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John J Moore - One of the best experts on this subject based on the ideXlab platform.

  • granulocyte macrophage colony stimulating factor gm csf the critical intermediate of inflammation induced Fetal Membrane weakening primarily exerts its weakening effect on the choriodecidua rather than the amnion
    Placenta, 2020
    Co-Authors: Anudeepa Sharma, Robert M Moore, Joseph M Mansour, Deepak Kumar, Brian M. Mercer, Aditi Deshmukh, John J Moore
    Abstract:

    Abstract Introduction We have previously demonstrated two associations of PPROM, (1) inflammation/infection (modeled by tumor necrosis factor (TNF) and (2) decidual bleeding (modeled by thrombin), both decrease Fetal Membrane (FM) rupture strength in-vitro. Furthermore, Granulocyte-Macrophage-Colony-Stimulating-Factor (GM-CSF) induced by both TNF and thrombin is a critical intermediate, necessary and sufficient for weakening by either agent. The amnion is the strength component of FM and must weaken for FM to rupture. It is unclear whether GM-CSF weakens amnion (AM) directly, or initially targets choriodecidua (CD) which secondarily releases agents to act on amnion. Methods Full thickness FM fragments were treated with/without GM-CSF. Some were preincubated with alpha-lipoic acid (LA), a known inhibitor of FM weakening. The FM fragments were then strength-tested. Separately, FM fragments were initially separated to AM and CD. AM fragments were cultured with Medium ± GM-CSF and then strength-tested. In other experiments, CD fragments were cultured with Medium, GM-CSF, LA, or LA + GM-CSF. Conditioned medium from each group was then incubated with AM. AM was then strength-tested. Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Matrix Metalloproteinases (TIMPs) were analyzed by Mutiplex Elisa. Results GM-CSF weakened intact FM which was blocked by LA. GM-CSF did not weaken isolated AM. However, GM-CSF conditioned CD media weakened AM and this weakening was inhibited by LA. GM-CSF treatment of CD increased MMPs 2, 9, and 10, and decreased TIMPs 1–3. LA reversed these effects. Conclusions GM-CSF does not weaken amnion directly; GM-CSF acts on CD to increase proteases and decrease anti-proteases which secondarily weaken the amnion.

  • in an in vitro model using human Fetal Membranes α lipoic acid inhibits inflammation induced Fetal Membrane weakening
    Placenta, 2018
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Anudeepa Sharma, John J Moore
    Abstract:

    Abstract Introduction We established an in-vitro model for the study of human Fetal Membrane (FM) weakening leading to pPROM. In this model, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both tumor necrosis factor-α (TNF; modeling infection/inflammation) and thrombin (modeling decidual bleeding/abruption)-induced weakening. Thus, inhibitors of FM weakening can be categorized as targeting GM-CSF production, GM-CSF downstream action, or both. Most progestogens inhibit both, except 17-α hydroxyprogesterone caproate which inhibits FM weakening at only one point, GM-CSF production. α-lipoic acid (LA), an over-the-counter dietary supplement, has also been previously shown to inhibit TNF and thrombin induced FM weakening. Objective To determine the point of action of LA inhibition of FM weakening. Methods FM fragments were mounted in Transwell inserts and preincubated with/without LA/24 h, then with/without addition of TNF, thrombin or GM-CSF. After 48 h, medium was assayed for GM-CSF, and FM fragments were rupture-strength tested. Results TNF and thrombin both weakened FM and increased GM-CSF levels. GM-CSF also weakened FM. LA inhibited both TNF and thrombin induced FM weakening and concomitantly inhibited the increase in GM-CSF in a concentration-dependent manner. In addition, LA inhibited GM-CSF induced FM weakening in a concentration dependent manner. Conclusions LA blocks TNF and thrombin induced FM weakening at two points, inhibiting both GM-CSF production and downstream action. Thus, we speculate that LA may be a potential standalone therapeutic agent, or supplement to current therapy for prevention of pPROM related spontaneous preterm birth, if preclinical studies to examine feasibility and safety during pregnancy are successfully accomplished.

  • in an in vitro model using human Fetal Membranes 17 α hydroxyprogesterone caproate is not an optimal progestogen for inhibition of Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2017
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, John J Moore
    Abstract:

    Background The progestogen 17-α hydroxyprogesterone caproate (17-OHPC) is 1 of only 2 agents recommended for clinical use in the prevention of spontaneous preterm delivery, and studies of its efficacy have been conflicting. We have developed an in-vitro model to study the Fetal Membrane weakening process that leads to rupture in preterm premature rupture of the Fetal Membranes (pPROM). Inflammation/infection associated with tumor necrosis factor-α (TNF-α) induction and decidual bleeding/abruption associated thrombin release are leading causes of preterm premature rupture of the Fetal Membranes. Both agents (TNF-α and thrombin) cause Fetal Membrane weakening in the model system. Furthermore, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both TNF-α and thrombin-induced Fetal Membrane weakening. In a previous report, we demonstrated that 3 progestogens, progesterone, 17-alpha hydroxyprogesterone (17-OHP), and medroxyprogesterone acetate (MPA), each inhibit both TNF-α– and thrombin-induced Fetal Membrane weakening at 2 distinct points of the Fetal Membrane weakening pathway. Each block both the production of and the downstream action of the critical intermediate granulocyte-macrophage colony-stimulating factor. Objective The objective of the study was to characterize the inhibitory effects of 17-OHPC on TNF-α– and thrombin-induced Fetal Membrane weakening in vitro. Study Design Full-thickness human Fetal Membrane fragments from uncomplicated term repeat cesarean deliveries were mounted in 2.5 cm Transwell inserts and cultured with/without 17-alpha hydroxyprogesterone caproate (10 –9 to 10 –7 M). After 24 hours, medium (supernatant) was removed and replaced with/without the addition of tumor necrosis factor-alpha (20 ng/mL) or thrombin (10 U/mL) or granulocyte-macrophage colony-stimulating factor (200 ng/mL). After 48 hours of culture, medium from the maternal side compartment of the model was assayed for granulocyte-macrophage colony-stimulating factor and the Fetal Membrane fragments were rupture strength tested. Results Tumor necrosis factor-alpha and thrombin both weakened Fetal Membranes (43% and 62%, respectively) and increased granulocyte-macrophage colony-stimulating factor levels (3.7- and 5.9-fold, respectively). Pretreatment with 17-alpha hydroxyprogesterone caproate inhibited both tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening and concomitantly inhibited the induced increase in granulocyte-macrophage colony-stimulating factor in a concentration-dependent manner. However, contrary to our prior reports regarding progesterone and other progestogens, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced Fetal Membrane weakening. Conclusion 17-Alpha hydroxyprogesterone caproate blocks tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening by inhibiting the production of granulocyte-macrophage colony-stimulating factor. However, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced weakening. We speculate that progestogens other than 17-alpha hydroxyprogesterone caproate may be more efficacious in preventing preterm premature rupture of the Fetal Membranes–related spontaneous preterm birth.

