Curcumin Sustained Release with a Hybrid Chitosan-Silk Fibroin Nanofiber Containing Silver Nanoparticles as a Novel Highly Efficient Antibacterial Wound Dressing.

Drug loading in electrospun nanofibers has gained a lot of attention as a novel method for direct drug release in an injury site to accelerate wound healing. The present study deals with the fabrication of silk fibroin (SF)-chitosan (CS)-silver (Ag)-curcumin (CUR) nanofibers using the electrospinning method, which facilitates the pH-responsive release of CUR, accelerates wound healing, and improves mechanical properties. Response surface methodology (RSM) was used to investigate the effect of the solution parameters on the nanofiber diameter and morphology. The nanofibers were characterized via Fourier Transform Infrared Spectroscopy (FTIR), X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), zeta potential, and Dynamic Light Scattering (DLS). CS concentration plays a crucial role in the physical and mechanical properties of the nanofibers. Drug loading and entrapment efficiencies improved from 13 to 44% and 43 to 82%, respectively, after the incorporation of Ag nanoparticles. The application of CS hydrogel enabled a pH-responsive release of CUR under acid conditions. The Minimum Inhibitory Concentration (MIC) assay on E. coli and S. aureus bacteria showed that nanofibers with lower CS concentration cause stronger inhibitory effects on bacterial growth. The nanofibers do not have any toxic effect on cell culture, as revealed by in vitro wound healing test on NIH 3T3 fibroblasts.

Design and development of a self-assembling protein nanoparticle displaying PfHAP2 antigenic determinants recognized by natural acquired antibodies.

BACKGROUNDS: In order to move towards the elimination and eradication of malaria in the world, the development of vaccines is inevitable. Many modern vaccines are based on recombinant technology; however, they may not provide a fully protective, long-lasting immune response. One of the strategies to improve recombinant vaccines is designing the nanovaccines such as self-assembling protein nanoparticles (SAPNs). Hence, the presentation of epitopes in a repeat array and correct conformation should be considered. P. falciparum generative cell-specific 1 (PfGCS1) is a main transmission-blocking vaccine candidate with two highly conserved fragments, HAP2-GCS1 and cd loop, inducing partial malaria transmission inhibitory antibodies. Therefore, to design an effective malaria vaccine, we used cd loop and HAP2-GCS1 fragments at the amino and carboxy terminuses of the SAPN-forming amino acid sequence, respectively. METHODOLOGY/PRINCIPAL FINDINGS: The SAPN monomer (PfGCS1-SAPN) sequence was designed, and the three-dimensional (3D) structure was predicted. The result of this prediction ensured the presence of antigens on the SAPN surface. Then the accuracy of the predicted 3D structure and its stability were confirmed by 100 ns molecular dynamics (MD) simulation. The designed SAPN substructure sequence was synthesized, cloned, and expressed in Escherichia coli. With a gradual decrease in urea concentration in dialysis solutions, the purified proteins progressed to the final desired structure of the SAPN, which then was confirmed by Dynamic Light Scattering (DLS) and Field Emission Scanning Electron Microscopy (FESEM) tests. According to the Enzyme-Linked Immunosorbent Assay (ELISA), antigenic determinants were presented on the SAPN surface and interacted with antibodies in the serum of malaria patients. CONCLUSIONS/SIGNIFICANCE: Our results show that the SAPN formed by PfGCS1-SAPN has produced the correct shape and size, and the antigenic determinants are presented on the surface of the SAPN, which indicates that the designed SAPN has great potential to be used in the future as a malaria vaccine.

Unraveling Cancer Metastatic Cascade Using Microfluidics-based Technologies.

Cancer has long been a leading cause of death. The primary tumor, however, is not the main cause of death in more than 90% of cases. It is the complex process of metastasis that makes cancer deadly. The invasion metastasis cascade is the multi-step biological process of cancer cell dissemination to distant organ sites and adaptation to the new microenvironment site. Unraveling the metastasis process can provide great insight into cancer death prevention or even treatment. Microfluidics is a promising platform, that provides a wide range of applications in metastasis-related investigations. Cell culture microfluidic technologies for in vitro modeling of cancer tissues with fluid flow and the presence of mechanical factors have led to the organ-on-a-chip platforms. Moreover, microfluidic systems have also been exploited for capturing and characterization of circulating tumor cells (CTCs) that provide crucial information on the metastatic behavior of a tumor. We present a comprehensive review of the recent developments in the application of microfluidics-based systems for analysis and understanding of the metastasis cascade from a wider perspective.

