Evaluation of adhesion formation using Interceed (TC7) absorbable adhesion barrier on ovarian surgical wounds in the rabbit model.

OBJECTIVE: To determine whether an Interceed patch would decrease adhesion formation in ovarian surgical wounds healing by second intention in the rabbit model. METHODS: Twenty-five New Zealand White female rabbits underwent midline laparotomy. Using microsurgical technique, each ovary was bivalved longitudinally with a no. 15 blade scalpel, and hemostasis was achieved with bipolar electrocautery. One ovary was then covered with an Interceed patch in a randomized fashion, while the other ovary served as a control. The animals were then allowed to recover routinely. Four weeks later, the rabbits were euthanized and the adhesions were graded by a blinded observer. Adhesion scores between the Interceed-treated ovaries and the control ovaries were compared by Wilcoxon signed-rank test. RESULTS: The mean adhesion score of the Interceed-treated ovaries (1.65 +/- 0.87) was not significantly different from the mean adhesion score of the control ovaries (1.5 +/- 1.0) (P = .51). Some degree of atrophy was noted in 14 of the 50 ovaries. The mean adhesion score in the atrophied ovaries was significantly higher than in the nonatrophied ovaries, regardless of whether the ovaries had been treated with Interceed (2.21 versus 1.40; P = .01). There was no difference in the incidence of atrophy between the Interceed-treated and untreated sides. Exclusion of the ovaries with atrophy again demonstrated no difference in the mean adhesion scores between the treated and untreated sides. CONCLUSION: We were not able to demonstrate any benefit from using Interceed to decrease postoperative adhesion formation of ovarian surgical wounds in the rabbit model.

A comparative evaluation of Poloxamer 407 and oxidized regenerated cellulose (Interceed [TC7]) to reduce postoperative adhesion formation in the rat uterine horn model.

OBJECTIVES: To determine if Poloxamer 407 (poloxamer, Pluronic F-127; BASF Wyandotte Corp., Parsippany, NJ) is as effective as Interceed(TC7) (Ethicon, a Johnson and Johnson company, Sommerville, NJ) in preventing postoperative adhesion formation using the rat uterine horn model and to determine if the presence of blood or lactated Ringer's solution affects the effectiveness of Poloxamer 407. DESIGN, SETTING, PARTICIPANTS: Sprague-Dawley white rats, weighing 225 to 250 g in a conventional laboratory setting. The left or right sidewall was randomly assigned to receive no treatment (control), Interceed(TC7), or poloxamer. INTERVENTIONS: Each uterine horn and ipsilateral sidewall was subjected to a standardized lesion of denudation. To evaluate the barrier agents in the presence of blood, a sidewall vessel was ligated and a thrombus allowed to form. To evaluate the effectiveness of lactated Ringer's solution, 10 mL was injected intraperitoneally after abdominal closure. MAIN OUTCOME MEASURES: Degree of adhesion formation was evaluated 14 days after surgery. RESULTS: The adhesion score for the poloxamer-treated animals was significantly lower than its control. Interceed(TC7) did not reduce adhesion formation as compared with its control. In animals that received both poloxamer and Interceed(TC7) on either side, the poloxamer-treated sides had a significantly lower adhesion score than Interceed(TC7)-treated sides. The presence of blood and lactated Ringer's reduced the adhesion-reducing properties of poloxamer. CONCLUSION: In this model, poloxamer is more effective than Interceed(TC7) in the prevention of postoperative adhesion formation. These findings also suggest that the presence of blood compromises the effectiveness of poloxamer to prevent postoperative adhesion formation, therefore requiring complete hemostasis before poloxamer's application. Lactated Ringer's instillation was ineffective in reducing adhesion formation on the control or poloxamer-treated sidewall.

Review: cytokine involvement in ovarian processes.

