Production of letrozole-loaded alginate oxide-gelatin microgels using microfluidic systems for drug delivery applications.

Microfluidic systems are capable of producing microgels with a monodisperse size distribution and a spherical shape due to their laminar flow and superior flow. A significant challenge in producing these drug-carrying microgels is simultaneous drug loading into microgels. Various factors such as the type of polymer, the type of drug, the volume ratio of the drug to the polymer, and the geometry of the microfluidic system used to generate microgels can effectively address these challenges. The overall goal of this study was to produce mono-disperse drug-carrying microgels capable of controlled drug release. To achieve this goal, this study used a stream-focused microfluidic chip containing a coating current to prevent chip clogging. Alginate oxide was synthesized with a 30 % oxidation percentage. Alginate oxide, gelatin, and compositions of them with volume ratios of 50-50, 70-30, and 30-70, by determining their appropriate weight percentage, were used for the controlled release of letrozole. The properties of the produced microgels were measured through various tests such as drug release test, loading percentage, SEM, FTIR, swelling ratio, and dimensional stability. It was found that microgels made of a combination of alginate oxide-gelatin with volume ratios of 70-30 had a good swelling ratio and structural stability. The drug loading percentages for alginate, alginate oxide, and alginate oxide-gelatin with volume ratios of 50-50 and 30-70, respectively, were 56 %, 68 %, and 66 %, 61 % and the alginate oxide-gelatin with a volume ratio of 70-30 compared to other samples had over 70 % drug loading percentages. Furthermore, samples of alginate, alginate oxide, and alginate oxide-gelatin with volume ratios of 50-50 and 30-70 had 94 %, 63 %, 56 %, and 68 % drug release in 13 days, respectively. However, alginate oxide-gelatin with a volume ratio of 70-30 had a release rate of about 50 % in 13 days, which is a more controlled release for letrozole compared to the volume ratios of 50-50 and 30-70. Examining the drug release profile, it was concluded that drug release follows the Higuchi model and therefore follows Fick's first law of diffusion. It can be concluded that the combination of alginate oxide-gelatin produces more suitable microgels than alginate and alginate oxide for the controlled-release of letrozole. A comparison of microgels of alginate oxide and gelatin with volume ratios of 50-50 and 70-30 had better results for the cytotoxicity study compared to other samples.

Cell encapsulation in alginate-based microgels using droplet microfluidics; a review on gelation methods and applications.

Cell encapsulation within the microspheres using a semi-permeable polymer allows the two-way transfer of molecules such as oxygen, nutrients, and growth factors. The main advantages of cell encapsulation technology include controlling the problems involved in transplanting rejection in tissue engineering applications and reducing the long-term need for immunosuppressive drugs following organ transplantation to eliminate the side effects. Cell-laden microgels can also be used in 3D cell cultures, wound healing, and cancerous clusters for drug testing. Since cell encapsulation is used for different purposes, several techniques have been developed to encapsulate cells. Droplet-based microfluidics is one of the most valuable techniques in cell encapsulating. This study aimed to review the geometries and the mechanisms proposed in microfluidic systems to precisely control cell-laden microgels production with different biopolymers. We also focused on alginate gelation techniques due to their essential role in cell encapsulation applications. Finally, some applications of these microgels and researches will be explored.

An electro-conductive hybrid scaffold as an artificial Bruch's membrane.

Many research groups have investigated the various kinds of scaffolds to mimic the natural Bruch's membrane (BM) and support the retinal pigmented epithelial cells to form an organized cellular monolayer. While using prosthetic BM is identified as a promising treatment of age-related macular degeneration (AMD), a degenerative and progressive retinal disease, the effects of different signals such as electrical and morphological cues on the retinal pigmented epithelial (RPE) cells are still unknown. In this study, a laminated and conductive hydrogel/fiber composite scaffold by adding conductive polyaniline (PANi) to the scaffold's nanofibrous phase was prepared. This hybrid scaffold offers the closest morphology to the native structure of the human Bruch's membrane by imitating the inner and outer collagenous layer and induces the electrical signal to the scaffold to assess the electrical cue on behaviors of polarized retinal pigmented epithelial cells in the retina. The electrospun nanofibrous phase consisted of gelatin-Polyaniline in different ratios incorporated into the hydrogel precursor, a blend of gelatin and 4-armed PEG. We used a novel dual crosslinking process by incorporating the exposure of gamma irradiation and glutaraldehyde vapor treatment to construct the scaffold's hydrogel phase. The results showed the best composition was the sample which included the 40/60, Polyaniline/gelatin nanofiber sheets ratio because this scaffold revealed a 2.66 ± 0.33 MPa, Young's modulus and 1.84 ± 0.21 S/cm, electrochemical conductivity, which are close to the main features of native Bruch's membrane. In addition, this scaffold showed good biocompatibility by reaching 83.47% cell viability.

