Accelerating skin regeneration and wound healing by controlled ROS from photodynamic treatment.

Cellular metabolisms produce reactive oxygen species (ROS) which are essential for cellular signaling pathways and physiological functions. Nevertheless, ROS act as "double-edged swords" that have an unstable redox balance between ROS production and removal. A little raise of ROS results in cell proliferation enhancement, survival, and soft immune responses, while a high level of ROS could lead to cellular damage consequently protein, nucleic acid, and lipid damages and finally cell death. ROS play an important role in various pathological circumstances. On the contrary, ROS can show selective toxicity which is used against cancer cells and pathogens. Photodynamic therapy (PDT) is based on three important components including a photosensitizer (PS), oxygen, and light. Upon excitation of the PS at a specific wavelength, the PDT process begins which leads to ROS generation. ROS produced during PDT could induce two different pathways. If PDT produces control and low ROS, it can lead to cell proliferation and differentiation. However, excess production of ROS by PDT causes cellular photo damage which is the main mechanism used in cancer treatment. This review summarizes the functions of ROS in living systems and describes role of PDT in production of controllable ROS and finally a special focus on current ROS-generating therapeutic protocols for regeneration and wound healing.

An update on molecular mechanisms of curcumin effect on diabetes.

Owing to its prevalent nature, diabetes mellitus has become one of the most serious endocrine illnesses affecting a patient's quality of life due to the manifestation of side effects such as cardiovascular diseases, retinopathy, neuropathy, and nephropathy. Curcumin ((1E, 6E) 21, 7-bis (4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione), a major compound of turmeric, has been used in conventional medicine because of its safe nature and cost-effectiveness to meliorate diabetes and its comorbidities. These effects have also been observed in rodent models of diabetes resulting in a reduction of glycemia and blood lipids. Both the preventive and therapeutic activities of this compound are due to its antioxidant and anti-inflammatory characteristics. Furthermore, preclinical outcomes and clinical investigation demonstrate that the use of curcumin neutralizes insulin resistance, obesity, and hyperglycemia. Despite the many benefits of curcumin, its two limiting factors, solubility and bioavailability, remain a challenge for researchers; therefore, several methods such as drug formulation, nano-drug delivery, and the use of curcumin analogs have been developed to deliver curcumin and increase its bioavailability. PRACTICAL APPLICATIONS: The rise of people with type 2 diabetes has become a major concern at the global healthcare level. The best diabetes treatments today are anti-diabetic drug administration, lifestyle-related interventions (such as healthy eating and daily physical activity), arterial pressure detection, and fat control. The polyphenol curcumin, found in turmeric, can promote health by acting on a variety of cellular signaling pathways. This review article discusses curcumin and its role in the treatment of diabetes.

Spectroscopic investigation on molecular aspects and structural and functional effects of tetraethyl pyrophosphate organophosphorus insecticide interaction with adult human hemoglobin.

Organophosphates are extremely toxic compounds that use extensively in agriculture and household as insecticides. However, their binding mechanism to bio-macromolecules especially blood proteins is not clearly understood. In this research, various spectroscopic techniques utilized to analyze the effect of Tetraethyl Pyrophosphate (TEPP), as an organophosphorus insecticide, on the structure, function, stability, and aggregation of adult human hemoglobin and also hemolysis potential of the TEPP on red blood cells (RBCs) examined. Molecular docking was used for TEPP binding to human Hemoglobin (Hb), too. The results demonstrated that the TEPP insecticide has the potential for lysing RBCs. UV-Vis experiment indicated that globin part and heme group influenced by TEPP. Oxygen affinity measurements revealed the formation of deoxy-Hb and met-Hb, also decreased in oxygen affinity of Hb upon interaction with TEPP that is due to heme destruction. Fluorescence spectroscopy confirmed the production of heme degradation species after interaction of Hb with TEPP, which is inconsistent with oxygen affinity measurements. Thermal and aggregation studies indicated that TEPP induced aggregation of Hb in a concentration manner and Tm of protein reduced to lower temperatures. Docking's study also showed that TEPP interacts with Hb through hydrophobic interactions, which confirms UV-Vis results. ATR-FTIR study also revealed that TEPP can induce changes in the alpha helix element of Hb's secondary structure. Totally, Experimental and theoretical results indicate that tetraethyl pyrophosphate has unfavorable effects on hemoglobin structure and function.Communicated by Ramaswamy H. Sarma.

