Design of chemobrionic and biochemobrionic scaffolds for bone tissue engineering.

Chemobrionic systems have attracted great attention in material science for development of novel biomimetic materials. This study aims to design a new bioactive material by integrating biosilica into chemobrionic structure, which will be called biochemobrionic, and to comparatively investigate the use of both chemobrionic and biochemobrionic materials as bone scaffolds. Biosilica, isolated from Amphora sp. diatom, was integrated into chemobrionic structure, and a comprehensive set of analysis was conducted to evaluate their morphological, chemical, mechanical, thermal, and biodegradation properties. Then, the effects of both scaffolds on cell biocompatibility and osteogenic differentiation capacity were assessed. Cells attached to the scaffolds, spread out, and covered the entire surface, indicating the absence of cytotoxicity. Biochemobrionic scaffold exhibited a higher level of mineralization and bone formation than the chemobrionic structure due to the osteogenic activity of biosilica. These results present a comprehensive and pioneering understanding of the potential of (bio)chemobrionics for bone regeneration.

Comprehensive Analysis of Lutein and Loroxanthin in Scenedesmus obliquus: From Quantification to Isolation.

Carotenoids are hydrophobic pigments produced exclusively by plants, fungi, and specific microbes. Microalgae are well suited for the production of valuable carotenoids due to their rapid growth, efficient isoprenoid production pathway, and ability to store these compounds within their cells. The possible markets for bio-products range from feed additives in aquaculture and agriculture to pharmaceutical uses. The production of carotenoids in microalgae is affected by several environmental conditions, which can be utilized to enhance productivity. The current study focused on optimizing the extraction parameters (time, temperature, and extraction number) to maximize the yield of carotenoids. Additionally, the impact of various nitrogen sources (ammonia, nitrate, nitrite, and urea) on the production of lutein and loroxanthin in Scenedesmus obliquus was examined. To isolate the carotenoids, 0.20 g of biomass was added to 0.20 g of CaCO3 and 10.0 mL of ethanol solution containing 0.01% (w/v) pyrogallol. Subsequently, the extraction was performed using an ultrasonic bath for a duration of 10 min at a temperature of 30 °C. This was followed by a four-hour saponification process using a 10% methanolic KOH solution. The concentration of lutein and loroxanthin was measured using HPLC-DAD at 446 nm, with a flow rate of 1.0 mL/min using a Waters YMC C30 Carotenoid column (4.6 × 250 mm, 5 µm). The confirmation of carotenoids after their isolation using preparative chromatography was achieved using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with an atmospheric pressure chemical ionization (APCI) probe and UV-vis spectroscopy. In summary, S. obliquus shows significant promise for the large-scale extraction of lutein and loroxanthin. The findings of this study provide strong support for the application of this technology to other species.

Induction of antioxidant activities of Arthrospira platensis and Chlorella vulgaris by modified culture conditions.

Microalgae are considered a promising source for obtaining natural compounds with strong antioxidant activity. Despite the great progress made in this field, there is still need for further studies applying simple and cost-effective modifications to reveal their full potential and enhance antioxidant properties. Arthrospira platensis and Chlorella vulgaris are some of the most common cells studied for this purpose. In this study, it was aimed to develop a bioprocess for the enhancement of antioxidant properties of these two microalgae by evaluating the effect of different culture conditions. With this aim, the impacts of light intensity/reactive oxygen species and nitrogen sources/reactive oxygen species were evaluated for the A. platensis and C. vulgaris cells, respectively. Results showed that the antioxidant potential of A. platensis was found to be correlated with the phycocyanin and total phenolic content of cells, and 80 µmol photons m-2 s-1 light intensity induced antioxidant activity in a two-step cultivation mode. For C. vulgaris cells, maximum antioxidant activities of 68.10 ± 1.51% and 75.68 ± 0.66% were obtained in cultures with NH4Cl (0.016% (w/v)) for DPPH and ABTS assays, respectively. The applied oxidative stress factors exhibited different effects on the antioxidant activities of the cells because of their cellular morphologies and changing mechanisms of reactive oxygen species. These outcomes show the potential of applied modifications on cells and suggest a promising route to enhance antioxidant activities of microalgae for further research.

Comparative Evaluation of Chemical Garden Growth Techniques.

