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Wettability interpretation of oxygen plasma modified poly(methyl methacrylate).

Poly(methyl methacrylate) (PMMA) has been modified via a dc pulsed oxygen plasma for different treatment times. The modified surfaces were characterized by X-ray photoelectron spectroscopy (XPS), optical profilometer, zeta potential, and advancing contact angle measurements. The measured advancing contact angles of water decreased considerably as a function of discharge. Several oxygen-based functionalities (carbonyl, carboxyl, carbonate, etc.) were detected by XPS, while zeta potential measurements confirmed an increase in negative charge for the treated PMMA surface. Evaluating the correlation between the concentration of polar chemical species and zeta potential, we found that increase in surface hydrophilicity results from the coeffect due to incorporation of oxygen functional groups and creation of charge states. The electrical double layer (EDL) effect was also considered in contact angle interpretation by introducing an additional surface tension term into Young's equation. We also found that EDL contribution to the solid-liquid interfacial tension is negligible and can be safely ignored for the systems considered here.

Molecular Structure↗

Assessing the role of tryptophan residues in the binding site.

Instead of looking at the interfacial area as a measure of the extent of a protein--protein recognition site, a new procedure has been developed to identify the importance of a specific residue, namely tryptophan, in the binding process. Trp residues which contribute more towards the free energy of binding have their accessible surface area reduced, on complex formation, for both the main-chain and side-chain atoms, whereas for the less important residues the reduction is restricted only to the aromatic ring of the side chain. The two categories of residues are also distinguished by the presence or absence of hydrogen bonds involving the Trp residue in the complex. A comparison of the observed change in the accessible surface area with the value calculated using an analytical expression provides another way of characterizing the Trp residues critical for binding and this has been used to identify such residues involved in binding non-proteinaceous molecules in protein structures.

Amino Acid Sequence↗

Surface engineering of titanium by collagen immobilization. Surface characterization and in vitro and in vivo studies.

Collagen was covalently linked to the surface of Titanium (Ti) by a surface modification process involving deposition of a thin film from hydrocarbon plasma followed by acrylic acid grafting. The composition and properties of surface-modified Ti were investigated by a number of surface sensitive techniques: XPS, ATR-IR, atomic force microscopy and AFM force-separation curves. In vitro tests were performed to check samples cytotoxicity and the behavior of osteoblast-like SaOS-2 cells. In vivo experiments involved 12 weeks implants in rabbit muscle as general biocompatibility assessment and 1-month implants in rabbit bone to evaluate the effect of surface modification on osteointegration rate. Results of XPS measurements show how surface chemistry is affected throughout each step of the surface modification process, finally leading to a complete and homogeneous collagen overlayer on top of the Ti samples. AFM data clearly display the modification of the surface topography and of the surface area of the samples as a consequence of the grafting and coupling process. AFM force-distance curves show that the interfacial structure responds by shrinking or swelling to variations of ionic force of the surrounding aqueous environment, suggesting that the aqueous interface of the biochemically modified Ti samples has enhanced degrees of freedom as compared to the inorganic surface of plain Ti. As to biological evaluations, the biochemically modified Ti samples are safe in terms of cytotoxicity and in vivo biocompatibility assessment. SaOS-2 cells growth rate is lower on collagen modified surfaces, and no significant difference is detected in terms of alkaline phosphatase production as compared to control Ti. Importantly, implants in rabbit femur show a significant increase of bone growth and bone-to-implant contact in the case of the collagen modified samples, confirming that biochemical modifications of Ti surface can enhance the rate of bone healing as compared to plain Ti.

Adsorption↗

Biophysical characterization of the interaction of the beta-lactamase TEM-1 with its protein inhibitor BLIP.

BLIP is a secreted protein from Streptomyces clavuligerus that inhibits a wide range of beta-lactamases. Here we investigate the tight interaction of BLIP, expressed heterologousely in E. coli, with TEM-1. Kinetic and thermodynamic constants were determined using methods with the proteins either in a homogeneous or in a heterogeneous phase. While values of Delta DeltaG(mut-wt) are similar whether measured by fluorescence quench, enzyme inhibition, or surface plasmon resonance, absolute values of DeltaG and kinetic constants vary. Association and dissociation rate constants of 10(5) M-1 s-1 and 10(-)4 s-1, respectively, and a nanomolar affinity were determined for the wild-type proteins. The highest affinity is measured at pH 7.5, with a decreasing association rate constant at higher pH values, and an increasing dissociation rate constant at lower pH values. The marginal effect of salt on the kinetics of binding, as well as the calculated surface potentials, suggests a limited role for electrostatic forces in guiding this reaction. Still, mutations of interfacial residues affect the rate of association significantly, so that an increase in the net negative charge on either protein reduces the association rate constant. We show that simple electrostatic rules can explain this behavior. BLIP inhibits the catalytic activity of TEM-1 by binding its active site. Yet, mutations of active site residues on TEM-1 only have a moderate though cooperative effect on the binding energy. This can be explained in light of the peripheral location of the active site in the interface between the two proteins.