Preparation of cerium-curcumin and cerium-quercetin complexes and their LEDs irradiation assisted anticancer effects on MDA-MB-231 and A375 cancer cell lines.

Cancer remains common and often is difficult to eradicate. In particular resistant forms like triple negative breast cancer and melanoma generally allow for very short survival. Curcumin and quercetin as two important polyphenols from plants which have different biological roles, potentially including anti-cancer effect. But their clinical application is limited due to poor solubility in aqueous medium. Photodynamic therapy (PDT) is a cancer treatment using select chemical compounds as photosensitizers, which when activated by light create toxic singlet oxygen. Studies done on plant based photosensitizers such as curcumin and quercetin have shown the ability to ablate tumors. Here we discuss using them as improved PS by making their complex with cerium ions as a delivery system for MDA-MB-231 and A375 cancer cell lines treatment. For this purpose, the MDA-MB-231 human breast cancer cell line exposed to red light irradiation (as pretreatment) then treated with curcumin and quercetin alone and also their complex with cerium. In another study the cells treated with curcumin-cerium and quercetin-cerium complex and then irradiated with blue light (photodynamic treatment). Cell survival and apoptosis were determined using MTT and fluorescence microscopy. The result showed that curcumin and quercetin in complex with cerium ions have better toxic effect against both breast and melanoma cancer cells as compared to each compound alone. The finding revealed that curcumin and quercetin in cerium complex could be considered as a new approach in the photodynamic treatment of breast and melanoma cancer cells.

A mechanistic perspective on targeting bacterial drug resistance with nanoparticles.

Bacterial infections are an important cause of mortality worldwide owing to the prevalence of drug resistant bacteria. Bacteria develop resistance against antimicrobial drugs by several mechanisms such as enzyme inactivation, reduced cell permeability, modifying target site or enzyme, enhanced efflux because of high expression of efflux pumps, biofilm formation or drug-resistance gene expression. New and alternative ways such as nanoparticle (NP) applications are being established to overcome the growing multidrug-resistance in bacteria. NPs have unique antimicrobial characteristics that make them appropriate for medical application to overcome antibiotic resistance. The proposed antibacterial mechanisms of NPs are cell membrane damage, changing cell wall penetration, reactive oxygen species (ROS) production, effect on DNA and proteins, and impact on biofilm formation. The present review mainly focuses on discussing various mechanisms of bacterial drug resistance and the applications of NPs as alternative antibacterial systems. Combination therapy of NPs and antibiotics as a novel approach in medicine towards antimicrobial resistance is also discussed.

Nanomaterials as drug delivery systems with antibacterial properties: current trends and future priorities.

Introduction:Despite extensive advances in the production and synthesis of antibiotics, infectious diseases are one of the main problems of the 21st century due to multidrug-resistant (MDR) distributing in organisms. Therefore, researchers in nanotechnology have focused on new strategies to formulate and synthesis the different types of nanoparticles (NPs) with antimicrobial properties.Areas covered:The present review focuses on nanoparticles which are divided into two groups, organic (micelles, liposomes, polymer-based and lipid-based NPs) and inorganic (metals and metal oxides). NPs can penetrate the cell wall then destroy permeability of cell membrane, the structure and function of cell macromolecules by producing of reactive oxygen species (ROS) and eventually kill the bacteria. Moreover, their characteristics and mechanism in various bacteria especially MDR bacteria and finally their biocompatibility and the factors affecting their activity have been discussed.Expert opinion:Nanotechnology has led to higher drug absorption, targeted drug delivery and fewer side effects. NPs can overcome MDR through affecting several targets in the bacteria cell and synergistically increase the effectiveness of current antibiotics. Moreover, organic NPs with regard to their biodegradability and biocompatibility characteristics can be suitable agents for medical applications. However, they are less stable in environment in comparison to inorganic NPs.

