Osteoblast cell response to β-tricalcium phosphate scaffolds with controlled architecture in flow perfusion culture system

Porous β-tricalcium phosphate (β-TCP) scaffolds with controlled architecture and improved mechanical properties were fabricated by combining the gel-casting and rapid prototyping techniques. The pore morphology, size, and distribution of the β-TCP scaffolds were characterized using a scanning electron microscope. The porosity of the resulting scaffolds with pore size range from 300 to 500 μm was 46%. The average compressive strength was 16.1 MPa. X-ray diffraction was used to determine the crystal structure and chemical composition of scaffolds. The result indicated that the sintering process has not changed the composition of β-TCP. Flow perfusion culture system was developed in our lab to improve mass transfer for seeded cells. For scaffold/cell constructs cultured under flow perfusion for 4, 8, and 16 days, there was greater scaffold cellularity and alkaline phosphatase activity compared with static culture condition. These results indicated that flow perfusion culture system had evident effects on osteoblast viability and functions in vitro.     

Static and dynamic cultivation of bone marrow stromal cells on biphasic calcium phosphate scaffolds derived from an indirect rapid prototyping technique

The adequate regeneration of large bone defects is still a major problem in orthopaedic surgery. Synthetic bone substitute materials have to be biocompatible, biodegradable, osteoconductive and processable into macroporous scaffolds tailored to the patient specific defect. Hydroxyapatite (HA) and tricalcium phosphate (TCP) as well as mixtures of both phases, biphasic calcium phosphate ceramics (BCP), meet all these requirements and are considered to be optimal synthetic bone substitute materials. Rapid prototyping (RP) can be applied to manufacture scaffolds, meeting the criteria required to ensure bone ingrowth such as high porosity and defined pore characteristics. Such scaffolds can be used for bone tissue engineering (BTE), a concept based on the cultivation of osteogenic cells on osteoconductive scaffolds. In this study, scaffolds with interconnecting macroporosity were manufactured from HA, TCP and BCP (60 wt% HA) using an indirect rapid prototyping technique involving wax ink-jet printing. ST-2 bone marrow stromal cells (BMSCs) were seeded onto the scaffolds and cultivated for 17 days under either static or dynamic culture conditions and osteogenic stimulation. While cell number within the scaffold pore system decreased in case of static conditions, dynamic cultivation allowed homogeneous cell growth even within deep pores of large (1,440 mm³) scaffolds. Osteogenic cell differentiation was most advanced on BCP scaffolds in both culture systems, while cells cultured under perfusion conditions were generally more differentiated after 17 days. Therefore, scaffolds manufactured from BCP ceramic and seeded with BMSCs using a dynamic culture system are the method of choice for bone tissue engineering.     

Stem cell engineered bone with calcium-phosphate coated porous titanium scaffold or silicon hydroxyapatite granules for revision total joint arthroplasty

Aseptic loosening in total joint replacements (TJRs) is mainly caused by osteolysis which leads to a reduction of the bone stock necessary for implant fixation in revision TJRs. Our aim was to develop bone tissue-engineered constructs based on scaffolds of clinical relevance in revision TJRs to reconstitute the bone stock at revision operations by using a perfusion bioreactor system (PBRS). The hypothesis was that a PBRS will enhance mesenchymal stem cells (MSCs) proliferation and osteogenic differentiation and will provide an even distribution of MSCs throughout the scaffolds when compared to static cultures. A PBRS was designed and implemented. Scaffolds, silicon substituted hydroxyapatite granules and calcium-phosphate coated porous TiAl6V4 cylinders, were seeded with MSCs and cultured either in static conditions or in the PBRS at 0.75 mL/min. Statistically significant increased cell proliferation and alkaline phosphatase activity was found in samples cultured in the PBRS. Histology revealed a more even cell distribution in the perfused constructs. SEM showed that cells arranged in sheets. Long cytoplasmic processes attached the cells to the scaffolds. We conclude that a novel tissue engineering approach to address the issue of poor bone stock at revision operations is feasible by using a PBRS.     