Chemical gardens are an exciting area of self-organized precipitation structures that form nano- and micro-sized structures in different shapes. This field has attracted great interest from researchers due to the specific characteristics and potential applications of these structures. Today, research on chemical gardens has provided deeper information regarding the formation mechanisms of these structures, and several techniques have been developed for chemical garden growth. However, they all show different growth patterns and lead to the formation of structures with a variety of morphological, chemical, or physical properties. This study aimed to evaluate the effects of different production techniques on chemical garden growth, taking into consideration the growth patterns, morphology, microstructure, and chemical composition. The chemical garden structures obtained in seed and injection experiments, two common methods, showed highly similar surface structures, void formation, and chemical composition. The membrane growth method has a small number of applications; thus, it was comprehensively evaluated to add new insights to the existing limited data. It produced the most stable and standard structures in a flat sheet-like shape and showed different morphologies than those observed in other two methods. Overall, this study presented significant results about the effect of growth techniques on chemical garden structures and similar systems.

Manipulation in Culture Conditions of Nanofrustulum shiloi for Enhanced Fucoxanthin Production and Isolation by Preparative Chromatography.

Microalgae produce a variety of high-value chemicals including carotenoids. Fucoxanthin is also a carotenoid that has many physiological functions and biological properties. For this reason, the cost-effective production of fucoxanthin at an industrial scale has gained significant attention. In the proposed study, fucoxanthin production was aimed to be increased by altering the culture conditions of N. shiloi. The effect of light intensity aeration rate, different nitrogen sources, and oxidative stress on the biomass and fucoxanthin productivity have been discussed. Based on these results, the fucoxanthin increased to 97.45 ± 2.64 mg/g by adjusting the light intensity to 50 µmol/m2s, and aeration rate at 5 L/min using oxidative stress through the addition of 0.1 mM H2O2 and 0.1 mM NaOCl to the culture medium. Fucoxanthin was then purified with preparative HPLC using C30 carotenoid column (10 mm × 250 mm, 5 µm). After the purification procedure, Liquid chromatography tandem mass spectrometry (LC-MS/MS) and UV-vis spectroscopy were employed for the confirmation of fucoxanthin. This study presented a protocol for obtaining and purifying considerable amounts of biomass and fucoxanthin from diatom by manipulating culture conditions. With the developed methodology, N. shiloi could be evaluated as a promising source of fucoxanthin at the industrial scale for food, feed, cosmetic, and pharmaceutical industries.

Aligned with sustainable development goals: microwave extraction of astaxanthin from wet algae and selective cytotoxic effect of the extract on lung cancer cells.

Astaxanthin is one of the most attractive carotenoid in the cosmetic, food, pharmaceutical, and aquaculture industries due to its strong bioactive properties. Among the various sources, several algae species are considered as rich sources of astaxanthin. Downstream processing of algae involves the majority of the total processing costs. Thus, elimination of high energy involved steps is imperative to achieve cost-effective scale in industry. This study aimed to determine operation conditions for astaxanthin extraction from wet Haematococcus pluvialis using microwave-assisted extraction. The isolated astaxanthin extract was evaluated for cytotoxicity on human lung cancer cells. The microwave-assisted extraction process at 75 °C under the power of 700 Watt for 7 min gave the highest astaxanthin yield (12.24 ± 0.54 mg astaxanthin/g wet cell weight). Based on MTT cell viability and Hoechst 33342 nuclear staining assays on A549 lung cancer cells, astaxanthin inhibited cell growth in dose- and time-dependent manners, where IC50 value was determined as 111.8 ± 14.8 µg/mL and apoptotic bodies were observed along with positive control group at 72 hr. These results showed that the treatment with astaxanthin extracted from wet H. pluvialis by microwave-assisted extraction exhibited anti-cancer activity on lung cancer cells indicating a newly potential to be utilized in industry.

Development of a Controlled Injection Method Using Support Templates for the Production of Chemobrionic Materials.

