Protein Matrices for Improved Wound Healing: Elastase Inhibition by a Synthetic Peptide Model
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The unique properties of silk fibroin were combined with keratin to develop new wound-dressing materials. Silk fibroin/keratin (SF/K) films were prepared to reduce high levels of elastase found on chronic wounds. This improved biological function was achieved by the incorporation of a small peptide synthesized based on the reactive-site loop of the Bowman−Birk Inhibitor (BBI) protein. In vitro degradation and release were evaluated using porcine pancreatic elastase (PPE) solution as a model of wound exudate. It was found that biological degradation and release rate are highly dependent on film composition. Furthermore, the level of PPE activity can be tuned by changing the film composition, thus showing an innovative way of controlling the elastase−antielastase imbalance found on chronic wounds.Arterial wall
Pancreatic elastase
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The aim of this study was to investigate the presence of free elastase in GCF Samples were taken from inflamed sites in 12 subjects with gingivitis alone and from inflamed sites with and without tissue destruction in 19 patients having periodontitis. Elastase activity was measured with a low molecular weight substrate. To distinguish between free elastase and elastase bound to alpha-2-macroglobulin (A2MG), an excess of alpha-1-antitrypsin (A1AT) was added to the samples. The activity that could be inhibited by A1AT was considered as free elastase, and the uninhibited activity as derived from the elastase-A2MG complex. The elastase-A1AT complex was measured with an ELISA. Free elastase was found in almost all samples, but both the total amount and the proportion of free elastase were higher in samples from sites showing destruction. The elastase-A2MG complex was also increased in sites with tissue destruction, while there was no significant difference in the amount of A1AT complex between the 3 categories of sites. In conclusion, our study clearly reveals free elastase in GCF The elevated levels of free elastase in sites showing tissue destruction seem to be due to a combination of increased release of elastase and an inactivation of A1AT.
Pancreatic elastase
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The effect of elastase on dermal elastic fibers from three age groups (children, young and old adults) has been investigated using ultrathin sections embedded in Epon. There were significant differences in the effect of elastase among the age groups; younger elastin was more rapidly digested by elastase than older elastin. In children and young adults, digested areas by elastase were first recognized as tiny round holes scattered in the amorphous material of the elastic fiber, extending to the large part of the fiber except for microfibrils. In old adults, digested areas were limited to the only small part of the elastic fiber.
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Arterial wall
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Azurophilic granule
Proteinase inhibitor
Neutrophil elastase
Protease inhibitor (pharmacology)
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Sheep are often used to study tissue damage following shock after traumatic injury and in the course of other diseases. The processes involved are thought to be caused at least in part by elastase released from polymorphonuclear leukocytes (PMNs). Since little is known about elastase and its role as a mediator of tissue damage in sheep, we studied the biochemical properties and release characteristics to sheep leukocyte elastase (SLE) in comparison of those of human leukocyte elastase (HLE). Both enzymes showed similar molecular masses, amino-acid compositions, N-terminal amino-acid sequences, and abilities to digest elastin substrates. Differences, however, were found in kinetic parameters measured with the elastase-specific substrate N-methoxysuccinyl-(L-alanyl)2-L-prolyl-L- valine-4-nitroanilide (MeoSuc-AAPV-pNa). The Michaelis constant (Km) of ovine elastase was nearly 10 times higher (1.82 mM) than the Km of HLE (0.21 mM). Values of SLE calculated for kcat were 70% and for kcat/Km 8% of corresponding values determined for HLE. In addition, significant differences between sheep and human PMNs were found in in vitro stimulation experiments. In contrast to human PMNs, sheep neutrophils released no active elastase, and only 50 to 70% of the H2O2 produced by human PMNs. This failure to release active elastase could not be explained by a lower elastase content of sheep PMNs, as there were no significant differences found between the elastase contents of sheep and human PMNs. We conclude that elastase liberated by stimulated sheep PMNs is inactivated by a concomitantly released proteinase inhibitor also located within the sheep PMNs.(ABSTRACT TRUNCATED AT 250 WORDS)
Neutrophil elastase
Pancreatic elastase
Enzyme Kinetics
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To develop a new anti-skin aging cosmetics or functional foods by using elastase inhibitor, a potent elastase inhibitor was screened from various extracts of medicinal plants and its optimal extraction condition was investigated. Methanol extracts of Rubi fructus showed the highest elastase inhibitory activity of 85%. The elastase inhibitor of Rubi fructus was maximally extracted when it was treated with 80% methanol at for 12hr and its elastase inhibitory activity was 0.52 mg.
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