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J R Bellare

Publications and source records attributed to J R Bellare.

6 recordsLinked to original sources

Comparison of in vitro surface properties of clove oil-phospholipid suspensions with those of ALEC, Exosurf and Survanta.

Dipalmitoyl phosphatidylcholine, the main component of lung surfactant is ineffective as a replacement surfactant due to its poor adsorption. We studied clove oil as a possible additive for improving the surface activity of protein-free phospholipid suspensions. We added low doses of clove oil, to phospholipid suspensions and studied the surface properties by in vitro analysis using a pulsating bubble surfactometer and a Wilhelmy balance. Survanta, ALEC and Exosurf were used as controls for comparison. The test surfactants, which were phospholipid-oil suspensions at 1% concentration, in buffer containing either 2 or 5 mM calcium, were pulsated at 40 cpm in a pulsating bubble surfactometer. The phospholipids studied were dipalmitoyl phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), binary mixtures of PC:PE (2:3) and PC:PG (2:3). The addition of clove oil (CO) to each of the above phospholipids was in the ratio of nine parts of phospholipid to one part of oil. The presence of CO caused a significant improvement in the adsorption and minimum surface tension of all the phospholipid suspensions studied. The mixtures PC with CO, both in the presence of 2 and 5 mM calcium, and PCPE with CO at 2 mM calcium concentration had surface properties significantly better than those of ALEC and Exosurf and equivalent to those of Survanta. The addition of clove oil helps improve the surface properties of phospholipids.

Adsorption↗

In vitro evaluation of surfactants with eucalyptus oil for respiratory distress syndrome.

The effects of low doses of eucalyptus oil (EO) were studied on the surface properties of phospholipid suspensions as exogenous surfactants, by in vitro analysis using a pulsating bubble surfactometer and a Wilhelmy balance. Survanta, ALEC and Exosurf, commonly used surfactants in therapy of respiratory distress syndrome were used as controls for comparison. The test surfactants, in Ringer's lactate at 1%, were pulsated at 40 cpm in the surfactometer. EO caused a significant improvement of adsorption of the surfactants. In the case of the binary mixture of dipalmitoylphosphatidylcholine and phosphatidylethanolamine (2:3), EO significantly improved the adsorption, stability and minimum surface tension obtained. This combination performed better than ALEC and Exosurf and was comparable to Survanta with respect to minimum surface tension attained. The re-spreading of a surface excess film of this mixture in a Wilhelmy balance was higher than that of ALEC and Exosurf. The ultrastructure of the EO enriched surfactants using cryogenic scanning electron microscopy showed easy facturability and formation of open membranous structures, which could have been associated with the improved surface activity.

Eucalyptus↗

Ultrastructure of exogenous surfactants using cryogenic scanning electron microscopy.

Therapy with specialised biomaterials, exogenous surfactants, is known to significantly decrease the mortality rates in Respiratory Distress Syndrome (RDS). Surfactants available commercially vary widely in composition and biophysical properties. The present paper studies the ultrastructure of three exogenous surfactants used for the treatment of Respiratory Distress Syndrome, namely, Survanta, ALEC and Exosurf Neonatal with respect to their ability to form liposomes using cryogenic scanning electron microscopy. Liposomal organisation is more obvious in Exosurf than in Survanta and is most pronounced in ALEC. ALEC forms closed regular liposomes with an onion-ring-like internal bilayer arrangement. Survanta forms open membranous structures with wavy ribbon-like membranes. The complex membrane-like structures seen with Survanta may be due to the interaction of lipids with surfactant-specific proteins present in this surfactant which is derived from natural lung extracts and might indicate superior spreading at the lipid-water interface. Artificial protein-free surfactants (ALEC and Exosurf) did not appear to form these open membranous structures. Further study of the ultrastructure of possible biomaterials as surfactants could help in the development of new, improved artificial protein-free surfactants with open membranous structures that might facilitate spreading at the air-liquid interface of lungs.

Biocompatible Materials↗

Scoring of surface parameters of physiological relevance to surfactant therapy in respiratory distress syndrome.