  • the physiology of Fetal Membrane weakening and rupture insights gained from the determination of physical properties revisited
    Placenta, 2016
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Raymond W Redline, John J Moore
    Abstract:

    Rupture of the Fetal Membranes (FM) is precipitated by stretch forces acting upon biochemically mediated, pre-weakened tissue. Term FM develop a para-cervical weak zone, characterized by collagen remodeling and apoptosis, within which FM rupture is thought to initiate. Preterm FM also have a weak region but are stronger overall than term FM. Inflammation/infection and decidual bleeding/abruption are strongly associated with preterm premature FM rupture (pPROM), but the specific mechanisms causing FM weakening-rupture in pPROM are unknown. There are no animal models for study of FM weakening and rupture. Over a decade ago we developed equipment and methodology to test human FM strength and incorporated it into a FM explant system to create an in-vitro human FM weakening model system. Within this model TNF (modeling inflammation) and Thrombin (modeling bleeding) both weaken human FM with concomitant up regulation of MMP9 and cellular apoptosis, mimicking the characteristics of the spontaneous FM rupture site. The model has been enhanced so that test agents can be applied directionally to the choriodecidual side of the FM explant consistent with the in-vivo situation. With this enhanced system we have demonstrated that the pathways involving inflammation/TNF and bleeding/Thrombin induced FM weakening overlap. Furthermore GM-CSF production was demonstrated to be a critical common intermediate step in both the TNF and the Thrombin induced FM weakening pathways. This model system has also been used to test potential inhibitors of FM weakening and therefore pPROM. The dietary supplement α-lipoic acid and progestogens (P4, MPA and 17α-hydroxyprogesterone) have been shown to inhibit both TNF and Thrombin induced FM weakening. The progestogens act at multiple points by inhibiting both GM-CSF production and GM-CSF action. The use of a combined biomechanical/biochemical in-vitro human FM weakening model system has allowed the pathways of Fetal Membrane weakening to be delineated, and agents that may be of clinical use in inhibiting these pathways to be tested.

  • progesterone inhibits in vitro Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2015
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, Edward Springel, Elliot Philipson, John J Moore
    Abstract:

    Objective Inflammation/infection and abruption are leading causes of preterm premature rupture of the Membranes. Recently, we identified granulocyte-macrophage colony-stimulating factor (GM-CSF) as a critical mediator of both tumor necrosis factor-α– (TNF; modeling inflammation) and thrombin-induced (modeling abruption) weakening of the Fetal Membranes. We found that (1) TNF and thrombin both induced GM-CSF in the choriodecidua, (2) blockade of GM-CSF action with neutralizing antibodies inhibited both TNF- and thrombin-induced Fetal Membrane weakening, and (3) GM-CSF alone induced Fetal Membrane weakening. GM-CSF is thus part of an overlap of the inflammation and abruption-induced Fetal Membrane weakening pathways. The effects of progesterone analogs on the pathways by which Fetal Membranes are weakened have not been investigated. We examined the effects of progesterone, medroxyprogesterone acetate (MPA) and 17α-hydroxyprogesterone (HP) on TNF- and thrombin-induced Fetal Membrane weakening. Study Design Full-thickness Fetal Membranes from uncomplicated term repeat cesarean deliveries were mounted in Transwell inserts in Minimum Essential Medium alpha and incubated at 37°C in 5% CO2. The choriodecidua side of the Fetal Membrane fragments were preincubated with progesterone, MPA, HP, or vehicle for 24 hours. Fetal Membranes were then exposed to TNF, thrombin, or GM-CSF on the choriodecidua side for an additional 48 hours. The Fetal Membrane tissues were then strength tested, and medium from the choriodecidua and amnion compartments was assayed for GM-CSF content. Results TNF and thrombin both weakened Fetal Membranes and elevated media GM-CSF levels on the choriodecidua side of the Fetal Membrane. Pretreatment with progesterone, MPA, or HP inhibited both TNF- and thrombin-induced Fetal Membrane weakening and also inhibited the induced increase in GM-CSF. GM-CSF decreased Fetal Membrane rupture strength by 68%, which was inhibited by progestogen pretreatment with a potency order: progesterone Conclusion Progestogen pretreatment blocks TNF- and thrombin-induced Fetal Membrane weakening by inhibiting both the production and action of GM-CSF. These findings are consistent with the administration of progestogens in the prevention of preterm premature rupture of the Membranes.

Deepak Kumar - One of the best experts on this subject based on the ideXlab platform.

  • granulocyte macrophage colony stimulating factor gm csf the critical intermediate of inflammation induced Fetal Membrane weakening primarily exerts its weakening effect on the choriodecidua rather than the amnion
    Placenta, 2020
    Co-Authors: Anudeepa Sharma, Robert M Moore, Joseph M Mansour, Deepak Kumar, Brian M. Mercer, Aditi Deshmukh, John J Moore
    Abstract:

    Abstract Introduction We have previously demonstrated two associations of PPROM, (1) inflammation/infection (modeled by tumor necrosis factor (TNF) and (2) decidual bleeding (modeled by thrombin), both decrease Fetal Membrane (FM) rupture strength in-vitro. Furthermore, Granulocyte-Macrophage-Colony-Stimulating-Factor (GM-CSF) induced by both TNF and thrombin is a critical intermediate, necessary and sufficient for weakening by either agent. The amnion is the strength component of FM and must weaken for FM to rupture. It is unclear whether GM-CSF weakens amnion (AM) directly, or initially targets choriodecidua (CD) which secondarily releases agents to act on amnion. Methods Full thickness FM fragments were treated with/without GM-CSF. Some were preincubated with alpha-lipoic acid (LA), a known inhibitor of FM weakening. The FM fragments were then strength-tested. Separately, FM fragments were initially separated to AM and CD. AM fragments were cultured with Medium ± GM-CSF and then strength-tested. In other experiments, CD fragments were cultured with Medium, GM-CSF, LA, or LA + GM-CSF. Conditioned medium from each group was then incubated with AM. AM was then strength-tested. Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Matrix Metalloproteinases (TIMPs) were analyzed by Mutiplex Elisa. Results GM-CSF weakened intact FM which was blocked by LA. GM-CSF did not weaken isolated AM. However, GM-CSF conditioned CD media weakened AM and this weakening was inhibited by LA. GM-CSF treatment of CD increased MMPs 2, 9, and 10, and decreased TIMPs 1–3. LA reversed these effects. Conclusions GM-CSF does not weaken amnion directly; GM-CSF acts on CD to increase proteases and decrease anti-proteases which secondarily weaken the amnion.