Amino Acid Substitution↗

Preparation and characterization of high-surface-area polymer substrates for microcalorimetry.

The preparation and characterization of high-surface-area polymeric substrates suitable for the microcalorimetry of protein adsorption are described. High-surface-area polystyrene, poly(styrene-co-butyl methacrylate) and poly(styrene-co-allyl alcohol) were prepared by adsorbing polymer from solution onto fumed silica. Verification of adsorption of polystyrene by silica was determined by noting peak shifts of the surface silanol group in the infrared. The amount of polymer adsorbed was determined from adsorption isotherms. The minimum thickness of polystyrene required to mask silicon oxide properties was found to be that thickness at which contact angles became constant, about 35 A. Polymer densities were measured. Water contact angles on each polymer surface indicate that poly(styrene-co-allyl alcohol) has the surface most wettable by water. Polymer-water interfacial energies were estimated from pendant drop results and a harmonic mean equation along with contact angles. Two methods were used to estimate the polar and dispersion components of the three polymers. Both methods predicted polystyrene to have the highest interfacial energy against water, and one method predicted poly(styrene-co-allyl alcohol) to have the lowest. A Wilhelmy plate study verified the change in interfacial properties as a function of contact time with water. A study of the heats of adsorption of lysozyme by each substrate using a modified Tien-Calvet microcalorimeter demonstrated the suitability of the substrates for microcalorimetry.

Adsorption↗

Initial characterization of new bacteria degrading high-molecular weight polycyclic aromatic hydrocarbons isolated from a 2-year enrichment in a two-liquid-phase culture system.

AIMS: To characterize some polycyclic aromatic hydrocarbons (PAH)-degrading microorganisms isolated from an enriched consortium degrading high molecular weight (HMW) PAHs in a two-liquid-phase (TLP) soil slurry bioreactor, and to determine the effect of low molecular weight (LMW) PAH on their growth and HMW PAH-degrading activity. METHODS AND RESULTS: Several microorganisms were isolated from a HMW-PAH (pyrene, chrysene, benzo[a]pyrene and perylene) degrading consortium enriched in TLP cultures using silicone oil as the organic phase. From 16S rRNA analysis, four isolates were identified as Mycobacterium gilvum B1 (99% identity),Bacillus pumilus B44 (99% identity), Microbacterium esteraromaticum B21 (98% identity), and to the genus Porphyrobacter B51 (96% identity). The two latter isolates have not previously been associated with PAH degradation. Isolate B51 grew strongly in the interfacial fraction in the presence of naphthalene vapours and phenanthrene compared with cultures without LMW PAHs. Benzo[a]pyrene was degraded in cultures containing a HMW PAH mixture but pyrene had no effect on its degradation. The growth of isolates B1 and B21 was improved in the aqueous phase than in the interfacial fraction for cultures with naphthalene vapours. Pyrene was required for benzo[a]pyrene degradation by isolate B1. For isolate B21, pyrene and chrysene were degraded only in cultures without naphthalene vapours. CONCLUSION: Consortium enriched in a TLP culture is composed of microorganisms with different abilities to grow at the interface or in the aqueous phase according to the culture conditions and the PAH that are present. Naphthalene vapours increased the growth of the microorganisms in TLP cultures but did not stimulate the HMW PAH degradation. SIGNIFICANCE AND IMPACT OF THE STUDY: New HMW PAH-degrading microorganisms and a better understanding of the mechanisms involved in HMW PAH degradation in TLP cultures.

Bacteria↗

Interactions of myelin basic protein with palmitoyllysophosphatidylcholine: characterization of the complexes and conformations of the protein.

