Highly resilient, biocompatible, and antibacterial carbon nanotube/hydroxybutyl chitosan sponge dressing for rapid and effective hemostasis.

Uncontrolled hemorrhage is the leading cause of trauma death. The development of safe and efficient hemostatic agents that can rapidly and effectively control bleeding is of great significance to rescue the injured. However, the mechanical, absorptive, and antibacterial properties of conventional two-dimensional hemostatic agents are not satisfactory. Herein, a series of effective three-dimensional hemostatic dressings (JWCNT/HBC sponges) are developed by chemical modification of joint-welded carbon nanotube (JWCNT) sponges with hydroxybutyl chitosan (HBC) for hemorrhage hemostasis. The JWCNT/HBC sponges exhibit high elasticity, porous structure, and suitable blood-absorption and blood-maintaining performance. Moreover, the introduction of HBC endows the JWCNT/HBC sponges with favorable blood compatibility and good antibacterial activity. The sponge treated with 0.5% HBC (JWCNT/0.5%HBC sponge) displays better antiseptic capability, faster blood clotting ability in vitro and shorter hemostasis time in vivo than the commercial gelatin sponge. The JWCNT/HBC sponges combine the advantages of JWCNT sponges and HBC in the adhesion and activation of platelets and red blood cells, thus becoming a good medical material for trauma hemostasis.

Chitosan Augments Tetramycin against Soft Rot in Kiwifruit and Enhances Its Improvement for Kiwifruit Growth, Quality and Aroma.

In this study, the co-application of chitosan and tetramycin against kiwifruit soft rot and its effects on the disease resistance, growth, quality and aroma of kiwifruit were investigated. The results show that chitosan could effectively enhance tetramycin against soft rot of kiwifruit with the field control efficacy of 85.33% for spraying chitosan 100 time + 0.3% tetramycin AS 5000-time dilution liquid, which was higher than 80.99% for 0.3% tetramycin AS 5000-time dilution liquid and significantly (p < 0.01) higher than 40.66% for chitosan 100-time dilution liquid. Chitosan could significantly (p < 0.05) improve the promoting effects of tetramycin on total phenolics, total flavonoids, SOD activity of kiwifruit compared to tetramycin during storage for 0-28 days and enhance the disease resistance of kiwifruit. Moreover, the co-application of chitosan and tetramycin was more effective than tetramycin or chitosan alone in enhancing fruit growth, improving fruit quality and increasing fruit aroma. This study highlights that chitosan can be used as an adjuvant to enhance tetramycin against soft rot of kiwifruit and promote tetramycin's improvement for the single fruit volume and weight, vitamin C, soluble sugar, soluble solid, dry matter, soluble protein, titratable acidity and aroma of kiwifruit.

Chitosan-based nanoparticle co-delivery of docetaxel and curcumin ameliorates anti-tumor chemoimmunotherapy in lung cancer.

The application of traditional chemotherapy drugs for lung cancer has obvious limitations, such as toxic side effects, uncontrolled drug-release, poor bioavailability, and drug-resistance. Thus, to address the limitations of free drugs and improve treatment effects, we developed novel T7 peptide-modified nanoparticles (T7-CMCS-BAPE, CBT) based on carboxymethyl chitosan (CMCS), which is capable of targeted binding to the transferrin receptor (TfR) expressed on lung cancer cells and precisely regulating drug-release according to the pH value and reactive oxygen species (ROS) level. The results showed that the drug-loading content of docetaxel (DTX) and curcumin (CUR) was approximately 7.82% and 6.48%, respectively. Good biosafety was obtained even when the concentration was as high as 500 µg/mL. More importantly, the T7-CMCS-BAPE-DTX/CUR (CBT-DC) complexes exhibited better in vitro and in vivo anti-tumor effects than DTX monotherapy and other nanocarriers loaded with DTX and CUR alone. Furthermore, we determined that CBT-DC can ameliorate the immunosuppressive micro-environment to promote the inhibition of tumor growth. Collectively, the current findings help lay the foundation for combinatorial lung cancer treatment.

Fabrication of self-healing pectin/chitosan hybrid hydrogel via Diels-Alder reactions for drug delivery with high swelling property, pH-responsiveness, and cytocompatibility.

