Elastase-induced matrix degradation in arterial organ cultures: An in vitro model of aneurysmal disease Andrew Wills, BSc, Matthew M. Thompson, MD, FRCS,

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Presentation transcript:

Elastase-induced matrix degradation in arterial organ cultures: An in vitro model of aneurysmal disease Andrew Wills, BSc, Matthew M. Thompson, MD, FRCS, Matthew Crowther, MSc, Nicholas P. Brindle, PhD, Ahktar Nasim, FRCS, Robert D. Sayers, MD, FRCS, Peter R.F. Bell, MD, FRCS Journal of Vascular Surgery Volume 24, Issue 4, Pages 667-679 (October 1996) DOI: 10.1016/S0741-5214(96)70083-6 Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Graph illustrating elastin (□), collagen (▨), and smooth-muscle cell (■) concentration in freshly fixed aorta, aorta cultured for 14 days without any elastase, and in aortas cultured for 14 days that were preincubated with pancreatic elastase at concentration of 1, 10, and 100 U/ml for 24 hours. Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Light micrographs illustrating control (a) and elastase (100 U/ml)-pretreated aortic samples stained with Miller's elastin and van Gieson's stain after 24 hours (b), 4 days (c), 9 days (d), and 14 days (e) of organ culture. Original magnification ×40. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Light micrographs illustrating control (a) and elastase (100 U/ml)-pretreated aortic samples stained with Miller's elastin and van Gieson's stain after 24 hours (b), 4 days (c), 9 days (d), and 14 days (e) of organ culture. Original magnification ×40. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Light micrographs illustrating control (a) and elastase (100 U/ml)-pretreated aortic samples stained with Miller's elastin and van Gieson's stain after 24 hours (b), 4 days (c), 9 days (d), and 14 days (e) of organ culture. Original magnification ×40. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Light micrographs illustrating control (a) and elastase (100 U/ml)-pretreated aortic samples stained with Miller's elastin and van Gieson's stain after 24 hours (b), 4 days (c), 9 days (d), and 14 days (e) of organ culture. Original magnification ×40. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Light micrographs illustrating control (a) and elastase (100 U/ml)-pretreated aortic samples stained with Miller's elastin and van Gieson's stain after 24 hours (b), 4 days (c), 9 days (d), and 14 days (e) of organ culture. Original magnification ×40. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Graph illustrating relative change in concentration of elastin, collagen, and smooth-muscle cells between two groups of aortic organ cultures, one preincubated with pancreatic elastase (100 U/ml) for 24 hours (elastase—solid squares—) and one cultured in standard conditions (control—open circles—). Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Graph illustrating relative change in concentration of elastin, collagen, and smooth-muscle cells between two groups of aortic organ cultures, one preincubated with pancreatic elastase (100 U/ml) for 24 hours (elastase—solid squares—) and one cultured in standard conditions (control—open circles—). Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Graph illustrating relative change in concentration of elastin, collagen, and smooth-muscle cells between two groups of aortic organ cultures, one preincubated with pancreatic elastase (100 U/ml) for 24 hours (elastase—solid squares—) and one cultured in standard conditions (control—open circles—). Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Photograph of gelatin-substrate zymogram comparing elastase-pretreated (elastase) and control aortic samples (control) after 1, 4, 9, and 14 days in organ culture. Equal quantities of protein were loaded into each lane. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Photograph of Western blot to MMP-1 in elastase pretreated and control samples after 4, 9, and 14 days in tissue culture. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 Light micrograph illustrating inflammatory infiltrate in aortic segment preincubated with elastase and autologous leukocytes after 14 days in culture. Original magnification ×200. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 7 Graph illustrating relative change in concentration of elastin, collagen, and smooth-muscle cells between two groups of aortic samples cultured in presence of autologous leukocytes, one preincubated with pancreatic elstase (100 U/ml) for 24 hours (elastase + ly—solid squares— ) and one cultured in standard conditions (control + ly—open circles— ). Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 7 Graph illustrating relative change in concentration of elastin, collagen, and smooth-muscle cells between two groups of aortic samples cultured in presence of autologous leukocytes, one preincubated with pancreatic elstase (100 U/ml) for 24 hours (elastase + ly—solid squares— ) and one cultured in standard conditions (control + ly—open circles— ). Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 7 Graph illustrating relative change in concentration of elastin, collagen, and smooth-muscle cells between two groups of aortic samples cultured in presence of autologous leukocytes, one preincubated with pancreatic elstase (100 U/ml) for 24 hours (elastase + ly—solid squares— ) and one cultured in standard conditions (control + ly—open circles— ). Values are medians with interquartile ranges. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 8 Photograph of gelatin-substrate zymogram comparing elastase-pretreated (Elastase + Ly) and control aortic samples (Control) cocultured with autologous leukocytes after 3, 5, 10, and 14 days in organ culture. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 9 Photograph of gelatin-substrate zymogram comparing elastase-pretreated aortic segments in presence (Elastase + Ly) and absence (Elastase) of autologous leukocytes after 14 days in organ culture. Journal of Vascular Surgery 1996 24, 667-679DOI: (10.1016/S0741-5214(96)70083-6) Copyright © 1996 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions