Repair of elastase-digested elastic fibers in acellular matrices by replating with neonatal rat-lung lipid interstitial fibroblasts or other elastogenic cell types.
Disruption of elastic fibers is a major factor in the pathogenesis of pulmonary emphysema. Elastic fibers in culture, injured by exposure to elastase, undergo repair in the presence of elastogenic cells that restores the fibers toward normal as determined by biochemical and ultrastructural methods. The repair appears to be the result of both salvage and de novo repair mechanisms. The evidence for salvage repair is that hot-alkali resistance, lost as a result of elastase treatment, is restored to previously radiolabeled elastic fibers. This repair mechanism has been shown in aortic smooth muscle cell cultures. In order to determine the potential relevance of this mechanism for elastic fiber repair in the lungs, experiments were carried out using neonatal rat lung lipid interstitial fibroblasts (LIF). Treatment of the LIF cultures with elastase, in the absence of serum, caused solubilization of 12% of elastin; however, 81% of the elastin protein and 80% of the elastin-associated radioactivity (EAR) were solubilized by subsequent hot-alkali treatment, indicating that most of the elastin was retained in the matrix but was damaged. Ultrastructurally, the elastic fibers were frayed. After 6 additional wk in culture, hot-alkali resistant elastin protein and EAR were restored to 88 and 62% of control values, respectively, and the ultrastructural appearance of elastic fibers was restored to normal. We calculate that about 42% of the restored elastin represented salvage repair; the remainder was new elastin. No repair occurred in matrices rendered acellular by azide treatment; however, when acellular matrices were replated with LIF, repair was complete at 6 wk. No repair was seen when acellular matrices were replated with a transformed mouse macrophage cell line. We conclude that lung LIF are capable of mounting a robust repair process after elastolytic injury of elastin and that the repair is the result of both salvage and de novo repair mechanisms.
Stone
P J
PJ
Department of Biochemistry, Boston University School of Medicine, Massachusetts 02118, USA. stone@med-biochm.bu.edu
Morris
S M
SM
Thomas
K M
KM
Schuhwerk
K
K
Mitchelson
A
A
D016475:3T3 Cells / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D000818:Animals; D000831:Animals, Newborn; D001386:Azides / Q000494:pharmacology; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D004548:Elasticity; D004549:Elastin / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D005347:Fibroblasts / Q000737:chemistry / Q000201:enzymology / Q000648:ultrastructure; D008168:Lung / Q000737:chemistry* / Q000166:cytology*; D008239:Lysine / Q000378:metabolism / Q000494:pharmacology; D016676:Macrophages, Alveolar / Q000166:cytology; D051379:Mice; D008854:Microscopy, Electron; D010196:Pancreatic Elastase / Q000494:pharmacology*; D008249:Protein-Lysine 6-Oxidase / Q000378:metabolism; D051381:Rats; D013997:Time Factors
D001386:Azides; D002250:Carbon Radioisotopes; D004549:Elastin; D008249:Protein-Lysine 6-Oxidase; D010196:Pancreatic Elastase; D008239:Lysine