Photoinitiated cross-linking of the biodegradable polyester poly(propylene fumarate). Part II. In vitro degradation

This study investigated the in vitro degradation of both solid PPF networks and porous PPF scaffolds formed by photoinitiated cross-linking of PPF polymer chains. Three formulations of scaffolds of differing porosity and pore size were constructed by varying porogen size and content. The effects of pore size and pore volume on scaffold mass, geometry, porosity, mechanical properties, and water absorption were then examined. Throughout the study, the solid networks and porous scaffolds exhibited continual mass loss and slight change in length. Porogen content appeared to have the greatest effect upon physical degradation. For example, scaffolds initially fabricated with 80 wt % porogen content lost approximately 30% of their initial PPF content after 32 weeks of degradation, whereas scaffolds fabricated with 70 wt % porogen content lost approximately 18% after 32 weeks of degradation. For all scaffold formulations, water absorption capacity, porosity, and compressive modulus were maintained at constant values following porogen leaching. These results indicate the potential of photo-cross-linked PPF scaffolds in tissue engineering applications which require maintenance of scaffold structure, strength, and porosity during the initial stages of degradation.     

Porogen Effect on Structural and Physical Properties of β-TCP Scaffolds for Bone Tissue Regeneration

Scaffolds for bone tissue applications have been an outstanding alternative to repair and regenerate bone tissue defects caused by traumas or illness. There are many methods available to fabricate porous scaffold such as solvent casting, gas bubble, phase separation, electrospinning, particle-leaching, among others. The particle-leaching technique has shown advantages in bone tissue regeneration applications, the main benefit of this technique is related to the porogen particle size and the porogen content in the manufacture of scaffolds. Tricalcium phosphate is one calcium phosphate that presented appropriated characteristic to be used for bone tissue engineering due to the chemical properties similar to the human bones. Scaffolds of tricalcium phosphate β phase were made using sugar particles. The porogen was varied in amounts of 50, 60 and 70 wt.% of two commercial sugars with the remainder of the composition made up of tricalcium phosphate powders. The pore sizes in all the scaffolds were in the range of 90 to 600 μm with an irregular pore morphology and the porosity was in the range of 63 to 77%.     

Sodium alginate/Na(+)-rectorite composite microspheres: Preparation, characterization, and dye adsorption

Sodium alginate/Na(+)-rectorite (SA/Na(+)REC) intercalated nano-composite microspheres were prepared in an inverse suspension system. The effect of the preparation conditions of SA/Na(+)REC composite microspheres on adsorption capacity for Basic Blue 9 was investigated. The structure and morphology were analyzed by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscope (SEM). The results showed that the optimal condition was that the amount of Na(+)-rectorite was 2wt%, the amount of cross-linker was 0.384% and the amount of the initiator was 8%. SEM showed that it is porous products with spherical particulate surface. XRD showed that intercalation is formed between Na(+)-rectorite and sodium alginate. The adsorption capacity of SA/Na(+)REC was investigated in comparison with Na(+)-rectorite and sodium alginate using different cationic dyes. The SA/Na(+)REC composite microspheres showed the highest adsorption capacity. The reason lies in the existence of intercalated sodium alginate. It could enlarge the pore structure of microspheres, facilitating the penetration of macromolecular dyes.     

Pore-Scale Modeling of Pore Structure Effects on P-Wave Scattering Attenuation in Dry Rocks

Underground rocks usually have complex pore system with a variety of pore types and a wide range of pore size. The effects of pore structure on elastic wave attenuation cannot be neglected. We investigated the pore structure effects on P-wave scattering attenuation in dry rocks by pore-scale modeling based on the wave theory and the similarity principle. Our modeling results indicate that pore size, pore shape (such as aspect ratio), and pore density are important factors influencing P-wave scattering attenuation in porous rocks, and can explain the variation of scattering attenuation at the same porosity. From the perspective of scattering attenuation, porous rocks can safely suit to the long wavelength assumption when the ratio of wavelength to pore size is larger than 15. Under the long wavelength condition, the scattering attenuation coefficient increases as a power function as the pore density increases, and it increases exponentially with the increase in aspect ratio. For a certain porosity, rocks with smaller aspect ratio and/or larger pore size have stronger scattering attenuation. When the pore aspect ratio is larger than 0.5, the variation of scattering attenuation at the same porosity is dominantly caused by pore size and almost independent of the pore aspect ratio. These results lay a foundation for pore structure inversion from elastic wave responses in porous rocks.     

