin V-PE/7-ADD (7-aminoactinomycin D) staining as well as DAPI (4', 6-diamidino-2-phenylindole) staining, and the results indicated that the cells with apoptotic characteristics were significantly increased in groups given combined therapy. Rhodamine-123 staining and cytochrome c release revealed more serious damage of mitochondria after combined treatment. The generation of reactive oxygen species detected by flow cytometry was greatly increased in cells treated with the combination therapy, and the loss in cell viability could be effectively rescued with the reactive oxygen species inhibitor N-acetylcysteine. Moreover, enhancement of cell membrane permeability after ultrasound treatment was evaluated using FD-500, and it was found that the much higher uptake of Ce6 might be involved in PDT therapy with pre-treatment ultrasound. These results suggest that ultrasound enhances the cytotoxicity of Ce6-mediated PDT, possibly because of the increased intracellular Ce6 level and ROS formation by ultrasound pre-treatment.

Topics: Apoptosis; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Chlorophyllides; Electroporation; Humans; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Treatment Outcome; Ultrasonic Therapy

Photodynamic therapy (PDT) and sonodynamic therapy (SDT) are therapeutic modalities for tumors. In this study we investigated the combined cytotoxic effect of 0.36W/cm(2) and 0.72W/cm(2) ultrasound with various Ce6 concentrations (1, 2, 5, 10μg/ml), and that of 1μg/ml Ce6 with different laser light dose (650nm; 10.4mW/cm(2); 0.3, 0.6, 1.2 and 2.5J/cm(2)) on MDA-MB-231 cells. Both high reactive oxygen species (ROS) production and a decline in mitochondrial membrane potential (MMP) were detected with high Ce6 concentrations (5 and 10μg/ml) combined with 0.72W/cm(2) ultrasound and 1.2, 2.5J/cm(2) laser light with 1μg/ml Ce6. In addition, cell membrane integrity was evaluated by using propidium iodide (PI), revealing membrane damage was aggravated with the increasing ultrasound intensity, but no significant difference on cell membrane integrity could be observed after PDT treatment. These results suggest ROS may play an important role both in SDT and PDT. Besides, mitochondria may be an initial target in PDT while SDT can cause multi-site damages in MDA-MB-231 cells.

Topics: Breast Neoplasms; Cell Line, Tumor; Cell Membrane; Chlorophyllides; Humans; Mitochondria; Photochemotherapy; Photosensitizing Agents; Porphyrins; Reactive Oxygen Species; Ultrasonic Therapy

A novel iodinated chitosan-backboned conjugate (GC-I-Ce6) was designed and prepared to fabricate self-assembled biopolymeric nanoparticles with heavy atom-effected enhanced singlet oxygen generation as well as biological merits. The heavy atom-rich nature of the hydrophobic particle interior was characterized with X-ray absorption and the modified photophysical properties of a chemically embedded photosensitizer, chlorin e6 (Ce6). From the comparative spectroscopic studies as well as cellular and animal experiments, it has been shown that the self-assembled GC-I-Ce6 nanoparticles have enhanced capability of singlet oxygen generation by the intraparticle heavy-atom effect, along with high tumor targetability in vitro and in vivo thanks to the glycol chitosan-surfaced exterior with biocompatible, positively charged and tumor-homing characteristics. Actual efficacy improvement in the photodynamic therapy of a human breast cancer cell line (MDA-MB-231) demonstrates potential of our photophysically and pharmaceutically motivated hybrid bioconjugate approach for nanomedicine applications.