The growth of stem cells within β-TCP scaffolds in a fluid-dynamic environment

A three-dimensional dynamic perfusion system was developed to provide mass transport and nutrient supply to permit the cell proliferation during the long-term culture inside a β-tricalcium phosphate (β-TCP) scaffold. Also the flow field throughout the scaffold was studied. The porous cylindrical scaffold with a central channel was seeded with the sheep mesenchymal stem cells (MSCs). Then the cell-seeded scaffolds were continuously perfused with the complete α-MEM medium by a peristaltic pump for 7, 14 and 28 days, respectively. Histological study showed that the cell proliferation rates were different throughout the whole scaffolds and the different cell coverage was shown in different positions of the scaffold. Unoccupied spaces were found in many macropores. A computational fluid dynamics (CFD) modeling was used to simulate the flow conditions within perfused cell-seeded scaffolds to give an insight into the mechanisms of these cell growth phenomena. Relating the simulation results to perfusion experiments, the even fluid velocity (approximately 0.52 mm/s) and shear stress (approximately 0.0055 Pa) were found to correspond to increased cell proliferation within the cell–scaffold constructs. Flow speeds were between 0.25 and 0.75 mm/s and shear stresses were between 0.003 and 0.008 Pa in approximately 75% of the regions. This method exhibits novel capabilities to compare the results obtained for different perfusion rates or different scaffold microarchitectures. It may allow specific fluid velocities and shear stresses to be determined to optimize the perfusion flow rate, porous scaffold architecture and distribution of in vitro tissue growth.     

Effect of oxidized low-density lipoprotein concentration polarization on human smooth muscle cells' proliferation,cycle, apoptosis and oxidized low-density lipoprotein uptake

To clarify the effect of concentration polarization of oxidative modification of low-density lipoproteins (ox-LDLs) on human smooth muscle cells (SMCs), the proliferation, ox-LDL uptake and apoptosis with SMCs cultured on permeable (the permeable group) or non-permeable membranes (the non-permeable group) were analysed by 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay, spectrofluorometry and flow cytometry using a parallel-plate flow chamber technique. The concentration polarization of ox-LDLs at the surface of the cultured cell monolayer was assessed by confocal laser microscopy. The results showed that concentration polarization of ox-LDLs could indeed occur at the cultured cell monolayer surface of the permeable group, leading to an enhanced wall concentration of ox-LDLs that was over 15 per cent higher than the bulk concentration of the perfusion solution at a pressure of 100 mmHg. When concentration of ox-LDLs in the perfusion solution was less than or equal to 100 µg ml^–1, SMCs'' proliferation was induced, while cell apoptosis was induced when its concentration was above 150 µg ml^–1. The uptake of ox-LDLs by the cultured cells was significantly higher for the permeable group than for the non-permeable group. In addition, the ox-LDL-induced cell death and apoptosis were much more severe in the permeable group than that in the non-permeable group. Therefore, the experimental study suggests that concentration polarization of ox-LDLs plays an adverse role in the vascular system owing to its toxicity to vascular cells, in turn enhance ox-LDL infiltration into the arterial wall and accelerate SMC apoptosis.     

3-D Nanofibrous electrospun multilayered construct is an alternative ECM mimicking scaffold

Extra cellular matrix (ECM) is a natural cell environment, possesses complicated nano- and macro- architecture. Mimicking this three-dimensional (3-D) web is a challenge in the modern tissue engineering. This study examined the application of a novel 3-D construct, produced by multilayered organization of electrospun nanofiber membranes, for human bone marrow-derived mesenchymal stem cells (hMSCs) support. The hMSCs were seeded on an electrospun scaffold composed of poly ε-caproloactone (PCL) and collagen (COL) (1:1), and cultured in a dynamic flow bioreactor prior to in vivo implantation. Cell viability after seeding was analyzed by AlamarBlue™ Assay. At the various stages of experiment, cell morphology was examined by histology, scanning electron microscopy (SEM) and confocal microscopy. Results: A porous 3-D network of randomly oriented nanofibers appeared to support cell attachment in a way similar to traditionally used tissue culture polysterene plate. The following 6 week culture process of the tested construct in the dynamic flow system led to massive cell proliferation with even distribution inside the scaffold. Subcutaneous implantation of the cultured construct into nude mice demonstrated good integration with the surrounding tissues and neovascularization. Conclusion: The combination of electrospinning technology with multilayer technique resulted in the novel 3-D nanofiber multilayered construct, able to contain efficient cell mass necessary for a successful in vivo grafting. The success of this approach with undifferentiated cells implies the possibility of its application as a platform for development of constructs with cells directed into various tissue types.     