Effect of protein adsorption on the transport characteristics of asymmetric ultrafiltration membranes.

The effects of bovine serum albumin adsorption on the transport characteristics of asymmetric poly(ether sulfone) ultrafiltration membranes were determined using polydisperse dextrans with gel permeation chromatography. Actual dextran sieving coefficients were evaluated from observed sieving data for both the clean and preadsorbed membranes using a stagnant film model. The flux dependence of the actual dextran sieving coefficients was used to evaluate the intrinsic membrane hindrance factors for convective (i.e., sieving) and diffusive transport for the different molecular weight dextrans using classical membrane transport theory. Protein adsorption caused a reduction in both dextran sieving and diffusion, with the magnitude of the reduction a function of the dextran molecular weight and pore size. The effects of adsorption on the specific pore area and the membrane porosity were then determined using a recent model for solute transport through asymmetric ultrafiltration membranes. The data indicate that protein adsorption occurs preferentially in the larger membrane pores, causing a greater reduction in solute sieving compared to the membrane hydraulic permeability and porosity than would be predicted on the basis of either a simple pore blockage or pore constriction model.     

Emulsan, a tailorable biopolymer for controlled release

Microsphere hydrogels made with emulsan-alginate were used as carrier for the microencapsulation of blue dextran in order to study the effect of emulsan on the alginate bead stability. Blue dextran release studies indicated an increase of microsphere stability in presence of emulsan, as a coating agent. BSA adsorption by emulsan-alginate microspheres is also enhanced 40% compared to alginate alone. XPS studies confirm the presence of BSA adsorbed on emulsan microsphere surfaces. These results are in agreement with the equilibrium adsorption model of Freundlich. Studies of BSA adsorption using non-equilibrium Lagergren second-order and intraparticle models, are suggesting a complex mechanisms of protein adsorption by chemisorption and intraparticle diffusion. Also, enzymatic release of BSA from emulsan microspheres containing azo-BSA under physiological conditions is suggests the possibility of using microspheres as a depot for pre-proteins of medical interest.     

Cellulose morphology and enzymatic reactivity: A modified solute exclusion technique

An expeditious and accurate simplification of Stone and Scallan's solute exclusion technique was developed, thereby avoiding several sources of experimental error coupled with the determination of cellulose pore volume. Using this method, it is shown that cellulolytic enzymes do not enter into the micropores of five studied celluloses. These results suggestes that hydrolysis occurs initially at the external surface of the fibers. This surface area was calculated with the help of adsorption isotherms of bovine serum albumin. The obtained values for the different samples agree with the microscopically observed cellulose morphology. The correlation obtained by several authors relating cellulose porosity and its digestibility is explained as a consequence of the lower crystallinity and easier fragmentation of the more porous celluloses during hydrolysis. (c) 1994 John Wiley & Sons, Inc.     

Separation of ursodeoxycholic acid from its isomeric mixture using core–shell molecular imprinting polymer

In order to separate ursodeoxycholic acid (UDCA) from its isomeric mixture, the molecular imprinting polymers (MIPs) were synthesized by using core–shell emulsion polymerization. In the porous imprinting polymer, ursodeoxycholic acid was used as imprinting molecule, acrylamide (AM) and α-methacrylic acid (MAA) were functional monomers, and CaCO₃ was used for the porogen in the polymerization to obtain large pore. Characterization of the MIP structure with IR spectra demonstrated the expected MIPs. Through adsorption and selectivity assays, AM as the functional monomer showed better separation efficiency than MAA, and nonspecific and specific adsorption capacities of MIP with AM were 43.52 and 13.93 mg/g, respectively. The separation factor of MIP with AM for UDCA was 2.20. Furthermore, MIP with AM could be applied to separate UDCA from the isomeric mixture by column chromatography successfully.     