Self-healing hydrogels with pH-responsiveness could protect loaded drugs from being destroyed till it arrives to the target. The pectin-based hydrogel is a candidate due to the health benefit, anti-inflammation, antineoplastic activity, nontoxicity, and biospecific degradation, et al. However, the abundant existence of water-soluble branched heteropolysaccharide chains influenced its performance resulting in limitation of the potential. In the present study, we prepared a series of self-healing pectin/chitosan hydrogels via the Diels-Alder reaction. Moreover, pectin/chitosan composite hydrogel was prepared as a contrast. By comparison, it can be seen that the Diels-Alder reaction greatly improved the cross-linking density of hydrogels. The self-healing experiments showed excellent self-healing performance. In different swelling mediums, significant transformation in the swelling ratio was shown, indicating well-swelling property, pH- and thermo-responsiveness. The drug loading and release studies presented high loading efficiency and sustained release performance. The cytotoxicity assay that showed a high cell proliferation ratio manifested great cytocompatibility.

Biocidal chitosan-magnesium oxide nanoparticles via a green precipitation process.

In the present scenario, the development of eco-friendly multifunctional biocidal substances with low cost and high efficiency, has become the center of focus. This study is, focused on the synthesis of magnesium oxide (MgO) and chitosan-modified magnesium oxide (CMgO) nanoparticles (NPs), via a green precipitation process. In this process, leaves extract of Plumbago zeylanica L was, used as a nucleating agent. The MgO and CMgO NPs exhibit face-centered cubic structures, as confirmed by XRD studies. Morphologically, the FESEM and TEM images showed that the MgO and CMgO NPs were spherical, with an average particle size of ~40±2 and ~37±2 nm, respectively. EDX spectra were used to identify the elemental compositions of the nanoparticles. By using FTIR spectra, the Mg-O stretching frequency of MgO and CMgO NPs were observed at 431 and 435 cm-1, respectively. The photoluminescence (PL) spectra of MgO and CMgO NPs, revealed oxygen vacancies at 499 nm and 519 nm, respectively, due to the active radicals generated, which were responsible for their biocidal activities. The toxicity effects of the nanoparticles developed, on cell viability (antibacterial and anticancer), were measured on the MCF-7 cell line and six different types of gram-negative bacteria. The antibacterial activities of the nanoparticles on: Klebsiella pneumoniae, Escherichia coli, Shigella dysenteriae, Pseudomonas aeruginosa, Proteus vulgaris and Vibrio cholerae bacteria, were studied with the well diffusion method. The MgO and CMgO NPs were tested on breast cancer cell line (MCF-7) via an MTT assay and it proved that CMgO NPs possess higher anticancer properties than MgO NPs. Overall, CMgO NPs showed a higher amount of cytotoxicity for both the bacterial and cancer cells when compared to the MgO NPs. Toxicity studies of fibroblast L929 cells revealed that the CMgO NPs were less harmful to the healthy cells when compared to the MgO NPs. These results suggest that biopolymer chitosan-modified MgO NPs can be used for healthcare industrial applications in order to improve human health conditions.

Inherent Safety Assessment of Industrial-Scale Production of Chitosan Microbeads Modified with TiO2 Nanoparticles.

In this study, the inherent safety analysis of large-scale production of chitosan microbeads modified with TiO2 nanoparticles was developed using the Inherent Safety Index (ISI) methodology. This topology was structured based on two main stages: (i) Green-based synthesis of TiO2 nanoparticles based on lemongrass oil extraction and titanium isopropoxide (TTIP) hydrolysis, and (ii) Chitosan gelation and modification with nanoparticles. Stage (i) is divided into two subprocesses for accomplishing TiO2 synthesis, lemongrass oil extraction and TiO2 production. The plant was designed to produce 2033 t/year of chitosan microbeads, taking crude chitosan, lemongrass, and TTIP as the primary raw materials. The process was evaluated through the ISI methodology to identify improvement opportunity areas based on a diagnosis of process risks. This work used industrial-scale process inventory data of the analyzed production process from mass and energy balances and the process operating conditions. The ISI method comprises the Chemical Inherent Safety Index (CSI) and Process Inherent Safety Index (PSI) to assess a whole chemical process from a holistic perspective, and for this process, it reflected a global score of 28. Specifically, CSI and PSI delivered scores of 16 and 12, respectively. The analysis showed that the most significant risks are related to TTIP handling and its physical-chemical properties due to its toxicity and flammability. Insights about this process's safety performance were obtained, indicating higher risks than those from recommended standards.

Antibacterial and Hemostatic Thiol-Modified Chitosan-Immobilized AgNPs Composite Sponges.

