Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O

Björn O. Backhaus, Martin Kaefer, Karen M. Haberstroh, Karen Hile, Jiro Nagatomi, Richard C. Rink, Mark P. Cain, Anthony J. Casale, Rena Bizior

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

Purpose: Bladder outlet obstruction with intravesical pressures exceeding 40 cm. H2O often results in irreversible renal damage. Bladder outlet obstruction also results in alterations in bladder physiology, including wall thickening, reduced compliance and decreased capacity. If unchecked these changes may lead to the subsequent need for bladder augmentation. From a biomechanical standpoint, compliance is primarily related to extracellular matrix deposition, which in turn is dependent on the balanced activity of proteolytic enzymes (that is matrix metalloproteinases [MMPs]) and their endogenous inhibitors (that is tissue inhibitors of metalloproteinases [TIMPs]). To date, the threshold pressure above which alterations in these key determinants of bladder compliance occur has not been determined. Therefore, using a novel device of our own design, we applied hydrostatic pressures in the physiological range to human bladder smooth muscle cells to determine the effect on MMPs, TIMP-1 and transcription of the major structural collagens (types I and III). Materials and Methods: Human bladder smooth muscle cells (staining positive for α-smooth muscle actin) were plated at a density of 100,000 cells per 10 cm.2 and cultured for 2 days in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum. Cells were subsequently exposed to pressures of 0.3, 20 and 40 cm. H2O for 1, 3, 7 and 24 hours in serum-free DMEM. A computer interface maintained pressure levels for the duration of the experimentr and collected pressure data. MMP,1 and 3, and TIMP-1 immunoassay and zymography for MMP-2 and 9 were performed. Polymerase chain reaction for human collagen types I and III was performed following reverse transcription of total purified mRNA. All experiments were repeated 3 times and statistical analysis was performed using a 2-tailed Student t test. Results: Exposure of bladder smooth muscle cells to a sustained hydrostatic pressure of 20 cm. H2O for 7 hours in serum-free DMEM resulted in a time dependent decrease in MMP-1, 2 and 9 activity (15%, 37% and 25%) compared to controls maintained at atmospheric pressure (p <0.01). TIMP-1 levels increased an average of 10% after exposure to 20 cm. H2O. These changes became statistically significant when the cells were exposed to 40 cm. H2O pressure for 3, 7 and 24 hours (+14%, +21% and +50%, respectively). No statistically significant differences in MMP-3 and collagen type I or III mRNA levels were observed. Conclusions: Our results reveal that MMP-1, 2 and 9 are significantly down-regulated in a time and pressure dependent fashion following exposure of bladder smooth muscle cells to 20 cm. H2O for as little as 7 hours. TIMP-1 levels increased under similar conditions. These alterations in MMPs and TIMP-1 favor accumulation of extracellular matrix, structural components associated with bladder wall thickness and decreased compliance. These results are consistent with previous data from animal models of complete outlet obstruction. Our results support the concept that pressures 40 cm. H2O or less contribute to molecular changes consistent with decreased compliance associated with bladder dysfunction.

Original languageEnglish (US)
Pages (from-to)2600-2604
Number of pages5
JournalJournal of Urology
Volume168
Issue number6
StatePublished - Dec 1 2002
Externally publishedYes

Fingerprint

Hydrostatic Pressure
Compliance
Urinary Bladder
Tissue Inhibitor of Metalloproteinase-1
Pressure
Eagles
Matrix Metalloproteinase 1
Smooth Muscle Myocytes
Collagen Type III
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Collagen Type I
Urinary Bladder Neck Obstruction
Matrix Metalloproteinase 3
Matrix Metalloproteinase Inhibitors
Extracellular Matrix
Serum
Tissue Inhibitor of Metalloproteinases
Messenger RNA
Atmospheric Pressure

Keywords

  • Hydrostatic pressure, matrix metalloproteinases, bladder
  • Muscle, smooth

ASJC Scopus subject areas

  • Urology

Cite this

Backhaus, B. O., Kaefer, M., Haberstroh, K. M., Hile, K., Nagatomi, J., Rink, R. C., ... Bizior, R. (2002). Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O. Journal of Urology, 168(6), 2600-2604.

Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O. / Backhaus, Björn O.; Kaefer, Martin; Haberstroh, Karen M.; Hile, Karen; Nagatomi, Jiro; Rink, Richard C.; Cain, Mark P.; Casale, Anthony J.; Bizior, Rena.

In: Journal of Urology, Vol. 168, No. 6, 01.12.2002, p. 2600-2604.

Research output: Contribution to journalArticle

Backhaus, BO, Kaefer, M, Haberstroh, KM, Hile, K, Nagatomi, J, Rink, RC, Cain, MP, Casale, AJ & Bizior, R 2002, 'Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O', Journal of Urology, vol. 168, no. 6, pp. 2600-2604.
Backhaus BO, Kaefer M, Haberstroh KM, Hile K, Nagatomi J, Rink RC et al. Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O. Journal of Urology. 2002 Dec 1;168(6):2600-2604.
Backhaus, Björn O. ; Kaefer, Martin ; Haberstroh, Karen M. ; Hile, Karen ; Nagatomi, Jiro ; Rink, Richard C. ; Cain, Mark P. ; Casale, Anthony J. ; Bizior, Rena. / Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O. In: Journal of Urology. 2002 ; Vol. 168, No. 6. pp. 2600-2604.
@article{7b67386c861e4af783fc66a19c4295a2,
title = "Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O",
abstract = "Purpose: Bladder outlet obstruction with intravesical pressures exceeding 40 cm. H2O often results in irreversible renal damage. Bladder outlet obstruction also results in alterations in bladder physiology, including wall thickening, reduced compliance and decreased capacity. If unchecked these changes may lead to the subsequent need for bladder augmentation. From a biomechanical standpoint, compliance is primarily related to extracellular matrix deposition, which in turn is dependent on the balanced activity of proteolytic enzymes (that is matrix metalloproteinases [MMPs]) and their endogenous inhibitors (that is tissue inhibitors of metalloproteinases [TIMPs]). To date, the threshold pressure above which alterations in these key determinants of bladder compliance occur has not been determined. Therefore, using a novel device of our own design, we applied hydrostatic pressures in the physiological range to human bladder smooth muscle cells to determine the effect on MMPs, TIMP-1 and transcription of the major structural collagens (types I and III). Materials and Methods: Human bladder smooth muscle cells (staining positive for α-smooth muscle actin) were plated at a density of 100,000 cells per 10 cm.2 and cultured for 2 days in Dulbecco's modified Eagle's medium (DMEM) with 10{\%} fetal bovine serum. Cells were subsequently exposed to pressures of 0.3, 20 and 40 cm. H2O for 1, 3, 7 and 24 hours in serum-free DMEM. A computer interface maintained pressure levels for the duration of the experimentr and collected pressure data. MMP,1 and 3, and TIMP-1 immunoassay and zymography for MMP-2 and 9 were performed. Polymerase chain reaction for human collagen types I and III was performed following reverse transcription of total purified mRNA. All experiments were repeated 3 times and statistical analysis was performed using a 2-tailed Student t test. Results: Exposure of bladder smooth muscle cells to a sustained hydrostatic pressure of 20 cm. H2O for 7 hours in serum-free DMEM resulted in a time dependent decrease in MMP-1, 2 and 9 activity (15{\%}, 37{\%} and 25{\%}) compared to controls maintained at atmospheric pressure (p <0.01). TIMP-1 levels increased an average of 10{\%} after exposure to 20 cm. H2O. These changes became statistically significant when the cells were exposed to 40 cm. H2O pressure for 3, 7 and 24 hours (+14{\%}, +21{\%} and +50{\%}, respectively). No statistically significant differences in MMP-3 and collagen type I or III mRNA levels were observed. Conclusions: Our results reveal that MMP-1, 2 and 9 are significantly down-regulated in a time and pressure dependent fashion following exposure of bladder smooth muscle cells to 20 cm. H2O for as little as 7 hours. TIMP-1 levels increased under similar conditions. These alterations in MMPs and TIMP-1 favor accumulation of extracellular matrix, structural components associated with bladder wall thickness and decreased compliance. These results are consistent with previous data from animal models of complete outlet obstruction. Our results support the concept that pressures 40 cm. H2O or less contribute to molecular changes consistent with decreased compliance associated with bladder dysfunction.",
keywords = "Hydrostatic pressure, matrix metalloproteinases, bladder, Muscle, smooth",
author = "Backhaus, {Bj{\"o}rn O.} and Martin Kaefer and Haberstroh, {Karen M.} and Karen Hile and Jiro Nagatomi and Rink, {Richard C.} and Cain, {Mark P.} and Casale, {Anthony J.} and Rena Bizior",
year = "2002",
month = "12",
day = "1",
language = "English (US)",
volume = "168",
pages = "2600--2604",
journal = "Journal of Urology",
issn = "0022-5347",
publisher = "Elsevier Inc.",
number = "6",