The stoichiometry of palmitoyllysophosphatidylcholine/myelin basic protein (PLPC/MBP) complexes, the location of the protein in the lysolipid micelles, and the conformational changes occurring in the basic protein and peptides derived from it upon interaction with lysolecithin micelles were investigated by circular dichroic spectropolarimetry, ultracentrifugation, electron paramagnetic resonance (EPR) and 31P, 13C, and 1H nuclear magnetic resonance spectroscopy (NMR), and electron magnetic resonance spectroscopy (NMR), and electron microscopy. Ultracentrifugation measurements indicated that well-defined complexes were formed by the association of one protein molecule with approximately 141 lysolipid molecules. Small-angle X-ray scattering data indicated that the PLPC/MBP complexes form particles with a radius of gyration of 3.8 nm. EPR spectral parameters of the spin labels 5-, and 16-doxylstearate incorporated into lysolecithin/basic protein aggregates, and 13C- and 1H-NMR relaxation times of PLPC indicated that the addition of the protein did not affect the environment and location of the labels and the organization of the lysolipid micelles. The data suggested that MBP lies primarily near the surface of the micelles, with segments penetrating beyond the interfacial region into the hydrophobic interior, but without any part of the protein being protected against rapid exchange of its amide groups with the aqueous environment. The basic protein acquired about 20% alpha-helix when bound to lysolipid micelles. Circular dichroic spectra of sequential peptides derived by cleavage of the protein revealed the formation of alpha-helical regions in the association with lysolecithin. Specific residues in myelin basic protein that participated in binding to the micelles were identified from magnetic resonance data on changes in the chemical shifts and intensities of assigned resonances, and line broadening of peaks by fatty acid spin-labels incorporated into the micelles.

Animals↗

Photosensitized electron transfer processes in SiO(2) colloids and sodium lauryl sulfate micellar systems: Correlation of quantum yields with interfacial surface potentials.

The effectiveness of negatively charged colloidal SiO(2) particles in controlling photosensitized electron transfer reactions has been studied and compared with that of the negatively charged sodium lauryl sulfate (NaLauSO(4)) micellar system. In particular, the photosensitized reduction of the zwitterionic electron acceptor propylviologen sulfonate (PVS(0)) with tris(2,2'-bipyridinium)ruthenium(II) [Ru(bipy)(3) (2+)] as the sensitizer and triethanolamine as the electron donor is found to have a quantum yield of 0.033 for formation of the radical anion (PVS([unk])) in the SiO(2) colloid compared with 0.005 in the homogeneous system and 0.0086 in a NaLauSO(4) micellar solution. The higher quantum yields obtained with the SiO(2) colloidal system are attributed to substantial stabilization against back reaction of the intermediate photoproducts-i.e., Ru(bipy)(3) (3+) and PVS([unk])-by electrostatic repulsion of the reduced electron acceptor from the negatively charged particle surface. The binding properties of the SiO(2) particles and NaLauSO(4) micelles were investigated by flow dialysis. The results show that the sensitizer binds to both interfaces and that the SiO(2) interface is characterized by a much higher surface potential than the micellar interface ( approximately -170 mV vs. -85 mV). The effect of ionic strength on the surface potential was estimated from the Gouy-Chapman theory, and the measured quantum yields of photosensitized electron transfer were correlated with surface potential at different ionic strengths. This correlation shows that the quantum yield is not affected by surface potentials smaller than approximately -40 mV. At larger potentials, the quantum yield increases rapidly. The quantum yield obtained in the micellar system at different strengths fits nicely on the correlation curve for the colloid SiO(2) system. These results indicate that the surface potential is the dominant factor in the quantum yield improvement for PVS(0) reduction.

Journal Article↗

Osseointegration of sintered porous-surfaced and plasma spray-coated implants: An animal model study of early postimplantation healing response and mechanical stability.