  • in an in vitro model using human Fetal Membranes α lipoic acid inhibits inflammation induced Fetal Membrane weakening
    Placenta, 2018
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Anudeepa Sharma, John J Moore
    Abstract:

    Abstract Introduction We established an in-vitro model for the study of human Fetal Membrane (FM) weakening leading to pPROM. In this model, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both tumor necrosis factor-α (TNF; modeling infection/inflammation) and thrombin (modeling decidual bleeding/abruption)-induced weakening. Thus, inhibitors of FM weakening can be categorized as targeting GM-CSF production, GM-CSF downstream action, or both. Most progestogens inhibit both, except 17-α hydroxyprogesterone caproate which inhibits FM weakening at only one point, GM-CSF production. α-lipoic acid (LA), an over-the-counter dietary supplement, has also been previously shown to inhibit TNF and thrombin induced FM weakening. Objective To determine the point of action of LA inhibition of FM weakening. Methods FM fragments were mounted in Transwell inserts and preincubated with/without LA/24 h, then with/without addition of TNF, thrombin or GM-CSF. After 48 h, medium was assayed for GM-CSF, and FM fragments were rupture-strength tested. Results TNF and thrombin both weakened FM and increased GM-CSF levels. GM-CSF also weakened FM. LA inhibited both TNF and thrombin induced FM weakening and concomitantly inhibited the increase in GM-CSF in a concentration-dependent manner. In addition, LA inhibited GM-CSF induced FM weakening in a concentration dependent manner. Conclusions LA blocks TNF and thrombin induced FM weakening at two points, inhibiting both GM-CSF production and downstream action. Thus, we speculate that LA may be a potential standalone therapeutic agent, or supplement to current therapy for prevention of pPROM related spontaneous preterm birth, if preclinical studies to examine feasibility and safety during pregnancy are successfully accomplished.

  • in an in vitro model using human Fetal Membranes 17 α hydroxyprogesterone caproate is not an optimal progestogen for inhibition of Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2017
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, John J Moore
    Abstract:

    Background The progestogen 17-α hydroxyprogesterone caproate (17-OHPC) is 1 of only 2 agents recommended for clinical use in the prevention of spontaneous preterm delivery, and studies of its efficacy have been conflicting. We have developed an in-vitro model to study the Fetal Membrane weakening process that leads to rupture in preterm premature rupture of the Fetal Membranes (pPROM). Inflammation/infection associated with tumor necrosis factor-α (TNF-α) induction and decidual bleeding/abruption associated thrombin release are leading causes of preterm premature rupture of the Fetal Membranes. Both agents (TNF-α and thrombin) cause Fetal Membrane weakening in the model system. Furthermore, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both TNF-α and thrombin-induced Fetal Membrane weakening. In a previous report, we demonstrated that 3 progestogens, progesterone, 17-alpha hydroxyprogesterone (17-OHP), and medroxyprogesterone acetate (MPA), each inhibit both TNF-α– and thrombin-induced Fetal Membrane weakening at 2 distinct points of the Fetal Membrane weakening pathway. Each block both the production of and the downstream action of the critical intermediate granulocyte-macrophage colony-stimulating factor. Objective The objective of the study was to characterize the inhibitory effects of 17-OHPC on TNF-α– and thrombin-induced Fetal Membrane weakening in vitro. Study Design Full-thickness human Fetal Membrane fragments from uncomplicated term repeat cesarean deliveries were mounted in 2.5 cm Transwell inserts and cultured with/without 17-alpha hydroxyprogesterone caproate (10 –9 to 10 –7 M). After 24 hours, medium (supernatant) was removed and replaced with/without the addition of tumor necrosis factor-alpha (20 ng/mL) or thrombin (10 U/mL) or granulocyte-macrophage colony-stimulating factor (200 ng/mL). After 48 hours of culture, medium from the maternal side compartment of the model was assayed for granulocyte-macrophage colony-stimulating factor and the Fetal Membrane fragments were rupture strength tested. Results Tumor necrosis factor-alpha and thrombin both weakened Fetal Membranes (43% and 62%, respectively) and increased granulocyte-macrophage colony-stimulating factor levels (3.7- and 5.9-fold, respectively). Pretreatment with 17-alpha hydroxyprogesterone caproate inhibited both tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening and concomitantly inhibited the induced increase in granulocyte-macrophage colony-stimulating factor in a concentration-dependent manner. However, contrary to our prior reports regarding progesterone and other progestogens, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced Fetal Membrane weakening. Conclusion 17-Alpha hydroxyprogesterone caproate blocks tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening by inhibiting the production of granulocyte-macrophage colony-stimulating factor. However, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced weakening. We speculate that progestogens other than 17-alpha hydroxyprogesterone caproate may be more efficacious in preventing preterm premature rupture of the Fetal Membranes–related spontaneous preterm birth.

  • the physiology of Fetal Membrane weakening and rupture insights gained from the determination of physical properties revisited
    Placenta, 2016
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Raymond W Redline, John J Moore
    Abstract:

    Rupture of the Fetal Membranes (FM) is precipitated by stretch forces acting upon biochemically mediated, pre-weakened tissue. Term FM develop a para-cervical weak zone, characterized by collagen remodeling and apoptosis, within which FM rupture is thought to initiate. Preterm FM also have a weak region but are stronger overall than term FM. Inflammation/infection and decidual bleeding/abruption are strongly associated with preterm premature FM rupture (pPROM), but the specific mechanisms causing FM weakening-rupture in pPROM are unknown. There are no animal models for study of FM weakening and rupture. Over a decade ago we developed equipment and methodology to test human FM strength and incorporated it into a FM explant system to create an in-vitro human FM weakening model system. Within this model TNF (modeling inflammation) and Thrombin (modeling bleeding) both weaken human FM with concomitant up regulation of MMP9 and cellular apoptosis, mimicking the characteristics of the spontaneous FM rupture site. The model has been enhanced so that test agents can be applied directionally to the choriodecidual side of the FM explant consistent with the in-vivo situation. With this enhanced system we have demonstrated that the pathways involving inflammation/TNF and bleeding/Thrombin induced FM weakening overlap. Furthermore GM-CSF production was demonstrated to be a critical common intermediate step in both the TNF and the Thrombin induced FM weakening pathways. This model system has also been used to test potential inhibitors of FM weakening and therefore pPROM. The dietary supplement α-lipoic acid and progestogens (P4, MPA and 17α-hydroxyprogesterone) have been shown to inhibit both TNF and Thrombin induced FM weakening. The progestogens act at multiple points by inhibiting both GM-CSF production and GM-CSF action. The use of a combined biomechanical/biochemical in-vitro human FM weakening model system has allowed the pathways of Fetal Membrane weakening to be delineated, and agents that may be of clinical use in inhibiting these pathways to be tested.

  • progesterone inhibits in vitro Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2015
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, Edward Springel, Elliot Philipson, John J Moore
    Abstract:

    Objective Inflammation/infection and abruption are leading causes of preterm premature rupture of the Membranes. Recently, we identified granulocyte-macrophage colony-stimulating factor (GM-CSF) as a critical mediator of both tumor necrosis factor-α– (TNF; modeling inflammation) and thrombin-induced (modeling abruption) weakening of the Fetal Membranes. We found that (1) TNF and thrombin both induced GM-CSF in the choriodecidua, (2) blockade of GM-CSF action with neutralizing antibodies inhibited both TNF- and thrombin-induced Fetal Membrane weakening, and (3) GM-CSF alone induced Fetal Membrane weakening. GM-CSF is thus part of an overlap of the inflammation and abruption-induced Fetal Membrane weakening pathways. The effects of progesterone analogs on the pathways by which Fetal Membranes are weakened have not been investigated. We examined the effects of progesterone, medroxyprogesterone acetate (MPA) and 17α-hydroxyprogesterone (HP) on TNF- and thrombin-induced Fetal Membrane weakening. Study Design Full-thickness Fetal Membranes from uncomplicated term repeat cesarean deliveries were mounted in Transwell inserts in Minimum Essential Medium alpha and incubated at 37°C in 5% CO2. The choriodecidua side of the Fetal Membrane fragments were preincubated with progesterone, MPA, HP, or vehicle for 24 hours. Fetal Membranes were then exposed to TNF, thrombin, or GM-CSF on the choriodecidua side for an additional 48 hours. The Fetal Membrane tissues were then strength tested, and medium from the choriodecidua and amnion compartments was assayed for GM-CSF content. Results TNF and thrombin both weakened Fetal Membranes and elevated media GM-CSF levels on the choriodecidua side of the Fetal Membrane. Pretreatment with progesterone, MPA, or HP inhibited both TNF- and thrombin-induced Fetal Membrane weakening and also inhibited the induced increase in GM-CSF. GM-CSF decreased Fetal Membrane rupture strength by 68%, which was inhibited by progestogen pretreatment with a potency order: progesterone Conclusion Progestogen pretreatment blocks TNF- and thrombin-induced Fetal Membrane weakening by inhibiting both the production and action of GM-CSF. These findings are consistent with the administration of progestogens in the prevention of preterm premature rupture of the Membranes.

Brian M. Mercer - One of the best experts on this subject based on the ideXlab platform.

  • granulocyte macrophage colony stimulating factor gm csf the critical intermediate of inflammation induced Fetal Membrane weakening primarily exerts its weakening effect on the choriodecidua rather than the amnion
    Placenta, 2020
    Co-Authors: Anudeepa Sharma, Robert M Moore, Joseph M Mansour, Deepak Kumar, Brian M. Mercer, Aditi Deshmukh, John J Moore
    Abstract:

    Abstract Introduction We have previously demonstrated two associations of PPROM, (1) inflammation/infection (modeled by tumor necrosis factor (TNF) and (2) decidual bleeding (modeled by thrombin), both decrease Fetal Membrane (FM) rupture strength in-vitro. Furthermore, Granulocyte-Macrophage-Colony-Stimulating-Factor (GM-CSF) induced by both TNF and thrombin is a critical intermediate, necessary and sufficient for weakening by either agent. The amnion is the strength component of FM and must weaken for FM to rupture. It is unclear whether GM-CSF weakens amnion (AM) directly, or initially targets choriodecidua (CD) which secondarily releases agents to act on amnion. Methods Full thickness FM fragments were treated with/without GM-CSF. Some were preincubated with alpha-lipoic acid (LA), a known inhibitor of FM weakening. The FM fragments were then strength-tested. Separately, FM fragments were initially separated to AM and CD. AM fragments were cultured with Medium ± GM-CSF and then strength-tested. In other experiments, CD fragments were cultured with Medium, GM-CSF, LA, or LA + GM-CSF. Conditioned medium from each group was then incubated with AM. AM was then strength-tested. Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Matrix Metalloproteinases (TIMPs) were analyzed by Mutiplex Elisa. Results GM-CSF weakened intact FM which was blocked by LA. GM-CSF did not weaken isolated AM. However, GM-CSF conditioned CD media weakened AM and this weakening was inhibited by LA. GM-CSF treatment of CD increased MMPs 2, 9, and 10, and decreased TIMPs 1–3. LA reversed these effects. Conclusions GM-CSF does not weaken amnion directly; GM-CSF acts on CD to increase proteases and decrease anti-proteases which secondarily weaken the amnion.

  • in an in vitro model using human Fetal Membranes α lipoic acid inhibits inflammation induced Fetal Membrane weakening
    Placenta, 2018
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Anudeepa Sharma, John J Moore
    Abstract:

    Abstract Introduction We established an in-vitro model for the study of human Fetal Membrane (FM) weakening leading to pPROM. In this model, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both tumor necrosis factor-α (TNF; modeling infection/inflammation) and thrombin (modeling decidual bleeding/abruption)-induced weakening. Thus, inhibitors of FM weakening can be categorized as targeting GM-CSF production, GM-CSF downstream action, or both. Most progestogens inhibit both, except 17-α hydroxyprogesterone caproate which inhibits FM weakening at only one point, GM-CSF production. α-lipoic acid (LA), an over-the-counter dietary supplement, has also been previously shown to inhibit TNF and thrombin induced FM weakening. Objective To determine the point of action of LA inhibition of FM weakening. Methods FM fragments were mounted in Transwell inserts and preincubated with/without LA/24 h, then with/without addition of TNF, thrombin or GM-CSF. After 48 h, medium was assayed for GM-CSF, and FM fragments were rupture-strength tested. Results TNF and thrombin both weakened FM and increased GM-CSF levels. GM-CSF also weakened FM. LA inhibited both TNF and thrombin induced FM weakening and concomitantly inhibited the increase in GM-CSF in a concentration-dependent manner. In addition, LA inhibited GM-CSF induced FM weakening in a concentration dependent manner. Conclusions LA blocks TNF and thrombin induced FM weakening at two points, inhibiting both GM-CSF production and downstream action. Thus, we speculate that LA may be a potential standalone therapeutic agent, or supplement to current therapy for prevention of pPROM related spontaneous preterm birth, if preclinical studies to examine feasibility and safety during pregnancy are successfully accomplished.

  • in an in vitro model using human Fetal Membranes 17 α hydroxyprogesterone caproate is not an optimal progestogen for inhibition of Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2017
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, John J Moore
    Abstract:

    Background The progestogen 17-α hydroxyprogesterone caproate (17-OHPC) is 1 of only 2 agents recommended for clinical use in the prevention of spontaneous preterm delivery, and studies of its efficacy have been conflicting. We have developed an in-vitro model to study the Fetal Membrane weakening process that leads to rupture in preterm premature rupture of the Fetal Membranes (pPROM). Inflammation/infection associated with tumor necrosis factor-α (TNF-α) induction and decidual bleeding/abruption associated thrombin release are leading causes of preterm premature rupture of the Fetal Membranes. Both agents (TNF-α and thrombin) cause Fetal Membrane weakening in the model system. Furthermore, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both TNF-α and thrombin-induced Fetal Membrane weakening. In a previous report, we demonstrated that 3 progestogens, progesterone, 17-alpha hydroxyprogesterone (17-OHP), and medroxyprogesterone acetate (MPA), each inhibit both TNF-α– and thrombin-induced Fetal Membrane weakening at 2 distinct points of the Fetal Membrane weakening pathway. Each block both the production of and the downstream action of the critical intermediate granulocyte-macrophage colony-stimulating factor. Objective The objective of the study was to characterize the inhibitory effects of 17-OHPC on TNF-α– and thrombin-induced Fetal Membrane weakening in vitro. Study Design Full-thickness human Fetal Membrane fragments from uncomplicated term repeat cesarean deliveries were mounted in 2.5 cm Transwell inserts and cultured with/without 17-alpha hydroxyprogesterone caproate (10 –9 to 10 –7 M). After 24 hours, medium (supernatant) was removed and replaced with/without the addition of tumor necrosis factor-alpha (20 ng/mL) or thrombin (10 U/mL) or granulocyte-macrophage colony-stimulating factor (200 ng/mL). After 48 hours of culture, medium from the maternal side compartment of the model was assayed for granulocyte-macrophage colony-stimulating factor and the Fetal Membrane fragments were rupture strength tested. Results Tumor necrosis factor-alpha and thrombin both weakened Fetal Membranes (43% and 62%, respectively) and increased granulocyte-macrophage colony-stimulating factor levels (3.7- and 5.9-fold, respectively). Pretreatment with 17-alpha hydroxyprogesterone caproate inhibited both tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening and concomitantly inhibited the induced increase in granulocyte-macrophage colony-stimulating factor in a concentration-dependent manner. However, contrary to our prior reports regarding progesterone and other progestogens, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced Fetal Membrane weakening. Conclusion 17-Alpha hydroxyprogesterone caproate blocks tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening by inhibiting the production of granulocyte-macrophage colony-stimulating factor. However, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced weakening. We speculate that progestogens other than 17-alpha hydroxyprogesterone caproate may be more efficacious in preventing preterm premature rupture of the Fetal Membranes–related spontaneous preterm birth.

