Zinc pthalocyanine loaded poly (lactic acid) nanoparticles by double emulsion methodology for photodynamic therapy against 9 L/LacZ gliosarcoma cells.

Development delivery systems, such as nanoparticles, represent a growing area in biomedical research. Nanoparticles (NP) were prepared using a double-emulsion method to load zinc(II) phthalocyanine (ZnPc). NP were obtained using poly (lactic acid) (PLA). ZnPc is a second generation of photosensitizer used in photodynamic therapy (PDT). ZnPc loaded PLA nanoparticles (NPLA-ZnPc) were prepared by double-emulsion method, characterized and available in cellular culture. The mean nanoparticle size presented particle size was 384.7 ± 84.2 nm with polydispersity index (PDI) of 0.150 ± 0.015, and the encapsulation efficiency was of 83%. The nanoparticle formulations presented negative zeta potential values (-27.5 ± 1.0 mV), explaining their colloidal stability. ZnPc loaded nanoparticles maintain its photophysical behavior after encapsulation. Photosensitizer release from nanoparticles was sustained over 168 h with a biphasic ZnPc release profile. An in vitro phototoxic effect in range of 80% was observed in 9 L/LacZ gliosarcoma cells at laser light doses (10 J cm-2) with 3.0 µg mL-1 of NPLA-ZnPc. All the physical-chemical, photophysical and photobiological measurements performed allow us to conclude that ZnPc loaded PLGA nanoparticles is a promising drug delivery system for PDT.

Gelatin nanoparticles via template polymerization for drug delivery system to photoprocess application in cells.

Photodynamic therapy (PDT) is a clinical treatment based on the activation of light-absorbing photosensitizers (PS) to generate reactive oxygen species, which are toxic to the targeted disease cells. Because most PS are hydrophobic with poor water solubility, it is necessary to encapsulate and solubilize PS in aqueous conditions to improve the photodynamic action for this compound. In this work, gelatin-poly(acrylic acid) nanoparticles (PAA/gelatin nanoparticles) via template polymerization for incorporation aluminum chloride phthalocyanine (ClAlPc) as a model drug for PDT application were developed. Biocompatible core-shell polymeric nanoparticles were fabricated via template polymerization using gelatin and acrylic acid as a reaction system. The nanoparticulate system was studied by scanning electron microscopy, steady-state, and their biological activity was evaluated using in vitro cancer cell lines by classical MTT assay. The obtained nanoparticles had a spherical shape and DLS particle size were determined further and was found to be around 170 nm. The phthalocyanine-loaded-nanoparticles maintained their photophysical behaviour after encapsulation. It is found that ClAlPc can be released from the nanoparticles in a sustained manner with a small initial burst release. In vitro cytotoxicity revealed that ClAlPc-loaded nanoparticles had similar cytotoxicity to free ClAlPc with mouse melanoma cancer cell line (B16-F10). In vitro photoeffects assay indicated that the nanoparticle formulation was superior in anticancer effect to free ClAlPc on mouse melanoma cancer cell line B16-F10. The results indicate that ClAlPc encapsulated in gelatin-poly(acrylic acid) nanoparticles are a successful delivery system for improving photodynamic activity in the target tissue.

Effect of diode-laser and AC magnetic field of bovine serum albumin nanospheres loaded with phthalocyanine and magnetic particles.

This study reports on the development and characterization of bovine serum albumin (BSA) nanospheres containing Silicon(IV) phthalocyanine (NzPc) and/or maghemite nanoparticles (MNP), the latter introduced via ionic magnetic fluid (MF). The nanosized BSA-loaded samples were designed for synergic application while combining Photodynamic Therapy and Hyperthermia. Incorporation of MNP in the albumin-based template, allowing full control of the magnetic content, was accomplished by adding a highly-stable ionic magnetic fluid sample to the albumin suspension, following heat denaturing. The material's evaluation was performed using Zeta potential measurements and scanning electron microscopy. The samples were characterized by steady-state techniques and time-resolved fluorescence. The in vitro assay, using human fibroblasts, revealed no cytotoxic effect in all samples investigated, demonstrating the potential of the tested system as a synergistic drug delivery system.

BSA nanoparticles loaded-methylene blue for photodynamic antimicrobial chemotherapy (PACT): effect on both growth and biofilm formation by Candida albicans.