Effect of low-level laser irradiation on cytotoxicity of benzene in human normal fibroblast cells.

Benzene is volatile organic hydrocarbon which is widely used in a wide range of industries. Studies have shown that exposure to benzene consequences serious health risks for human. Understanding the effect and risks of environmental hazard materials in the laser therapy of skin is interesting which can show useful or harmful role of these effects in therapies. In this study, the effect of low-level laser therapy was investigated on benzene-induced cytotoxicity on human skin fibroblast cells (HU02). Human skin fibroblast cells (HU02) were exposed to various concentrations of benzene (0-100 µg/mL) and incubated for 2 h. Then the effect of low-level laser therapy (LLLT) at 660-nm wavelength with 3 J/cm2 energy for 90 s was investigated on the viability of the cells exposed to benzene using MTT assay and inverted light microscope. The effect of low-level laser therapy on the viability of the cells was positive at concentrations 0-15 µg/mL but negative at higher concentrations than 15 µg/mL. Low-level laser therapy in low concentrations of benzene decreases the cytotoxicity caused by benzene and maintains cell viability. At high concentrations and in the presence of low-level laser therapy, the cell viability decreased compared to dark experiment. The morphology study of the cells using inverted light microscopy has confirmed the MTT results.

Curcumin-silica nanocomplex preparation, hemoglobin and DNA interaction and photocytotoxicity against melanoma cancer cells.

Melanoma is a malignant cancer of the skin associated with a high mortality. Early medical diagnosis and surgical intervention are essential for the treatment of melanoma. The use of plant-based compounds is an important strategy for the prevention and treatment of different types of cancers. Curcumin is a promising natural anticancer compound used towards treatment for various kinds of cancers. Studies have shown that curcumin could be applied as a photosensitizer in cancer photodynamic therapy (PDT). PDT uses light and a photosensitizing agent which produce reactive oxygen species leading to cancer cell death. The main obstacle for using curcumin as photosensitizer is its low solubilization ability in an aqueous environment. To improve its application in cancer treatment, we synthetized curcumin-silica nanoparticles as photosensitizer for photodynamic treatment of human melanoma cancer cells. Scanning electron microscopy, Transmission electron microscopy, Powder X-ray diffraction and Thermo geometric analysis indicated that curcumin was loaded on silica. The solubility of curcumin in water increased by using silica nanoparticles which wasconfirmed by spectroscopy results. The spectroscopy study confirmed the interaction of curcumin-silica nanocomplex with double strand DNA and no interaction with hemoglobin. The curcumin-silica nanocomplex and curcumin photodynamic effect was investigated on human melanoma cancer cells (A375) and also human fibroblast cells. The cell toxicity experiments showed that the curcumin-silica nanocomplex had greater photodynamic effects on cancer cell death as compared to free curcumin. The apoptotic assay by acridine orange/ethidium bromide (AO/EB) dual staining and colony forming ability confirmed the MTT results. Therefore, these results suggest that the curcumin-silica nanocomplex has great potential to be employed in photodynamic treatment of melanoma cancer.

Molecular interaction and cellular studies on combination photodynamic therapy with rutoside for melanoma A375 cancer cells: an in vitro study.