  • the physiology of Fetal Membrane weakening and rupture insights gained from the determination of physical properties revisited
    Placenta, 2016
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Raymond W Redline, John J Moore
    Abstract:

    Rupture of the Fetal Membranes (FM) is precipitated by stretch forces acting upon biochemically mediated, pre-weakened tissue. Term FM develop a para-cervical weak zone, characterized by collagen remodeling and apoptosis, within which FM rupture is thought to initiate. Preterm FM also have a weak region but are stronger overall than term FM. Inflammation/infection and decidual bleeding/abruption are strongly associated with preterm premature FM rupture (pPROM), but the specific mechanisms causing FM weakening-rupture in pPROM are unknown. There are no animal models for study of FM weakening and rupture. Over a decade ago we developed equipment and methodology to test human FM strength and incorporated it into a FM explant system to create an in-vitro human FM weakening model system. Within this model TNF (modeling inflammation) and Thrombin (modeling bleeding) both weaken human FM with concomitant up regulation of MMP9 and cellular apoptosis, mimicking the characteristics of the spontaneous FM rupture site. The model has been enhanced so that test agents can be applied directionally to the choriodecidual side of the FM explant consistent with the in-vivo situation. With this enhanced system we have demonstrated that the pathways involving inflammation/TNF and bleeding/Thrombin induced FM weakening overlap. Furthermore GM-CSF production was demonstrated to be a critical common intermediate step in both the TNF and the Thrombin induced FM weakening pathways. This model system has also been used to test potential inhibitors of FM weakening and therefore pPROM. The dietary supplement α-lipoic acid and progestogens (P4, MPA and 17α-hydroxyprogesterone) have been shown to inhibit both TNF and Thrombin induced FM weakening. The progestogens act at multiple points by inhibiting both GM-CSF production and GM-CSF action. The use of a combined biomechanical/biochemical in-vitro human FM weakening model system has allowed the pathways of Fetal Membrane weakening to be delineated, and agents that may be of clinical use in inhibiting these pathways to be tested.

  • progesterone inhibits in vitro Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2015
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, Edward Springel, Elliot Philipson, John J Moore
    Abstract:

    Objective Inflammation/infection and abruption are leading causes of preterm premature rupture of the Membranes. Recently, we identified granulocyte-macrophage colony-stimulating factor (GM-CSF) as a critical mediator of both tumor necrosis factor-α– (TNF; modeling inflammation) and thrombin-induced (modeling abruption) weakening of the Fetal Membranes. We found that (1) TNF and thrombin both induced GM-CSF in the choriodecidua, (2) blockade of GM-CSF action with neutralizing antibodies inhibited both TNF- and thrombin-induced Fetal Membrane weakening, and (3) GM-CSF alone induced Fetal Membrane weakening. GM-CSF is thus part of an overlap of the inflammation and abruption-induced Fetal Membrane weakening pathways. The effects of progesterone analogs on the pathways by which Fetal Membranes are weakened have not been investigated. We examined the effects of progesterone, medroxyprogesterone acetate (MPA) and 17α-hydroxyprogesterone (HP) on TNF- and thrombin-induced Fetal Membrane weakening. Study Design Full-thickness Fetal Membranes from uncomplicated term repeat cesarean deliveries were mounted in Transwell inserts in Minimum Essential Medium alpha and incubated at 37°C in 5% CO2. The choriodecidua side of the Fetal Membrane fragments were preincubated with progesterone, MPA, HP, or vehicle for 24 hours. Fetal Membranes were then exposed to TNF, thrombin, or GM-CSF on the choriodecidua side for an additional 48 hours. The Fetal Membrane tissues were then strength tested, and medium from the choriodecidua and amnion compartments was assayed for GM-CSF content. Results TNF and thrombin both weakened Fetal Membranes and elevated media GM-CSF levels on the choriodecidua side of the Fetal Membrane. Pretreatment with progesterone, MPA, or HP inhibited both TNF- and thrombin-induced Fetal Membrane weakening and also inhibited the induced increase in GM-CSF. GM-CSF decreased Fetal Membrane rupture strength by 68%, which was inhibited by progestogen pretreatment with a potency order: progesterone Conclusion Progestogen pretreatment blocks TNF- and thrombin-induced Fetal Membrane weakening by inhibiting both the production and action of GM-CSF. These findings are consistent with the administration of progestogens in the prevention of preterm premature rupture of the Membranes.

Robert M Moore - One of the best experts on this subject based on the ideXlab platform.

  • granulocyte macrophage colony stimulating factor gm csf the critical intermediate of inflammation induced Fetal Membrane weakening primarily exerts its weakening effect on the choriodecidua rather than the amnion
    Placenta, 2020
    Co-Authors: Anudeepa Sharma, Robert M Moore, Joseph M Mansour, Deepak Kumar, Brian M. Mercer, Aditi Deshmukh, John J Moore
    Abstract:

    Abstract Introduction We have previously demonstrated two associations of PPROM, (1) inflammation/infection (modeled by tumor necrosis factor (TNF) and (2) decidual bleeding (modeled by thrombin), both decrease Fetal Membrane (FM) rupture strength in-vitro. Furthermore, Granulocyte-Macrophage-Colony-Stimulating-Factor (GM-CSF) induced by both TNF and thrombin is a critical intermediate, necessary and sufficient for weakening by either agent. The amnion is the strength component of FM and must weaken for FM to rupture. It is unclear whether GM-CSF weakens amnion (AM) directly, or initially targets choriodecidua (CD) which secondarily releases agents to act on amnion. Methods Full thickness FM fragments were treated with/without GM-CSF. Some were preincubated with alpha-lipoic acid (LA), a known inhibitor of FM weakening. The FM fragments were then strength-tested. Separately, FM fragments were initially separated to AM and CD. AM fragments were cultured with Medium ± GM-CSF and then strength-tested. In other experiments, CD fragments were cultured with Medium, GM-CSF, LA, or LA + GM-CSF. Conditioned medium from each group was then incubated with AM. AM was then strength-tested. Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Matrix Metalloproteinases (TIMPs) were analyzed by Mutiplex Elisa. Results GM-CSF weakened intact FM which was blocked by LA. GM-CSF did not weaken isolated AM. However, GM-CSF conditioned CD media weakened AM and this weakening was inhibited by LA. GM-CSF treatment of CD increased MMPs 2, 9, and 10, and decreased TIMPs 1–3. LA reversed these effects. Conclusions GM-CSF does not weaken amnion directly; GM-CSF acts on CD to increase proteases and decrease anti-proteases which secondarily weaken the amnion.

