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Fabrication of Thymoquinone‐Loaded Albumin Nanoparticles by Microfluidic Particle Synthesis and Their Effect on Planarian Regeneration

Aslihan Kazan, Ozlem Yesil‐Celiktas, Yu Shrike Zhang

Thymoquinone is the main bioactive component of the plant Nigella sativa, which is commonly known as black seeds and has several therapeutic effects. However, clinical applications of thymoquinone are limited due to its hydrophobic nature. In this study, thymoquinone is encapsulated in albumin nanoparticles by using a microfluidic platform to overcome this limitation. The mean particle sizes of empty and thymoquinone‐loaded nanoparticles are determined as 271.3 and 315.6 nm, respectively, with polydispersity index values both lower than 0.25. In addition to particle size distribution measurements, characterizations of the prepared nanoparticles such as zeta potential measurements, in vitro release studies, as well as scanning electron microscopy, Fourier‐transform infrared, and differential scanning calorimetry analyses are also carried out. To determine the effect of thymoquinone on neural regeneration, planarians are used as the model organism. After application of free and encapsulated thymoquinone, planarians are amputated and the fragments are observed in terms of head and tail regeneration, swimming pattern, and behavior. The results indicate that thymoquinone affects their behavior and primarily enhances head regeneration of planarians. In addition, it is shown that encapsulation of thymoquinone not only enhances the thermal stability of the molecule but also decreases its toxicity.

Antimicrobial activity of propolis and gentamycin against methicillin-resistant Staphylococcus aureus in a 3D thermo-sensitive hydrogel

Yuksel Gezgin, Aslihan Kazan, Fulden Ulucan, Ozlem Yesil-Celiktas

The antibiotic resistance has become a major public health problem globally. Combinatorial administration of natural compounds and antibiotics is an alternative approach, particularly if synergistic effects can be elicited against multi-drug resistant bacteria. The aim of this study was to investigate the combinatorial activity of propolis and gentamycin against methicillin-resistant Staphylococcus aureus (MRSA) ATCC 43300 in three dimensional (3D) thermo-sensitive hydrogel (Poloxamer 407 25% (w/w)) to mimic the microenvironment realistically. Rheological properties, sol-gel transition temperature and the porosity of the hydrogel were determined. Subsequently, thermo-sensitive gel was loaded with propolis and gentamycin for evaluation of the combined effect. The fractional inhibitory concentration indices showed marked synergy and partial synergy without and within 3D termo-sensitive hydrogel at a range of 0.38 to 0.56, respectively. This 3D platform can be utilized to screen potential antimicrobial compounds and readily expanded to other microbial species.

A novel niosome formulation for encapsulation of anthocyanins and modelling intestinal transport

Melike Fidan-Yardimci, Seref Akay, Fatemeh Sharifi, Canan Sevimli-Gur, Gaye Ongen, Ozlem Yesil-Celiktas

Bioprinting covers the precise deposition of cells, biological scaffolds and growth factors to produce desired tissue models. The main focus of bioprinting is the creation of functional three-dimensional (3D) biomimetic composites for various application areas. Successful creations of model tissues depend on certain parameters such as determination of optimum microenvironment conditions, selection of appropriate scaffold, and cell source. As the cell culture-based assays have vital roles in the biomedical field, bioprinted tissue analogs would provide unprecedented chances to study, screen, and treat diseases. Today’s 3D bioprinting technology is able to print cells and scaffolds simultaneously, which provides the opportunity for disease modeling. This paper presents a general overview of the current state of the art in bioprinting technologies and potential 3D cell culture systems now being developed to model microbial infections, host-pathogen interactions, niches for microbiota, biofilm formation, and assess microbial resistance to antibiotics.