Silk fibroin hydrogel/dexamethasone sodium phosphate loaded chitosan nanoparticles as a potential drug delivery system.

The application of nanoparticles-loaded hydrogel as a novel formulation has gotten much attention for a potential drug delivery method for desire drug controlling and targeting. This study prepared a sustained release formulation using dexamethasone sodium phosphate-loaded chitosan nanoparticles embedded in silk fibroin hydrogel. Dexamethasone sodium phosphate-loaded chitosan nanoparticles (DEX-CSNPs) was developed using the ionotropic-gelation technique and inserted in the silk fibroin hydrogel (SFH). Mean particle size, polydispersity index (PDI), and zeta potential of DEX-CSNPs were 488.05±38.69 nm, 0.15±0.07, 32.12±2.42 mV, respectively. The encapsulation efficiency (EE), drug loading capacity (LC), and the cumulative amount of released drug of DEX-loaded CSNPs, which detected in phosphate buffer saline (PBS) solution, were 67.6±6.7%, 15.7±5.7%, and 75.84%, respectively. The DEX-CSNPs were then mixed with silk fibroin (SF) solution and induced gelation by sonication to prepare a drug-releasing system. As a result, the scanning electron microscopy (SEM) image shows that the prepared drug delivery system had a properly interconnected porous structure. Smaller pore size, greater porosity, higher water uptake, and swelling ratio were achieved by incorporating CSNPs and DEX-loaded CSNPs. The cytotoxicity study was performed for the L929 fibroblast cell line. The drug release kinetics study was performed on a prepared drug delivery system. Finally, the release test results showed a suitable extended-release of DEX from the carrier over 16 days. Overall, the developed drug-releasing system can be a promising candidate for drug delivery applications.

Microfluidic on-chip production of microgels using combined geometries.

Microfluidic on-chip production of microgels using external gelation can serve numerous applications that involve encapsulation of sensitive cargos. Nevertheless, on-chip production of microgels in microfluidic devices can be challenging due to problems induced by the rapid increase in precursor solution viscosity like clogging. Here, a novel design incorporating a step, which includes a sudden increase in cross-sectional area, before a flow-focusing nozzle was proposed for microfluidic droplet generators. Besides, a shielding oil phase was utilized to avoid the occurrence of emulsification and gelation stages simultaneously. The step which was located before the flow-focusing nozzle facilitated the full shielding of the dispersed phase due to 3-dimensional fluid flow in this geometry. The results showed that the microfluidic device was capable of generating highly monodispersed spherical droplets (CV < 2% for step and CV < 5% for flow-focusing nozzle) with an average diameter in the range of 90-190 µm, both in step and flow-focusing nozzle. Moreover, it was proved that the device could adequately create a shelter for the dispersed phase regardless of the droplet formation locus. The ability of this microfluidic device in the production of microgels was validated by creating alginate microgels (with an average diameter of ~ 100 µm) through an external gelation process with on-chip calcium chloride emulsion in mineral oil.

Conductive chitosan/polyaniline hydrogel with cell-imprinted topography as a potential substrate for neural priming of adipose derived stem cells.

Biophysical characteristics of engineered scaffolds such as topography and electroconductivity have shown potentially beneficial effects on stem cell morphology, proliferation, and differentiation toward neural cells. In this study, we fabricated a conductive hydrogel made from chitosan (CS) and polyaniline (PANI) with induced PC12 cell surface topography using a cell imprinting technique to provide both topographical properties and conductivity in a platform. The engineered hydrogel's potential for neural priming of rat adipose-derived stem cells (rADSCs) was determined in vitro. The biomechanical analysis revealed that the electrical conductivity, stiffness, and hydrophobicity of flat (F) and cell-imprinted (CI) substrates increased with increased PANI content in the CS/PANI scaffold. The conductive substrates exhibited a lower degradation rate compared to non-conductive substrates. According to data obtained from F-actin staining and AFM micrographs, both CI(CS) and CI(CS-PANI) substrates induced the morphology of rADSCs from their irregular shape (on flat substrates) into the elongated and bipolar shape of the neuronal-like PC12 cells. Immunostaining analysis revealed that both CI(CS) and CI (CS-PANI) significantly upregulated the expression of GFAP and MAP2, two neural precursor-specific genes, in rADSCs compared with flat substrates. Although the results reveal that both cell-imprinted topography and electrical conductivity affect the neural lineage differentiation, some data demonstrate that the topography effects of the cell-imprinted surface have a more critical role than electrical conductivity on neural priming of ADSCs. The current study provides new insight into the engineering of scaffolds for nerve tissue engineering.

Neural priming of adipose-derived stem cells by cell-imprinted substrates.

Cell-imprinting technology is a novel method for directing stem cell fate using substrates molded from target cells. Here, we fabricated and studied cell-imprinted substrates for neural priming in human adipose-derived stem cells in the absence of chemical cues. We molded polydimethylsiloxane silicone substrates on fixed differentiated neural progenitor cells (ReNcellTMVM). The ReNcellTMcell line consists of immortalized human neural progenitor cells that are capable to differentiate into neural cells. The fabricated cell-imprinted silicone substrates represent the geometrical micro- and nanotopology of the target cell morphology. During the molding procedure, no transfer of cellular proteins was detectable. In the first test with undifferentiated ReNcellTMVM cells, the cell-imprinted substrates could accelerate neural differentiation. With adipose-derived stem cells cultivated on the imprinted substrates, we observed modifications of cell morphology, shifting from spread to elongated shape. Both immunofluorescence and quantitative gene expression analysis showed upregulation of neural stem cell and early neuronal markers. Our study, for the first time, demonstrated the effectiveness of cell-imprinted substrates for neural priming of adipose-derived stem cells for regenerative medicine applications.

The Effects of Hydroxyethyl Starch 6% and Crystalloid on Volume Preloading Changes following Spinal Anesthesia.

BACKGROUND: Hypotension is one of the most common complications after spinal anesthesia for cesarean delivery. Normally, preloading with fluids, especially crystalloids, is used to prevention of hypotension. METHODS: In the present randomized clinical trial study, 120 parturients presenting for elective cesarean section with the American Society of Anesthesiologists Class I and II received either 15 cc normal saline or 7 cc/kg hydroxyethyl starch 6% (Voluven) fluid. Information regarding to systolic, diastolic, mean arterial pressure, and heart rate, incidence of hypotension, adverse effects, the total dose of atropine, and ephedrine were recorded in before and 3, 6, 9, 15, and 20 min after spinal anesthesia. Furthermore, Apgar score of newborn at the 1st and 5th min after birth was recorded. RESULTS: There was no significant difference in mean arterial pressure at different stages such as: Exactly after spinal and 3, 6, 15, and 20 min after spinal anesthesia between two groups (P > 0.05). Total dose of ephedrine and atropine were similar between groups (P > 0.05), respectively. There was no significant difference in Apgar score at the 1st and 5th min after birth between two groups. There were not any adverse effects of drugs in two groups. CONCLUSIONS: The results of this study show that hydroxyethyl starch 6% compared to normal saline are similar to prevent hypotension during spinal anesthesia for cesarean delivery.

Fabrication and characterization of SrAl2O4: Eu(2+)Dy(3+)/CS-PCL electrospun nanocomposite scaffold for retinal tissue regeneration.

Millions of people around the world become blind due to losing a part of the retina cells. In tissue engineering field one way to address this issue is to develop a retina tissue by scaffolds based on structure and signals received These scaffolds can play an essential role in repair and reformation of the damaged retina tissue. Here, SrAl2O4: Eu(2+), Dy(3+) nanophosphor were prepared by sol-gel method and then coated with PEG to become biocompatible. Next 10%, 30% and 50% concentration of the coated nanophosphors were dispersed in CS-PCL copolymer and electrospuned to form SrAl2O4: Eu(2+), Dy(3+)/CS-PCL scaffolds. The aforementioned photo -luminescence-scaffolds were studied for their optical, mechanical and morphological characteristics finally the effect of these scaffolds on the mice RPCs cells' proliferation and differentiation was observed. The 30% nanophosphor dispersion scaffold while providing adequate mechanical flexibility and integrity, and exhibiting superior proliferation rates and acceptable differentiation into retinal neural cells (particularly photo receptors retinal) is suggested as a promising choice in retinal tissue repair.