  • in an in vitro model using human Fetal Membranes α lipoic acid inhibits inflammation induced Fetal Membrane weakening
    Placenta, 2018
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Anudeepa Sharma, John J Moore
    Abstract:

    Abstract Introduction We established an in-vitro model for the study of human Fetal Membrane (FM) weakening leading to pPROM. In this model, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both tumor necrosis factor-α (TNF; modeling infection/inflammation) and thrombin (modeling decidual bleeding/abruption)-induced weakening. Thus, inhibitors of FM weakening can be categorized as targeting GM-CSF production, GM-CSF downstream action, or both. Most progestogens inhibit both, except 17-α hydroxyprogesterone caproate which inhibits FM weakening at only one point, GM-CSF production. α-lipoic acid (LA), an over-the-counter dietary supplement, has also been previously shown to inhibit TNF and thrombin induced FM weakening. Objective To determine the point of action of LA inhibition of FM weakening. Methods FM fragments were mounted in Transwell inserts and preincubated with/without LA/24 h, then with/without addition of TNF, thrombin or GM-CSF. After 48 h, medium was assayed for GM-CSF, and FM fragments were rupture-strength tested. Results TNF and thrombin both weakened FM and increased GM-CSF levels. GM-CSF also weakened FM. LA inhibited both TNF and thrombin induced FM weakening and concomitantly inhibited the increase in GM-CSF in a concentration-dependent manner. In addition, LA inhibited GM-CSF induced FM weakening in a concentration dependent manner. Conclusions LA blocks TNF and thrombin induced FM weakening at two points, inhibiting both GM-CSF production and downstream action. Thus, we speculate that LA may be a potential standalone therapeutic agent, or supplement to current therapy for prevention of pPROM related spontaneous preterm birth, if preclinical studies to examine feasibility and safety during pregnancy are successfully accomplished.

  • in an in vitro model using human Fetal Membranes 17 α hydroxyprogesterone caproate is not an optimal progestogen for inhibition of Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2017
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, John J Moore
    Abstract:

    Background The progestogen 17-α hydroxyprogesterone caproate (17-OHPC) is 1 of only 2 agents recommended for clinical use in the prevention of spontaneous preterm delivery, and studies of its efficacy have been conflicting. We have developed an in-vitro model to study the Fetal Membrane weakening process that leads to rupture in preterm premature rupture of the Fetal Membranes (pPROM). Inflammation/infection associated with tumor necrosis factor-α (TNF-α) induction and decidual bleeding/abruption associated thrombin release are leading causes of preterm premature rupture of the Fetal Membranes. Both agents (TNF-α and thrombin) cause Fetal Membrane weakening in the model system. Furthermore, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both TNF-α and thrombin-induced Fetal Membrane weakening. In a previous report, we demonstrated that 3 progestogens, progesterone, 17-alpha hydroxyprogesterone (17-OHP), and medroxyprogesterone acetate (MPA), each inhibit both TNF-α– and thrombin-induced Fetal Membrane weakening at 2 distinct points of the Fetal Membrane weakening pathway. Each block both the production of and the downstream action of the critical intermediate granulocyte-macrophage colony-stimulating factor. Objective The objective of the study was to characterize the inhibitory effects of 17-OHPC on TNF-α– and thrombin-induced Fetal Membrane weakening in vitro. Study Design Full-thickness human Fetal Membrane fragments from uncomplicated term repeat cesarean deliveries were mounted in 2.5 cm Transwell inserts and cultured with/without 17-alpha hydroxyprogesterone caproate (10 –9 to 10 –7 M). After 24 hours, medium (supernatant) was removed and replaced with/without the addition of tumor necrosis factor-alpha (20 ng/mL) or thrombin (10 U/mL) or granulocyte-macrophage colony-stimulating factor (200 ng/mL). After 48 hours of culture, medium from the maternal side compartment of the model was assayed for granulocyte-macrophage colony-stimulating factor and the Fetal Membrane fragments were rupture strength tested. Results Tumor necrosis factor-alpha and thrombin both weakened Fetal Membranes (43% and 62%, respectively) and increased granulocyte-macrophage colony-stimulating factor levels (3.7- and 5.9-fold, respectively). Pretreatment with 17-alpha hydroxyprogesterone caproate inhibited both tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening and concomitantly inhibited the induced increase in granulocyte-macrophage colony-stimulating factor in a concentration-dependent manner. However, contrary to our prior reports regarding progesterone and other progestogens, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced Fetal Membrane weakening. Conclusion 17-Alpha hydroxyprogesterone caproate blocks tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening by inhibiting the production of granulocyte-macrophage colony-stimulating factor. However, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced weakening. We speculate that progestogens other than 17-alpha hydroxyprogesterone caproate may be more efficacious in preventing preterm premature rupture of the Fetal Membranes–related spontaneous preterm birth.

  • the physiology of Fetal Membrane weakening and rupture insights gained from the determination of physical properties revisited
    Placenta, 2016
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Raymond W Redline, John J Moore
    Abstract:

    Rupture of the Fetal Membranes (FM) is precipitated by stretch forces acting upon biochemically mediated, pre-weakened tissue. Term FM develop a para-cervical weak zone, characterized by collagen remodeling and apoptosis, within which FM rupture is thought to initiate. Preterm FM also have a weak region but are stronger overall than term FM. Inflammation/infection and decidual bleeding/abruption are strongly associated with preterm premature FM rupture (pPROM), but the specific mechanisms causing FM weakening-rupture in pPROM are unknown. There are no animal models for study of FM weakening and rupture. Over a decade ago we developed equipment and methodology to test human FM strength and incorporated it into a FM explant system to create an in-vitro human FM weakening model system. Within this model TNF (modeling inflammation) and Thrombin (modeling bleeding) both weaken human FM with concomitant up regulation of MMP9 and cellular apoptosis, mimicking the characteristics of the spontaneous FM rupture site. The model has been enhanced so that test agents can be applied directionally to the choriodecidual side of the FM explant consistent with the in-vivo situation. With this enhanced system we have demonstrated that the pathways involving inflammation/TNF and bleeding/Thrombin induced FM weakening overlap. Furthermore GM-CSF production was demonstrated to be a critical common intermediate step in both the TNF and the Thrombin induced FM weakening pathways. This model system has also been used to test potential inhibitors of FM weakening and therefore pPROM. The dietary supplement α-lipoic acid and progestogens (P4, MPA and 17α-hydroxyprogesterone) have been shown to inhibit both TNF and Thrombin induced FM weakening. The progestogens act at multiple points by inhibiting both GM-CSF production and GM-CSF action. The use of a combined biomechanical/biochemical in-vitro human FM weakening model system has allowed the pathways of Fetal Membrane weakening to be delineated, and agents that may be of clinical use in inhibiting these pathways to be tested.