Topics: Animals; Breast; Breast Neoplasms; Cell Line, Tumor; Chitosan; Chlorophyllides; Drug Delivery Systems; Female; Halogenation; HeLa Cells; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Nanoparticles; Photochemotherapy; Photosensitizing Agents; Porphyrins; Singlet Oxygen

The cell surface receptor tissue factor (TF) is regarded as a common but specific target on angiogenic tumor vascular endothelial cells (VECs) and tumor cells in many types of cancer including breast cancer. The purpose of this study is to develop a selective and effective TF-targeting photodynamic therapy (PDT) by using its natural ligand factor VII (fVII)-conjugated Sn(IV) chlorin e6 (SnCe6) for the treatment of breast cancer. A cross linker EDC was used to covalently conjugate fVII protein to SnCe6, and the binding activity and phototoxicity was confirmed by ELISA and in vitro PDT. The efficacy of fVII-tPDT was assessed in vitro by crystal violet staining assay and in vivo by measuring tumor size in mice carrying murine or human breast cancer xenografts. We show that active site-mutated (K341A) fVII protein can be internalized into breast cancer cells and vascular endothelial growth factor (VEGF)-stimulated human umbilical vein endothelial cells (HUVECs) as angiogenic VECs. fVII-tPDT not only enhances 12-fold the in vitro efficacy but also selectively and effectively kills angiogenic HUVECs and breast cancer cells via specifically binding of fVII to TF and inducing apoptosis and necrosis as the underlying mechanism. Furthermore, fVII-tPDT can significantly inhibit the tumor growth of murine and human breast cancer without obvious toxicities in mice. We conclude that fVII-tPDT using fVII-SnCe6 conjugate can selectively and effectively kill angiogenic VECs and breast cancer cells in vitro and significantly inhibit the tumor growth of murine and human breast cancer in mice.

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Cross-Linking Reagents; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Factor VII; Female; Humans; Mammary Neoplasms, Animal; Mice; Neovascularization, Pathologic; Photochemotherapy; Porphyrins; Treatment Outcome

In the current study, semi-permeable alginate-oligochitosan microcapsules for multicellular tumor spheroids (MTS) generation were elaborated and tested, to estimate a response of the microencapsulated MTS (MMTS) to photodynamic therapy (PDT). The microcapsules (mean diameter 600 μm) with entrapped human breast adenocarcinoma MCF-7 cells were obtained using an electrostatic bead generator, and MMTS were generated by in vitro long-term cell cultivation. The formed MMTS were incubated in Chlorin e6 photosensitizer solution and then irradiated using 650-nm laser light. The cell viability was measured by MTT-assay in 24 h after irradiation, and histological analysis was performed. The proposed MTS-based model was found to be more resistant to the PDT than the two-dimensional monolayer cell culture model. Thus, MMTS could be considered as a promising three-dimesional in vitro model to estimate the doses of drugs or parameters for PDT in vitro before carrying out preclinical tests.

Topics: Alginates; Biocompatible Materials; Breast Neoplasms; Capsules; Cell Culture Techniques; Cell Line, Tumor; Chitosan; Chlorophyllides; Female; Humans; Materials Testing; Models, Biological; Particle Size; Photochemotherapy; Photosensitizing Agents; Porphyrins; Spheroids, Cellular; Static Electricity

The purpose of this study was to test a novel, dual tumour vascular endothelial cell (VEC)- and tumour cell-targeting factor VII-targeted Sn(IV) chlorin e6 photodynamic therapy (fVII-tPDT) by targeting a receptor tissue factor (TF) as an alternative treatment for chemoresistant breast cancer using a multidrug resistant (MDR) breast cancer line MCF-7/MDR.. The TF expression by the MCF-7/MDR breast cancer cells and tumour VECs in MCF-7/MDR tumours from mice was determined separately by flow cytometry and immunohistochemistry using anti-human or anti-murine TF antibodies. The efficacy of fVII-tPDT was tested in vitro and in vivo and was compared with non-targeted PDT for treatment of chemoresistant breast cancer. The in vitro efficacy was determined by a non-clonogenic assay using crystal violet staining for monolayers, and apoptosis and necrosis were assayed to elucidate the underlying mechanisms. The in vivo efficacy of fVII-tPDT was determined in a nude mouse model of subcutaneous MCF-7/MDR tumour xenograft by measuring tumour volume.. To our knowledge, this is the first presentation showing that TF was expressed on tumour VECs in chemoresistant breast tumours from mice. The in vitro efficacy of fVII-tPDT was 12-fold stronger than that of ntPDT for MCF-7/MDR cancer cells, and the mechanism of action involved induction of apoptosis and necrosis. Moreover, fVII-tPDT was effective and safe for the treatment of chemoresistant breast tumours in the nude mouse model.. We conclude that fVII-tPDT is effective and safe for the treatment of chemoresistant breast cancer, presumably by simultaneously targeting both the tumour neovasculature and chemoresistant cancer cells. Thus, this dual-targeting fVII-tPDT could also have therapeutic potential for the treatment of other chemoresistant cancers.