Wound bleeding and infection are two of the major threats to patients' lives, but developing safe materials with high hemostasis efficiency and antibacterial activity remains a major challenge. Silver nanoparticles (AgNPs) are suitable as antibacterial agents in the hemostatic process, but the application is hampered because of easy accumulation of toxicity. Herein, thiol-modified chitosan (TMC) was prepared by modifying with mercaptosuccinic acid and then was used to immobilize AgNPs to obtain composite sponges (TMC/AgNPs) for stemming the bleeding and preventing infection. TMC/AgNPs sponges had complex interlaced tubular porous structure with high porosity (99.42%), indicating high absorption. TMC had high immobilization efficiency for AgNPs-the release rate of AgNPs was 14.35% after 14 days-but the TMC/AgNPs sponge still had excellent antibacterial activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. In vitro and in vivo experiments confirm that the TMC/AgNPs sponge had fast and efficient hemostatic performance in comparison with the PVF sponge, and its possible mechanism was the synergistic effect of high blood absorption capacity and the interaction between amino, sulfydryl, and blood cells. Furthermore, the TMC/AgNPs sponge can promote wound healing by preventing wound infection, while the PVF sponge cannot. More importantly, the sponges had good safety due to the immobilization of TMC for AgNPs.

Quaternized Chitosan-Coated Montmorillonite Interior Antimicrobial Metal-Antibiotic in Situ Coordination Complexation for Mixed Infections of Wounds.

Conventional drug delivery systems for natural clay materials still face critical challenges in their practical application, including multiple bacterial infections, combined infection of bacteria and fungi, and low sterilization efficiency. In this work, we address these challenges using the multifunctional montmorillonite nanosheet-based (MMT-based) drug nanoplatform, which involves the antibiotic 5-fluorocytosine (5-FC), antibacterial metal copper ions, and quaternized chitosan (QCS). Composite material QCS/MMT/5-FCCu can can strongly inhibit Staphylococcus aureus (a typical Gram-positive bacterium), Escherichia coli (a typical Gram-negative bacterium), and Candida albicans (a fungus) because 5-FC coordinates with copper ions in situ and due to the deposition of QCS. The subsequent drug release behavior of 5-FCCu was studied, and the results show an initial high concentration kills microorganisms and long-acting sustained release inhibition. Moreover, in vivo wound experiments and toxicity experiments show the promotion of wound healing and excellent biocompatibility. As a demonstration of the utility of the latter, we have shown that the MMT-based smart platform can be used for the treatment of mixed infections of wounds.

Folate-chitosan nanoparticles triggered insulin cellular uptake and improved in vivo hypoglycemic activity.

The aim of this study was to prove a prolonged control of glucose levels, in rats, by the oral use of insulin folate-chitosan nanoparticles (FA-CS NPs). It was possible to prepare positively charged NPs with an average particle size of 288 ±â€¯5.11 nm and >80% entrapment efficiency. The system was able to enhance the stability of insulin in presence of GIT enzymes, with less than 10% release at pH 1.2 and an 8 hr released amount of 38.92 ±â€¯4.52% in PBS pH 7.4. A 5 fold enhancement in insulin intestinal permeability and cellular uptake over insulin solution was proven. The cellular uptake pathways was found to occur by several mechanisms. Besides, cell compatibility and absence of histopathological alterations was also demonstrated. Finally, a controlled blood glucose level for 8 h in rats. These results anticipated FA-CS NPs as a promising oral insulin candidate.

Comparative study on complexes formed by chitosan and different polyanions: Potential of chitosan-pectin biomaterials as scaffolds in tissue engineering.

Polyelectrolyte complexes of chitosan (Ch) and pectin (Pc) or alginate (Alg) were produced in the presence or absence of the silicone gel Silpuran® 2130 A/B (Sil) and the surfactant Kolliphor® P188 (Kol). Ch-Pc-Kol-based formulations presented higher porosity (up to 83.3%) and thickness (maximum of 2273.5 µm in PBS). Lower water contact angle was observed for Ch-Alg formulations (minimum of 36.8°) and these formulations presented higher swelling and mass loss in PBS (reaching up to 21.7 g/g and 80.4%, respectively). The addition of Sil to the matrices improved their elastic moduli, reaching a maximum of 4-fold change at 40% strain. The use of pectin instead of alginate augmented the elastic moduli, reaching 66 and 4-fold changes for dense and porous formulations, respectively. Pectin-containing scaffolds presented poroviscoelasticity, a typical mechanical feature of many soft tissues. The suitability of the materials for tissue engineering applications was demonstrated in terms of stability upon degradation in culture medium or lysozyme solution, as well as lack of cytotoxicity. This study evidences the potential of Ch-Pc-based materials to be further explored for this purpose, especially to improve the mechanical properties of chitosan-based scaffolds aiming medical applications.

pH-responsive selenium nanoparticles stabilized by folate-chitosan delivering doxorubicin for overcoming drug-resistant cancer cells.

Herein, we first report pH-responsive SeNPs stabilized with modified folic acid-N-trimethyl chitosan (TMC-FA) as nanocarriers for delivery of doxorubicin (DOX) to overcome drug-resistant cancer cells, which could enhance the activity of DOX by approximately 10-fold for a reduced IC50 value compared to free DOX. When nanoparticles were taken up by cells, the DOX-loaded SeNPs@TMC-FA demonstrated a faster release rate under acidic conditions. The cumulative release amount of DOX at pH 5.3 was 54.1% within 2h and 95.5% at 6h, whereas the release rate at pH 7.4 was 12.3% in 2h and 42.2% for 6h; release was not completed at the end of the study, 72h. Mechanistic studies suggested that DOX-SeNPs@TMC-FA induced cell death through the apoptosis pathway by involvement of caspase-3 and PARP proteins. The results demonstrated that pH-responsive SeNPs@TMC-FA, as targeted nanocarriers, promoted the efficacy of DOX and overcame drug resistance in NCI/ADR-RES cells.

Cytotoxicity of chitosan/streptokinase nanoparticles as a function of size: An artificial neural networks study.

Predicting the size and toxicity of chitosan/streptokinase nanoparticles at various values of processing parameters was the aim of this study. For the first time, a comprehensive model could be developed to determine the cytotoxicity of the nanoparticles as a function of their size. Then, artificial neural networks were used for identifying main factors influencing self-assembly prepared nanoparticles size and cytotoxicity. Three variables included polymer concentration; pH and stirring time were used for a modeling study. A second modeling was performed to evaluate the influence of particles' size on toxicity. Experimentally data modeled using ANNs was validated against unseen data. The response surfaces generated from the software demonstrated that chitosan concentration is the dominant factor with a direct effect on size. Results also showed that the most important factor in determining the particles' toxicity is size--smaller particles showed more toxic effects, regardless of the effect of other input parameters. From the Clinical Editor: The understanding of toxicity of nanoparticles is of prime importance. In this article, the authors generated a model to visualize the relationship between nanoparticle size and its cellular toxicity, using chitosan/streptokinase nanoparticles. The data generated here would help the design of future nanoparticles of appropriate sizes for the application in the clinical setting.

Chitosan nanoparticles loaded the herbicide paraquat: the influence of the aquatic humic substances on the colloidal stability and toxicity.

Polymeric nanoparticles have been developed for several applications, among them as carrier system of pesticides. However, few studies have investigated the fate of these materials in the environment in relation to colloidal stability and toxicity. In nature, humic substances are the main agents responsible for complexation with metals and organic compounds, as well as responsible for the dynamics of these nanoparticles in aquatic and terrestrial environments. In this context, the evaluation of the influence of aquatic humic substances (AHS) on the colloidal stability and toxicity of polymeric nanoparticles of chitosan/tripolyphosphate with or without paraquat was performed. In this study, the nanoparticles were prepared by the ionic gelation method and characterized by size distribution measurements (DLS and NTA), zeta potential, infrared and fluorescence spectroscopy. Allium cepa genotoxicity studies and ecotoxicity assays with the alga Pseudokirchneriella subcapitata were used to investigate the effect of aquatic humic substances (AHS) on the toxicity of this delivery system. No changes were observed in the physical-chemical stability of the nanoparticles due to the presence of AHS using DLS and NTA techniques. However some evidence of interaction between the nanoparticles and AHS was observed by infrared and fluorescence spectroscopies. The ecotoxicity and genotoxicity assays showed that humic substances can decrease the toxic effects of nanoparticles containing paraquat. These results are interesting because they are important for understanding the interaction of these nanostructured carrier systems with species present in aquatic ecosystems such as humic substances, and in this way, opening new perspectives for studies on the dynamics of these carrier systems in the ecosystem.

Chitosan/tripolyphosphate nanoparticles loaded with paraquat herbicide: an environmentally safer alternative for weed control.