BACKGROUND: Melanoma as a type of skin cancer, is associated with a high mortality rate. Therefore, early diagnosis and efficient surgical treatment of this disease is very important. Photodynamic therapy (PDT) involves the activation of a photosensitizer by light at specific wavelength that interacts with oxygen and creates singlet oxygen molecules or reactive oxygen species (ROS), which can lead to tumor cell death. Furthermore, one of the main approches in the prevention and treatment of various cancers is plant compounds application. Phenolic compounds are essential class of natural antioxidants, which play crucial biological roles such as anticancer effects. It was previously suggested that flavonoid such as rutoside could acts as pro-oxidant or antioxidant. Hence, in this study, we aimed to investigate the effect of rutoside on the combination therapy with methylene blue (MB) assisted by photodynamic treatment (PDT) using red light source (660 nm; power density: 30 mW/cm2) on A375 human melanoma cancer cells. METHODS: For this purpose, the A375 human melanoma cancer cell lines were treated by MB-PDT and rutoside. Clonogenic cell survival, MTT assay, and cell death mechanisms were also determined after performing the treatment. Subsequently, after the rutoside treatment and photodynamic therapy (PDT), cell cycle and intracellular reactive oxygen species (ROS) generation were measured. RESULTS: The obtained results showed that, MB-PDT and rutoside had better cytotoxic and antiprolifrative effects on A375 melanoma cancer cells compared to each free drug, whereas the cytotoxic effect on HDF human dermal fibroblast cell was not significant. MB-PDT and rutoside combination induced apoptosis and cell cycle arrest in the human melanoma cancer cell line. Intracellular ROS increased in A375 cancer cell line after the treatment with MB-PDT and rutoside. CONCLUSION: The results suggest that, MB-PDT and rutoside could be considered as novel approaches as the combination treatment of melanoma cancer.

Effect of rutin as flavonoid compound on photodynamic inactivation against P. aeruginosa and S. aureus.

Antimicrobial photodynamic therapy (aPDT) has drawn increasing attention for its potential to effectively kill multidrug-resistant pathogenic bacteria and also for its low tendency to induce drug resistance. Antimicrobial photodynamic therapy (aPDT) is the application of photoactive dye followed by light irradiation that leads to the death of microbial cells mainly by reactive oxygen species (ROS) production in the presence of oxygen molecules. Methylene Blue (MB) as a photosensitizer is a hydrophobic drug molecule and prone to aggregation and dimer formation which lead to its low phototoxicity. Rutin, a flavonoid compound which is derived from plants such as wheat, apple, and tea has many properties such as antibacterial activity. In this study, we investigated the effect of rutin as a flavonoid compound on photodynamic inactivation by MB on Pseudomonas aeruginosa and Staphylococcus aureus. After performing the Minimum Inhibitory Concentration (MIC) assay (to measure minimum inhibitory concentration) and the MTT assay (to evaluate methylene blue toxicity), the effect of aPDT at 660 nm and pretreatment or post treatment with rutin on bacteria in the forms of planktonic and biofilm were investigated. The results showed that by a combination of rutin (800 µg/mL) with methylene blue (MB 8 µg/mL) as a photosensitizer and aPDT (660 nm, 5 min), there is a more reduction in the number of bacteria in the planktonic condition and bacterial biofilm production in comparison to MB alone. MB-aPDT showed no toxic effect against human dermal fibroblast with the proposed strategy which could suggest its application with rutin as a novel approach in the treatment of bacteria in wound infection.

Biological Responses of Stem Cells to Photobiomodulation Therapy.

BACKGROUND: Stem cells have attracted the researchers interest, due to their applications in regenerative medicine. Their self-renewal capacity for multipotent differentiation, and immunomodulatory properties make them unique to significantly contribute to tissue repair and regeneration applications. Recently, stem cells have shown increased proliferation when irradiated with low-level laser therapy or Photobiomodulation Therapy (PBMT), which induces the activation of intracellular and extracellular chromophores and the initiation of cellular signaling. The purpose of this study was to evaluate this phenomenon in the literature. METHODS: The literature investigated the articles written in English in four electronic databases of PubMed, Scopus, Google Scholar and Cochrane up to April 2019. Stem cell was searched by combining the search keyword of "low-level laser therapy" OR "low power laser therapy" OR "low-intensity laser therapy" OR "photobiomodulation therapy" OR "photo biostimulation therapy" OR "LED". In total, 46 articles were eligible for evaluation. RESULTS: Studies demonstrated that red to near-infrared light is absorbed by the mitochondrial respiratory chain. Mitochondria are significant sources of reactive oxygen species (ROS). Mitochondria play an important role in metabolism, energy generation, and are also involved in mediating the effects induced by PBMT. PBMT may result in the increased production of (ROS), nitric oxide (NO), adenosine triphosphate (ATP), and cyclic adenosine monophosphate (cAMP). These changes, in turn, initiate cell proliferation and induce the signal cascade effect. CONCLUSION: The findings of this review suggest that PBMT-based regenerative medicine could be a useful tool for future advances in tissue engineering and cell therapy.