Topics: Adult; Aged; Animals; Apoptosis; Blotting, Western; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; CHO Cells; Cricetinae; Cricetulus; Drug Resistance, Neoplasm; Endothelial Cells; Factor VII; Female; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Mammary Neoplasms, Experimental; Mice; Mice, Nude; Middle Aged; Necrosis; Neovascularization, Pathologic; Photochemotherapy; Photosensitizing Agents; Porphyrins; Thromboplastin; Treatment Outcome; Tumor Burden; Xenograft Model Antitumor Assays

The photodynamic therapy is a well-known method of treatment of both malignant tumors and non-tumor lesions in human patients. In the present study, we aimed at evaluating the in vitro efficacy of the new photosensitizing agents, vacataporphyrin (VP), and divacataporphyrin (DVP).. The effectiveness of VP and DVP was compared to well-known photosensitizers, that is, hematoporphyrin derivative (HPD) and chlorin e6 (Ce6) in identical in vitro conditions. The experiment was performed on a well-established breast cancer cell line, MCF-7 and compared to HCV 29T11-2-D1 cell line. Cells were incubated in standard conditions and they were exposed to different concentrations of VP, DVP, HPD, and Ce6, that is, 180, 90, 45, 22.5, and 10 µg/ml. After incubation with photosensitizers, the cells were washed, medium was exchanged and the cells were subjected to irradiation at the proper wavelengths, light intensity (100 mW/sq cm), and total light doses 4.5 and 9 J/sq cm.. Our results showed that the VP and DVP are potent photosensitizers and the photocytotoxic effect after the incubation with DVP was much better than that of VP. The cytotoxic effects of VP and DVP were less intensive than these of HPD and Ce6. VP and DVP also accumulated well in the tumor cells. Our results also indicated that the VP and DVP effectiveness on MCF-7 cells was photosensitizer dose and light dose dependent.. The overall properties revealed by both new porphyrins and particularly a possibility for excitation at a higher wavelength and thus a deeper tissue penetration, make them promising candidates for further in vivo experiments.

Topics: Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Dose-Response Relationship, Drug; Female; Hematoporphyrin Derivative; Humans; Photochemotherapy; Photosensitizing Agents; Porphyrins

To evaluate the effect of adriamycin (ADM) in enhancing the sonodynamic effect of chlorin e6 against the proliferation of human breast cancer MDA-MB-231 cells in vitro.. MDA-MB-231 cells were treated with ultrasound/Chlorin e6 alone or in combination with ADM, and the changes in the cell proliferation was determined by MTT assay.. Ultrasound (1.0 MHz) at the power intensity of 0.5-2.0 W/cm2 inhibited the proliferation of MDA-MB-231 cells in an intensity-dependent manner, and chlorin-e6 (0.05-1.6 mg/ml) and ADM (0.1-0.4 g/ml) alone both inhibited the proliferation of MDA-MB-231 cells dose-dependently. Compared with ultrasound (0.5 W/cm2, 1.0 MHz, 60 s) or chlorin-e6 (0.05-0.2 mg/ml) alone, a combined treatment with ultrasound and chlorin e6 significantly enhanced the inhibitory effect on the proliferation of MDA-MB-231 cells (P<0.05). ADM significantly enhanced the sonodynamic effect of chlorin e6 (0.1 mg/ml) against the cell proliferation of MDA-MB-231 cells (P<0.05), and the effect was schedule-dependent, which was greater when ADM was added after the sonodynamic treatment (P<0.05).. ADM can enhance the sonodynamic effect of chlorin e6 against the proliferation of MDA-MB-231 cells in vitro.

