fibrin and Fetal-Membranes--Premature-Rupture

To investigate whether damaged human amniotic epithelial cells (HAEC) could be repaired on the matrix of formulated fibrin clot in vitro and the effects of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and transforming growth factor beta(1) (TGF-beta(1)) on the proliferation of HAEC.. Ring drill was used to drill the HAEC layer on culture sheets to make quantified models of damaged HAEC, on which the lacks were then covered with fibrin clot. Subsequently, EGF (EGF group), bFGF (bFGF group) and TGF-beta(1) (TGF-beta(1) group) of different concentration were added into the sheets respectively. After the predesigned culturing time, the growing and transiting conditions of HAEC were observed under inverted microscope after Giemsa stain. Also, the proliferating conditions of HAEC were detected by using 5-bromodeoxyuridine (BrdU).. In all groups, HAEC could transit toward damaged area on fibrin clot and grow there. Higher transiting speed and larger cell numbers were observed in the EGF and bFGF groups followed by the control group, while the TGF-beta(1) group showed the relatively poorer results. Proliferating rates of HAEC were 17.8%, 28.0%, 35.3%, 51.6%, 34.1%, 34.2% and 26.0% respectively by EGF of different cultured concentration (1.0 ng/ml, 5.0 ng/ml, 10.0 ng/ml, 20.0 ng/ml, 40.0 ng/ml, 80.0 ng/ml and 160.0 ng/ml). Proliferating rates of HAEC were 18.0%, 35.7%, 43.0%, 52.7%, 67.4%, 43.6% and 30.5% respectively by bFGF of different cultured concentration (1.0 ng/ml, 5.0 ng/ml, 10.0 ng/ml, 20.0 ng/ml, 40.0 ng/ml, 80.0 ng/ml and 160.0 ng/ml). Compared with the control group, EGF groups (EGF concentration ranging from 10 ng/ml to 80 ng/ml) and bFGF groups (bFGF concentration ranging from 5 ng/ml to 80 ng/ml) showed better proliferating effects of HAEC (P < 0.05), especially the 20 ng/ml EGF group and 40 ng/ml bFGF group had the best proliferating results among their own respective groups (P < 0.05). Proliferating rates of HAEC were 17.1%, 15.1%, 9.3%, 6.2%, 4.8%, 3.6%, 2.0% and 1.2% respectively by TGF-beta(1) of different cultured concentration (0.1 ng/ml, 0.2 ng/ml, 0.4 ng/ml, 0.8 ng/ml, 1.6 ng/ml, 3.2 ng/ml, 6.4 ng/ml and 12.8 ng/ml). Proliferating rates of HAEC in TGF-beta(1) groups (TGF-beta(1) concentration ranging from 0.8 ng/ml to 12.8 ng/ml) were significantly lower than that in the control group (P < 0.05).. HAEC could transit and grow on the matrix of fibrin clot and repair the damaged area. EGF and bFGF could obviously stimulate HAEC proliferation, while TGF-beta(1) might have the inhibitive effects.

Topics: Amnion; Cell Movement; Cell Proliferation; Cells, Cultured; Cytokines; Epidermal Growth Factor; Epithelial Cells; Female; Fetal Membranes, Premature Rupture; Fibrin; Fibroblast Growth Factor 2; Humans; Pregnancy; Transforming Growth Factor beta

Recent evidence has linked preterm premature rupture of the fetal membranes (PPROM) to placental abruption. Because neutrophils are a rich source of proteases that can degrade extracellular matrix in abruption-associated PPROM, we examined whether decidual neutrophil infiltration complicates abruption-associated PPROM. Accordingly, immunostaining for the neutrophil marker CD15 was performed in placentas obtained after overt abruption (decidual hemorrhage) with or without PPROM and in control placentas. Abruptions were associated with a marked decidual neutrophil infiltration that peaked after PPROM, whereas decidua from gestational age-matched controls were virtually devoid of neutrophils. Neutrophil infiltrates co-localized with fibrin deposition. Because abruptions elicit intense decidua-enhanced thrombin production, we examined the regulation of abruption-induced neutrophil infiltration. Expression of the primary neutrophil chemoattractant interleukin-8 (IL-8) was evaluated in leukocyte-free term decidual cells incubated with estradiol (E2; control) or with E2+medroxyprogesterone acetate (to mimic pregnancy)+/-thrombin. After 24 hours, enzyme-linked immunosorbent assay measurements indicated that thrombin (0.1 to 2.5 U/ml) elicited a dose-dependent elevation in secreted IL-8 (P<0.05) with 2.5 U/ml of thrombin increasing IL-8 levels by >14-fold in E2 and E2+medroxyprogesterone incubations. Results were validated by Western blot and quantitative reverse transcriptase-polymerase chain reaction. In summary, thrombin-enhanced IL-8 expression in term decidual cells may explain how abruption-associated PPROM promotes decidual neutrophil infiltration.

Topics: Abruptio Placentae; Adult; Blotting, Western; Cells, Cultured; Decidua; Enzyme-Linked Immunosorbent Assay; Estradiol; Female; Fetal Membranes, Premature Rupture; Fibrin; Humans; Immunohistochemistry; Interleukin-8; Lewis X Antigen; Medroxyprogesterone Acetate; Neutrophil Infiltration; Neutrophils; Pregnancy; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Thrombin

Operative fetoscopy may be limited by its relatively high associated risk of preterm prelabour rupture of membranes. The objective of this study was to study closure techniques of the access site for fetoscopy in the mid-gestational rabbit. A total of 32 does (288 amniotic sacs) at 22 days gestational age (GA; term = 32 days) underwent 14 gauge needle fetoscopy, by puncture through surgically exposed amnion. Entry site was randomly allocated to four closure technique groups: myometrial suture (n = 14), fibrin sealant (n = 15), autologous maternal blood plug (n = 13), collagen plug (n = 14); 16 sacs were left unclosed (positive controls), and the unmanipulated 216 sacs were negative controls. Membrane integrity, presence of amniotic fluid and fetal lung to body weight ratio (FLBWR) were evaluated at 31 days GA. Following fetoscopy without an attempt to close the membranes, amniotic integrity was restored in 41% of cases (amniotic integrity in controls 94%; P = 0.00001). When the access site was surgically closed, the amnion resealed in 20-44% of cases, but none of the tested techniques was significantly better than the others or than positive controls. Permanent amniotic disruption was associated with a significantly lower FLBWR in all groups. In conclusion, the rate of fetoscopy-induced permanent membrane defects in this model did not improve by using any of the closure techniques tested here.

Topics: Animals; Blood; Collagen; Disease Models, Animal; Evaluation Studies as Topic; Female; Fetal Membranes, Premature Rupture; Fetoscopy; Fibrin; Gestational Age; Humans; Iatrogenic Disease; Pregnancy; Rabbits; Suture Techniques; Tissue Adhesives

Topics: Administration, Intravaginal; Adult; Female; Fetal Membranes, Premature Rupture; Fibrin; Humans; Pregnancy

Topics: Adult; Ampicillin; Bed Rest; Female; Fetal Membranes, Premature Rupture; Fibrin; Humans; Pregnancy; Tissue Adhesives

Topics: Female; Fetal Death; Fetal Membranes, Premature Rupture; Fibrin; Humans; Obstetric Labor, Premature; Pregnancy; Pregnancy Trimester, Second; Tissue Adhesives