It has been demonstrated an increase in resistance of Candida albicans to conventional therapies, probably, due the indiscriminate use of the conventional antifungal drugs. In this aspect, the nanotechnology generates the possibility of creating new therapeutic agents. Thus, the objective of this paper was to produce and characterize a bovine serum albumin (BSA) nanoparticle encapsulated with Methylene Blue (MB). In addition, the effect of BSA nanoparticles encapsulated with MB (BSA-MB) was evaluated on both growth and biofilm formation by C. albicans by Photodynamic Antimicrobial Chemotherapy (PACT) protocols. The BSA-MB nanoparticles were prepared by the desolvation process. The nanoparticulate system was studied by steady-state techniques, scanning electron microscopy and their biological activity was evaluated in vitro both growth and biofilm formation by C. albicans. The synthetized BSA-MB nanoparticles were spherical in shape exhibiting a 100-200 nm diameter with a low tendency to aggregate (PDI values < 0.2). MB photophysical properties were shown to be preserved after BSA encapsulation. A significant reduction in C. albicans growth, after PACT was observed, in a dependent manner on MB-loaded in BSA nanoparticles concentration used. It was observed an inhibition of 23, 65 and 83% in the presence of MB-loaded in BSA nanoparticles 0.1, 0.5 and 1.0 µg.mL-1, respectively. In addition, MB-loaded BSA nanoparticles 0.5 µg.mL-1 were able to reduce both biofilm formation (80%) and the transition from yeast to filamentous form by C. albicans. The results presented here demonstrated a potentiation of the phototoxic effect of MB after BSA encapsulation, since the concentrations of MB-loaded BSA nanoparticles necessary to inhibits ∼50% of C. albicans development was 10 times minor than that observed for free MB. Taken together, these results suggest the potential of PACT, using MB-loaded BSA nanoparticles in inhibiting C. albicans development. The synthesis and design of BSA nanoparticles can be successfully applied for MB encapsulation and offer the possibility to drive the toxicity effect to a specific target, as an evaluation on both growth and biofilm formation by Candida albicans.

Polyelectrolytic BSA nanoparticles containing silicon dihydroxide phthalocyanine as a promising candidate for drug delivery systems for anticancer photodynamic therapy.

Recently several scientific-technological advances in the health area have developed. Among them, we can highlight research addressing nanoscience and nanotechnology focusing on the development of formulations for the cancer treatment. This work describes the synthesis and characterization of bovine serum albumin (BSA) polyelectrolytic nanoparticles for controlled release using silicon dihydroxide phthalocyanine [SiPc (OH)2] as a photosensitizer model for application in Photodynamic Therapy (PDT). BSA nanoparticles were prepared by the one-step desolvation process and the nanoparticulate system was coated with polyelectrolytes using poly-(4-styrene sulfonate - PSS) as a strong polyanion and polyallylamine hydrochloride (PAH) as a weak polycation by the technique self-assembling layer-by-layer (LbL). The formulation was characterized and available in cellular culture. The profile of drug release was investigated and compared to that of free [SiPc (OH)2]. The nanoparticles have a mean diameter of 226.9 nm, a narrow size distribution with polydispersive index of 0.153, smooth surface and spherical shape. [SiPc(OH)2] loaded nanoparticles maintain its photophysical behaviour after encapsulation. The polyelectrolytic nanoparticles improved efficiency in release and photocytotoxicity assay when compared to pure drug. The results demonstrate that photosensitizer adsorption on BSA nanoparticles together with biopolymer layer-by-layer assembly provides a way to manufacture biocompatible nanostructured materials that are intended for use as biomaterials for Photodynamic Therapy applications.

Zinc phthalocyanine tetrasulfonate-loaded polyelectrolytic PLGA nanoparticles for photodynamic therapy applications.

BACKGROUND: Photodynamic Therapy (PDT) is a modality for the treatment of neoplastic tissues, which is based on the administration of a phototherapeutic agent and light irradiation at an appropriate wavelength, aiming to locate and destroy the target cell with the formation of reactive oxygen species. Nanoencapsulation technology presents itself as a tool for incorporation of bioactive substances aiming to improve their solubility in physiological environment, obtain a longer circulation time in the organism, administration of lower dosages and the minimization of side effects. The present work aimed at the development of poly (lactic acid-glycolic acid) (PLGA) nanoparticles coated with polyelectrolyte film layers for encapsulating zinc phthalocyanine tetrasulfonated (ZnPcSO4) as a bioactive substance model. METHODS: PLGA nanoparticles were produced by the double emulsion/solvent evaporation technique and polyelectrolytic coating was performed using polyalkylamine hydrochloride (PAH) as a weak polycation and poly (4-styrene sulfonate) (PSS) as a strong polyanion by layer-by-layer self-assembly technique (known as layer-by-layer-LbL). The nanoparticulate system was studied by scanning electron microscopy, steady-state, and their biological activity was evaluated using in vitro cancer cell lines by classical MTT assay. RESULTS: The polyelectrolytic PLGA nanoparticles had an average diameter of 384.7 ± 138.6 nm, restricted distribution size with a polydispersity index. The obvious change in zeta potential indicates successful alternation in polycation (PAH) and polyanion (PSS) deposition directly in PLGA nanoparticles. Scanning electron microscopy (SEM) analysis showed that the formed system had morphology spherical, typical of these release systems. The loading efficiency was 82.1 % ± 1.2 %. The polyelectrolytic nanoparticles loaded with phthalocyanine maintained their photophysical behavior after encapsulation. Cell viability was determined, obtaining 90 % cell death. CONCLUSIONS: Therefore, the presented work depicts ZnPcSO4-loaded polyelectrolytic PLGA nanoparticles as a promise drug delivery system for phototherapeutic agent, which are thus expected to have superior therapeutic efficiency than free drug.