Topics: Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Chlorophyllides; Doxorubicin; Female; Humans; Porphyrins; Ultrasonic Therapy

Photodynamic therapy has clinical indications in treatment of localized cancers and could be interesting for eradication of local recurrence of chemoresistant tumors. In the present study, the intracellular accumulation and distribution of chlorin e6 was investigated in MCF-7 and in P-glycoprotein overexpressing, doxorubicin resistant MCF-7/DXR cell lines. After 3-h incubation with chlorin e6 (1.7 micro mol.l(-1)), no significant difference in accumulation was observed between MCF-7 and MCF-7/DXR cells. Chlorin e6 cellular efflux did not differ in the two cell lines. The lack of influence of P-glycoprotein was confirmed since SDZ-PSC833 (PSC) had no influence in chlorin e6 accumulation and efflux in MCF-7/DXR cells. The subcellular distribution of chlorin e6 appeared different in MCF-7/DXR than in MCF-7 cells. Double staining colocalization fluorescence microscopy studies were performed to identify the subcellular localization sites for chlorin e6 using organelle probes for endoplasmic reticulum, Golgi apparatus, mitochondria and lysosomes. In MCF-7, chlorin e6 was distributed in all cytoplasmic organelles including endoplasmic reticulum and Golgi. In MCF-7/DXR, a diffuse cytoplasmic distribution was observed excepted for the endoplasmic reticulum and Golgi area in which less chlorin e6 was distributed. In MCF-7/DXR, PSC was found to restore the distribution of chlorin e6 in the endoplasmic reticulum and Golgi area while in MCF-7, no effect on the subcellular distribution of chlorin e6 was observed. Although the photodynamic activity of chlorin e6 (1.7 micro mol.l(-1), 650 nm, 8 mW.cm(-2)) was found to be lower in MCF-7/DXR than in MCF-7 cells, PSC was found to potentiate the photodynamic activity of chlorin e6 to similar extent in both cell lines. These results clearly demonstrate that PSC potentiates the photodynamic activity of Chlorin e6 independently of the expression of P-glycoprotein and further suggest that the photodynamic activity of chlorin e6 could be related to its intracellular distribution in the endoplasmic reticulum and the Golgi.

Topics: ATP Binding Cassette Transporter, Subfamily B, Member 1; Breast Neoplasms; Cell Line, Tumor; Chlorophyllides; Cyclosporins; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Endoplasmic Reticulum; Golgi Apparatus; Humans; Kinetics; Lasers; Light; Microscopy, Fluorescence; Porphyrins; Radiation-Sensitizing Agents; Spectrophotometry; Subcellular Fractions; Temperature; Time Factors

The transferrin receptor is often highly expressed in tumor cells whereas it is usually present at low levels in surrounding normal adult tissue. Here, a potential anti-cancer agent is described, which is directed at this receptor and consists of a toxin-modified transferrin, which is activated via photodynamic therapy. The porphyrin chlorin e6 was conjugated to transferrin using a procedure, which involved the preliminary binding of the protein to quaternary amino ethyl-sephadex. This maintained the natural activity of the transferrin, and the un-activated conjugate exhibited no in vitro cellular toxicity. The conjugate's singlet oxygen yield was estimated by assessment of its light-dependent oxidation of tetramethylbenzidine, where it displayed approximately 70% of the efficiency of native chlorin e6. When chlorin e6-transferrin treated human MCF7 and rat MTLn3 mammary adenocarcinoma cells were exposed to toxin-activating visible light, a tumor cell killing effect was achieved in normal (medium plus 10% FBS) culture conditions with an ED50 of approximately 10-20 microg/ml. A method for the synthesis of chlorin e6-transferrin was developed, and the conjugate was shown to exhibit a light-dependent killing of mammary adenocarcinoma cells in culture. The conjugate demonstrated potential as an anti-cancer agent.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Breast Neoplasms; Chlorophyllides; Drug Screening Assays, Antitumor; Female; Humans; Immunotoxins; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Rats; Receptors, Transferrin; Transferrin; Tumor Cells, Cultured