担载b-FGF的PLGA微球复合明胶支架的体外释放研究

目的:研究担载碱性成纤维细胞生长因子(b-FGF)微球复合明胶支架的外形特征、孔径、孔隙率及体外释放动力学,以期构建具有缓释功能、高孔隙率的担载细胞因子的新型复合明胶支架。方法:本文利用冷冻相分离法和S/O/W法先将b-FGF水溶液包裹于PLGA微球中,然后埋置于明胶溶液中制备为多孔复合明胶支架。分别对微球的形态和复合明胶支架的基本形态、孔径、孔隙率进行表征,通过Elisa法测定b-FGF在复合明胶支架中的体外释放行为。结果:制备成形态良好的三维复合明胶支架,其孔隙率为82.90%±1.45%,孔径范围为150~300μm,复合明胶支架中b-FGF在体外缓慢释放20余天。结论:担载蛋白微球复合明胶支架不仅满足组织工程支架的要求,还能有效缓释细胞因子,为细胞和组织生长提供良好的微环境,为进一步应用于组织工程领域提供了可能。     

Evaluation of the in vitro degradation of macroporous hydrogels using gravimetry,confined compression testing,and microcomputed tomography

This study investigated the in vitro degradation characteristics of macroporous hydrogels based on poly(propylene fumarate-co-ethylene glycol) (P(PF-co-EG)). Four formulations were fabricated to test the effect of porosity and cross-linking density on the degradation of the resulting macroporous hydrogels. Macroporosity was introduced by the addition of sodium bicarbonate and ascorbic acid, the precursors of the carbon dioxide porogen, in the initiation system for the hydrogel cross-linking. Macroporous hydrogels with porosities of 0.80 +/- 0.03 and 0.89 +/- 0.03 were synthesized by the addition of sodium bicarbonate of concentrations 40 and 80 mg/mL and ascorbic acid of concentrations 0.05 and 0.1 mol/L, respectively. Poly(ethylene glycol) diacrylate (PEG-DA) was utilized as a cross-linker. The molecular weight between cross-links had a significant effect on weight loss after 12 weeks, where samples with M(C) of 1,880 +/- 320 synthesized with a P(PF-co-EG):PEG-DA ratio of 3:1 had a significantly greater mass loss due to degradation than those with M(C) of 1,000 +/- 100 synthesized with a P(PF-co-EG):PEG-DA ratio of 1:1. In contrast, porosity played a minimal role in determining the weight loss. Mechanical testing of the hydrogels under confined compression showed a decrease in compressive modulus over the degradation time for all formulations. In addition, an increase in hydrogel equilibrium water content and pore wall thickness was observed with degradation time, whereas the hydrogel porosity and surface area density remained invariant. The results from microcomputed tomography corroborated with the rest of the measurements and indicated a bulk degradation mechanism of the macroporous hydrogels.     

Investigation of production of dextran and dextransucrase by Leuconostoc mesenteroides immobilized within porous stainless steel

Cells of Leuconostoc mesenteroides were immobilized within porus, stainless-steel (SS) supports and used for dextransucrase (DS) and dextran production. The pore size of the support significantly affected the dextran yields, which were greatest with average pore sizes of 2-5 mum. All immobilized-cell biocatalysts in porous stainless steel produced higher yields than free cells, with the exception of cells confined in submicrometer pores (0.5 mum). Coating supports of larger pore size (40 and 100 mum) with calcium alginate enhanced the cell-loading capacity of the supports and increased dextran and fructose yields in the cell-free broth. Controlled, fed-batch, DS production (activation), as a step preliminary to dextran production, significantly improved the subsequent dextran and fructose yields and shortened the time required to attain the maximum such yields. Scanning electron microscopy (SEM) of immobilized L. mesenteroides in stainless steel shows an irregular pattern of the microorganism inside the pores of the solid supports. Coating the porous solid supports with a cell-free calcium alginate layer led to an increase in the cell density inside the support. Cell growth inside the coated, porous stainless steel had no distinct growth form. (c) 1992 John Wiley & Sons, Inc.     