PROBLEM: Expression of tumor necrosis factor-alpha (TNF) and interleukins 1 alpha and 1 beta (IL-1) have been reported in ovaries of several species and humans and are implicated in ovarian follicular development and atresia, ovulation, steroidogenesis, and corpus luteum function (including formation, development, and regression). The principal abnormal processes affected by these cytokines are ovarian cancer and reduction of ovarian function during sepsis. METHODS: A literature review. RESULTS: Numerous studies indicate that TNF and IL-1 inhibit gonadotropin-stimulated steroidogenesis of undifferentiated ovarian cells due to inhibition of adenylyl cyclase and post-cAMP sites. In differentiated ovarian cells, these cytokines either stimulate progesterone synthesis or have little to no effect on steroidogenesis. Both cytokines participate in ovulation and levels of these cytokines increase during the preovulatory period. Endotoxin inhibits gonadotropin-stimulated ovarian steroidogenesis and follicular development and these effects are mediated, in part, by TNF and by direct effects of endotoxin on ovarian cells. IN newly formed corpora lutea, progesterone secretion is inhibited by TNF and IL-1, although each has proliferative effects. TNF also has been implicated in regression of corpora lutea because TNF stimulates prostaglandin synthesis and luteal TNF increases after initiation of the decline in progesterone secretion. TNF and IL-1 are secreted by some ovarian cancer cells and stimulate growth of these cells. CONCLUSIONS: Thus, TNF and IL-1 are multifunctional factors affecting various ovarian processes.

Changes in circulating and ovarian concentrations of bioactive tumour necrosis factor alpha during the first ovulation at puberty in rats and in gonadotrophin-treated immature rats.

Tumour necrosis factor alpha (TNF-alpha) concentrations were measured during periods of controlled and natural follicular development and ovulation in rat ovaries. Concentrations of bioactive TNF-alpha were determined in the ovaries and sera of rats during puberty (the period of vaginal opening and the first ovulation) and in immature rats after gonadotrophin treatment. Ovaries and sera were collected from 33-, 35-, 37-, 39-, 41- and 43-day-old rats (n = 6 or 7 per group); vaginal opening occurs on day 35. The presence of ovarian follicles and corpora lutea or ova in the oviducts was assessed. For gonadotrophin treatment, a single subcutaneous injection of 5 iu equine chorionic gonadotrophin (eCG) was administered at 08:00 h to 28-day-old rats to stimulate follicular development. A single subcutaneous injection of 10 iu human chorionic gonadotrophin (hCG) was administered 48 h later to induce ovulation. Ovaries and sera from three to six animals per group were collected 0, 3, 24, 48, 51, 54, 57, 60 and 72 h after injection of eCG. At puberty, ovarian concentrations of TNF-alpha were highest (approximately 1.1 fg micrograms-1 ovarian protein) before vaginal opening and the first ovulation. After vaginal opening and ovulation at day 37, ovarian concentrations of TNF-alpha were markedly reduced (0.091 fg microgram-1 ovarian protein) and remained low up to day 43. Serum concentrations of TNF-alpha remained low throughout the period of vaginal opening and the first ovulation (8-32 pg ml-1). In 43-day-old rats serum concentrations of TNF-alpha increased (105 pg ml-1). In the immature ovaries of 28-day-old rats TNF-alpha concentrations were highest before injection of eCG (approximately 1.2 fg micrograms-1 ovarian protein) and decreased to approximately 0.4 fg microgram-1 protein 3 h after injection. TNF-alpha concentrations decreased further 24 h after eCG injection (< 0.1 fg microgram-1 protein) and remained low until 48 h after eCG injection. Serum concentrations of TNF-alpha did not change during the 48 h period after injection of eCG. hCG was administered 48 h after eCG, and ovarian and serum TNF-alpha concentrations increased transiently. Serum TNF-alpha concentrations increased 3 h after hCG and remained elevated until 9 h after injection, after which concentrations decreased. Ovarian concentrations of TNF-alpha increased 6 h after hCG, peaked (approximately 0.5 fg microgram-1 protein), and then declined. These results indicate that during puberty and the first ovulation, circulating and ovarian TNF-alpha concentrations change. In addition, exogenous gonadotrophins alter circulating and ovarian TNF-alpha concentrations. These data suggest that TNF-alpha has a role in follicular development and ovulation during puberty and in immature rats treated with gonadotrophins to induce ovulation.