Photodynamic therapy with aluminum-chloro-phthalocyanine induces necrosis and vascular damage in mice tongue tumors.

In this paper we describe the efficacy of the liposomal-AlClPc (aluminum-chloro-phthalocyanine) formulation in PDT study against Ehrlich tumor cells proliferation in immunocompetent swiss mice tongue. Experiments were conduced in sixteen tumor induced mice that were divided in three control groups: (1) tumor without treatment; (2) tumor with 100J/cm(2) laser (670nm) irradiation; and (3) tumor with AlClPc peritumoral injection; and a PDT experimental group when tumors received AlClPc injection followed by tumor irradiation. Control groups present similar macroscopically and histological patterns after treatments, while PDT treatment induced 90% of Ehrlich tumor necrosis after 24h of one single application, showing the efficacy of liposome-AlClPc (aluminum-chloro-phthalocyanine) mediated PDT on the treatment of oral cancer.

Synthesis, photophysical and photobiological characterization of BSA nanoparticles loaded with chloroaluminium phthalocyanine by one-step desolvation technique for photodynamic therapy action.

This work describes the preparation of bovine serum albumin (BSA) nanoparticles by one-step desolvation method using acetone/ethanol as precipitating agent, glutaraldehyde as crosslinking agent, sodium azide as a preservative and water as reaction media for chloroaluminium phthalocyanine (ClAlPc) incorporation for photodynamic therapy action. The characterization of the nanoparticulate system was carried out using steady-state technique, scanning electron microscopy study and their biological activity was evaluated using in vitro macrophages cell lines by classical MTT assay. All the spectroscopy measurements demonstrated good photophysical properties and the in vitro cytotoxicity study showed that the system is not cytotoxic in the darkness, but it exhibits a substantial phototoxicity at 0.90 mol.L-1 of photosensitizer concentration and 10.0 J cm-2 of light. These conditions are sufficient to kill about 90% of the cells. The results allowed us to conclude that ClAlPc-loaded in BSA nanoparticles might have potential application on drug delivery system for PDT protocols.

Functionalized photosensitive gelatin nanoparticles for drug delivery application.

In this study, zinc phthalocyanine (ZnPc) was loaded onto gelatin nanoparticles functionalized with polyelectrolytes (polystyrene sulfonate/polyallylamine hydrochloride) by layer-by-layer (LbL) assembly. The process yield and the encapsulation efficiency were 76.0% ± 2.5 and 86.0% ± 1.8, respectively. The functionalized photosensitive gelatin nanoparticles (FPGN) had a mean diameter of 396.5 ± 45.8 nm, narrow distribution size with a polydispersity index of 0.106. The obvious switching of zeta potential indicates successful alternating deposition of the polyanion PSS and polycation PAH directly on the gelatin nanoparticles. The in vitro drug release investigation found that the LbL deposited polyelectrolyte bilayer is very efficient to reduce the release rate and assuage the initial burst for drugs loaded in gelatin nanoparticles. The photobiological activity of FPGN was evaluated on mouse macrophage carcinoma line J774 A-1. The cells viability decreased with the increase of the light dose in the range of 1-10.0 J.cm-2. ZnPc-loaded in functionalized gelatin nanoparticles are the release systems that promise photodynamic therapy use.

Gelatin nanoparticles loaded methylene blue as a candidate for photodynamic antimicrobial chemotherapy applications in Candida albicans growth.