Chemobrionics is a research field about the well-known self-organized inorganic structures. Numerous research works have focused on controlling their growth pattern and characteristic features. In the present study, a controlled injection method is proposed to produce more regular self-assembled chemobrionics compared to the standard direct injection technique. This method involves the injection of a metal salt solution into an agarose support template filled with an anionic solution. The obtained structures were studied by scanning electron microscopy, X-ray microtomography, X-ray photoelectron spectroscopy, Raman spectroscopy, Fourier-transform IR spectroscopy, and thermogravimetric analysis. Despite the complex mechanism and chemistry underlying the self-organization phenomena, the controlled injection method enabled the generation of regular standard chemobrionic structures with high experimental reproducibility. It provided the extraction of tubular structures from the reaction vessel without breakage, thus allowing comprehensive characterization. Furthermore, the morphological, chemical, and thermal features of these structures were highly correlated with the standard chemobrionics obtained in the direct injection method. The proposed controlled injection method holds great promise for understanding and controlling the properties of chemobrionics and related structures.

Induction of lutein production in Scenedesmus obliquus under different culture conditions prior to its semipreparative isolation.

Microalgae with their improved growth rates and accumulation of high-value-added products make their commercial production attractive. Among them, lutein, which is a carotenoid, plays a very important role due to its various applications in the food and pharmaceutical industry. Induction of its biosynthesis can be triggered by various stress conditions like light. In this study, three different light intensities (50,150 and 300 µmol photons/m2s) and aeration rates (1, 3, and 5 L/min) were utilized to induce the lutein biosynthesis and biomass productivity in Scenedesmus obliquus. Lutein was isolated by preparative chromatography using a semiprep C30 column (10 × 250 mm, 5µm) and its confirmation was made by LC-MS/MS. According to the results, Scenedesmus obliquus synthesized the maximum lutein (8.01 ± 0.1 mg/g) with biomass productivity of 1.698 g/L at 150 µmol photons/m2s light intensity using 3 L/min as aeration rate. To the best of the authors' knowledge, this was the first study that the lutein was isolated by preparative chromatography using semiprep C30 carotenoid column with a simple and rapid separation, which can be used as a reference methodology for the isolation of other carotenoids. Scenedesmus obliquus can be an important alternative source for commercial production of lutein, as it is indicated from the results of this study.

Effects of Extraction Methods and Conditions on Bioactive Compounds Extracted from Phaeodactylum tricornutum.

The effect of homogenization, ultrasound and microwave extraction methods and conditions on fucoxanthin content, total phenolic content and antioxidant activity of extracts obtained from Phaeodactylum tricornutum were investigated in this study. The solvent/biomass ratio was the most effective parameter on fucoxanthin content, total phenolic content and antioxidant activity. The maximum fucoxanthin content (5.60 ± 0.06 mg/g) and antioxidant activity (763.00 ± 15.88 EC50 ?g/mL extract) were obtained with the homogenization extraction method whose optimum conditions were 1.93% biomass/solvent ratio, ~5200 rpm homogenization rate and 14.2 min extraction time. Although the ultrasonic extraction method has reached the approximately same level of fucoxanthin content (5.24 ± 0.07 mg/g)), TPC (67.68 ± 1.58 mg gallic acid/L) and antioxidant activity (619.90 ± 17.16 EC50 ?g/mL extract) at an amplitude of 55.72%, a higher biomass/ solvent ratio (2.72%) and a longer extraction time (17.37 min) have been required. The lowest fucoxanthin content, total phenolic content and antioxidant activity were determined for the microwave extraction method.

Computational fluid dynamics simulation in scaling-up of airlift photobioreactor for astaxanthin production.

The unicellular green microalga Haematococcus pluvialis accumulates large amounts of the red ketocarotenoid astaxanthin. Aiming to cultivate these microalgae with high astaxanthin efficiency, cultivations were scaled-up from 1000 mL bottle to 2 L and 8 L airlift photobioreactor using volumetric power consumption rate (W/m3) as scale up strategy. After cultivations, computational fluid dynamics (CFD) simulation was used to investigate the flow patterns, mixing efficiency and gas holdup profile within the 2 L photobioreactor. At the end, astaxanthin content was enhanced with increasing the cultivation volume and highest astaxanthin amount of 49.39 ± 1.64 mg/g cell was obtained in 8 L photobioreactor. Hydrodynamic characteristics of photobioreactor was simulated and gas holdup showed difference between the riser and the downcomer regions. Velocity profiles of air and medium had higher values inside the draft tube than obtained in downcomer region. However liquid circulation was achieved from draft tube to the downcomer, mixing was not provided effectively considering the turbulence kinetic energy. For the further research, some developments about column configuration, sparger diameter may be necessary to enhance the mixing characteristics.