Production of cycloisomaltooligosaccharides from dextran using enzyme immobilized in multilayers onto porous membranes

Anion-exchange porous hollow-fiber membranes with a thickness of about 1.2 mm and a pore size of about 0.30 microm were used as a supporting matrix to immobilize cycloisomaltooligosaccharide glucanotransferase (CITase). CITase was immobilized to the membrane via anion-exchange adsorption and by subsequent enzymatic cross-linking with transglutaminase, the amount of which ranged from 3 to 110 mg per gram of the membrane. The degree of enzyme multilayer binding was equivalent to 0.3-9.8. Dextran, as the substrate, was converted into seven- to nine-glucose-membered cycloisomaltooligosaccharides (CI-7, -8, and -9) at a maximum yield of 28% in weight at a space velocity of 10 per hour during the permeation of 2.0% (w/w) dextran solution across the CITase-immobilized porous hollow-fiber membrane. The yield of CIs increased with increasing degree of CITase multilayering.     

Potential application of hydrogel-based strong anion-exchange membrane for plasmid DNA purification

The potential application of a hydrogel-based strong anion-exchange (Q) membrane to purify plasmid DNAs was evaluated. The maximum binding capacity of plasmid DNA was estimated to be 12.4 mg/ml of membrane volume with a plasmid recovery yield of ～90%. The effect of the inherent properties of plasmid DNA, membrane adsorbent, and the ionic environment on membrane performance was systematically investigated. Plasmid DNAs with smaller tertiary structure tended to have a better recovery than those with larger tertiary structure. Environmental Scanning Electron Microscopy (ESEM) revealed that the hydrogel structure is more porous on one side of membrane than the other. Membrane pre-treatment significantly improved pore distribution and increased membrane porosity resulting in a better adsorption, recovery, and higher flux. The selection of proper operating pH led to further improvement. The relative contribution of these factors to improve membrane chromatography of plasmid DNAs was analyzed using statistical modeling. It was found that the adsorption of plasmid DNA was mainly affected by the available adsorptive area associated with membrane porosity, whereas the recovery of plasmid DNAs was mainly affected by the environmental pH.     

Ethylcellulose-Based Matrix-Type Microspheres: Influence of Plasticizer RATIO as Pore-Forming Agent

In this study, ethylcellulose (EC)-based microsphere formulations were prepared without and with triethyl citrate (TEC) content of 10% and 30% by water-in-oil emulsion-solvent evaporation technique. Diltiazem hydrochloride (DH) was chosen as a hydrophilic model drug and used at different drug/polymer ratios in the microspheres. The aim of the work was to evaluate the influence of plasticizer ratio on the drug release rate and physicochemical characteristics of EC-based matrix-type microspheres. The resulting microspheres were evaluated for encapsulation efficiency, particle size and size distribution, surface morphology, total pore volume, thermal characteristics, drug release rates, and release mechanism. Results indicated that the physicochemical properties of microspheres were strongly affected by the drug/polymer ratio investigated and the concentration of TEC used in the production technique. The surface morphology and pore volume of microspheres significantly varied based on the plasticizer content in the formulation. DH release rate from EC-based matrix-type microspheres can be controlled by varying the DH to polymer and plasticizer ratios. Glass transition temperature values tended to decrease in conjunction with increasing amounts of TEC. Consequently, the various characteristics of the EC microspheres could be modified based on the plasticized ratio of TEC.     