Expansion and preservation of multipotentiality of rabbit bone-marrow derived mesenchymal stem cells in dextran-based microcarrier spin culture

The use of mesenchymal stem cells (MSCs) in tissue repair and regeneration despite their multipotentiality has been limited by their cell source quantity and decelerating proliferative yield efficiency. A study was thus undertaken to determine the feasibility of using microcarrier beads in spinner flask cultures for MSCs expansion and compared to that of conventional monolayer cultures and static microcarrier cultures. Isolation and characterization of bone marrow derived MSCs were conducted from six adult New Zealand white rabbits. Analysis of cell morphology on microcarriers and culture plates at different time points (D0, D3, D10, D14) during cell culture were performed using scanning electron microscopy and bright field microscopy. Cell proliferation rates and cell number were measured over a period of 14 days, respectively followed by post-expansion characterization. MTT proliferation assay demonstrated a 3.20 fold increase in cell proliferation rates in MSCs cultured on microcarriers in spinner flask as compared to monolayer cultures (p < 0.05). Cell counts at day 14 were higher in those seeded on stirred microcarrier cultures (6.24 ± 0.0420 cells/ml) × 10⁵ as compared to monolayer cultures (0.22 ± 0.004 cells/ml) × 10⁵ and static microcarrier cultures (0.20 ± 0.002 cells/ml) × 10⁵. Scanning electron microscopy demonstrated an increase in cell colonization of the cells on the microcarriers in stirred cultures. Bead-expanded MSCs were successfully differentiated into osteogenic and chondrogenic lineages. This system offers an improved and efficient alternative for culturing MSCs with preservation to their phenotype and multipotentiality.     

Micro-computed tomography (μ -CT) as a potential tool to assess the effect of dynamic coating routes on the formation of biomimetic apatite layers on 3D-plotted biodegradable polymeric scaffolds

This work studies the influence of dynamic biomimetic coating procedures on the growth of bone-like apatite layers at the surface of starch/polycaprolactone (SPCL) scaffolds produced by a 3D-plotting technology. These systems are newly proposed for bone Tissue Engineering applications. After generating stable apatite layers through a sodium silicate-based biomimetic methodology the scaffolds were immersed in Simulated Body Fluid solutions (SBF) under static, agitation and circulating flow perfusion conditions, for different time periods. Besides the typical characterization techniques, Micro-Computed Tomography analysis (μ -CT) was used to assess scaffold porosity and as a new tool for mapping apatite content. 2D histomorphometric analysis was performed and 3D virtual models were created using specific softwares for CT reconstruction. By the proposed biomimetic routes apatite layers were produced covering the interior of the scaffolds, without compromising their overall morphology and interconnectivity. Dynamic conditions allowed for the production of thicker apatite layers as consequence of higher mineralizing rates, when comparing with static conditions. μ -CT analysis clearly demonstrated that flow perfusion was the most effective condition in order to obtain well-defined apatite layers in the inner parts of the scaffolds. Together with SEM, this technique was a useful complementary tool for assessing the apatite content in a non-destructive way.     

Investigation of in vitro bone cell adhesion and proliferation on Ti using direct current stimulation

Our objective was to establish an in vitro cell culture protocol to improve bone cell attachment and proliferation on Ti substrate using direct current stimulation. For this purpose, a custom made electrical stimulator was developed and a varying range of direct currents, from 5 to 25 μA, was used to study the current stimulation effect on bone cells cultured on conducting Ti samples in vitro. Cell–material interaction was studied for a maximum of 5 days by culturing with human fetal osteoblast cells (hFOB). The direct current was applied in every 8 h time interval and the duration of electrical stimulation was kept constant at 15 min for all cases. In vitro results showed that direct current stimulation significantly favored bone cell attachment and proliferation in comparison to nonstimulated Ti surface. Immunochemistry and confocal microscopy results confirmed that the cell adhesion was most pronounced on 25 μA direct current stimulated Ti surfaces as hFOB cells expressed higher vinculin protein with increasing amount of direct current. Furthermore, MTT assay results established that cells grew 30% higher in number under 25 μA electrical stimulation as compared to nonstimulated Ti surface after 5 days of culture period. In this work we have successfully established a simple and cost effective in vitro protocol offering easy and rapid analysis of bone cell–material interaction which can be used in promotion of bone cell attachment and growth on Ti substrate using direct current electrical stimulation in an in vitro model.     