The osseointegration and long-term success of bone-interfacing implants are dependent on mechanical stability of the implant relative to host bone during the early healing period. The geometric design of an implant surface may play an important role in affecting early implant stabilization, possibly by influencing tissue healing dynamics. In this study, we compared the early tissue healing response and resulting implant stability for two surface designs by characterizing the histological and mechanical properties of the healing tissue around Ti6Al4V sintered porous-surfaced and Ti plasma-sprayed implants. The implants were inserted transversely in rabbit femoral condyles and evaluated at 0, 4, 8, and 16 days postimplantation. At 4 and 8 days after implantation, the early healing tissue (fibrin and collagenous matrix) was more extensively integrated with the three-dimensional interconnected structure of the sintered porous surface than with the irregular geometry of the plasma-sprayed coating. In addition, histological examination indicated that initial matrix mineralization leading to osseointegration occurred more rapidly with the porous-surfaced implants. The more extensive tissue integration and more rapid matrix mineralization with the porous-surfaced implants were reflected in the mechanical test data, which demonstrated greater attachment strength and interfacial stiffness for the porous-surfaced implants 4 and 8 days postimplantation (p <.05). Sixteen days after implantation, both implant designs were osseointegrated and had comparable attachment characteristics. These data demonstrate that appropriate surface design selection can improve early implant stability and induce an accelerated healing response, thereby improving the potential for implant osseointegration.

Animals↗

Preparation of montmorillonite/titania nanocomposite and enhanced electrorheological activity.

We prepared a new type of electrorheological particle composed of TiO2 nanocrystallites-coated montmorillonite (MMT/TiO2) nanocomposite by the sol-gel technique. The characterizations including TGA, XRD, TEM, SEM, EDS, and FTIR showed that TiO2 was deposited on the surface of the MMT flakes with anatase nanocrystallite. An obviously enhanced ER effect was found in the MMT/TiO2 nanocomposites based ER fluids compared with pure MMT and TiO2. Furthermore, the temperature and sedimentation stabilities of the MMT/TiO2 ER fluids had also been improved greatly. Interestingly, the content of TiO2 was demonstrated to have an important influence on the ER effect. When the content of TiO2 was about 20 wt%, the ER effect of MMT/TiO2 ER fluid reached its maximum, which was about 5 times that of pure MMT ER fluid and 27 times that of pure TiO2 ER fluid. Based on dielectric analysis, the significant ER enhancement by formation nanocrystallites-coated montmorillonite was attributed to the enhanced interfacial polarization in this nanocomposite particle due to the effective limitation of the long-range drift of active ions in montmorillonite particles.

Journal Article↗

Photophysical study of 3-acetyl-4-oxo-6,7-dihydro-12H-indolo[2,3-a]quinolizine in biomimetic reverse micellar nanocavities: a spectroscopic approach.

Photophysical properties of 3-acetyl-4-oxo-6,7-dihydro-12H-indolo[2,3-a]quinolizine (AODIQ), a bioactive molecule, has been investigated in well-characterized, monodispersed biomimicking nanocavities formed by sodium bis(2-ethylhexyl)sulfosuccinate (AOT) in heptane using steady-state and picosecond time resolved fluorescence and fluorescence anisotropy. The emission behavior of AODIQ is very much dependent upon the water/surfactant mole ratio (W), i.e., on the water pool size of the reverse micellar core. AODIQ exhibits a sharp decrease in fluorescence anisotropy with increasing W, implying that the overall motional restriction experienced by the molecule is decreased with increased hydration. Some of the depth-dependent relevant fluorescence parameters, namely, fluorescence maxima and fluorescence anisotropy (r), have been monitored for exploiting the distribution and microenvironment around the probe in the reverse micelles. Fluorescence spectral position and fluorescence quenching studies suggest that the probe does not penetrate into the reverse micellar core; rather it binds at the interfacial region. Quantitaive estimates of the micropolarity and microviscosity at the binding sites of the probe molecule have been determined as a function of W.

Biomimetic Materials↗

Effect of urea, dimethylurea, and tetramethylurea on the phase behavior of dioleoylphosphatidylethanolamine.

The phase behavior of dioleoylphosphatidylethanolamine in aqueous solutions of urea, N,N'-dimethylurea (DMU), and N,N,N',N'-tetramethylurea (TMU) has been characterized by synchrotron X-ray diffraction and differential scanning calorimetry. All three solutes stabilize the lamellar liquid-crystalline phase at the expense of lamellar-gel phase and inverted hexagonal phase of the phospholipid when present in concentrations up to 3 M. X-ray diffraction data demonstrated that the repeat spacing of DOPE increased with increasing urea concentration, but decreased as the DMU and TMU concentrations increased. The repeat spacing of DOPE in the liquid-crystal phase dispersed in the three solutes is d(urea)>d(DMU)>d(TMU). The molecular mechanisms underlying these observations are discussed in terms of either membrane Hofmeister effect, where urea acts as a water structure breaker, or a direct insertion effect of the amphiphilic DMU and TMU molecules into the lipid head groups in the interfacial region of the phospholipid bilayer.