Tumour necrosis factor-alpha inhibits follicle-stimulating hormone-induced granulosa cell oestradiol secretion in the human: dependence on size of follicle.

This study investigated the effects of tumour necrosis factor-alpha (TNF) on human granulosa cells taken from ovaries of 11 premenopausal women undergoing oophorectomy during the luteal phase of the cycle for reasons unrelated to ovarian pathology. Granulosa cells from follicles ranging from 5-10 mm diameter (small) and from >10-25 mm (large) were subjected to culture for 48 and 96 h. Granulosa cells were cultured with human FSH (1 ng/ml), testosterone (1 microM) and human TNF (10 ng/ml), each alone, and in various combinations. In granulosa cells of small follicles, FSH alone increased progesterone and cAMP accumulation and the conversion of testosterone to oestradiol. In granulosa cells of large follicles, FSH increased progesterone and cAMP accumulation but not the conversion of testosterone to oestradiol. Only in granulosa cells of small follicles did TNF significantly inhibit FSH-induced conversion of testosterone to oestradiol but it was not apparent until the second 48 h of culture and concomitantly TNF did not alter the ability of FSH to stimulate progesterone and cAMP accumulation. In granulosa cells of large follicles, TNF did not alter FSH-stimulated oestradiol, progesterone or cAMP accumulation. Interestingly, progesterone accumulation in the presence of TNF and FSH was significantly greater in granulosa cells of large follicles than in granulosa cells of small follicles. The results indicate that TNF suppresses FSH-induced oestradiol secretion in granulosa cells from small follicles and this modulatory effect of TNF appears to be independent of decreases in progesterone and cAMP. The potential physiological significances of these findings is that TNF may be a relevant cytokine in suppressing FSH-induced oestradiol secretion and follicular growth during the luteal phase of the cycle.

Herbimycin, a tyrosine kinase inhibitor with Src selectivity, reduces progesterone and estradiol secretion by human granulosa cells.

The purpose of the present study was to investigate whether the tyrosine kinase inhibitor herbimycin with some selectivity to block Src would alter the stimulatory effects of follicle-stimulating hormone (FSH) and cyclic adenosine monophosphate (cAMP) on estradiol secretion by human granulosa cells. Granulosa cells were taken from ovaries of premenopausal women undergoing oophorectomy for reasons unrelated to ovarian pathology. Granulosa cells from follicles ranging from 5-20 mm in diameter were subjected to culture. Granulosa cells were cultured with human FSH (2 ng/mL) or cAMP (0-1 mM) and testosterone (1 microM) in the presence and absence of herbimycin (0-2 pM). Media were collected at 24, 48, and 72 h. Accumulation of cAMP, progesterone, and estradiol in the media was determined by radioimmunoassay. Herbimycin dose dependently inhibited the ability of FSH to induce increases in progesterone and estradiol secretion. Although herbimycin increased (p < 0.0001) the accumulation of cAMP in response to FSH, this was evident only at the high concentrations of herbimycin (2 microM). To determine whether herbimycin would inhibit the ability of exogenous cAMP to induce estradiol and progesterone secretion, granulosa cells were incubated with 0-1 mM cAMP in the presence and absence of various doses of herbimycin. Herbimycin inhibited cAMP-induced estradiol and progesterone secretion in granulosa cells. The results from seven experiments indicate that herbimycin inhibits FSH stimulation of estradiol and progesterone secretion and that this inhibition may be, in part, at post-cAMP site(s).