Effects of dynamic loading on solute transport through the human cartilage endplate

Nutrient and metabolite transport through the cartilage endplate (CEP) is important for maintaining proper disc nutrition, but the mechanisms of solute transport remain unclear. One unresolved issue is the role of dynamic loading. In comparison to static loading, dynamic loading is thought to enhance transport by increasing convection. However, the CEP has a high resistance to fluid flow, which could limit solute convection. Here we measure solute transport through site-matched cadaveric human lumbar CEP tissues under static vs. dynamic loading, and we determine how the degree of transport enhancement from dynamic loading depends on CEP porosity and solute size. We found that dynamic loading significantly increased small and large solute transport through the CEP: on average, dynamic loading increased the transport of sodium fluorescein (376 Da) by a factor of 1.85 ± 0.64 and the transport of a large dextran (4000 Da) by a factor of 4.97 ± 3.05. Importantly, CEP porosity (0.65 ± 0.07; range: 0.47–0.76) strongly influenced the degree of transport enhancement. Specifically, for both solutes, transport enhancement was greater for CEPs with low porosity than for CEPs with high porosity. This is because the CEPs with low porosity were susceptible to larger improvements in fluid flow under dynamic loading. The CEP becomes less porous and less hydrated with aging and as disc degeneration progresses. Together, these findings suggest that as those changes occur, dynamic loading has a greater effect on solute transport through the CEP compared to static loading, and thus may play a larger role in disc nutrition.     

Insulin adsorption into porous charged membranes: Effect of the electrostatic interaction

Insulin adsorption into a series of porous charged membranes was investigated by batch adsorption experiments, and the experimental results were analyzed by the homogeneous diffusion model. The membranes used in this study were prepared by pore‐surface modification of porous poly(acrylonitrile) (PAN) membranes by grafting with weak acidic and basic functional groups. The amount of insulin adsorbed into the membrane was determined from the material balance of insulin. The insulin partition coefficient K between the membrane and solution was estimated from the equilibrium adsorption amount, and the effective diffusion coefficient D was estimated by matching the model with the experimental data as a fitting parameter. The dependence of K and D on the charge properties of the insulin and membrane is observed and discussed. The partition coefficient K increased when the insulin and the membrane carried opposite charges, on the other hand, the effective diffusion coefficient D was reduced. These results indicate that the electrostatic interaction between the insulin and the membranes played an important role in the insulin adsorption. © 2009 American Institute of Chemical Engineers Biotechnol. Prog. 2009     

Alteration of the structural attributes of a compact clay loam soil by growth of a perennial grass crop

Soil aggregate stability, organic matter content, pH, bulk density, strength, and macropore attributes were assessed in order to evaluate the influence of grass root growth in a field sward. The amount of grass grown was varied by varying the quantity of applied nitrogen fertilizer: following one year with a uniform application rate, nitrogen fertilizer was applied over the subsequent three years to a compact soil at zero (N0), moderate (N1) and high (N2) rates. Differences in herbage production were evident in the three years of the contrasting nitrogen treatments. An index of soil aggregate stability increased in response to the increased grass growth promoted by heavier applications of nitrogen, but both bulk density and vane shear strength were unchanged. Binary images of the soil solid and pore space showed that for each treatment the largest volume of macropores occurred close to the surface, particularly in N0 where there was more pore space than in either N1 or N2. Analysis of the pore structure attributes of the binary images revealed further differences between treatments, in particular, at 40–80 mm depth, the soil in treatment N0 had fewer and smaller pores, and greater distance between pores, than the soil in the N1 and N2 treatments. The larger macropore volume in N1 and N2 constituted a major portion of the air-filled porosity when the soil was relatively wet. It was concluded that the root growth in the intensively cropped grassland was conducive to maintenance of a relatively stable and porous soil structure. An attendant increase in soil acidity close to the soil surface was a disadvantage of the larger nitrogen inputs.     

Improvement of Coarse Sand Engineering Properties by Microbially Induced Calcite Precipitation

Abstract

This paper investigates the effectiveness of microbially induced calcite precipitation method in improving the strength and stiffness of coarse sands using treatments based on a four-phase percolation technique. An increase of biochemical treatment cycles was associated with increased deposition of calcium carbonate (CaCO₃) and consequently an increase in compressive strength. Furthermore, the bio-cemented coarse sand retained reasonable porosity and permeability, which should allow dissipation of pore water pressure if required. The results also establish a correlation between the strength gained and stiffness of the bio-cemented coarse sand with the increase in the amount of deposited CaCO₃, initial relative density and dry density. Scanning electron microscopy and electron dispersive spectroscopy analysis indicate that the inter-structure of the bio-cemented coarse sand tend to change in morphology based upon the number of biochemical treatments used.     