Evaluation of alginate hydrogels under in vivo–like bioreactor conditions for cartilage tissue engineering

Alginate hydrogels in forms of discs and packed beds of microbeads (~800 μm) were tested in a novel bioreactor at 10% strain using two regimes: at a loading rate of 337.5 μm/s and at sequential increments of 50 μm displacement every 30 min. Compressive strength increased with the increase in alginate concentration (1.5 vs. 2% w/w) and the content of guluronic residues (38.5 vs. 67%). Packed beds of microbeads exhibited significantly higher (~1.5–3.4 fold) compression moduli than the respective discs indicating the effects of gel form and entrapped water. Short-term cultivation of microbeads with immobilized bovine calf chondrocytes (1.5% w/w, 33 × 10⁶ cells/ml) under biomimetic conditions (dynamic compression: 1 h on/1 h off, 0.42 Hz, 10% strain) resulted in cell proliferation and bed compaction, so that the compression modulus slightly increased. Thus, the novel bioreactor demonstrated advantages in evaluation of biomaterial properties and cell-biomaterial interactions under in vivo–like settings.     

General aviation accidents related to exceedance of airplane weight/center of gravity limits

BackgroundObesity, affects a third of the US population and its corollary occupant weight adversely impacts safe flight operations. Increased aircraft weight results in longer takeoff/landing distances, degraded climb gradients and airframe failure may occur in turbulence. In this study, the rate, temporal changes, and lethality of accidents in piston-powered, general aviation aircraft related to exceeding the maximum aircraft weight/center of gravity (CG) limits were determined.MethodsNation-wide person body mass were from the National Health and Nutrition Examination Survey. The NTSB database was used to identify accidents related to operation of aircraft outside of their weight/CG envelope. Statistical analyses employed T-tests, proportion tests and a Poisson distribution.ResultsWhile the average body mass climbed steadily (p < 0.001) between 1999 and 2014 the rate of accidents related to exceedance of the weight/CG limits did not change (p = 0.072). However, 57% were fatal, higher (p < 0.001) than the 21% for mishaps attributed to other causes/factors. The majority (77%) of accidents were due to an overloaded aircraft operating within its CG limits. As to the phase of flight, accidents during takeoff and those occurring enroute carried the lowest (50%) and highest (85%) proportion of fatal accidents respectively.ConclusionWhile the rate of general aviation accidents related to operating an aircraft outside of its weight/CG envelope has not increased over the past 15 years, these types of accidents carry a high risk of fatality. Airmen should be educated as to such risks and to dispel the notion held by some that flights may be safely conducted with an overloaded aircraft within its CG limits.     

Impact of Scaffold Micro and Macro Architecture on Schwann Cell Proliferation under Dynamic Conditions in a Rotating Wall Vessel Bioreactor

Over the last decade tissue engineering has emerged as a powerful alternative to regenerate lost tissues owing to trauma or tumor. Evidence shows that Schwann cell containing scaffolds have improved performance in vivo as compared to scaffolds that depend on cellularization post implantation. However, owing to limited supply of cells from the patients themselves, several approaches have been taken to enhance cell proliferation rates to produce complete and uniform cellularization of scaffolds. The most common approach is the application of a bioreactor to enhance cell proliferation rate and therefore reduce the time needed to obtain sufficiently significant number of glial cells, prior to implantation.In this study, we show the application of a rotating wall bioreactor system for studying Schwann cell proliferation on nanofibrous spiral shaped scaffolds, prepared by solvent casting and salt leaching techniques. The scaffolds were fabricated from polycaprolactone (PCL), which has ideal mechanical properties and upon degradation does not produce acidic byproducts. The spiral scaffolds were coated with aligned or random nanofibers, produced by electrospinning, to provide a substrate that mimics the native extracellular matrix and the essential contact guidance cues.At the 4 day time point, an enhanced rate of cell proliferation was observed on the open structured nanofibrous spiral scaffolds in a rotating wall bioreactor, as compared to static culture conditions. However, the cell proliferation rate on the other contemporary scaffolds architectures such as the tubular and cylindrical scaffolds show reduced cell proliferation in the bioreactor as compared to static conditions, at the same time point. Moreover, the rotating wall bioreactor does not alter the orientation or the phenotype of the Schwann cells on the aligned nanofiber containing scaffolds, wherein, the cells remain aligned along the length of the scaffolds. Therefore, these open structured spiral scaffolds pre-cultured with Schwann cells, in bioreactors could potentially shorten the time needed for grafts for peripheral nerve regeneration.     

