The hernia sac-A suitable source for obtaining mesenchymal stem cells.

BACKGROUND: Inguinal hernia sac, extended tissue from peritoneum, gradually enlarged in size with hernia disease time and prolapsed tissue volume. We hypothesize that mesenchymal stem cells are present in the development of hernia sac. The current study aimed to test the hypothesis that hernia sac, which is often resected and discarded as medical waste, contains mesenchymal stem cells and thus might be a suitable source to harvest mesenchymal stem cells. METHODS: Between July 2019 and June 2020, 4 hernia sacs were resected during hernia surgery and then obtained for mesenchymal extraction using the Miltenyi gentleMACS Dissociator. The presence of mesenchymal stem cells was determined by the markers CD105, CD73, and CD90, with assessment of the expressions ≥ 95%, whereas markers CD45, CD34, CD11b, CD19, and HLA-DR were used to assess lack expression (≤ 2%). Moreover, von Kossa staining, Alcian blue staining, and Oil Red O staining were used to verify the cells' ability for differentiation. RESULTS: Cells retrieved from the hernia sacs displayed a spindle-shaped morphology and exhibited adherence to plastics. The cell surface immunophenotypic profile was confirmed using surface markers APC-A (CD73), FITC-A (CD90), and PerCP-Cy5-5-A (CD105), with results showing 100%, 100%, and 99.2%, respectively, strongly indicating the presence of mesenchymal stem cells. Moreover, staining of in vitro cell cultures showed in vitro differentiation of precursor cells into osteoblasts, adipocytes, and chondroblasts, suggesting positive differentiation ability and identification of mesenchymal stem cells. CONCLUSION: Inguinal hernia sac is a novel source of mesenchymal stem cells that can be easily obtained and stored for future usage.

Triple-combined herniorrhaphy for inguinal hernia repair: experience of 1411 cases.

BACKGROUND/AIMS: The optimum method for inguinal hernia repair has not yet been determined. Triple-Combined Herniorrhaphy, using the combined methods of McVay, Shouldice and Halsted repairs, was developed in our hospital over the past years in order to improve the overall results of treatment of inguinal hernia. The aim of this study was to verify the value of this surgical technique for primary inguinal hernia in a specialized hospital setting. METHODOLOGY: We describe our experience of 1411 consecutive patients for whom Triple-Combined Herniorrhaphy was performed for inguinal hernia repair at our hospital between September 2000 and August 2003, under local anesthesia with a "one-day surgery" regimen. RESULTS: The type of hernias included 342 direct inguinal hernias, 913 indirect inguinal hernias, and 156 pantaloon (mixed) type inguinal hernias. No mortality or major intraoperative complications were observed. The median duration of operation time was 25 min. Wound infection and hematoma formation requiring drainage was observed in 9 and 16 patients, respectively. Patients had fast convalescence with rapid resumption of working activity. The postoperative recurrence rate was 1.2%. CONCLUSIONS: Triple-Combined Herniorrhaphy is a simple, safe, comfortable, and effective method with low early and later morbidity and recurrence rate. The good results of this procedure constitute a good alternative to mesh or other open inguinal repairs.

Effect of bilateral in vivo ischemia/reperfusion on the activities of superoxide dismutase and catalase: response to a standardized grape suspension.

PURPOSE: Ischemia/reperfusion (I/R) is a major etiological factor in the bladder dysfunctions observed in men with lower tract obstruction, women with postmenopausal incontinence and with aging. A standardized grape suspension protects the rabbit urinary bladder from both the contractile dysfunctions and the morphologic changes mediated by I/R. Using a model of in vivo bilateral ischemia/reperfusion, the current study investigated the effect of this grape suspension on the endogenous antioxidant defense systems. MATERIALS AND METHODS: 24 NZW rabbits were separated into 6 groups of 4. Groups 1-3 were treated by gavage with aqueous grape suspensions; groups 4-6 received sugar-water vehicle. Groups 3 and 6 were controls. Groups 1 and 4 were subjected to bilateral ischemia for 2 h (I). Groups 2 and 5 underwent bilateral ischemia for 2 h and reperfusion for 1 week (I/R). For all rabbit bladders, the muscle and mucosa were separated by blunt dissection and analyzed separately. The effects of the various treatments on bladder antioxidant systems of cytoplasmic superoxide dismutase (Cu-Zn superoxide dismutase; SOD), and catalase (CAT) were evaluated. RESULTS: The standardized grape suspension up-regulated both SOD and CAT activity of bladder muscle and mucosa in control animals. There were few differences in the grape suspension treated animals after ischemia, and in general the activities decreased following I/R. CONCLUSIONS: Increases of SOD and CAT activity in control animals as a result of grape suspension suggest a greater antioxidant capacity. This increase in the antioxidant defense system may explain the increased protection of grape suspension in the face of ischemia and I/R. However, the activities of both enzyme systems decreased in the smooth muscle subjected to I/R showing that reperfusion damages these systems probably via oxidation damage to the enzymes themselves.

Alteration of contractile and regulatory proteins in estrogen-induced hypertrophy of female rabbit bladder.