Anti-cancer effect of gallic acid in presence of low level laser irradiation: ROS production and induction of apoptosis and ferroptosis.

BACKGROUND: There are different treatments for breast cancer and melanoma that mostly have some side effects. One of the therapeutic strategies is the use of natural components. Phenol components are a class of antioxidants in plants that have many biological functions like anticancer effects. Gallic acid (GA) is a natural polyhydroxy phenolic compound and commonly found in various foods. In the present study, GA effects alone and in combination with low-level laser irradiation on human dermal fibroblast cell line (HDF), human non-tumorigenic breast epithelial cell line (MCF10A), breast cancer cell line (MDA-MB-231) and melanoma cancer cell line (A375) was under the investigation. METHODS: The normal and cancerous cell lines were exposed to 660 nm low-level laser with 3 J/cm2 for 90 s. Then, the cells were treated with different concentrations of GA for 24 h. In another study, the cell lines firstly were treated with GA and then exposed to low-level laser irradiation. The effects of GA and low-level laser on cell survival and apoptosis were examined using MTT assay, light microscopy, ROS production assay, fluorescence microscopy (AO/EB double staining) and flow cytometry. RESULTS: The results showed that pre-treatment with low-level laser and then GA reduced the survival of breast cancer cells and melanoma more than the first treatment with GA and then low-level laser irradiation. Our findings showed that ROS production in cells treated with both low-level laser and GA was more than the cells treated with GA alone. The apoptosis and ferroptosis assays confirmed the MTT results which combination treatment with low-level laser and then GA increase the cell death probably via apoptosis and ferroptosis cell death mechanisms compared to GA alone. CONCLUSIONS: This study suggests that low-level laser irradiation alone is not able to cause death in human normal and cancerous cells. Preirradiation followed by GA treatment did not change the cell viability in human normal significantly but reduces the cell survival of cancer cells more than GA alone.

Antimicrobial photodynamic and wound healing activity of curcumin encapsulated in silica nanoparticles.

The multidrug resistance of pathogenic bacteria has become a serious problem to public health. Therefore finding novel approaches to combat multidrug resistant bacteria have therefore become increasingly important. One promising approach is antimicrobial photodynamic therapy (aPDT) which involves the use of photosensitizer (nontoxic dyes) that are excited by visible light and produce oxygen free radicals in the presence of oxygen. In this study, we investigated the antimicrobial photodynamic effect of free curcumin and curcumin silica nanoparticle as photosensitizer on planktonic and biofilm forms of Pseudomonas aeruginosa, Staphylococcus aureus. After performing MTT assay (to evaluate curcumin nano complex toxicity on human fibroblast cells), the effect of aPDT at 465 nm on bacteria in the planktonic and biofilm forms were investigated. The results showed that by using curcumin in the form of curcumin-silica nanoparticle as photosensitizer, there is a reduction in the number of bacteria in planktonic condition and bacterial biofilm production. Curcumin-silica nanoparticle not only did not show any significant cytotoxic effect against human normal fibroblast but also showed wound healing properties as confirmed with in vitro scratch assay. Therefore, aPDT based curcumin-silica nanoparticle could be suggested as novel approach in the treatment of multi drug resistant bacteria in chronic wound infection condition.

Low-level laser irradiation potentiates anticancer activity of p-coumaric acid against human malignant melanoma cells.