}

TY - JOUR

T1 - Alterations in the molecular determinants of bladder compliance at hydrostatic pressures less than 40 cm. H2O

AU - Backhaus, Björn O.

AU - Kaefer, Martin

AU - Haberstroh, Karen M.

AU - Hile, Karen

AU - Nagatomi, Jiro

AU - Rink, Richard C.

AU - Cain, Mark P.

AU - Casale, Anthony J.

AU - Bizior, Rena

PY - 2002/12/1

Y1 - 2002/12/1

N2 - Purpose: Bladder outlet obstruction with intravesical pressures exceeding 40 cm. H2O often results in irreversible renal damage. Bladder outlet obstruction also results in alterations in bladder physiology, including wall thickening, reduced compliance and decreased capacity. If unchecked these changes may lead to the subsequent need for bladder augmentation. From a biomechanical standpoint, compliance is primarily related to extracellular matrix deposition, which in turn is dependent on the balanced activity of proteolytic enzymes (that is matrix metalloproteinases [MMPs]) and their endogenous inhibitors (that is tissue inhibitors of metalloproteinases [TIMPs]). To date, the threshold pressure above which alterations in these key determinants of bladder compliance occur has not been determined. Therefore, using a novel device of our own design, we applied hydrostatic pressures in the physiological range to human bladder smooth muscle cells to determine the effect on MMPs, TIMP-1 and transcription of the major structural collagens (types I and III). Materials and Methods: Human bladder smooth muscle cells (staining positive for α-smooth muscle actin) were plated at a density of 100,000 cells per 10 cm.2 and cultured for 2 days in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum. Cells were subsequently exposed to pressures of 0.3, 20 and 40 cm. H2O for 1, 3, 7 and 24 hours in serum-free DMEM. A computer interface maintained pressure levels for the duration of the experimentr and collected pressure data. MMP,1 and 3, and TIMP-1 immunoassay and zymography for MMP-2 and 9 were performed. Polymerase chain reaction for human collagen types I and III was performed following reverse transcription of total purified mRNA. All experiments were repeated 3 times and statistical analysis was performed using a 2-tailed Student t test. Results: Exposure of bladder smooth muscle cells to a sustained hydrostatic pressure of 20 cm. H2O for 7 hours in serum-free DMEM resulted in a time dependent decrease in MMP-1, 2 and 9 activity (15%, 37% and 25%) compared to controls maintained at atmospheric pressure (p <0.01). TIMP-1 levels increased an average of 10% after exposure to 20 cm. H2O. These changes became statistically significant when the cells were exposed to 40 cm. H2O pressure for 3, 7 and 24 hours (+14%, +21% and +50%, respectively). No statistically significant differences in MMP-3 and collagen type I or III mRNA levels were observed. Conclusions: Our results reveal that MMP-1, 2 and 9 are significantly down-regulated in a time and pressure dependent fashion following exposure of bladder smooth muscle cells to 20 cm. H2O for as little as 7 hours. TIMP-1 levels increased under similar conditions. These alterations in MMPs and TIMP-1 favor accumulation of extracellular matrix, structural components associated with bladder wall thickness and decreased compliance. These results are consistent with previous data from animal models of complete outlet obstruction. Our results support the concept that pressures 40 cm. H2O or less contribute to molecular changes consistent with decreased compliance associated with bladder dysfunction.