Paraquat is a fast acting nonselective contact herbicide that is extensively used worldwide. However, the aqueous solubility and soil sorption of this compound can cause problems of toxicity in nontarget organisms. This work investigates the preparation and characterization of nanoparticles composed of chitosan and sodium tripolyphosphate (TPP) to produce an efficient herbicidal formulation that was less toxic and could be used for safer control of weeds in agriculture. The toxicities of the formulations were evaluated using cell culture viability assays and the Allium cepa chromosome aberration test. The herbicidal activity was investigated in cultivations of maize (Zea mays) and mustard (Brassica sp.), and soil sorption of the nanoencapsulated herbicide was measured. The efficiency association of paraquat with the nanoparticles was 62.6 ± 0.7%. Encapsulation of the herbicide resulted in changes in its diffusion and release as well as its sorption by soil. Cytotoxicity and genotoxicity assays showed that the nanoencapsulated herbicide was less toxic than the pure compound, indicating its potential to control weeds while at the same time reducing environmental impacts. Measurements of herbicidal activity showed that the effectiveness of paraquat was preserved after encapsulation. It was concluded that the encapsulation of paraquat in nanoparticles can provide a useful means of reducing adverse impacts on human health and the environment, and that the formulation therefore has potential for use in agriculture.

Micropatterning of bioactive glass nanoparticles on chitosan membranes for spatial controlled biomineralization.

Bioactive glass nanoparticles (BG-NPs) capable of inducing apatite precipitation upon immersion in simulated body fluid (SBF) were patterned on free-standing chitosan membranes by microcontact printing using a poly(dimethylsiloxane) (PDMS) stamp inked in a BG-NPs pad. Formation of the patterns was characterized by scanning electron microscopy (SEM). Mineralization of the bioactive glass patterns was induced in vitro by soaking the samples in SBF over different time points up to 7 days. The confined apatite deposition in the patterned regions with diameters of 50 µm was confirmed by Fourier-transformed infrared spectroscopy (FTIR), energy-dispersive X-ray (EDX) analysis, and SEM. In vitro tests confirmed the preferential attachment and proliferation of L929 cells to the areas printed with BG-NPs of the membranes. This approach permits one to spatially control the properties of biomaterials at the microlevel and could be potentially used in guided tissue regeneration for skin, vascular, articular, and bone tissue engineering and in cellular cocultures or to develop substrates able to confine cells in regions with controlled geometry at the cell's length scale.

Hemocompatibility of poly(ɛ-caprolactone) lipid-core nanocapsules stabilized with polysorbate 80-lecithin and uncoated or coated with chitosan.

The hemocompatibility of nanoparticles is of critical importance for their systemic administration as drug delivery systems. Formulations of lipid-core nanocapsules, stabilized with polysorbate 80-lecithin and uncoated or coated with chitosan (LNC and LNC-CS), were prepared and characterized by laser diffraction (D[4,3]: 129 and 134 nm), dynamic light scattering (119 nm and 133 nm), nanoparticle tracking (D50: 124 and 139 nm) and particle mobility (zeta potential: -15.1 mV and +9.3 mV) analysis. In vitro hemocompatibility studies were carried out with mixtures of nanocapsule suspensions in human blood at 2% and 10% (v/v). The prothrombin time showed no significant change independently of the nanocapsule surface potential or its concentration in plasma. Regarding the activated partial thromboplastin time, both suspensions at 2% (v/v) in plasma did not influence the clotting time. Even though suspensions at 10% (v/v) in plasma decreased the clotting times (p<0.05), the values were within the normal range. The ability of plasma to activate the coagulation system was maintained after the addition of the formulations. Suspensions at 2% (v/v) in blood showed no significant hemolysis or platelet aggregation. In conclusion, the lipid-core nanocapsules uncoated or coated with chitosan are hemocompatible representing a potential innovative nanotechnological formulation for intravenous administration.

Evaluation of the genotoxicity of chitosan nanoparticles for use in food packaging films.