Analytical Grade Purification of Phycocyanin from Cyanobacteria.

Phycocyanin is a blue-colored pigment-protein complex that exhibits numerous biofunctions such as anti-inflammation, antioxidation, antitumor, neuroprotective effect, and immunological enhancement. Purified phycocyanin has pharmaceutical and nutraceutical applications. In addition, as a nontoxic and non-carcinogenic natural coloring agent, phycocyanin has many applications in the food and cosmetic industries. This chapter describes a protocol for extraction and analytical grade purification of phycocyanin from cyanobacteria. The purification steps include (1) extraction of phycocyanin from biomass, (2) ammonium sulfate precipitation of phycocyanin and dialysis, and (3) purification of phycocyanin by gel filtration and ion-exchange chromatography.

Growth Kinetics of Nanofrustulum Shiloi Under Different Mixing Conditions in Flat-plate Photobioreactor

Abstract Diatoms are the major group of microalgae which have been utilized by the potential applications as food industries, aquatic feeds, cosmetics, biofuels, and pharmaceuticals. In this study, current approaches were made in order to determine growth rate, biomass productivity, protein, carbohydrate, lipid and fatty acid composition for Nanofrustulum shiloi cultures using both aeration and mixing conditions in flat-plate photobioreactor (PBR). Physical (the intensity of aeration, mixing, light intensity etc.) and chemical (nutritional materials) factors are affecting the growth and bioproduct contents of a diatom. Biomass and lipid productivities of N. shiloi were measured as 31.29 and 36.9622±0.0598 mg L-1 day-1 in flat-plate PBR having the combination of aeration and stirring system, respectively. A slightly higher amount of saturated fatty acids was detected in PBR having only bubbling system while the increase of mono- and poly- unsaturated fatty acids were found in PBR having the combination of aeration and stirring system. Flat-plate PBR design was also investigated for improving not only biomass but also the lipid productivity of N. shiloi.

Enhancement of biomass and phycocyanin content of Spirulina platensis.

Response surface methodology (RSM) based on the 23 factorial central composite design (CCD) was employed to evaluate optimum culture conditions (temperature, light irradiance and agitation) to enhance biomass and phycocyanin content of Spirulina platensis. The predicted maximum biomass and phycocyanin content by RSM was 1.06 g L-1 and 107 mg L-1, respectively, whereas maximum biomass and phycocyanin content of 1.32 g L-1 and 127 mg L-1 was obtained in the validation experiments under optimized conditions after 10 days of cultivation. Further, influence of optimized conditions (temperature 33±2 ºC, light irradiance 44 µmol photons m-2 s-1 and a flow rate of 2.5 L min-1) on growth and phycocyanin content of S. platensis in 7L Panel photobioreactor (PPBR) cultivation was investigated. A 15 days production was carried out and it was observed that a maximum biomass yield of 2.42 g L-1 with a specific growth rate 0.202 day-1 and phycocyanin content of 228 mg L-1 was obtained in the PPBR. The optimum culture conditions obtained through response surface methodology were successfully determined to maximize the biomass and phycocyanin.

Large-scale bioprospecting of cyanobacteria, micro- and macroalgae from the Aegean Sea.

Marine organisms constitute approximately one-half of the total global biodiversity, being rich reservoirs of structurally diverse biofunctional components. The potential of cyanobacteria, micro- and macroalgae as sources of antimicrobial, antitumoral, anti-inflammatory, and anticoagulant compounds has been reported extensively. Nonetheless, biological activities of marine fauna and flora of the Aegean Sea have remained poorly studied when in comparison to other areas of the Mediterranean Sea. In this study, we screened the antimicrobial, antifouling, anti-inflammatory and anticancer potential of in total 98 specimens collected from the Aegean Sea. Ethanol extract of diatom Amphora cf capitellata showed the most promising antimicrobial results against Candida albicans while the extract of diatom Nitzschia communis showed effective results against Gram-positive bacterium, S. aureus. Extracts from the red alga Laurencia papillosa and from three Cystoseira species exhibited selective antiproliferative activity against cancer cell lines and an extract from the brown alga Dilophus fasciola showed the highest anti-inflammatory activity as measured in primary microglial and astrocyte cell cultures as well as by the reduction of proinflammatory cytokines. In summary, our study demonstrates that the Aegean Sea is a rich source of species that possess interesting potential for developing industrial applications.