AB - Purpose: Bladder outlet obstruction with intravesical pressures exceeding 40 cm. H2O often results in irreversible renal damage. Bladder outlet obstruction also results in alterations in bladder physiology, including wall thickening, reduced compliance and decreased capacity. If unchecked these changes may lead to the subsequent need for bladder augmentation. From a biomechanical standpoint, compliance is primarily related to extracellular matrix deposition, which in turn is dependent on the balanced activity of proteolytic enzymes (that is matrix metalloproteinases [MMPs]) and their endogenous inhibitors (that is tissue inhibitors of metalloproteinases [TIMPs]). To date, the threshold pressure above which alterations in these key determinants of bladder compliance occur has not been determined. Therefore, using a novel device of our own design, we applied hydrostatic pressures in the physiological range to human bladder smooth muscle cells to determine the effect on MMPs, TIMP-1 and transcription of the major structural collagens (types I and III). Materials and Methods: Human bladder smooth muscle cells (staining positive for α-smooth muscle actin) were plated at a density of 100,000 cells per 10 cm.2 and cultured for 2 days in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum. Cells were subsequently exposed to pressures of 0.3, 20 and 40 cm. H2O for 1, 3, 7 and 24 hours in serum-free DMEM. A computer interface maintained pressure levels for the duration of the experimentr and collected pressure data. MMP,1 and 3, and TIMP-1 immunoassay and zymography for MMP-2 and 9 were performed. Polymerase chain reaction for human collagen types I and III was performed following reverse transcription of total purified mRNA. All experiments were repeated 3 times and statistical analysis was performed using a 2-tailed Student t test. Results: Exposure of bladder smooth muscle cells to a sustained hydrostatic pressure of 20 cm. H2O for 7 hours in serum-free DMEM resulted in a time dependent decrease in MMP-1, 2 and 9 activity (15%, 37% and 25%) compared to controls maintained at atmospheric pressure (p <0.01). TIMP-1 levels increased an average of 10% after exposure to 20 cm. H2O. These changes became statistically significant when the cells were exposed to 40 cm. H2O pressure for 3, 7 and 24 hours (+14%, +21% and +50%, respectively). No statistically significant differences in MMP-3 and collagen type I or III mRNA levels were observed. Conclusions: Our results reveal that MMP-1, 2 and 9 are significantly down-regulated in a time and pressure dependent fashion following exposure of bladder smooth muscle cells to 20 cm. H2O for as little as 7 hours. TIMP-1 levels increased under similar conditions. These alterations in MMPs and TIMP-1 favor accumulation of extracellular matrix, structural components associated with bladder wall thickness and decreased compliance. These results are consistent with previous data from animal models of complete outlet obstruction. Our results support the concept that pressures 40 cm. H2O or less contribute to molecular changes consistent with decreased compliance associated with bladder dysfunction.

KW - Hydrostatic pressure, matrix metalloproteinases, bladder

KW - Muscle, smooth

UR - http://www.scopus.com/inward/record.url?scp=0036895216&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036895216&partnerID=8YFLogxK

M3 - Article

VL - 168

SP - 2600

EP - 2604

JO - Journal of Urology

JF - Journal of Urology

SN - 0022-5347

IS - 6

ER -