  • progesterone inhibits in vitro Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2015
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, Edward Springel, Elliot Philipson, John J Moore
    Abstract:

    Objective Inflammation/infection and abruption are leading causes of preterm premature rupture of the Membranes. Recently, we identified granulocyte-macrophage colony-stimulating factor (GM-CSF) as a critical mediator of both tumor necrosis factor-α– (TNF; modeling inflammation) and thrombin-induced (modeling abruption) weakening of the Fetal Membranes. We found that (1) TNF and thrombin both induced GM-CSF in the choriodecidua, (2) blockade of GM-CSF action with neutralizing antibodies inhibited both TNF- and thrombin-induced Fetal Membrane weakening, and (3) GM-CSF alone induced Fetal Membrane weakening. GM-CSF is thus part of an overlap of the inflammation and abruption-induced Fetal Membrane weakening pathways. The effects of progesterone analogs on the pathways by which Fetal Membranes are weakened have not been investigated. We examined the effects of progesterone, medroxyprogesterone acetate (MPA) and 17α-hydroxyprogesterone (HP) on TNF- and thrombin-induced Fetal Membrane weakening. Study Design Full-thickness Fetal Membranes from uncomplicated term repeat cesarean deliveries were mounted in Transwell inserts in Minimum Essential Medium alpha and incubated at 37°C in 5% CO2. The choriodecidua side of the Fetal Membrane fragments were preincubated with progesterone, MPA, HP, or vehicle for 24 hours. Fetal Membranes were then exposed to TNF, thrombin, or GM-CSF on the choriodecidua side for an additional 48 hours. The Fetal Membrane tissues were then strength tested, and medium from the choriodecidua and amnion compartments was assayed for GM-CSF content. Results TNF and thrombin both weakened Fetal Membranes and elevated media GM-CSF levels on the choriodecidua side of the Fetal Membrane. Pretreatment with progesterone, MPA, or HP inhibited both TNF- and thrombin-induced Fetal Membrane weakening and also inhibited the induced increase in GM-CSF. GM-CSF decreased Fetal Membrane rupture strength by 68%, which was inhibited by progestogen pretreatment with a potency order: progesterone Conclusion Progestogen pretreatment blocks TNF- and thrombin-induced Fetal Membrane weakening by inhibiting both the production and action of GM-CSF. These findings are consistent with the administration of progestogens in the prevention of preterm premature rupture of the Membranes.

Joseph M Mansour - One of the best experts on this subject based on the ideXlab platform.

  • granulocyte macrophage colony stimulating factor gm csf the critical intermediate of inflammation induced Fetal Membrane weakening primarily exerts its weakening effect on the choriodecidua rather than the amnion
    Placenta, 2020
    Co-Authors: Anudeepa Sharma, Robert M Moore, Joseph M Mansour, Deepak Kumar, Brian M. Mercer, Aditi Deshmukh, John J Moore
    Abstract:

    Abstract Introduction We have previously demonstrated two associations of PPROM, (1) inflammation/infection (modeled by tumor necrosis factor (TNF) and (2) decidual bleeding (modeled by thrombin), both decrease Fetal Membrane (FM) rupture strength in-vitro. Furthermore, Granulocyte-Macrophage-Colony-Stimulating-Factor (GM-CSF) induced by both TNF and thrombin is a critical intermediate, necessary and sufficient for weakening by either agent. The amnion is the strength component of FM and must weaken for FM to rupture. It is unclear whether GM-CSF weakens amnion (AM) directly, or initially targets choriodecidua (CD) which secondarily releases agents to act on amnion. Methods Full thickness FM fragments were treated with/without GM-CSF. Some were preincubated with alpha-lipoic acid (LA), a known inhibitor of FM weakening. The FM fragments were then strength-tested. Separately, FM fragments were initially separated to AM and CD. AM fragments were cultured with Medium ± GM-CSF and then strength-tested. In other experiments, CD fragments were cultured with Medium, GM-CSF, LA, or LA + GM-CSF. Conditioned medium from each group was then incubated with AM. AM was then strength-tested. Matrix Metalloproteinases (MMPs) and Tissue Inhibitors of Matrix Metalloproteinases (TIMPs) were analyzed by Mutiplex Elisa. Results GM-CSF weakened intact FM which was blocked by LA. GM-CSF did not weaken isolated AM. However, GM-CSF conditioned CD media weakened AM and this weakening was inhibited by LA. GM-CSF treatment of CD increased MMPs 2, 9, and 10, and decreased TIMPs 1–3. LA reversed these effects. Conclusions GM-CSF does not weaken amnion directly; GM-CSF acts on CD to increase proteases and decrease anti-proteases which secondarily weaken the amnion.

  • in an in vitro model using human Fetal Membranes α lipoic acid inhibits inflammation induced Fetal Membrane weakening
    Placenta, 2018
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Anudeepa Sharma, John J Moore
    Abstract:

    Abstract Introduction We established an in-vitro model for the study of human Fetal Membrane (FM) weakening leading to pPROM. In this model, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both tumor necrosis factor-α (TNF; modeling infection/inflammation) and thrombin (modeling decidual bleeding/abruption)-induced weakening. Thus, inhibitors of FM weakening can be categorized as targeting GM-CSF production, GM-CSF downstream action, or both. Most progestogens inhibit both, except 17-α hydroxyprogesterone caproate which inhibits FM weakening at only one point, GM-CSF production. α-lipoic acid (LA), an over-the-counter dietary supplement, has also been previously shown to inhibit TNF and thrombin induced FM weakening. Objective To determine the point of action of LA inhibition of FM weakening. Methods FM fragments were mounted in Transwell inserts and preincubated with/without LA/24 h, then with/without addition of TNF, thrombin or GM-CSF. After 48 h, medium was assayed for GM-CSF, and FM fragments were rupture-strength tested. Results TNF and thrombin both weakened FM and increased GM-CSF levels. GM-CSF also weakened FM. LA inhibited both TNF and thrombin induced FM weakening and concomitantly inhibited the increase in GM-CSF in a concentration-dependent manner. In addition, LA inhibited GM-CSF induced FM weakening in a concentration dependent manner. Conclusions LA blocks TNF and thrombin induced FM weakening at two points, inhibiting both GM-CSF production and downstream action. Thus, we speculate that LA may be a potential standalone therapeutic agent, or supplement to current therapy for prevention of pPROM related spontaneous preterm birth, if preclinical studies to examine feasibility and safety during pregnancy are successfully accomplished.