Certain tumor cells, such as squamous carcinoma cells, express an increased number of epidermal growth factor (EGF) receptors. Therefore, we studied the targeted delivery of the photocytotoxic compound Sn-(IV)chlorin e6 monoethylenediamine [SnCe6(ED)] to tumors that overexpress the EGF receptor. EGF was conjugated to SnCe6(ED) through a carrier, such as dextran (Dex) and human serum albumin (HSA), and the photocytotoxicity on the EGF receptor-overexpressing MDA-MB-468 breast adenocarcinoma cell line was evaluated. The photobiological activities of these EGF conjugates, of the conjugates of the photosensitizer to HSA or Dex, or of the photosensitizer alone were compared. The affinity of EGF for its receptor was substantially impaired when conjugated in EGF-Dex-SnCe6(ED), in contrast to EGF-HSA-SnCe6(ED). In corresponding results, EGF-HSA-SnCe6(ED) displayed a high photocytotoxicity (IC50, 63 nM) on MDA-MB-468 cells at a light dose of 27 kJ/m2, whereas EGF-Dex-SnCe6(ED) showed very limited photocytotoxicity. EGF-HSA-SnCe6(ED) was no longer photocytotoxic in the presence of a competing EGF concentration. The high photocytotoxicity of EGF-HSA-SnCe6(ED) was shown to be the result of a high intracellular concentration in MDA-MB-468 cells, which could be lowered dramatically by incubating the conjugate with a competing EGF concentration. In contrast, EGF-Dex-SnCe6(ED) accumulated poorly in MDA-MB-468 cells, in agreement with its low EGF receptor affinity and photocytotoxicity. EGF-HSA-SnCe6(ED) produced much more intracellular reactive oxygen species on light irradiation than EGF-Dex-SnCe6(ED). It is concluded that the photodynamic activity of the EGF-HSA conjugate of SnCe6(ED) on MDA-MB-468 breast adenocarcinoma cells is EGF specific and is much more potent than EGF-Dex-SnCe6(ED) or free SnCe6.

Topics: Adenocarcinoma; Binding, Competitive; Biological Transport; Breast Neoplasms; Cell Division; Chlorophyllides; Dextrans; Drug Synergism; Epidermal Growth Factor; ErbB Receptors; Humans; Inhibitory Concentration 50; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Reactive Oxygen Species; Recombinant Fusion Proteins; Serum Albumin; Substrate Specificity; Transfection; Tumor Cells, Cultured