A new insight to deformability correlation of circulating tumor cells with metastatic behavior by application of a new deformability-based microfluidic chip.

Isolation and characterization of circulating tumor cells (CTCs) found in blood samples of cancer patients have been considered as a reliable source for cancer prognosis and diagnosis. A new continuous microfluidic platform has been designed in this investigation for simultaneous capture and characterization of CTCs based on their deformability. The deformability-based chip (D-Chip) consists of two sections of separation and characterization where slanted weirs with a gap of 7 µm were considered. Although sometimes CTCs and leukocytes have the same size, the deformability differs in such a way that can be exploited for enrichment purposes. MCF7 and MDA-MB-231 cell lines were used for the initial evaluation of the D-Chip performance. In the separation section, cancer cells were isolated based on deformability differences with an efficiency of higher than 93% (∼average capturing capacity of 2085 out of 2200 cancer cells ml-1) and with significantly high purity (15-40 WBCs ml-1; ∼5 log depletion of WBCs). Cancer cells were categorized based on the deformability difference in the characterization section. Subsequently, 15 clinical blood samples from breast cancer patients were analyzed by the D-Chip. Suggest 'The chip detected CTCs in all patient samples, processed the blood sample at a high throughput of 5.3 ml/h, and properly categorized CTCs based on deformability differences. Further characterization showed that the highly deformable breast cancer CTCs in our patient samples also showed higher potential of metastasis in support of a broader correlation between deformability of CTCs and metastatic behavior.

Potential effects of alginate-pectin biocomposite on the release of folic acid and their physicochemical characteristics.

Potential effects of folates on the treatment of several human diseases like cognitive function, neural tube defects, coronary heart disease and certain kinds of cancers have been discovered. However, the stability of folic acid against adverse conditions is a great concern. The present study investigates various alginate (A)-pectin (P) gastrointestinal-resistant hydrogel to immobilize folic acid. This involves evaluating different compositions of alginate-pectin to achieve higher encapsulation efficiency and stability during simulated gastric (SG) and simulated intestinal (SI) conditions. Coated alginate hydrogels with pectin resulted significant (p < 0.05) better protection of folic acid compared to non-coated alginate hydrogel when exposed to SG condition and when exposed to SI condition, sustained release behavior obtained with the ratio of A70-P30. The structural and physicochemical properties of blended A-P hydrogel were characterized using scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray diffractometer, indicating the presence of folic acid into the matrix and signified no covalent reaction between components. Therefore, this adequate composition of alginate-pectin showed to be a potential carrier for folic acid stability.

A porous hydrogel-electrospun composite scaffold made of oxidized alginate/gelatin/silk fibroin for tissue engineering application.

In the article, a bilayer nanocomposite scaffold made of oxidized alginate (OAL), gelatin (G), and silk fibroin (SF) has been prepared via combining electrospinning, in situ gas foaming, in situ crosslinking and freeze drying methods. The physicochemical and mechanical properties, as well as thermal stability of the proposed composite, have been investigated by SEM, FTIR, XRD, tensile, and TGA analysis. The data indicate that structure and degree of crosslinking play a vital role in adjusting the physical and mechanical properties of composite scaffolds. Further, the authors find a favorable adipose-derived mesenchymal stem cell's (AMSC) attachment and distribution within this novel hydrogel-electrospun composite. Such a nanocomposite structure with its promising properties and cell-material interaction may be considered as a new scaffold for different tissue engineering applications.

Ag-doped magnetic metal organic framework as a novel nanostructured material for highly efficient antibacterial activity.