Process optimization and modeling for the cultivation of Nannochloropsis sp. and Tetraselmis striata via response surface methodology.

The aim of this study was to determine the optimal physical process conditions for the cultivation of locally isolated strains of Nannochloropsis sp. and Tetraselmis striata to achieve maximum growth rate. It was essential to evaluate biomass production at different agitation rates, light intensities, and temperature levels. Central composite design and response surface methodology were applied to design the experiments and optimize the cultivation process for Nannochloropsis sp. and T. striata. The specific growth rate of 0.250 d(-1) was obtained for Nannochloropsis sp. cells under the light intensity of 54 µmol photons · m(-2) · s(-1) , at the agitation rate of 151 rpm in 24.5°C. The optimal physical process conditions for T. striata were obtained under the light intensity of 56 µmol photons · m(-2) · s(-1) in 25.5°C at the agitation rate of 151 rpm in 25.5°C, resulting in a specific growth rate of 0.226 d(-1) . The predicted values were justified by the verification tests. Good agreement between the predicted values and the experimental values confirmed the validity of the models for the cultivation of microalgal strains. In this article, the noteworthy result was that temperature was a dominant factor in obtaining high chl-a content for Nannochloropsis sp., whereas the growth of T. striata strongly depended on light exposure.

Carotenoid and fatty acid compositions of an indigenous Ettlia texensis isolate (Chlorophyceae) under phototrophic and mixotrophic conditions.

Ettlia oleoabundance (formerly known as Neochloris oleoabundance) is an attractive candidate for biodiesel production because of its high lipid accumulation, and it's taking the majority of the attention among the strains of Ettlia genus; however, potential of the other genus members is unknown. An indigenous strain from Salda Lake (South West Turkey) identified by 18S rDNA sequencing as Ettlia texensis (GenBank accession no: JQ038221), and its fatty acid and carotenoid compositions under phototrophic and mixotrophic conditions was investigated to evaluate the potential of the strain for commercial uses. A threefold increase was observed in total lipid content (total fatty acids; from 13% to 37%) in mixotrophic culture respect to the phototrophic growth conditions. The oleic acid (C18:1) and alpha-linolenic acid (18:3) were the major unsaturated fatty acids accounting for 40% and 13.2% of total fatty acids in mixotrophic culture, respectively. Carotenoid analyses of the mixotrophic culture revealed the metabolite canthaxanthin, a commercially valuable carotenoid used mainly for food coloring, was the major constituent among other pigments. The possible use of E. texensis in biotechnological applications is discussed.

Equilibrium, thermodynamic and kinetic studies for the biosorption of aqueous lead(II), cadmium(II) and nickel(II) ions on Spirulina platensis.

The biosorption of lead(II), cadmium(II) and nickel(II) ions from aqueous solution by Spirulina platensis was studied as a function of time, concentration, temperature, repetitive reactivity, and ionic competition. The kinetic results obeyed well the pseudo second-order model. Freundlich, Dubinin Radushkevich and Temkin isotherm models were applied in describing the equilibrium partition of the ions. Freundlich isotherm was applied to describe the design of a single-stage batch sorption system. According to the thermodynamic parameters such as DeltaG degrees, DeltaH degrees and DeltaS degrees calculated, the sorption process was endothermic and largely driven towards the products. Sorption activities in a three metal ion system were studied which indicated that there is a relative selectivity of the biosorbent towards Pb2+ ions. The measurements of the repetitive reusability of S. platensis indicated a large capacity towards the three metal ions.

Agricultural fertilizers as economical alternative for cultivation of Haematococcus pluvialis.

A Haematococcus pluvialis strain isolated from the ruins of Ephesus in Turkey was investigated as regards its adaptation to laboratory conditions and maximum growth rate. In the first stage of the experiment, the growth of H. pluvialis was compared in common culture media. Furthermore, in an effort to minimize the culture costs, the second stage of the experiment compared the growth rate in the culture medium selected in the first stage with that in commercial plant fertilizers. The results demonstrated that the maximum cell concentration of 0.90 g/l, corresponding to a growth rate of 0.150 d(-1), was found with an N-P-K 20:20:20 fertilizer under a light intensity of 75 micromol photons m(-2) s(-1) on the 12th day of cultivation.