Cytotoxicity screening of supercritical fluid extracted seaweeds and phenylpropanoids

Canan Sevimli-Gur, Ozlem Yesil-Celiktas

Detached leaves of Posidonia oceanica and Zostera marina creating nuisance at the shores were extracted by means of supercritical CO2 enriched with a co-solvent, compared with that of soxhlet extraction. The extracts and their active compounds which are phenylpropanoids (chicoric, p-coumaric, rosmarinic, benzoic, ferulic and caffeic acids) were screened for cytotoxicity in cancer cell lines including human breast adenocarcinoma (MCF-7, MDA-MB-231, SK-BR-3), human colon adenocarcinoma (HT-29), human cervix adenocarcinoma (HeLa), human prostate adenocarcinoma (PC-3), Mus musculus neuroblastoma (Neuro 2A) cell lines and African green monkey kidney (VERO) as healthy cell line. Supercritical CO2 extracts proved to be more active than soxhlet counterparts. Particularly, Zostera marina extract obtained by supercritical CO2 at 250 bar, 80 °C, 20% co-solvent and a total flow rate of 15 g/min revealed the best IC50 values of 25, 20, 8 μg/ml in neuroblastoma, colon and cervix cancer cell lines. Among the major compounds tested, p-coumaric acid exhibited the highest cytotoxic against colon and cervix cell lines by with IC50 values of 25, 11 μg/ml. As for the effects on healthy cells, the extract was not cytotoxic indicating a selective cytotoxicity. Obtained supercritical CO2 extracts can be utilized as a supplement for preventive purposes.

Design of a new generation wound dressing with pine bark extract

Pelin Secim Karakaya, Ayberk Oktay, Necdet Seventekin, Ozlem Yesil-Celiktas

Medical textiles are one of the fastest growing sectors in the technical textile market. Wound dress is one of the significant applications with the largest share in medical textiles. Active molecules doped in the dressings may be therapeutic agents, vitamins, antibiotics, minerals, and growth factors, which contribute to wound healing. Medical plants have a great potential with positive effects in wound care and accelerate the rate of wound healing. Pine bark, which is known to exhibit wound healing properties, is also used in the medical field. The purpose of this study is to design a new wound dressing enriched with Pinus brutia extract. Microwave-assisted extraction which is an environmentally friendly method was carried out at 70℃, 900 W for 10 min to obtain the extracts. Subsequently, P. brutia bark extract was embedded to the alginate gel dressing and characterized and evaluated by in vivo studies on rats. According to the results, the extract was rapidly released from the alginate gel in the first 6 h, whereas the release was slowly increased to 24 h and then reached a steady state. Therefore, P. brutia extract-embedded alginate gel dressings applied for in vivo studies were changed every 24 h, reaching a healing rate of 75.7%, whereas the control group showed a healing rate of 48.6% indicating the superiority of the newly designed wound healing dress enriched with pine bark extract.

High-Yield Biocatalysis of Baicalein 7-O-β-d-Glucuronide to Baicalein Using Soluble Helix pomatia-Derived β-Glucuronidase in a Chemically Defined Acidic Medium

Cahit Muderrisoglu, Ozlem Yesil-Celiktas

Baicalein, showing stronger pharmacological activity, can be obtained by removal of the distal glucuronic acid (GluA) from
baicalein 7-O-β-D-glucuronide (baicalin). In the present study, a chemically defned reaction medium comprised of mildly
acidic (pH 4.5, 37 °C) aqueous solution, was formulated for biotransformation of baicalin to baicalein using acidic Helix
pomatia derived beta-glucuronidase (HP-GUS), an untested biocatalyst source. The biotransformation was carried out as a
batchwise process within an optimised reaction cocktail (with 5% dimethylformamide, v/v) by a 4-h HP-GUS (250 unit/ml)
incubation of baicalin (60 ppm) and resulted in a promising conversion ratio of 99% without any by-product formation. The
formulated reaction system may ofer a novel and efcient alternative for bioproduction of baicalein, which can be vital for
pharmaceutical applications.