In the last decades, numerous attempts have been made to prevent microbial pollution spreading, using antibacterial agents. Zeolitic imidazolate framework-8 (ZIF-8) belongs to a subgroup of metal organic frameworks (MOFs) merits of attention due to the zinc ion clusters and its effective antibacterial activity. In this work, Ag-doped magnetic microporous γ-Fe2O3@SiO2@ZIF-8-Ag (FSZ-Ag) was successfully synthesized by a facile methodology in room temperature and used as an antibacterial agent against the growth of the Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus bacteria. Several characterization methods were applied to analyze the properties of the materials, and the results confirmed the accuracy of the synthesis procedure. Silver ions have employed to enhance the efficiency of antibacterial activity. As the results illustrated, FSZ-Ag nanostructured material had superior performance to inactive E. coli and S. aureus in growth inhibition test in liquid media. The best antibacterial activity as minimum inhibitory concentration (MIC) was 100 mg/L of FSZ-Ag against both bacteria. Leaching rates of silver ions showed that 80% of Ag released in the solutions, which was responsible for inhibiting the growth of bacteria. Also, fluorescence microscopy was used to investigate bacterial viability after 20 h contacting FSZ-Ag to distinguish live and dead bacteria by staining with DAPI and PI fluorescence stains. This novel magnetic nanostructured material is an excellent promising candidate to use in biological applications as high potential bactericidal materials.

Examination of chondroitinase ABC I immobilization onto dextran-coated Fe3O4 nanoparticles and its in-vitro release.

Chondroitinase ABC I (cABC I) has received notable attention in treatment of spinal cord injuries and its application as therapeutics has been limited due to low thermal stability at physiological temperature. In this study, cABC I enzyme was immobilized on the dextran-coated Fe3O4 nanoparticles through physical adsorption to improve the thermal stability. The nanoparticles were characterized using XRD, SEM, VSM, and FTIR analyses. Response surface methodology and central composite design were employed to assess factors affecting the activity of immobilized cABC I. Experimental results showed that pH 6.3, temperature 24 °C, enzyme/support mass ratio 1.27, and incubation time 5.7 h were the optimal immobilization conditions. It was found that thermal stability of immobilized cABC I was significantly improved. In-vitro cABC I release was studied under pH 7.5 and temperature 37 °C and the results indicated that 70 % release occurred after 9 h and the release mechanism was first-order kinetic model.

Optimization of Alginate-Whey Protein Isolate Microcapsules for Survivability and Release Behavior of Probiotic Bacteria.

The present study aimed to improve the survivability of L. acidophilus encapsulated in alginate-whey protein isolate (AL-WPI) biocomposite under simulated gastric juice (SGJ) and simulated intestinal juice (SIJ). Microcapsules were prepared based on emulsification/internal gelation technique. Optimal compositions of AL and WPI and their ratio in the aqueous phase were evaluated based on minimizing mean diameter (MD) of the microcapsules and maximizing encapsulation efficiency (EE), survivability of cells under SGJ (Viability), and release of viable cells under SIJ (Release) using Box-Behnken experimental design. Optimal composition comprising 4.54% (w/v) AL, 10% (w/v) WPI, and 10% (v/v) AL-WPI gum in the aqueous phase was determined statistically. Physicochemical characteristics of the optimized matrix were investigated by SEM, FTIR, and XRD analysis to determine surface morphology, molecular bonds, and crystalline nature of such hydrocolloid. It could be concluded that the proposed biocomposite is a good promise for nutrients encapsulation in the food industry.

Magnetite nanoparticle as a support for stabilization of chondroitinase ABCI.

Chondroitinase ABCI (cABCI) is a drug enzyme that can be used to treat spinal cord injuries. Due to low thermal stability of cABCI, this enzyme was immobilized on Fe3O4 nanoparticle to increase its thermal stability. The size and morphology, structure and magnetic property of the Fe3O4 nanoparticles were characterized by the analyses of SEM, XRD and VSM, respectively, and FTIR spectroscopy was employed to confirm the immobilization of cABCI on the surface of Fe3O4 nanoparticles. The results indicated that the optimum conditions for pH, temperature, cABCI-to-Fe3O4 mass ratio and incubation time in immobilization process were 6.5, 15 °C, 0.75 and 4.5 h, respectively, and about 0.037 mg cABCI was bound to 1 mg of Fe3O4 nanoparticles at these conditions. The value of Vmax was the same for free and immobilized cABCI, but Km value for immobilized cABCI was 1.6 times higher than that for free one. The storage stability of immobilized cABCI was significantly enhanced at low temperatures, e.g. free cABCI retained 19% of its activity after six days at -20 °C, while the immobilized one retained 96% of its activity. In vitro release of cABCI from Fe3O4 particles showed that about 94% of the enzyme was released after 6 h.