Gelatin nanoparticles (GN) with an intrinsic antimicrobial activity maybe a good choice to improve the effectiveness of photodynamic antimicrobial chemotherapy (PACT). The aim of this study was to development gelatin nanoparticles loaded methylene blue (GN-MB) and investigate the effect of GN-MB in the Candida albicans growth by PACT protocols. The GN and GN-MB were prepared by two-step desolvation. The nanoparticulate systems were studied by scanning electron microscopy and steady-state techniques, the in vitro drug release was investigated, and we studied the effect of PACT on C. albicans growth. Satisfactory yields and encapsulation efficiency of GN-MB were obtained (yield = 76.0% ± 2.1 and EE = 84.0% ± 1.3). All the spectroscopic results presented here showed excellent photophysical parameters of the studied drug. Entrapment of MB in GN significantly prolongs it's in vitro release. The results of PACT experiments clearly demonstrated that the photosensitivity of C. albicans was higher when GN-MB was used. Gelatin nanoparticles loaded methylene blue-mediated photodynamic antimicrobial chemotherapy may be used against Candida albicans growth.

Photosensitizer-loaded magnetic nanoemulsion for use in synergic photodynamic and magnetohyperthermia therapies of neoplastic cells.

In this study a magnetic nanoemulsion (MNE) was developed from a mixture of two components, namely biodegradable surfactants and biocompatible citrate-coated cobalt ferrite-based magnetic fluid, for entrapment of Zn(II)-Phthalocyanine (ZnPc), the latter a classical photosensitizer (PS) species used in photodynamic therapy (PDT) procedures. The sample's stability was evaluated as a function of time using photocorrelation spectroscopy (PCS) for determination of the average hydrodynamic diameter, diameter dispersion and zeta potential. The ZnPc-loaded magneto nanoemulstion (ZnPc/MNE) formulation was evaluated in vitro assays to access the phototoxicity and the effect of application of AC magnetic fields (magnetohyperthermia damage) after incubation with J774-A1 macrophages cells. Darkness toxicity, phototoxicity and AC magnetic field exposures revealed an enhancement response for combined photodynamic and magnetohyperthermia (MHT) processes, indicating the presence of the synergic effect.

Layer-by-layer hollow photosensitizer microcapsule design via a manganese carbonate hard template for photodynamic therapy in cells.

BACKGROUND: Microcapsules fabricated using layer-by-layer self-assembly have unique properties, making them attractive for drug delivery applications. The technique has been improved, allowing the deposition of multiple layers of oppositely charged polyelectrolytes on spherical, colloidal templates. These templates can be decomposed by coating multiple layers, resulting in hollow shells. In this paper, we describe a novel drug delivery system for loading photosensitizer drugs into hollow multilayered microcapsules for photoprocess applications. METHODS: Manganese carbonate particles were prepared by mixing NH4HCO3 and MnSO4 and performing consecutive polyelectrolyte adsorption processes onto these templates using poly-(sodium 4-styrene sulfonate) and poly-(allylamine hydrocholoride). A photosensitizer was also incorporated into the layers. Hollow spheres were fabricated by removing the cores in the acidic solution. The hollow, multilayered microcapsules were studied by scanning electron microscopy, steady-state, and time-resolved techniques. Their biological activity was evaluated in vitro with cancer cells using a conventional MTT assay. RESULTS: The synthesized CaCO3 microparticles were uniform, non-aggregated, and highly porous spheres. The phthalocyanine derivatives loaded in the microcapsules maintained their photophysical behaviour after encapsulation. The spectroscopic results presented here showed excellent photophysical behaviour of the studied drug. We observed a desirable increase in singlet oxygen production, which is favourable for the PDT protocol. Cell viability after treatment was determined and the proposed microcapsules caused 80% cell death compared to the control. CONCLUSIONS: The results demonstrate that photosensitizer adsorption into the CaCO3 microparticle voids together with the layer-by-layer assembly of biopolymers provide a method for the fabrication of biocompatible microcapsules for use as biomaterials.

Preparation of gelatin nanoparticles by two step desolvation method for application in photodynamic therapy.

Gelatin nanoparticles have recently been receiving considerable attention because they offer a good option as release systems due to their low cost, biocompatibility, biodegradability and its application in several types of formulations. This study aim was to evaluate the potential application of gelatin nanoparticles entrapping a photosensitizer in Photodynamic Therapy. Gelatin nanoparticles were studied by steady-state techniques and the biological activity evaluated by in vitro MTT assay. The particles were spherical in shape exhibiting a 273 nm diameter with a low tendency to aggregate. The loading efficiency was 76%. Photosensitizer photophysical properties were shown to be preserved after GN encapsulation. The cells viability obtaining 85% cells death compared with control. The results demonstrate that gelatin nanoparticles can be successfully applied for photosensitizers encapsulation or other active drugs and be used as an optimal medium for a variety of bioactive materials, which can also be encapsulated by the proposed method.