Landfill site selection for municipal solid wastes in mountainous areas with landslide susceptibility.

Several cities across the world are located in mountainous and landslide prone areas. Any landfill siting without considering landslide susceptibility in such regions may impose additional environmental adversity. This study was aimed to propose a practical method for selecting waste disposal site that accounts for landslide exposure. The proposed method was applied to a city which is highly proneness to landslide due to its geology, morphology, and climatic conditions. First, information on the previously occurred landslides of the region was collected. Based on this information, proper landslide causative factors were selected and their thematic maps were prepared. Factors' classes were then standardized in 0-1 domain, and thematic layers were weighted by using analytical hierarchy process (AHP). The landslide susceptibility map was prepared afterwards. Unsuitable areas for landfill location were masked in GIS environment by Boolean method, retaining sufficient areas for further evaluation. Nine remaining alternatives were selected through comprehensive field visits and were ranked by using AHP. Consequently, 17 factors in three environmental, economical, and social perspectives were employed. Sensitivity analyses were performed to assess the stability of the alternatives ranking with respect to variations in criterion weights. Based on the obtained landslide susceptible map, nearly 36 % of the entire region is proneness to landslide. The prepared Boolean map indicates that potential areas for landfill construction cover 11 % of the whole region. The results further indicated that if landslide susceptible areas are not considered in landfill site selection, the potential landfill sites would become more than twice. It can be concluded that if any of these landslide prone sites are selected for landfilling, further environmental disaster would be terminated in the future. It can be further concluded that the proposed method could reasonably well be adjusted to consider landslide exposure when siting a solid waste landfill.

An integrated multi criteria approach for landfill siting in a conflicting environmental, economical and socio-cultural area.

Landfill site selection is a complicated multi criteria land use planning that should convince all related stakeholders with different insights. This paper addresses an integrating approach for landfill siting based on conflicting opinions among environmental, economical and socio-cultural expertise. In order to gain optimized siting decision, the issue was investigated in different viewpoints. At first step based on opinion sampling and questionnaire results of 35 experts familiar with local situations, the national environmental legislations and international practices, 13 constraints and 15 factors were built in hierarchical structure. Factors divided into three environmental, economical and socio-cultural groups. In the next step, the GIS-database was developed based on the designated criteria. In the third stage, the criteria standardization and criteria weighting were accomplished. The relative importance weights of criteria and subcriteria were estimated, respectively, using analytical hierarchy process and rank ordering methods based on different experts opinions. Thereafter, by using simple additive weighting method, the suitability maps for landfill siting in Marvdasht, Iran, was evaluated in environmental, economical and socio-cultural visions. The importance of each group of criteria in its own vision was assigned to be higher than two other groups. In the fourth stage, the final suitability map was obtained after crossing three resulted maps in different visions and reported in five suitability classes for landfill construction. This map indicated that almost 1224 ha of the study area can be considered as best suitable class for landfill siting considering all visions. In the last stage, a comprehensive field visit was performed to verify the selected site obtained from the proposed model. This field inspection has confirmed the proposed integrating approach for the landfill siting.

A gaussian model for substrates of entangled cross-linked poly(ethylene glycol) in biomedical applications.

Cells usually spread on a synthetic substrate through bonds between receptors and chemical groups on the substrate (ligands). Therefore, it is valuable to study the effects of the average number density of these chemical groups and the average distance between them to model and predict the cell behavior. Poly(ethylene glycol) [PEG] modified with peptide groups has been used widely in biomedical applications as a substrate material. In this study, a coarse-grained model is proposed for PEG to predict the average number density of ligands and the average distance between them. Molecular information such as initial molecular weight distribution, average molecular weight between cross-links, and average molecular weight between entanglements is used as input parameters. Based on simulation results, it is concluded that both entanglement and cross-link densities are required to create a network structure. The results suggest that an average initial molecular weight 2-3 times the average molecular weight between entanglements and a moderate cross-link density are sufficient to create a closed network structure with a high ligand density and a small average distance between them.