Fabrication of hierarchically porous silk fibroin-bioactive glass composite scaffold via indirect 3D printing: Effect of particle size on physico-mechanical properties and in vitro cellular behavior.

In order to regenerate bone defects, bioactive hierarchically scaffolds play a key role due to their multilevel porous structure, high surface area, enhanced nutrient transport and diffusion. In this study, novel hierarchically porous silk fibroin (SF) and silk fibroin-bioactive glass (SF-BG) composite were fabricated with controlled architecture and interconnected structure, by combining indirect three-dimensional (3D) inkjet printing and freeze-drying methods. Further, the effect of 45S5 Bioactive glass particles of different sizes (<100 nm and 6 µm) on mechanical strength and cell behavior was investigated. The results demonstrated that the hierarchical structure in this scaffold was composed of two levels of pores in the order of 500-600 µm and 10-50 µm. The prepared SF-BG composite scaffolds utilized by nano and micro particles possessed mechanical properties with a compressive strength of 0.94 and 1.2 MPa, respectively, in dry conditions. In a wet condition, the hierarchically porous scaffolds did not exhibit any fluctuation after compression load cell and were incredibly flexible, with excellent mechanical stability. The SF-BG composite scaffold with nanoparticles presented a significant 50% increase in attachment of human bone marrow stem cells in comparison with SF and SF-BG scaffold with microparticles. Moreover, SF-BG scaffolds promoted alkaline phosphatase activity as compared to SF scaffolds without BG particles on day 14. In brief, the 3D porous silk fibroin-based composites containing BG nanoparticles with excellent mechanical properties are promising scaffolds for bone tissue regeneration in high load-bearing applications.

Design and Fabrication of a Novel Microfluidic System for Enrichment of Circulating Tumor Cells with the Assistance of Computer Simulations.

BACKGROUND: Cancer is the first cause of death in developed countries. The heterogeneous nature of cancer requires patient-specified treatment plans. One reliable approach is collecting Circulating Tumour Cells (CTCs) and using them for prognosis and drug response assessment purposes. CTCs are rare and their separation from normal cell requires high-accuracy methods. METHODS: A microfluidic cell capture device to separate CTCs from peripheral blood is presented in this study. The CTC separation device applies hydrodynamic forces to categorize cells according to their sizes. The proposed device is designed and evaluated by numerical simulations and validated experimentally. The simulation modified design was fabricated by soft lithography which allows prototyping the device in a few hours. For experimental setup two solutions: 1) fixed cells spiked in Phosphate Buffered Saline (PBS), and 2) fixed cells in blood were used. The CTC separation device was validated by tracking the flow and separation of cancer cell lines in the solutions. RESULTS: It is demonstrated that the setup is capable of CTC enrichment up to 50 times. CONCLUSION: The presented CTC enrichment method reduces costs by eliminating the use of antibodies. The high-throughput method has the potential to be used in preclinical studies of cancer.

Co-microencapsulation of Lactobacillus plantarum and DHA fatty acid in alginate-pectin-gelatin biocomposites.

The aim of this study was to investigate the co-microencapsulation of Lactobacillus plantarum and DHA-rich oil in a novel gastrointestinal-resistant biocomposite composed of alginate, pectin and gelatin. The optimal biocomposite consisted of 1.06% alginate, 0.55% pectin and 0.39% gelatin showed 88.66% survivability of the microencapsulated cells compared to the free cells (50.36%). In addition, co-microencapsule containing probiotic and DHA fatty acid was synthesized and physicochemically analyzed using SEM, FTIR, TGA, XRD. The results from SEM clearly confirmed that cells were completely entrapped in the matrix and DHA increased smoothness and compactness of the surface of the particles. FTIR spectra revealed the formation of hydrogen and Van der Waals bonds between macromolecules and the core materials. X-ray pattern of co-microencapsules identified amorphous structure compared to capsules containing only DHA or probiotic. TGA analysis revealed the thermal stability of DHA-loaded capsules compared to un-loaded ones.

A Biomimetic Heparinized Composite Silk-Based Vascular Scaffold with sustained Antithrombogenicity.