p-Coumaric acid (PCA) is a kind of phenolic compound, and as one of the cinnamic acid derivatives, it has many biological functions such as antioxidants, anti-inflammatory, antiplatelet, and anticancer activity. Low-level laser irradiation has received increasing interest in the fields of tissue regeneration and wound healing. In this study, the effect of low-level laser irradiation on human fibroblast cells (human dermal fibroblast) and human melanoma cancer cells (A375 and SK-MEL-37) treated with PCA was investigated. The human dermal fibroblast, A375, and SK-MEL-37 cells were exposed to low-level laser at 660-nm wavelength with 3 J/cm for 90 s, and then the cells were treated with different concentrations of PCA (0-1000 µg/ml for 24 h), separately. In another experiment, first the cells were treated by PCA and then irradiated with low-level laser as described before. The effect of various irradiation energy (1-6 J/cm) on the melanoma cells, which were then treated by PCA, was studied. The cell viability using MTT assay and lactate dehydrogenase assay was determined. Morphological changes owing to apoptosis induction by irradiation and PCA were detected by fluorescence microscopy using acridine orange/ethidium bromide double staining. The results showed that pretreatment with low-level laser irradiation and then PCA reduced the survival and growth of melanoma cells more than the early treatment with PCA and then low-level laser irradiation. Lactate dehydrogenase activity was reduced significantly by preirradiation and then PCA treatment in comparison with the dark group in melanoma cells. The cell cytotoxicity at different irradiation energy and then IC50 concentration of PCA was increased up to 3 J/cm and then decreased following increasing irradiation energy. The morphology study with light microscopy and apoptotic assay using acridine orange/ethidium bromide dual staining confirmed the MTT results. This study showed that low-level laser irradiation alone is not able to kill human normal fibroblast and human melanoma cancer cells. Preirradiation followed by treatment with PCA did not change the cell viability in human fibroblast significantly but reduced the cell viability in melanoma cells presumably through the apoptosis pathway.

Photodynamic effect of Zirconium phosphate biocompatible nano-bilayers containing methylene blue on cancer and normal cells.

Pharmaceutical applications of methylene blue, especially as photosensitizer, have been limited due to its rapid enzymatic reduction in the biological systems. In this study nano-platelet zirconium phosphate was synthesized and its biocompatibility was evaluated. The synthesized material was considered as drug delivery vehicle for methylene blue to enhance the photodynamic therapy efficacy in human breast cancer cells. Zirconium phosphate-methylene blue nano-hybrids were characterized by X-Ray Powder Diffraction (XRPD), Scanning Electron Microscopy (SEM), and Thermo gravimetric Analysis (TGA). Biocompatibility of synthesized nano materials were studied on Hu02 human fibroblast normal cell and MDA-MB-231 human breast cancer cell. The results clarified that ZrP-MB nanoparticles could decrease the dark toxicity of free methylene blue. Photodynamic therapy using zirconium phosphate-methylene blue on MDA-MB-231 human breast cancer was evaluated by MTT assay, colony forming ability assay, AO/EB dual staining and flow cytometry detection of apoptosis. The results suggest that zirconium phosphate-methylene blue nano-hybrids significantly enhance photodynamic therapy efficacy probably via apoptosis cell death mechanism against human breast cancer cells. According to the results, zirconium phosphate nanoparticles could be suggested as a promising nano-carrier for photosensitizer delivery in photodynamic therapy.

Biophysical study on the interaction of cartap hydrochloride and hemoglobin: Heme degradation and functional changes of protein.

Cartap hydrochloride is a mildly perilous insecticide known as "Padan" which is used largely in agricultural farms to control weevil and caterpillars. The over use of cartap causes harmful effects on human health. Since the blood may acts as a target and carrier for insecticides, the effect of these compounds on blood in mammalian toxicology is very important. Hemoglobin is a tetramer protein that play critical role in oxygen transport. The aim of this study was to analyze and compare the function and structural changes of hemoglobin in the presence of different concentrations of cartap by employing different spectroscopic techniques. The obtained results show that cartap has a high hemolytic effect which is increased with cartap concentration and reduces the thermal midpoint of hemoglobin. Fluorescence measurements reveal heme degradation at different concentrations of cartap. In consequence of theoretical and experimental results, cartap has an undesirable effect on hemoglobin structure and function.

Comparative study of photodynamic activity of methylene blue in the presence of salicylic acid and curcumin phenolic compounds on human breast cancer.