A Foreign Body Response‐on‐a‐Chip Platform

Fatemeh Sharifi, Su Su Htwe, Martina Righi, Hua Liu, Anna Pietralunga, Ozlem Yesil‐Celiktas, Sushila Maharjan, Byung‐Hyun Cha, Su Ryon Shin, Mehmet Remzi Dokmeci, Nihal Engin Vrana, Amir M Ghaemmaghami, Ali Khademhosseini, Yu Shrike Zhang

Understanding the foreign body response (FBR) and desiging strategies to modulate such a response represent a grand challenge for implant devices and biomaterials. Here, the development of a microfluidic platform is reported, i.e., the FBR‐on‐a‐chip (FBROC) for modeling the cascade of events during immune cell response to implants. The platform models the native implant microenvironment where the implants are interfaced directly with surrounding tissues, as well as vasculature with circulating immune cells. The study demonstrates that the release of cytokines such as monocyte chemoattractant protein 1 (MCP‐1) from the extracellular matrix (ECM)‐like hydrogels in the bottom tissue chamber induces trans‐endothelial migration of circulating monocytes in the vascular channel toward the hydrogels, thus mimicking implant‐induced inflammation. Data using patient‐derived peripheral blood mononuclear cells further reveal inter‐patient differences in FBR, highlighting the potential of this platform for monitoring FBR in a personalized manner. The prototype FBROC platform provides an enabling strategy to interrogate FBR on various implants, including biomaterials and engineered tissue constructs, in a physiologically relevant and individual‐specific manner.

Supercritical Fluid Technology in Bioseperation

Ece Yildiz-Ozturk, Ozlem Yesil-Celiktas

The use of biotechnology for the production of primary and secondary metabolites has great promise for sustainable manufacturing of biochemical and pharmaceutical products. Integrated production and recovery processes allowing in situ separation of cells, microorganisms, and microalgae, followed by extraction and separation of target compounds are of paramount importance. However, this imposes the need for the design and development of green and cost effective bioseparation processes. Supercritical fluid processing is a promising technology, which can serve to fulfill these requirements and enable separation of a wide range of bioactive compounds. In particular, supercritical fluid extraction is a well-established and widely used technique for processing of oil seeds, coffee beans, hops and various plant materials, practiced on industrial scale. In this chapter, the properties of supercritical fluids and instrumentation of supercritical fluid process are elaborated. Subsequently, various applications, particularly the extraction of biologically active compounds from medical plants and microalgae, are discussed in more details as a bioseparation approach. The tendency towards high quality and healthy foods and pharmaceutical compounds and awareness in environmental aspects will be the main drivers for extending the application of supercritical fluids in the near future.

Synthesis of alginate‐silica hybrid hydrogel for biocatalytic conversion by β‐glucosidase in microreactor

Rabia Onbas, Ozlem Yesil‐Celiktas

The organic–inorganic hybrid materials have been used in different fields to immobilize biomolecules since they offer many advantages. The aim of this study was to optimize and characterize the alginate‐silica hybrid hydrogel as a stable and injectable form for microfluidic systems using internal gelation method and increase the stability and activity of immobilized enzyme for biocatalytic conversions as well. Characterization was carried out by scanning electron microscopy, energy dispersive spectroscopy/mapping, Brunauer–Emmett–Teller, Barrett–Joyner–Halenda, and Fourier‐transform infrared spectroscopy analyses, and the shrinkages of monoliths were evaluated. Subsequent to optimizing the enzyme concentration (40 μg), hydrolytic conversion of 4‐nitrophenyl β‐d‐glucopyranoside (pNPG) was performed to understand the behavior of the bioconversion in the microfluidic system. The yield was 94% which reached the equilibrium at 24 h indicating that the alginate‐silica gel derived microsystem overcome some drawbacks of monolithic systems. Additionally, bioconversion of Ruscus aculeatus saponins was carried out at the same setup in order to obtain aglycon part, which has pharmaceutical significance. Although pure aglycon could not be achieved, an intermediate compound was obtained based on the HPLC analysis. The developed formulation can be utilized for various life science applications.

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