Metabolic and histological analysis of mesenchymal stem cells grown in 3-D hyaluronan-based scaffolds

Sheep mesenchymal stem cells (MSCs) were isolated and expanded using the principle of plastic adherence. Their identity as progenitor cells was confirmed by induction along the osteoblastic lineage using osteogenic supplements and observation of calcific deposits by von Kossa staining. MSCs were seeded onto two types of hyaluronan-based cylindrical scaffolds in high concentrations and cultured for varying time points up to three weeks. Culture medium was supplied using the following conditions: statically, on a shaker, by stirring with a magnetic stirrer or by perfusion in a tubular flow circuit. Total cell metabolism was assessed by MTT assay and the quality of cell coverage and matrix formation observed by SEM and histological analysis of thin sections of the constructs. Perfusion culture was established as the most appropriate culturing conditions, with cell metabolism increasing by approximately 300% over three weeks. The coverage of the scaffold surface was very good and the deposition of collagenous matrix was superior in these conditions compared to the, static and other dynamic culture conditions.     

Electrical and optical properties of Al-doped ZnO films deposited by hollow cathode gas flow sputtering

Al-doped ZnO (AZO) films were deposited on glass by hollow cathode gas flow sputtering using Zn-Al alloy targets. Sputtering power for all the depositions was fixed at 1500 W. Resistivities of 0.81-1.1 × 10^− 3 Ω cm were obtained for AZO films deposited at room temperature with an O₂ flow from 38 to 50 standard cubic centimetre/minute (SCCM), while static deposition rates were almost constant at 270-300 nm/min. On the other hand, lower resistivities of 5.2-6.4 × 10^− 4 Ω cm were obtained for AZO films deposited at 200 °C with an O₂ flow from 25 to 50 SCCM, while the static deposition rates were almost constant at 200-220 nm/min. Average transmittances in the visible light region were above 80% for both sets of films.     

Preparation and characterization of chitosan/cashew gum beads loaded with Lippia sidoides essential oil

Beads based on chitosan (CH) and cashew gum (CG), were prepared and loaded with an essential oil with larvicide activity (Lippia sidoides – Ls). CH and CH–CG beads were characterized by scanning electron microscopy (SEM), infrared and UV–VIS spectroscopy, thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), as well as, regarding their larvicide loading, swelling, in vitro and in vivo release kinetics. The oil encapsulation was evidenced by FTIR analysis and LS loading ranges from 2.4% to 4.4%. CH beads duly showed swelling degree (Q) values from 4.0 to 6.7, reaching equilibrium after 30 min, whereas crosslinked CH–CG beads showed lower swelling values, from 0.4 to 3.8, exhibiting a longer equilibrium time. Liquid transport parameters have revealed diffusion coefficient for CH–CG beads, as low as 2 × 10^? 15 m²/s. TGA and DSC revealed that CH:CG crosslinked beads are more thermally stable than CH beads. In vitro release follows a non-Fickian diffusion profile for both bead types, however, and a prolonged release being achieved only after beads crosslinking. In vivo release showed that both CH and CH–CG presented a prolonged larvicide effect. These aforesaid results, indicate that CH–CG beads loaded with LS are efficient for A. aegypti larval control.     

Human-like collagen/nano-hydroxyapatite scaffolds for the culture of chondrocytes

Three dimensional (3D) biodegradable porous scaffolds play a key role in cartilage tissue repair. Freeze-drying and cross-linking techniques were used to fabricate a 3D composite scaffold that combined the excellent biological characteristics of human-like collagen (HLC) and the outstanding mechanical properties of nano-hydroxyapatite (nHA). The scaffolds were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and compression tests, using Relive® Artificial Bone (RAB) scaffolds as a control. HLC/nHA scaffolds displayed homogeneous interconnected macroporous structure and could withstand a compression stress of 2.67 ± 0.37 MPa, which was higher than that of the control group. Rabbit chondrocytes were seeded on the composite porous scaffolds and cultured for 21 days. Cell/scaffold constructs were examined using SEM, histological procedures, and biochemical assays for cell proliferation and the production of glycosaminoglycans (GAGs). The results indicated that HLC/nHA porous scaffolds were capable of encouraging cell adhesion, homogeneous distribution and abundant GAG synthesis, and maintaining natural chondrocyte morphology compared to RAB scaffolds. In conclusion, the presented data warrants the further exploration of HLC/nHA scaffolds as a potential biomimetic platform for chondrocytes in cartilage tissue engineering.     

Integrating an anaerobic Bio-nest and an aerobic EMMC process as pretreatment of dairy wastewater for reuse: a pilot plant study

A large dairy farm located on the island of Oahu, Hawaii was the site for an investigation for the potential integration of the existing facultative lagoon system with a cost effective pretreatment unit process. Based on the results from a laboratory study, a pilot plant was installed with two anaerobic bioreactors (10 m3 each) and one aerobic reactor (3.8 m3). Two layers of media “Bio-nest,” providing a void volume of 98%, were placed into each anaerobic bioreactor with 19% space-based on the bioreactor water volume. For better performance and reduction of shock-load, the equalization/settling tank was employed prior to the first anaerobic Bio-nest reactor. The intermediate holding tank settled effluent suspended solids from the Bio-nest reactor and adjusted the loading rate in order to improve the performance of the aerobic EMMC (entrapped mixed microbial cell) bioreactors. Based on the start-up operation of the Bio-nest system at an organic loading rate of about 1.5 g TCOD/l/day, the production rate of biogas from the first and second Bio-nest reactors was 0.64 and 0.15 l/l/day, respectively. This indicates that the anaerobic degradation of organics occurs mainly in the first Bio-nest reactor due to the low loading rate. The removal efficiency from the Bio-nest system shows TCOD removal of about 70%. The EMMC process provided further treatment to achieve a removal efficiency of TCOD at about 50% and a TN of about 35%. The cost for these pretreatments in order to be integrated with the existing lagoon system is US $1.1 per 1,000 gallons (3.8 m3) for dairy wastewater and $1.1 per 1,000 gallons (3.8 m3) for dairy wastewater and 91 for each ton of TCOD removal. This integration system provides a sustainable improvement of environment and agricultural production.     

Superelastic anisotropy characteristics of columnar-grained Cu–Al–Mn shape memory alloys and its potential applications

The columnar-grained (CG) Cu–Al–Mn shape memory alloy samples possess a strong < 001>-oriented texture along the solidification direction (SD) and straight low-energy grain boundaries fabricated by unidirectional solidification technique. When the angle between tensile direction and the SD ranged from 0° to 90° at the tensile tests, the superelasticity of samples changed in a “V” shape and showed a large anisotropy. Meanwhile, the martensite transformation critical stress of the CG Cu–Al–Mn samples increased from 258.5 MPa for 0° to 521.9 MPa for 45°, and then decreased to 324.3 MPa. The large anisotropy of the superelasticity was attributed to the combined effects of grain orientation and grain boundaries, wherein the influence of the grain boundaries had an obvious dependence on orientation. The potential applications of CG Cu–Al–Mn alloys as anisotropic shock isolators and dampers in high rise buildings and precision instruments were also proposed.     

Deformation behavior and microstructure evolution in multistage hot working of TA15 titanium alloy: on the role of recrystallization

Interrupted compression tests of TA15 titanium alloy with initially equiaxed microstructure were carried out at deformation temperatures between 1173 to 1273 K and strain rates between 0.001 to 0.1 s⁻¹ to investigate the deformation behavior and microstructure evolution under multistage deformation. The TA15 alloy exhibits significant flow softening in both β and (α + β) working. It is found that the flow softening relates to dynamic recrystallization of β phases under current experimental conditions. In multistage β working, metadynamic recrystallization is the main softening mechanism during inter-pass holding. The grain refinement by metadynamic recrystallization leads to the decrease in peak stress upon reloading. In multistage (α + β) working, static recrystallization is the main softening mechanism during inter-pass holding. The static recrystallization kinetics increases with temperature and strain rate. The inter-pass holding has little influence on the morphology of the primary α phases. The β grain size is determined by spacing of primary α phases, which is more affected by working temperature but less dependent on strain rate and inter-pass holding time.     

Effect of Parameters on Treatment of Industrial Effluents Using Inverse Fluidized Bed Bioreactor: Statistical Analysis

Phenolic wastewater (effluents from steel plant) was treated in an inverse fluidized bed (IFB) bioreactor. Effects of different system parameters (viz. residence time (t), settled bed to bioreactor volume ratio (Vb/Vr), water flow rate, static bed height, and gas flow rate) on removal of chemical oxygen demand (COD) were studied to determine the optimum conditions. Optimal conditions corresponding to the largest COD removal were obtained at Vb/Vr ratio of 0.55 and gas flow of 40 LPH with time period greater than 62 h. COD removal was also measured with the addition of mineral salts in the wastewater. The results thus obtained with and without the addition of mineral salt to the wastewater were compared. Addition of nutrient salts to the wastewater yielded better percentage of COD removal. The effect of different types of salts on COD removal was also studied with respect to the biomass growth. Experiments were also carried out by continuous culturing of biomass samples. Conversions greater than 68% and 84% were achieved with batch and continuous mode operation respectively, implying that IFB bioreactor can be used successfully for the treatment of industrial effluents.