Microporous silicon and biosensor development: structural analysis, electrical characterisation and biocapacity evaluation

An investigation of the fabrication of microporous silicon (MPS) layers as a material for the development of an electrolyte insulator semiconductor (EIS) capacitance sensor has been performed. The goal was to create a high surface area substrate for the immobilisation of biorecognition elements. Structural analysis of MPS layers as a function of key etch parameters, namely implant type (p or n), implant dose, hydrofluoric acid (HF) etch concentration and current density has been performed using scanning electron microscopy (SEM). It was possible to image porous layers with average pore diameter as low as 4 nm. n-type silicon samples had larger pore networks than p-type samples and reducing the silicon resistivity led to a reduction in the pores per microm2. It was found that increasing the HF etch concentration reduced the average pore diameter and increased the pores per microm2. Increasing the current density at which the etch was performed has the same effect. Understanding the effect of these parameters allows the MPS layer to be tuned to match specifications for optimum biocapacity. Different MPS layers were electrically characterised using capacitance-voltage and capacitance-frequency sweeps, in order to determine the effect of porosity on increases in surface area. The measured capacitance increased with increasing pores per microm2. p-type silicon with a boron implant in the back of the wafer, which had been etched in 25% HF in ethanol at a current density of 75 mA/cm2 yielded the highest capacitance signal per unit area. The effect of porosity and pore size on the biocapacity of the samples was also determined. For avidin immobilisation, with pores sizes above 5 nm, as the porosity increased the biocapacity increased. MPS fabricated in p-type silicon with a front and back implant etched in 25% HF at a current density of 25 mA/cm2 was used for the capacitance detection of synthetic oligonucleotides.     

Expanded bed adsorption of human serum albumin from very dense Saccharomyces cerevesiae suspensions on fluoride-modified zirconia.

The adsorption of proteins from high cell density yeast suspensions on mixed-mode fluoride-modified zirconia (FmZr) particles (38 to 75 microm, surface area of 29 m(2)/g and density of 2.8 g/cm(3)) was investigated using human serum albumin (HSA) added to Saccharomyces cerevesiae as the model expression host. Because of the high density of the porous zirconia particles, HSA (4 mg/mL) can be adsorbed from a 100 g dry cell weight (DCW)/L yeast suspension in a threefold-expanded bed of FmZr. The expanded bed adsorption of any protein from a suspension containing >50 g DCW/L cells has not been previously reported. The FmZr bed expansion characteristics were well represented by the Richardson-Zaki correlation with a particle terminal velocity of 3.1 mm/s and a bed expansion index of 5.4. Expanded bed hydrodynamics were investigated as a function of bed expansion using residence time distribution studies with sodium nitrite as the tracer. The adsorption of HSA on FmZr exhibited features of multicomponent adsorption due to the presence of dimers. The protein binding capacity at 5% breakthrough decreased from 22 mg HSA/mL settled bed void volume for 20 g DCW/L yeast to 15 mg HSA/mL settled bed void volume for 40 g DCW/L yeast and remained unchanged for the higher yeast concentrations (60 to 100 g DCW/L). However, the batch (or equilibrium) binding capacity decreased monotonically as a function of yeast concentration (20 to 100 g DCW/L) and the binding capacity at 100 g DCW/L yeast was fivefold lower compared with that at 20 g DCW/L yeast. The lower batch binding capacity at high cell concentrations resulted from the adsorption of cells at the surface of the particles restricting access of HSA to the intraparticle surface area. Batch (or equilibrium) and column HSA adsorption results indicated that the adsorption of HSA on FmZr occurred at a time scale that may be much faster than that of yeast cells. The zirconia particles were cleaned of adsorbed HSA and yeast with a total of 1500 to 2000 column volumes (over many cycles) of 0. 25 M NaOH, without any significant effect on the chromatographic performance.