Effects of Photodynamic Process (PDP) in Implant Osseointegration: A Histologic and Histometric Study in Dogs.

BACKGROUND: The combination between photosensitivity substances with laser or light-emitting diode (LED) form the photodynamic therapy basis that consists of photosensitivity drug activated by low-frequency light. This mechanism is used in soft tissue healing process to improve the oxygen tension leading to a fast revascularization. PURPOSE: The objective of this study was to evaluate the effects of photosensitivity drugs activated through LED on osseointegration process. MATERIALS AND METHODS: Eight mongrel dogs underwent implant therapy in four mandibular bone defects using 5.0 mm trephine drill on each side of the mandible. The defects were randomly filled up with (1) Nano emulsion, (2) liposome, (3) blood clot, and (4) autogenous bone. LED with visible and infrared light were applied after 48/72 postoperative hours on four dogs and after 96/120 postoperative hours in the other four dogs. All the animals were euthanized at 15 days after surgery. Ground sections slides were prepared from the experimental site for histomorphometry and histological analysis. RESULTS: No difference was detected in the following parameters: bone-implant contact, bone inside the defect and crest level on LED 48/72. Significant difference was detected inside the defect when filled with autogenous bone (p = .0238) on LED 96/120. When LED 48/72 and LED 96/120 were compared, significant higher bone formation was detected when autogenous bone on bone-implant contact (p = .0043) and bone inside the defect (p = .0008) was used. CONCLUSION: The use of photosensitivity drugs activated by LED demonstrated a tendency to stimulate bone formation, similar to autogenous bone graft on later time point.

Photodynamic therapy leads to complete remission of tongue tumors and inhibits metastases to regional lymph nodes.

In patients diagnosed with oral cancer, the most important prognostic indicator for patient survival after primary treatment is metastasis to the cervical lymph nodes or distal sites. Therefore, we evaluated the utility of photodynamic therapy (PDT) mediated by aluminum-chloride-phthalocyanine entrapped in liposomes for the prevention of metastasis to regional cervical lymph nodes in the Erhlich tongue cancer model. The PDT protocol led to complete remission of tongue tumours and prevented the occurrence of regional metastasis. The prevention of regional metastasis was confirmed by histopathological and immunohistochemical analyses. In addition, PDT treatment increased the overall survival and reduced weight loss relative to control tumour-bearing mice. Thus, PDT should be clinically evaluated for use in the prevention of cervical lymph node metastasis in patients with oral cancer.

Successful strategy for targeting the central nervous system using magnetic albumin nanospheres.

This study reports on the successful use of magnetic albumin nanosphere (MAN), consisting of maghemite nanoparticles hosted by albumin-based nanosphere, to target different sites within the central nervous system (CNS). Ultrastructural analysis by transmission electron microscopy (TEM) of the material collected from the mice was performed in the time window of 30 minutes up to 30 days after administration. Evidence found that the administered MAN was initially internalized and transported by erythrocytes across the blood-brain-barrier and transferred to glial cells and neuropils before internalization by neurons, mainly in the cerebellum. We hypothesize that the efficiency of MAN in crossing the BBB with no pathological alterations is due to the synergistic effect of its two main components, the iron-based nanosized particles and the hosting albumin-based nanospheres. We found that the MAN in targeting the CNS represents an important step towards the design of nanosized materials for clinical and diagnostic applications.

Muscle electrical activity during exercises with and without load executed on dry land and in an aquatic environment

Introduction Muscle activity in the aquatic environment was investigated using electromyographic analyses. The physical properties of water and the resistance used may influence the response of the muscle during exercise. The objective of this study was to evaluate the electrical activity in water and on the floor during flexion and knee extension exercises with and without load and aimed at understanding the muscular response while performing resistance exercises in water. Methods The sample consisted of 14 volunteers between 18 and 35 years old who were subjected to active exercises involving knee flexion and extension with and without load on the floor and in water. Electromyography was performed during the movement. Results A significant increase was found in the electrical activity of the rectus femoris muscle during exercises on the floor. The biceps femoris muscle showed increased electromyographic activity when resistance was used. A significant increase was found in the electrical activity of the rectus femoris muscle compared with exercises with and without load and the moment of rest in immersion. The electrical activity of the rectus and biceps femoris muscles was reduced in exercises with load and without load in a therapy pool compared with on the floor. Conclusion There was a reduction of the electromyographic activity in the aquatic environment compared with that on the ground, which could be attributed to the effects from hot water. Therefore, it is believed that resistance exercises can be performed early in a therapy pool, which will facilitate the prevention and treatment of musculoskeletal disorders.