Calorimetry, Differential Scanning↗

Softness of the bacterial cell wall of Streptococcus mitis as probed by microelectrophoresis.

Chemical and structural complexity of bacterial cell surfaces complicate accurate quantification of cell surfaces properties. The presence of fibrils, fimbriae or other surface appendages on bacterial cell surfaces largely influence those properties and would therefore play a major function in interfacial phenomena as aggregation and adhesion. The electrophoretic softness and fixed charge density in the polyelectrolyte layer of nine Streptococcus mitis strains, usually carrying long sparsely distributed fibrils, were determined by the soft particle analysis using measured electrophoretic mobilities as a function of the ionic strength. In general, S. mitis cell surfaces are electrophoretically soft (1.0-2.5 nm) with a fixed negative charge density of -1.2 to -4.3 x 10(6) Cm(-3). Further, a comparison with surfaces of other bacterial strains that are reported to be soft indicates that the Ohshima soft layer model does not provide information on the surface morphology causing the softness. The most likely reason is that the electroosmotic flow occurs only in the very outer region of thick extracellular surface layers. Nevertheless, determining the surface softness is essential for proper characterization of the cell surface electrostatics.

Cell Wall↗

Mechanical interlocking of engineered cartilage to an underlying polymeric substrate: towards a biohybrid tissue equivalent.

This study investigates the feasibility of engineering a biohybrid cartilage equivalent (BCE) with the long-term goal of restoring the mechanical integrity and interfacial characteristics of severely damaged cartilage. The BCE depends on the successful adhesion, via mechanical interlocking, of a cartilage layer to a nondegradable composite scaffold or prosthesis. The model scaffold, consisting of a nonwoven mesh bonded to a solid core, was seeded with bovine articular chondrocytes. High molecular weight poly(L-lactic acid), which has a slow degradation time, was used to model the nondegradable polymer. Biochemical and histological analysis demonstrate that the BCE can support the growth of a cartilaginous matrix for at least 6 weeks in culture. Mechanical testing of the BCE showed cartilage adhesion strength increased from 19.27+/-1.62 to 43.79+/-3.88 kPa between 35 and 50 days in culture. Nonmechanically interlocked cartilage achieved less than 5% of this adhesion strength. For the first time, atomic force microscopy (AFM) was used to characterize surface topography of tissue-engineered cartilage. Surface roughness of constructs after 8 and 10 weeks ranged from 153 to 171 nm, falling within the range of native cartilage (100-600 nm). This study demonstrates the feasibility of creating a biohybrid cartilage equivalent by mechanically interlocking a cartilaginous layer to an underlying polymeric matrix.

Animals↗

Different modes of interaction of pulmonary surfactant protein SP-B in phosphatidylcholine bilayers.

Pulmonary surfactant-associated protein B (SP-B) has been incorporated into vesicles of dipalmitoyl phosphatidylcholine (DPPC) or egg yolk phosphatidylcholine (PC) by two different procedures to characterize the dependence of lipid-protein interactions on the method of reconstitution. In method A the protein was dissolved in a small volume of either methanol or 60% (v/v) acetonitrile and injected into an aqueous phase containing phospholipid vesicles. In method B the vesicles were prepared by injection of a mixture of phospholipid and SP-B dissolved in methanol or aqueous acetonitrile. Both methods of reconstitution led to the extensive interaction of SP-B with PC bilayers as demonstrated by co-migration during centrifugation, marked protection against proteolysis, change in the fluorescence emission intensity of SP-B, and protection of SP-B tryptophan fluorescence from quenching by acrylamide. SP-B promoted the rapid adsorption of DPPC on an air/liquid interface irrespective of the method of protein reconstitution. However, the interfacial adsorption activity of SP-B reconstituted by method B remained stable for hours, but that of SP-B prepared by method A decreased with time. Electron microscopy showed that the injection of SP-B into an aqueous phase containing PC or DPPC vesicles (method A) induced a rapid aggregation of vesicles. By contrast, a much longer time was required for detecting vesicle aggregation when the protein was reconstituted by co-injection of SP-B and phospholipids (method B). The presence of 5% (w/w) SP-B in DPPC bilayers prepared by method B broadened the differential scanning calorimetry thermogram and decreased the enthalpy of the transition. In contrast, the injection of SP-B into preformed DPPC vesicles (method A) did not influence the gel-to-liquid phase transition of DPPC bilayers. Taken together, these results indicate that the mode and extent of interaction of SP-B with surfactant phospholipids depends on the conditions of preparation of lipid/protein samples, and that care should be taken in the interpretation of findings from reconstituted systems on the role of these surfactant proteins in the alveolar space.

