The effects of thrombin and cytokines upon the biomechanics and remodeling of isolated amnion membrane, in vitro

D. Kumar, F. Schatz, R. M. Moore, B. M. Mercer, N. Rangaswamy, J. M. Mansour, C. J. Lockwood, J. J. Moore

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Abruption-induced thrombin generation and inflammation/infection induced cytokine production have both been associated with fetal membrane (FM) weakening and preterm premature rupture of the fetal membranes (PPROM). Using our in vitro model system we have demonstrated that thrombin, and separately the cytokines, tumor necrosis factor-alpha (TNFα) and interleukin-1-beta (IL-1β), remodel and weaken full thickness FM. Additionally, we have reported that the anti-oxidant and NFκB inhibitor, alpha-lipoic acid (LA), blocks these thrombin and cytokine induced effects. The purpose of these studies was to determine whether thrombin and cytokines directly weaken the amnion membrane (AM), the major load-bearing component of FM. Isolated AM or full thickness FM fragments from unlabored Cesarean deliveries were incubated with thrombin, TNFα, or IL-1β, for 48 h. Rupture strength (breaking force) of each fragment was thereafter determined using our published methodology. Biochemical evidence of remodeling and apoptosis; immunoreactive Matrix Metalloproteinase 9 (MMP9), Tissue Inhibitor of Matrix Metalloproteinase 3 (TIMP3) and cleaved poly (ADP-ribose) polymerase (C-PARP) levels in tissue extracts, were determined by western blot and densitometry. Thrombin induced a dose-dependent weakening of isolated AM (P < 0.001) coupled with dose dependent increases in PARP cleavage, and reciprocal increases and decreases, respectively, in MMP9 and TIMP3 protein (all P < 0.01). Thrombin receptor activating peptide-6 (TRAP) also weakened isolated AM. Neither TNFα nor IL-1β weakened isolated AM. However, both cytokines weakened AM when it was incubated together with the choriodecidua as part of full thickness FM (P < 0.001). Cytokine-conditioned choriodecidua medium also weakened isolated AM (P < 0.001). Under conditions in which cytokines weakened the AM, the changes in MMP9, TIMP3 and PARP cleavage were consistent with those seen after thrombin incubation. LA blocked the FM weakening and remodeling effects. In summary, thrombin weakens AM directly whereas cytokines weaken AM indirectly by causing the release of soluble intermediates from the choriodecidua.

Original languageEnglish (US)
Pages (from-to)206-213
Number of pages8
JournalPlacenta
Volume32
Issue number3
DOIs
StatePublished - Mar 2011

Fingerprint

Amnion
Biomechanical Phenomena
Thrombin
Extraembryonic Membranes
Cytokines
Membranes
Tissue Inhibitor of Metalloproteinase-3
Matrix Metalloproteinase 3
Matrix Metalloproteinase 9
Interleukin-1beta
Thioctic Acid
Tumor Necrosis Factor-alpha
Premature Rupture Fetal Membranes
In Vitro Techniques
Tissue Extracts
Matrix Metalloproteinase Inhibitors
Poly(ADP-ribose) Polymerases
Densitometry
Weight-Bearing
Conditioned Culture Medium

Keywords

  • Amnion
  • Biomechanics
  • Cytokines
  • Fetal membranes
  • IL-1β
  • Lipoic acid
  • Model system
  • PPROM
  • Thrombin
  • TNF

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Reproductive Medicine
  • Developmental Biology

Cite this

Kumar, D., Schatz, F., Moore, R. M., Mercer, B. M., Rangaswamy, N., Mansour, J. M., ... Moore, J. J. (2011). The effects of thrombin and cytokines upon the biomechanics and remodeling of isolated amnion membrane, in vitro. Placenta, 32(3), 206-213. https://doi.org/10.1016/j.placenta.2011.01.006

The effects of thrombin and cytokines upon the biomechanics and remodeling of isolated amnion membrane, in vitro. / Kumar, D.; Schatz, F.; Moore, R. M.; Mercer, B. M.; Rangaswamy, N.; Mansour, J. M.; Lockwood, C. J.; Moore, J. J.

In: Placenta, Vol. 32, No. 3, 03.2011, p. 206-213.

Research output: Contribution to journalArticle

Kumar, D, Schatz, F, Moore, RM, Mercer, BM, Rangaswamy, N, Mansour, JM, Lockwood, CJ & Moore, JJ 2011, 'The effects of thrombin and cytokines upon the biomechanics and remodeling of isolated amnion membrane, in vitro', Placenta, vol. 32, no. 3, pp. 206-213. https://doi.org/10.1016/j.placenta.2011.01.006
Kumar, D. ; Schatz, F. ; Moore, R. M. ; Mercer, B. M. ; Rangaswamy, N. ; Mansour, J. M. ; Lockwood, C. J. ; Moore, J. J. / The effects of thrombin and cytokines upon the biomechanics and remodeling of isolated amnion membrane, in vitro. In: Placenta. 2011 ; Vol. 32, No. 3. pp. 206-213.
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KW - Thrombin

KW - TNF

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