Autologous grafts, as the gold standard for vascular bypass procedures, associated with several problems that limit their usability, so tissue engineered vessels have been the subject of an increasing number of works. Nevertheless, gathering all of the desired characteristics of vascular scaffolds in the same construct has been a big challenge for scientists. Herein, a composite silk-based vascular scaffold (CSVS) was proposed to consider all the mechanical, structural and biological requirements of a small-diameter vascular scaffold. The scaffold's lumen composed of braided silk fiber-reinforced silk fibroin (SF) sponge covalently heparinized (H-CSVS) using Hydroxy-Iron Complexes (HICs) as linkers. The highly porous SF external layer with pores above 60 µm was obtained by lyophilization. Silk fibers were fully embedded in scaffold's wall with no delamination. The H-CSVS exhibited much higher burst pressure and suture retention strength than native vessels while comparable elastic modulus and compliance. H-CSVSs presented milder hemolysis in vitro and significant calcification resistance in subcutaneous implantation compared to non-heparinized ones. The in vitro antithrombogenic activity was sustained for over 12 weeks. The cytocompatibility was approved using endothelial cells (ECs) and vascular smooth muscle cells (SMCs) in vitro. Therefore, H-CSVS demonstrates a promising candidate for engineering of small-diameter vessels.

Synthesis of amine-modified zeolitic imidazolate framework-8, ultrasound-assisted dye removal and modeling.

The present research is focused on the ultrasound assisted adsorption of Acid blue 92 (AB92) and Direct red 80 (DR80) as anionic dyes in single and binary systems onto zeolitic imidazolate framework (ZIF-8) functionalized with 3-Aminopropyltrimethoxysilane (APTES). Different techniques such as Fourier transform infrared (FTIR), scanning electron microscope (SEM), field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) and thermogravimetric analyses (TGA) were used to characterize the prepared adsorbent. The individual effects and possible interactions between the various parameters including adsorbent dosage, sonication time, initial dye concentrations and pH on dyes removal efficiency were investigated by response surface methodology (RSM). The optimized experimental conditions were fixed at adsorbent dosage 0.005g for AB92 and 0.01g for DR80, pH 2.1, sonication time 15min, and initial dyes concentration 15mgL-1 to get maximum removal percentage (>95.0%). A reliable and intelligent model based on least-squares support vector machine (LS-SVM) was developed to predict dye removal efficiency. The root mean square error (RMSE) of 0.604, 0.734 and 1.549 with high determination coefficient (R2) of 0.999, 0.996 and 0.997 for AB92, DR80 and binary system, respectively, were able to predict and model the adsorption process. The presented model illustrates better performance in predicting dye removal efficiency compared to the kinetic models. The results showed that the adsorption process had better conformation with pseudo-second order model. The adsorption equilibrium data was investigated by Langmuir, Freundlich, Tempkin and Dubinin-Radushkevich isotherm models and the data were well fitted by Langmuir model with maximum adsorption capacity of 633.4 and 500.2mgg-1 for AB92 and DR80 dyes, respectively. APTES@ZIF-8 was regenerated and found to be reusable after four successive cycles without considerable loss in adsorption capacity.

Evaluation of trichloroethylene degradation by starch supported Fe/Ni nanoparticles via response surface methodology.

In this study, degradation of trichloroethylene (TCE), a chlorinated hydrocarbon, using starch supported Fe/Ni nanoparticles was investigated. The scanning electron microscope images showed applying water soluble starch as a stabilizer for the Fe/Ni nanoparticles tended to reduce agglomeration and discrete particle. Also the mean particle diameter reduced from about 70 nm (unsupported Fe/Ni nanoparticle) to about 30 nm. Effects of three key independent operating parameters including initial TCE concentration (10.0-300.0 mg L(-1)), initial pH (4.00-10.00) and Fe(0) dosage (0.10-2.00) g L(-1) on TCE dechlorination efficiency in 1 hour were analysed by employing response surface methodology (RSM). Based on a five-level three-factor central composite design, TCE removal efficiency was examined and optimized. The obtained RSM model fitted the experimental data to a second order polynomial equation. The optimum dechlorination conditions at initial TCE concentration 100.0 mg L(-1) were initial pH 5.77, Fe(0) dosage 1.67 g L(-1). At these conditions TCE removal concentration reached 94.87%, which is in close acceptance with predicted value by the RSM model.

Optimization of operating parameters for efficient photocatalytic inactivation of Escherichia coli based on a statistical design of experiments.