Certain tumor cells, such as squamous carcinoma cells, express an increased number of epidermal growth factor (EGF) receptors. The goal of this study was the targeted delivery of the photocytotoxic compound Sn(IV)chlorine e6 monoethylenediamine++ (SnCe6(ED)) to tumors that overexpress the EGF receptor. Therefore EGF was conjugated to SnCe6(ED) through a carrier, such as dextran (Dex) and human serum albumin (HSA), followed by the evaluation of the photocytotoxicity on the EGF receptor overexpressing MDA-MB-468 cell line. The photobiologic activity of these conjugates was then compared to a conjugate of the photosensitizer to HSA or dextran, or to the photosensitizer alone. In contrast to EGF-HSA-SnCe6(ED), the affinity of EGF for its receptor was substantially impaired upon conjugation in EGF-Dex-SnCe6(ED). In correlation with these results, EGF-HSA-SnCe6(ED) displayed a high cytotoxicity (IC50, 63 nM) on MDA-MB-468 cells at a light dose of 27 kJ/m2, whereas EGF-Dex-SnCe6(ED) showed very limited photocytotoxicity. In the presence of a competing EGF concentration (10 microM), EGF-HSA-SnCe6(ED) was not cytotoxic anymore. The high photocytoxicity of EGF-HSA-SnCe6(ED) was shown to be a result of a high intracellular concentration in MDA-MB-468 cells, which could be lowered dramatically by incubating the conjugate with a competing EGF concentration. In contrast, EGF-Dex-SnCe6(ED) displayed very poor accumulation in MDA-MB-468 cells, in agreement with its low EGF receptor affinity and photocytoxicity. Besides, it could be demonstrated that EGF-HSA-SnCe6(ED) produced intracellularly ROS (reactive oxygen species) upon light irradiation, more than EGF-Dex-SnCe6(ED) did. It was concluded that, in contrast to EGF-Dex-SnCe6(ED) the photodynamic activity of the EGF-HSA conjugate of SnCe6(ED) on MDA-MB-468 breast adenocarcinoma cells is EGF-specific and more potent than free SnCe6(ED).

Topics: Adenocarcinoma; Animals; Binding, Competitive; Breast Neoplasms; Chlorophyllides; Dextrans; Epidermal Growth Factor; ErbB Receptors; Humans; Male; Mice; Photochemotherapy; Porphyrins; Radiation-Sensitizing Agents; Serum Albumin; Tumor Cells, Cultured

Patients with ovarian cancer that is clinically resistant to cisplatin-based chemotherapy have little hope of a cure of their disease. Photoimmunotherapy, which involves the antibody-targeted delivery of a nontoxic photosensitizer that is activated to a cytotoxic state with visible light, may offer a new treatment option. Photoimmunotherapy may be applied intraperitoneally to target disseminated tumor. We tested the hypothesis that this treatment in combination with cisplatin potentiates cytotoxicity in ovarian cancer cell lines and primary cultures of human tumors.. Five human cancer cell lines (ovarian and breast) and 19 primary cultures were studied. The primary cultures were from solid and ascites tumor samples obtained from 14 patients with ovarian cancer who were undergoing primary surgery. The photosensitizer chlorin e(6) was conjugated to the F(ab')(2) fragment of the murine monoclonal antibody OC-125, which is directed against the antigen CA 125. Cytotoxicity was measured by the microculture tetrazolium assay. Treatments consisted of cisplatin alone, photoimmunotherapy alone, and photoimmunotherapy followed by cisplatin. The fractional product method was used to assess synergy in treatment effects. Ex vivo cultured human cells exhibiting 80% or greater survival at cisplatin concentrations of 10 microM for 24 hours were defined as cisplatin resistant for this study.. When all cell types (cisplatin sensitive and cisplatin resistant) were considered together, combination treatment yielded cytotoxicity that was, on average, 6.9 times (95% confidence interval = 1.86-11.94) greater than that of cisplatin alone (two-sided P =.023). Cisplatin-resistant cells showed a synergistic effect of the two treatments (two-sided P =.044), while cisplatin-sensitive cells showed an additive effect.. These ex vivo data suggest that platinum resistance in human ovarian cancer cells may be reversible by pretreatment with OC-125-targeted photoimmunotherapy. Further studies are required to confirm the efficacy of this approach in vivo.

Topics: Antibodies, Monoclonal; Antineoplastic Agents; Breast Neoplasms; CA-125 Antigen; Chlorophyllides; Cisplatin; Combined Modality Therapy; Drug Resistance, Neoplasm; Female; Fluorescent Antibody Technique, Indirect; Humans; Immunotherapy; Ovarian Neoplasms; Photosensitizing Agents; Phototherapy; Porphyrins; Treatment Outcome; Tumor Cells, Cultured