Jilska M. Perera

The Influence of Extracellular Matrix on the Generation of Vascularized, Engineered, Transplantable Tissue

Abstract: In a recently described model for tissue engineering, an arteriovenous loop comprising the femoral artery and vein with interposed vein graft is fabricated in the groin of an adult male rat, placed inside a polycarbonate chamber, and incubated subcutaneously. New vascularized granulation tissue will generate on this loop for up to 12 weeks. In the study described in this paper three different extracellular matrices were investigated for their ability to accelerate the amount of tissue generated compared with a no‐matrix control. Poly‐d,l‐lactic‐co‐glycolic acid (PLGA) produced the maximal weight of new tissue and vascularization and this peaked at two weeks, but regressed by four weeks. Matrigel was next best. It peaked at four weeks but by eight weeks it also had regressed. Fibrin (20 and 80 mg/ml), by contrast, did not integrate with the generating vascularized tissue and produced less weight and volume of tissue than controls without matrix. The limiting factors to growth appear to be the chamber size and the capacity of the neotissue to integrate with the matrix. Once the sides of the chamber are reached or tissue fails to integrate, encapsulation and regression follow. The intrinsic position of the blood supply within the neotissue has many advantages for tissue and organ engineering, such as ability to seed the construct with stem cells and microsurgically transfer new tissue to another site within the individual. In conclusion, this study has found that PLGA and Matrigel are the best matrices for the rapid growth of new vascularized tissue suitable for replantation or transplantation.

Increasing the volume of vascularized tissue formation in engineered constructs: an experimental study in rats.

The authors have previously described a model of in vivo tissue generation based on an implanted, microsurgically created vessel loop in a plastic chamber (volume, 0.45 ml) containing a poly(DL-lactic-co-glycolic acid) (PLGA) scaffold. Tissue grew spontaneously in association with an intense angiogenic sprouting from the loop and almost filled the chamber, resulting in a mean amount of tissue in chambers of 0.23 g with no added matrix scaffold and 0.33 g of tissue in PLGA-filled chambers after 4 weeks of incubation. The aim of the present study was to investigate whether a greater volume of tissue could be generated when the same-size vessel loop was inserted into a larger (1.9 ml) chamber. In four groups of five rats, an arteriovenous shunt sandwiched between two disks of PLGA, used as a scaffold for structural support, was placed inside a large polycarbonate growth chamber. Tissue and PLGA weight and volume, as well as histological characteristics of the newly formed tissue, were assessed at 2, 4, 6, and 8 weeks. Tissue weight and volume showed a strong linear correlation. Tissue weight increased progressively from 0.13 +/- 0.04 g at 2 weeks to 0.57 +/- 0.06 g at 6 weeks (p < 0.0005). PLGA weight decreased progressively from 0.89 +/- 0.07 g at 2 weeks to 0.20 +/- 0.09 g at 8 weeks (p < 0.0005). Histological examination of the specimens confirmed increased tissue growth and maturation over time. It is concluded that larger quantities of tissue can be grown over a longer period of time by using larger-size growth chambers.

Solid phase extraction of zinc(II) using a PVC-based polymer inclusion membrane with di(2-ethylhexyl)phosphoric acid (D2EHPA) as the carrier

A polymer inclusion membrane (PIM) is reported consisting of 45% (m/m) di(2-ethylhexyl)phosphoric acid (D2EHPA) immobilized in poly(vinyl chloride) (PVC) for use as a solid phase absorbent for selectively extracting Zn(II) from aqueous solutions in the presence of Cd(II), Co(II), Cu(II), Ni(II) and Fe(II). Interference from Fe(III) in the sample is eliminated by precipitation with orthophosphate prior to the extraction of Zn(II). Studies using a dual compartment transport cell have shown that the Zn(II) flux (2.58 x 10(-6)mol m(-2)s(-1)) is comparable to that observed for supported liquid membranes. The stoichiometry of the extracted complex is shown to be ZnR(2).HR, where R is the D2EHPA anion.

Investigation of AOT reverse microemulsions in supercritical carbon dioxide

Abstract Bis(2-ethylhexyl) sodium sulphosuccinate (AOT) was successfully solubilised in supercritical carbon dioxide (scCO2), with ethanol or pentanol as co-solvent. Three molecular spectroscopic probes: methyl orange (MO), 8-hydroxy-1,3,6-pyrenetrisulphonic acid trisodium salt (HPTS), and riboflavin (RF) were used to examine the solubilisation characteristics of the water/scCO2 microemulsions formed with AOT. MO was extracted at various operating conditions, although the wavelength of its solvatochromic absorption maximum was not indicative of bulk water properties. Instead, the spectral results imply that MO may be located at the surfactant/water interface. The highly water-soluble dye HPTS was unable to be extracted into scCO2/AOT/water systems, suggesting that the water in the reverse micelle core was not as polar under supercritical conditions as those at ambient conditions. Finally, RF was extracted into the supercritical phase (40°C, 175 bar) with pentanol co-solvent, with an apparent enhanced uptake compared with the value at 40°C and ambient pressure in bulk water. This appears to be due to the presence of microcrystals dispersed in the supercritical phase.