1,2-Dipalmitoylphosphatidylcholine↗

Porcelain adherence vs force to failure for palladium-gallium alloys: a critique of metal-ceramic bond testing.

OBJECTIVES: The purpose of this study was to characterize the metal-ceramic bond of four commercial Pd-Ga alloys by two separate tests: porcelain adherence and bond failure force. A Pd-Ag alloy was the control. A secondary goal was to investigate a possible correlation between the percent porcelain retained and bond failure forces. METHODS: For adherence testing, five cast plates of each alloy were air abraded and oxidized. The porcelain was applied on a circular area in the center of each plate and fractured using constant-strain flexure. The area fraction of adherent porcelain (%) was calculated via a standardized spectrometric technique. Bond failure forces were measured in three-point bending using bar specimens. The modulus of elasticity of the five alloys was also measured. No attempt was made to calculate bond strength since the residual thermal stresses at the interfacial region were unknown. Data were compared via ANOVA and the Tukey multiple range test (p < 0.05). RESULTS: All failures occurred at the porcelain termination sites with no tensile fracture of the porcelain. Two Pd-Ga alloys exhibited porcelain adherence comparable with previously studied Pd-Cu-Ga alloys. There was no correlation between the porcelain adherence and the force to failure (r2 = 0.0159). Additionally, no statistically significant differences were found among the moduli of elasticity of the five alloys. SIGNIFICANCE: The new Pd-Ga alloys have been commercially introduced with little information on the quality of the bond with porcelain. The use of two different means for assessing the metal-ceramic bond provides information on the relationship of data using different experimental techniques.

Analysis of Variance↗

EPR investigation of the free radicals generated during the photosensitization of TiO2 colloid by hypocrellin B.

The cation radical of dye produced from the interfacial electron transfer from a surface chelated dye to the conduction band of the colloidal TiO2 was studied by laser flash photolysis and electron paramagnetic resonance (EPR) techniques. The study employed hypocrellin B (HB), a natural photodynamic pigment with strong absorption over the visible light region, as a sensitizer and titanium dioxide as a colloid semiconductor. HB formed a chelate with this colloid semiconductor and exhibited a red-shifted and strongly enhanced absorption in the visible spectrum. Laser photolysis indicated that the electron excitation in the visible absorption band of the chelate resulted in extremely rapid and efficient electron injection from the excited triplet state of the dye into the conduction band of the semiconductor. A transient absorption of cation radical of HB at 570 nm was observed. The appearance of cation radical of HB was characterized by EPR spectrometry: the photoinduced EPR signal was not quenched by oxygen and its intensity decreased in the presence of NaI, a typical hole scavenger. The generation of conduction band electrons in HB-sensitized TiO2 system was also verified by the spin elimination of a stable cyclic nitroxide, 2,2, 6,6-tetramethylpiperidine-1-oxyl (TEMPO), and by the reduction of methyl viologen (MV2+) to its radical MV+.

Animals↗

Alveolar volume-surface area relation in air- and saline-filled lungs fixed by vascular perfusion.

The influence of volume changes and interfacial forces on the geometry of peripheral air spaces was studied in excised rabbit lungs inflated with either air or saline and fixed by vascular perfusion at four points of the deflation limb of the pressure-volume curve corresponding to 100, 80, 60, and 40% of the total lung capacity (TLC). In air-filled lungs pleating and folding of alveolar septa were observed, especially in alveolar corners. However, the alveolar surfaces were smooth, except at low lung volumes where some surface crumpling occurred. In saline-filled lungs pleats were absent; the alveolar surface was irregular at all inflation levels due to undulating walls and bulging capillaries. Morphometry indicated that at all alveolar volumes (VA) the surface areas (SA) were larger in saline- than air-filled lungs. No simple mathematical function was found to characterize the relation between SA and VA over the entire volume range studied. Within the range of normal breaths (80 to 40% TLC) the best fit for n in the function SA = k.VnA was 0.58 for saline-filled lungs (r = 0.93) and 0.33 for air-filled lungs (r = 0.68), suggesting different and complex deflation patterns.

Animals↗