The use of nanoparticles in food packaging has been proposed on the basis that it could improve protection of foods by, for example, reducing permeation of gases, minimizing odor loss, and increasing mechanical strength and thermal stability. Consequently, the impacts of such nanoparticles on organisms and on the environment need to be investigated to ensure their safe use. In an earlier study, Moura and others (2008a) described the effect of addition of chitosan (CS) and poly(methacrylic acid) (PMAA) nanoparticles on the mechanical properties, water vapor, and oxygen permeability of hydroxypropyl methylcellulose films used in food packaging. Here, the genotoxicity of different polymeric CS/PMAA nanoparticles (size 60, 82, and 111 nm) was evaluated at different concentration levels, using the Allium cepa chromosome damage test as well as cytogenetic tests employing human lymphocyte cultures. Test substrates were exposed to solutions containing nanoparticles at polymer mass concentrations of 1.8, 18, and 180 mg/L. Results showed no evidence of DNA damage caused by the nanoparticles (no significant numerical or structural changes were observed), however the 82 and 111 nm nanoparticles reduced mitotic index values at the highest concentration tested (180 mg/L), indicating that the nanoparticles were toxic to the cells used at this concentration. In the case of the 60 nm CS/PMAA nanoparticles, no significant changes in the mitotic index were observed at the concentration levels tested, indicating that these particles were not toxic. The techniques used show promising potential for application in tests of nanoparticle safety envisaging the future use of these materials in food packaging.

[Studies on the nasal epithelium toxicity of adjuvants and recombination hirudin (rHV2) nasal spary].

OBJECTIVE: To investigate the nasal epithelium toxicity of adjuvants and rHV2 nasal spary(HVS). METHOD: Ciliary movement were evaluated with in situ toad palate model; The histology assessment of nasal epithelium were carried out after long-lasting and repeated use of HVS. RESULT AND CONCLUSION: Adjuvants included SDS, Brij 35, azone, lecithin, EDTA, menthol, nipagin and thiomersal were able to significantly inhibited the ciliary movement, while tween80, glycyrrhizic acid monoammonium salt, benzalkonium bromide, sodium benzoate and adhensive materials investigated had less influence on it. HVS was able to damaged the nasal epithelium, but this effect recovered soon after stopping administration. It was demonstrated that SDS, Brij 35, azone,lecithin, EDTA, menthol, nipagin and thiomersal. It had significant cilitoxity, while tween80, glycyrrhizic acid monoammonium salt, benzalkonium bromide, sodium benzoate and adhensive materials investigated had no significance; Chitosan co-administration with some adjuvants may make the cillitoxity severer; It is available that rHV2 be administered by nasal spary.

Biological evaluation of chitosan salts cross-linked to genipin as a cell scaffold for disk tissue engineering.

Degenerative disc disease has been implicated as a major component of spine pathology. However, although biological repair of the degenerate disk would be the ideal treatment, there is no universally accepted scaffold for tissue engineering of the intervertebral disk. To help remedy this, we investigated the gelation kinetics of various concentrations (2.5 to 10%) of two water-soluble chitosan chlorides (low molecular weight Protasan UP CL113 and high molecular weight Protasan UP CL213) and two chitosan glutamates (low molecular weight Protasan UP G113 and high molecular weight Protasan UP G213). Various concentrations (5 to 20%) of genipin, a naturally occurring cross-linking reagent used in herbal medicine and in the fabrication of food dyes, were used to prepare cross-linked chitosan hydrogels. The results show that 2.5% Protasan UP G213 cross-linked to 5% genipin was the best candidate. This formulation gelled fastest at 37 degrees C, and maintained 95% viability of encapsulated cultured disk cells. The gel did not produce an inflammatory reaction when injected subcutaneously into C57BL/6 mice and is therefore biocompatible. Most importantly, when injected into the degenerated nucleus pulposus of human cadaveric intervertebral disk, the gel flowed into the clefts without leakage. This study demonstrates that 2.5% Protasan UP G213 cross-linked to 5% genipin might be a promising scaffold for disk tissue engineering.

Biological control of Phytophthora ramorumon rhododendron.

Phytophthora ramorum was found in Poland in 2000 as the causal agent of rhododendron blight. Besides eradication of diseased plants and rhododendron growing around, chemical and biological control of the pathogen is necessary. In this study in vitro activity of grapefruit extract and chitosan in the inhibition of P. ramorum growth and sporulation and their efficacy in the control of leaf and stem rot development was evaluated. Amendment of V8 juice agar and soil leachate with grapefruit extract resulted in the inhibition of colony growth and sporulation of P. ramorum. Zoosporangia were more susceptible to the extract than pathogen hyphae and chlamydospores. Chitosan only slightly inhibited the colony growth and zoosporngia production. Spraying of rhododendron inoculated with P. ramorum with grapefruit extract at conc. 165 microg/cm3 inhibited 2-3 times the spread of necrosis on stems and leaves. Pre- and postinoculation spraying of rhododendrons with chitosan at conc. of 1000 microg/cm3 suppressed the disease spread about 40%.