In this work, the individual and interaction effects of three key operating parameters of the photocatalytic disinfection process were evaluated and optimized using response surface methodology (RSM) for the first time. The chosen operating parameters were: reaction temperature, initial pH of the reaction mixture and TiO2 P-25 photocatalyst loading. Escherichia coli concentration, after 90 minutes irradiation of UV-A light, was selected as the response. Twenty sets of photocatalytic disinfection experiments were conducted by adjusting operating parameters at five levels using the central composite design. Based on the experimental data, a semi-empirical expression was established and applied to predict the response. Analysis of variance revealed a strong correlation between predicted and experimental values of the response. The optimum values of the reaction temperature, initial pH of the reaction mixture and photocatalyst loading were found to be 40.3 °C, 5.9 g/L, and 1.0 g/L, respectively. Under the optimized conditions, E. coli concentration was observed to reduce from 10(7) to about 11 CFU/mL during the photocatalytic process. Moreover, all these results showed the great significance of the RSM in developing high performance processes for photocatalytic water disinfection.

Biodegradability of oily wastewater using rotating biological contactor combined with an external membrane.

BACKGROUND: A novel implementation of a hybrid membrane bioreactor (HMBR) has been studied in this paper. It is utilized as combination of rotating biological contractor (RBC) and an external membrane, as a new biological system for oily wastewater treatment. METHODS: Chemical oxygen demand (COD) and total petroleum hydrocarbon (TPH) as factors of Biodegradability has been evaluated. They are both compared together for different hydraulic retention times (HRTs) and petroleum pollution concentrations in RBC and HMBR. The ratio of TPH to COD of Molasses has been varied between 0.2 to 0.8 at two HRTs of 18 and 24 hours while the temperature, pH and dissolved oxygen were kept in the range of 20-25°C, 6.5-7.5, and 2-3.5 mg/l, respectively. RESULTS: The best TPH removal efficiency (99%) was observed in TPH/COD = 0.6 and HRT = 24 hr in HMBR and Removal efficiency was decreased in the ratios above 0.6 in both bioreactors. CONCLUSIONS: The experimental results showed that HMBR had higher treatment efficiency than RBC at all ratios and HRTs.

An efficient approach to cathode operational parameters optimization for microbial fuel cell using response surface methodology.

BACKGROUND: In the recent study, optimum operational conditions of cathode compartment of microbial fuel cell were determined by using Response Surface Methodology (RSM) with a central composite design to maximize power density and COD removal. METHODS: The interactive effects of parameters such as, pH, buffer concentration and ionic strength on power density and COD removal were evaluated in two-chamber microbial batch-mode fuel cell. RESULTS: Power density and COD removal for optimal conditions (pH of 6.75, buffer concentration of 0.177 M and ionic strength of cathode chamber of 4.69 mM) improve by 17 and 5%, respectively, in comparison with normal conditions (pH of 7, buffer concentration of 0.1 M and ionic strength of 2.5 mM). CONCLUSIONS: In conclusion, results verify that response surface methodology could successfully determine cathode chamber optimum operational conditions.

Nano reengineering of horseradish peroxidase with dendritic macromolecules for stability enhancement.

A simple bio-conjugation procedure to surround a single horseradish peroxidase (HRP) enzyme molecule with dendritic polyester macromolecules (polyester-32-hydroxyl-1-carboxyl bis-MPA dendron, generation 5) was proposed. The characterization of resultant nanoparticles entitled HRP dendrozyme, was performed by transmission electron microscopy, dynamic light scattering, gel permeation chromatography and Fourier transform infrared spectroscopy. The results showed that HRP nanoparticles were spherical in shape and have an average size of 14±2 nm in diameter. Furthermore, bio-conformational characterization of HRP dendrozyme was performed by means of circular dichroism and fluorescence spectroscopy to evaluate the secondary and tertiary structure changes after enzyme modification. These investigations revealed that protein conformation had small changes (in secondary and tertiary structures) after bio-conjugation. We also reported here that dendritic modification did not significantly affect the kinetic parameters of free HRP. The stabilization of HRP with dendron macromolecules as single enzyme nanoparticles resulted in improvement of half-life over 70 days storage at 4 °C as well as its tolerance under different elevated temperatures up to 80 °C and in the presence of organic solvents for 15 min. These significant results promise extensive applications of HRP particularly in harsh environmental conditions.