The Wilhelmy balance was used for in vitro testing of surface parameters of surfactants used for respiratory distress syndrome therapy. Two commercial protein-free surfactants, ALEC and Exosurf, were compared with pure forms of the three main phospholipids in natural surfactants, dipalmitoyl phosphatidylcholine (PC), phosphatidylglycerol (PG), and phosphatidylethanolamine (PE), and their binary mixtures, PC with PE and PG each in the ratio 2:3. Surface excess films (15 A2/molecule) were compressed at 1.2 cycles/min past collapse to a compression ratio of 4:1. The maximum surface pressure, spreading time, compressibility, respreading ratio, recruitment index, and hysteresis area were compared. A consolidated list of criteria for selection of suitable surfactants was compiled from the literature. A relative scoring system was devised for comparison based on these criteria. PC/PG (2:3) performed the best as it fulfilled all the criteria and obtained the highest relative score. Exosurf also performed well, except on the respreading criterion. ALEC and PC/PE were equivalent in their performance and performed well, except on two criteria: hysteresis area and recruitment index. Thus the scoring system proposed here proved valuable to rate the overall efficacy as well as relative merits of surfactant formulations.

Drug Combinations↗

Analysis of dynamic surface properties of therapeutic surfactants and lung phospholipids.

Exogenous surfactant is a specialized biomaterial used for substitution of the lipoprotein mixture normally present in lungs--pulmonary surfactant. Respiratory Distress Syndrome is a disease of preterm infants mainly caused by a deficiency of mature lung surfactant. Pulmonary surfactant is known to stabilize small alveoli and prevent them from collapsing during expiration due to its unique surface properties. A pulsating bubble surfactometer was used for in vitro analysis of surface parameters of therapeutic surfactants and of test formulations to be used for exogenous therapy in Respiratory Distress Syndrome. Surface parameters that were considered for comparison were minimum surface tension (gamma(min)) at three different frequencies (20, 40 and 60 cpm), adsorption at two extreme bubble radii (Rmin and Rmax), stability index at the three frequencies, recruitment index and the surface viscoelastic parameters. Survanta, ALEC and Exosurf were compared with formulations consisting of the main phospholipids of pulmonary surfactant, namely dipalmitoyl phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) as well as binary mixtures of these phospholipids in the ratio 2:3. Survanta performed much better than the non-protein therapeutic surfactants in all parameters and at all three frequencies. Exosurf had a very low stability index and a very low modulus of surface dilatational elasticity at all three frequencies. The test compounds showed a frequency dependence in their performance. At 20 cpm, PC:PG (2:3) was the best test combination. It achieved a gamma(min) and stability index equivalent to Survanta at this frequency. None of the test compounds were comparable to Survanta at 40 and 60 cpm. These findings may have important therapeutic implications for exogenous surfactants.

Humans↗

Controlled environment vitrification system: an improved sample preparation technique.

The controlled environment vitrification system (CEVS) permits cryofixation of hydrated biological and colloidal dispersions and aggregates from a temperature- and saturation-controlled environment. Otherwise, specimens prepared in an uncontrolled laboratory atmosphere are subject to evaporation and heat transfer, which may introduce artifacts caused by concentration, pH, ionic strength, and temperature changes. Moreover, it is difficult to fix and examine the microstructure of systems at temperatures other than ambient (e.g., biological systems at in vivo conditions and colloidal systems above room temperature). A system has been developed that ensures that a liquid or partially liquid specimen is maintained in its original state while it is being prepared before vitrification and, once prepared, is vitrified with little alteration of its microstructure. A controlled environment is provided within a chamber where temperature and chemical activity of volatile components can be controlled while the specimen is being prepared. The specimen grid is mounted on a plunger, and a synchronous shutter is opened almost simultaneously with the release of the plunger, so that the specimen is propelled abruptly through the shutter opening into a cryogenic bath. We describe the system and its use and illustrate the value of the technique with TEM micrographs of surfactant microstructures in which specimen preparation artifacts were avoided. We also discuss applications to other instruments like SEM, to other techniques like freeze-fracture, and to novel "on the grid" experiments that make it possible to freeze successive instants of dynamic processes such as membrane fusion, chemical reactions, and phase transitions.

Microscopy, Electron↗