  • in an in vitro model using human Fetal Membranes 17 α hydroxyprogesterone caproate is not an optimal progestogen for inhibition of Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2017
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, John J Moore
    Abstract:

    Background The progestogen 17-α hydroxyprogesterone caproate (17-OHPC) is 1 of only 2 agents recommended for clinical use in the prevention of spontaneous preterm delivery, and studies of its efficacy have been conflicting. We have developed an in-vitro model to study the Fetal Membrane weakening process that leads to rupture in preterm premature rupture of the Fetal Membranes (pPROM). Inflammation/infection associated with tumor necrosis factor-α (TNF-α) induction and decidual bleeding/abruption associated thrombin release are leading causes of preterm premature rupture of the Fetal Membranes. Both agents (TNF-α and thrombin) cause Fetal Membrane weakening in the model system. Furthermore, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a critical intermediate for both TNF-α and thrombin-induced Fetal Membrane weakening. In a previous report, we demonstrated that 3 progestogens, progesterone, 17-alpha hydroxyprogesterone (17-OHP), and medroxyprogesterone acetate (MPA), each inhibit both TNF-α– and thrombin-induced Fetal Membrane weakening at 2 distinct points of the Fetal Membrane weakening pathway. Each block both the production of and the downstream action of the critical intermediate granulocyte-macrophage colony-stimulating factor. Objective The objective of the study was to characterize the inhibitory effects of 17-OHPC on TNF-α– and thrombin-induced Fetal Membrane weakening in vitro. Study Design Full-thickness human Fetal Membrane fragments from uncomplicated term repeat cesarean deliveries were mounted in 2.5 cm Transwell inserts and cultured with/without 17-alpha hydroxyprogesterone caproate (10 –9 to 10 –7 M). After 24 hours, medium (supernatant) was removed and replaced with/without the addition of tumor necrosis factor-alpha (20 ng/mL) or thrombin (10 U/mL) or granulocyte-macrophage colony-stimulating factor (200 ng/mL). After 48 hours of culture, medium from the maternal side compartment of the model was assayed for granulocyte-macrophage colony-stimulating factor and the Fetal Membrane fragments were rupture strength tested. Results Tumor necrosis factor-alpha and thrombin both weakened Fetal Membranes (43% and 62%, respectively) and increased granulocyte-macrophage colony-stimulating factor levels (3.7- and 5.9-fold, respectively). Pretreatment with 17-alpha hydroxyprogesterone caproate inhibited both tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening and concomitantly inhibited the induced increase in granulocyte-macrophage colony-stimulating factor in a concentration-dependent manner. However, contrary to our prior reports regarding progesterone and other progestogens, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced Fetal Membrane weakening. Conclusion 17-Alpha hydroxyprogesterone caproate blocks tumor necrosis factor-alpha– and thrombin-induced Fetal Membrane weakening by inhibiting the production of granulocyte-macrophage colony-stimulating factor. However, 17-alpha hydroxyprogesterone caproate did not also inhibit granulocyte-macrophage colony-stimulating factor–induced weakening. We speculate that progestogens other than 17-alpha hydroxyprogesterone caproate may be more efficacious in preventing preterm premature rupture of the Fetal Membranes–related spontaneous preterm birth.

  • the physiology of Fetal Membrane weakening and rupture insights gained from the determination of physical properties revisited
    Placenta, 2016
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Brian M. Mercer, Raymond W Redline, John J Moore
    Abstract:

    Rupture of the Fetal Membranes (FM) is precipitated by stretch forces acting upon biochemically mediated, pre-weakened tissue. Term FM develop a para-cervical weak zone, characterized by collagen remodeling and apoptosis, within which FM rupture is thought to initiate. Preterm FM also have a weak region but are stronger overall than term FM. Inflammation/infection and decidual bleeding/abruption are strongly associated with preterm premature FM rupture (pPROM), but the specific mechanisms causing FM weakening-rupture in pPROM are unknown. There are no animal models for study of FM weakening and rupture. Over a decade ago we developed equipment and methodology to test human FM strength and incorporated it into a FM explant system to create an in-vitro human FM weakening model system. Within this model TNF (modeling inflammation) and Thrombin (modeling bleeding) both weaken human FM with concomitant up regulation of MMP9 and cellular apoptosis, mimicking the characteristics of the spontaneous FM rupture site. The model has been enhanced so that test agents can be applied directionally to the choriodecidual side of the FM explant consistent with the in-vivo situation. With this enhanced system we have demonstrated that the pathways involving inflammation/TNF and bleeding/Thrombin induced FM weakening overlap. Furthermore GM-CSF production was demonstrated to be a critical common intermediate step in both the TNF and the Thrombin induced FM weakening pathways. This model system has also been used to test potential inhibitors of FM weakening and therefore pPROM. The dietary supplement α-lipoic acid and progestogens (P4, MPA and 17α-hydroxyprogesterone) have been shown to inhibit both TNF and Thrombin induced FM weakening. The progestogens act at multiple points by inhibiting both GM-CSF production and GM-CSF action. The use of a combined biomechanical/biochemical in-vitro human FM weakening model system has allowed the pathways of Fetal Membrane weakening to be delineated, and agents that may be of clinical use in inhibiting these pathways to be tested.

  • progesterone inhibits in vitro Fetal Membrane weakening
    American Journal of Obstetrics and Gynecology, 2015
    Co-Authors: Deepak Kumar, Robert M Moore, Joseph M Mansour, Frederick Schatz, Charles J Lockwood, Brian M. Mercer, Sam Mesiano, Edward Springel, Elliot Philipson, John J Moore
    Abstract:

    Objective Inflammation/infection and abruption are leading causes of preterm premature rupture of the Membranes. Recently, we identified granulocyte-macrophage colony-stimulating factor (GM-CSF) as a critical mediator of both tumor necrosis factor-α– (TNF; modeling inflammation) and thrombin-induced (modeling abruption) weakening of the Fetal Membranes. We found that (1) TNF and thrombin both induced GM-CSF in the choriodecidua, (2) blockade of GM-CSF action with neutralizing antibodies inhibited both TNF- and thrombin-induced Fetal Membrane weakening, and (3) GM-CSF alone induced Fetal Membrane weakening. GM-CSF is thus part of an overlap of the inflammation and abruption-induced Fetal Membrane weakening pathways. The effects of progesterone analogs on the pathways by which Fetal Membranes are weakened have not been investigated. We examined the effects of progesterone, medroxyprogesterone acetate (MPA) and 17α-hydroxyprogesterone (HP) on TNF- and thrombin-induced Fetal Membrane weakening. Study Design Full-thickness Fetal Membranes from uncomplicated term repeat cesarean deliveries were mounted in Transwell inserts in Minimum Essential Medium alpha and incubated at 37°C in 5% CO2. The choriodecidua side of the Fetal Membrane fragments were preincubated with progesterone, MPA, HP, or vehicle for 24 hours. Fetal Membranes were then exposed to TNF, thrombin, or GM-CSF on the choriodecidua side for an additional 48 hours. The Fetal Membrane tissues were then strength tested, and medium from the choriodecidua and amnion compartments was assayed for GM-CSF content. Results TNF and thrombin both weakened Fetal Membranes and elevated media GM-CSF levels on the choriodecidua side of the Fetal Membrane. Pretreatment with progesterone, MPA, or HP inhibited both TNF- and thrombin-induced Fetal Membrane weakening and also inhibited the induced increase in GM-CSF. GM-CSF decreased Fetal Membrane rupture strength by 68%, which was inhibited by progestogen pretreatment with a potency order: progesterone Conclusion Progestogen pretreatment blocks TNF- and thrombin-induced Fetal Membrane weakening by inhibiting both the production and action of GM-CSF. These findings are consistent with the administration of progestogens in the prevention of preterm premature rupture of the Membranes.