Curcumin and salicylic acid are both phenolic compounds and they can both affect cancer treatment efficacy. In this study, the effects of methylene blue-curcumin (CU-MB) and methylene blue-salicylic acid (SA-MB) ion pair complexes on MDA-MB-231 human breast cancer cells are studied. According to the thermodynamic parameters, the stability of curcumin and salicylic acid complexes ion pair complexes was compared. The free energy of ion pair interactions was calculated based on binding constants. A comparison of the free energies of the complexes (CU-MB: ∆G°b1 = - 21.11 kJ/mol and ∆G°b2 = - 8.37 kJ/mol, SA-MB: ∆G°b1 = - 12.92 kJ/mol and ∆G°b2 = - 9.02 kJ/mol) indicates that the interaction of methylene blue in first binding interaction with curcumin is greater than that of methylene blue with salicylic acid. Electrostatic interactions are the main forces in the binding of both compounds to methylene blue. All forces are inter-molecular physical interactions. The results of cellular experiments show that ion pairing has enhanced the reduction of cell viability. By increasing molecular stability and prevention of dimerization of methylene blue, the cell killing potential of methylene blue increases and it subsequently causes enhancement of photodynamic efficacy.

Photodynamic treatment with anionic nanoclays containing curcumin on human triple-negative breast cancer cells: Cellular and biochemical studies.

Photodynamic treatment is a minimally invasive and clinically approved procedure for eliminating selected malignant cells with activation of a photosensitizer agent at a specific light. Little is known, however, about the phototoxic properties of curcumin, as a natural phenolic compound, against different types of cancers. It is generally accepted that cellular damage occurs during photo treatment. There is a limitation in using of curcumin as a drug due to its low solubility, but nanoparticles such as anionic nanoclays or layered double hydroxide (LDH) could overcome it. The aim of this study was to investigate cellular responses to curcumin-LDH nanoparticles after photodynamic treatment of MDA-MB-231 human breast cancer cells. For this purpose, the MDA-MB-231 human breast cancer cell line treated with curcumin-LDH nanoparticle and then irradiated (photodynamic treatment). After irradiation, lactate dehydrogenase assay, clonogenic cell survival, cell death mechanisms such as autophagy and apoptosis were determined. Cell cycle distribution after photodynamic therapy (PDT) and also intracellular reactive oxygen species (ROS) generation were measured. The result showed that curcumin-LDH-PDT has a cytotoxic and antiprolifrative effect on MDA-MB-231 human breast cancer cells. Curcumin-LDH-PDT induced autophagy, apoptosis, and G0/G1 cell cycle arrest in human breast cancer cell line. Intracellular ROS increased in MDA-MB-231 cancer cell line after treatment with curcumin-LDH along with irradiation. The results suggest that curcumin-LDH nanoparticle could be considered as a novel approach in the photodynamic treatment of breast cancer.

Rational affinity enhancement of fragmented antibody by ligand-based affinity improvement approach.

Antibody engineering is now a noteworthy area in biopharmaceuticals as the next generation of marketed antibodies is engineered antibodies such as affinity- or stability-improved antibodies, fragmented or fused antibodies, antibody drug conjugates (ADCs), and PEGylated antibody fragments. In the current study, affinity enhancement of Nb against PlGF was performed by an in silico affinity maturation and molecular dynamics (MD) simulation. First, 300 single-point mutants were designed by identifying the residues involved in interaction with PlGF and different energy distributions. An energy based screening was performed to select best single-point mutants. Additionally, one variant containing two mutations was designed based on the selected single-point mutants. Finally, mutants-PlGF complexes were analyzed in details by all atom MD simulation. Trajectory analysis revealed that in both single (L112H, S31D, A97K, and R45E) and double (S31D & R45E) mutants, the free binding energies and the stability of complexes were significantly improved. The highest increment in affinity was observed for S31D mutant due to substantial increase in polar and electrostatic interactions. The secondary structure of Nb was intact in all variants and a shrinkage of PlGF over Nb was observed in all mutant-PlGF complexes during simulation. In addition, contact area and hydrogen-bond analysis as well as distance measurement in mutants-PlGF complexes also confirmed the affinity enhancement of variants relative to the native form. Our study showed that ligand-based affinity improvement could be considered as a promising approach for designing high affinity fragmented antibodies.