Mobility of protein through a porous membrane

The electrophoretic mobility of proteins in membrane pores has been investigated experimentally. When the size of the protein is small relative to the pore size, the protein mobility is identical to the free protein mobility. As the pore radius approaches the protein radius the mobility of the protein is significantly reduced. This phenomenon has been explained in terms of electrokinetic theory. Using a method of reflections, and taking into account the effect of the back-flow, an approximation has been developed for the average mobility in a closed system of a spherical particle moving under electrophoresis parallel to the axis of a cylindrical pore. This approximation assumes that the surface potential of the particle is low, and is valid for arbitrary double layer thickness relative to particle size, provided that there is minimal overlap between the double layers at the pore surface and around the particle. It is also predicted that when the protein and the membrane have surface potentials of the same sign, there can be a significant increase in protein mobility for medium-sized pores.

A Study of the Mass Transfer of CO2 through Different Membrane Materials in the Membrane Gas Absorption Process

Abstract The mass transfer of carbon dioxide through hydrophobic membrane materials into aqueous solutions of monoethanolamine has been studied. Microporous polypropylene, polytetrafluoroethylene and polyvinylidene fluoride hollow fiber membranes were compared. Membranes were characterized before and after use and wetting studies showed that the mass transfer resistance increased by 15% for polypropylene after 45 hours. Wetting may be due to membrane degradation as a result of contact with the solvent. This study highlights the need to choose membrane‐solvent systems that utilize a low cost membrane that remains unwetted by the solvent over long periods and when subjected to reasonable solvent‐side pressures.

Synthesis and characterization of hydrophobic zeolite for the treatment of hydrocarbon contaminated ground water

Hydrophobic zeolite was synthesized, modified and characterized for its suitability as a permeable reactive barrier (PRB) material for treatment of hydrocarbons in groundwater. Batch sorption tests were performed along with a number of standard characterization techniques. High and low ionic strength and pH tests were also conducted to determine their impact on hydrocarbon uptake. Further ion exchange tests were conducted to determine the potential for the zeolite to act as both a hydrocarbon capture material and nutrient a delivery system for bioremediation. The zeolite was coated with octadecyltrichlorosilane (C18) to change its surface properties. The results of the surface characterization tests showed that the underlying zeolite structure was largely unaffected by the coating. TGA measurements showed a reactive carbon content of 1-2%. Hydrocarbon (o-xylene and naphthalene) sorption isotherms results compared well with the behaviour of similar materials investigated by other researchers. Ionic strength and pH had little effect on hydrocarbon sorption and the treated zeolite had an ion exchange capacity of 0.3 mequiv./g, indicating it could be utilised as a nutrient source in PRBs. Recycle tests indicated that the zeolite could be used cleaned and reused at least three times without significant reduction in treatment effectiveness.

An attenuated total internal reflectance spectroscopy study of ET(30) at the free oil-water interface

Abstract Visible attenuated total internal reflectance (ATR) spectroscopy has been used to measure the mean solvent microenvironment at free “oil”-water interfaces using the solvatochromic and acid-base properties of 2,6-diphenyl-4-(2,4,6-triphenyl-1-pyridino)phenoxide (E T (30)). From the wavelength maximum of the solvatochromic visible absorption band of E T (30), the effective interfacial dielectric constants of a range of oil-water interfaces have been determined to be ≈8. Titration of the interfacially located probe from high to low pH leads to the creation of a positive electrostatic surface potential which can be largely accounted for using simple Gouy-Chapman theory. Complete agreement, however, is not obtained due to the non-ideal behaviour of E T (30) at the interface as the pH of the bulk aqueous phase is lowered. The relevance of the results obtained to solvent extraction processes are discussed.

Adsorption Behavior of Cadmium(II) and Lead(II) on Mesoporous Silicate MCM-41

Abstract This investigation examines metal ion adsorption on mesoporous silicate, MCM‐41, synthesized from sodium silicate solution and cethyltrimethylammonium bromide (CTAB). MCM‐41 has potential as an adsorbent material, with a regular hexagonal pore structure, large specific surface area, and large pore volume. The MCM‐41 synthesized for this investigation is characterized using powder X‐ray diffraction and nitrogen adsorption and desorption isotherms data. The adsorption behavior for cadmium(II) and lead(II) onto MCM‐41 was studied by contacting the mesoporous silicate with an aqueous solution of metal salts and acetylacetone. Both Cd2+ and Pb2+ were found to quantitatively adsorb onto MCM‐41. The results of this study suggest that MCM‐41 may have applications in the recovery of toxic metals from waste waters.

AOT reverse microemulsions in scCO2 — a further investigation

Abstract The solubility of riboflavin (RF), potassium ferricyanide (K 3 Fe(CN) 6 ) and polar components from yeast extract (YX) in a sodium bis-(2-ethylhexyl) sulfosuccinate (AOT) modified supercritical carbon dioxide (scCO 2 ) system was investigated. The results from RF solubility studies indicate that the microemulsion phase formed by AOT contains a polar core with an effective static dielectric constant of 20±5. Although this core is not indicative of bulk water, the overall improvement in polarity from pure scCO 2 is approximately a factor of 10. K 3 Fe(CN) 6 , a highly ionic compound, could not be solubilised into the modified supercritical fluid, indicating that the core was not sufficiently polar to dissolve highly ionic compounds. Finally, successful extraction of polar compounds from YX demonstrated the potential industrial applications of AOT-modified scCO 2 .