OBJECTIVES: Estrogen is essential to mediate physiologic functions in female bladders. Deficiency of estrogen has been speculated to be an etiologic factor for bladder dysfunction in postmenopausal women. Our previous studies have demonstrated that estrogen supplementation in female rabbits induces a "functional hypertrophy" of the urinary bladder smooth muscle. The present study investigated the alterations in the contractile and regulatory proteins in this model. METHODS: Twenty New Zealand white female rabbits were separated into five groups of 4 rabbits each. Group 1 served as the control, groups 2 to 6 underwent ovariectomy (Ovx), and group 2 served as the Ovx without estradiol treatment group. Two weeks after Ovx, groups 3 to 5 were given 17-beta estradiol (1 mg/kg/day) by subcutaneous implant for 1, 3, and 7 days, respectively. The expression of the contractile and regulatory proteins, such as myosin light chain kinase, rho-kinase, and caldesmon, was analyzed by Western blotting. RESULTS: The expression of myosin light chain kinase was enhanced by estradiol supplementation. The expression of rho-kinase-alpha was increased significantly (20-fold) after Ovx, which was downregulated after estrogen supplementation. No significant change was seen in rho-kinase-beta after Ovx or estradiol supplementation. The expression of caldesmon isoforms was enhanced by 1-day estradiol supplementation but decreased to lower levels than those of the control group by 3 and 7 days of estrogen treatment. CONCLUSIONS: The results of the present study have provided more understanding about the role of the contractile and regulatory proteins in detrusor muscle, in both dysfunctional atrophy induced by Ovx and functional hypertrophy induced by estrogen supplementation.

Estrogen induces angiogenesis of the female rabbit bladder.

Postmenopausal bladder dysfunction has been speculated to involve decreased circulating estrogen levels. It is our hypothesis that estrogen induces bladder dysfunctions by modulating blood flow to the bladder, i.e. low estrogen reduces blood flow to the bladder, whereas high estrogen increases blood flow. Our previous studies have demonstrated that estrogen administration in female rabbits induces a 'functional hypertrophy' of the urinary bladder smooth muscle represented by increased smooth muscle mass, which corresponds to increased contractile responses to all forms of stimulation. The present study investigates the effect of estrogen on vasculature density and distribution. Twenty-four female New Zealand white rabbits were separated into six groups of four rabbits each. Group 1 served as controls. Groups 2-6 were ovariectomized. Two weeks after ovariectomy (Ovx), groups 3-6 were given 17-beta estradiol (1 mg/kg per day) by s.c. implant for 1, 3, 7, and 14 days respectively. Blood vessel density and distribution were evaluated by immunohistochemistry and quantitative image analyses. Ovx resulted in significant vascular degeneration and decreased density, whereas estradiol administration mediated a significant angiogenic effect characterized by increased vascular density, and distribution of new vasculature within the smooth muscle bundles of the detrusor. Estradiol-induced vasculogenesis corresponds with our previously demonstrated increase in blood flow to the bladder and increased contractility. The most interesting aspect of these studies is the increased vascularization localized within the muscle bundles rather than between the muscle bundles, which may be important in the link between estrogen and increased incidence of cancers.

Estrogen induced functional hypertrophy and increased force generation of the female rabbit bladder.

AIMS: Estrogen is essential for physiological maintenance of the female urogenital tract. It is believed that alterations in female sex hormones play a major role in the etiology and response to urinary tract dysfunctions. In animal studies, ovariectomy (Ovx) results in smooth muscle (SM) weakness and atrophy whereas estrogen supplementation reverses these effects. Our study seeks to establish the mechanisms by which estrogen augmentation results in increased contractility. METHODS: Twenty New Zealand White female rabbits were separated into five groups of four each. Group 1 served as control, rabbits of groups 2-5 were ovariectomized, group 2 ovariectomized received no estradiol, groups 3-5 were given 17-beta estradiol (1 mg/kg/day) by subcutaneous slow release tablet implant for 1, 3, and 7 days, respectively, beginning 2 weeks after Ovx. At the end of the experimental period, each rabbit was anesthetized and the urinary bladder was removed for contractile, histological, and biochemical studies. RESULTS: Ovx resulted in significantly decreased bladder contractile function, whereas bladders tested after estradiol administration showed increased contractility. Ovx resulted in a decrease in SM/collagen ratio, whereas estrogen resulted in an increase. The estrogen receptor (ER) density significantly increased following Ovx. After 1 day of estrogen treatment, the ER density decreased significantly below control levels, but rose progressively during the estrogen treatment. CONCLUSION: The present study demonstrates that estrogen supplementation mediates a "functional hypertrophy," that is a hypertrophy characterized by increased contractile responses to all forms of stimulation, and an increased ratio of SM/collagen.

Effect of partial bladder outlet obstruction on nitrotyrosine levels and their correlation with contractile function.

AIMS: It has been demonstrated that partial bladder outlet obstruction (PBOO) causes free radical generation that, in turn, results in cellular and subcellular damage. We tested the hypothesis that nitration of proteins is associated with contractile dysfunctions in obstructive bladder disease. METHODS: Thirty rabbits were subjected to 1-28 days of partial outlet obstruction. Sham operated rabbits served as controls. Western blotting was used to determine the amount of nitrotyrosine level at the protein level. At each time point, isolated strips of bladder body were mounted in individual baths and the contractile response to field stimulation (FS), carbachol, and KCl determined. RESULTS: Bladder weight increased rapidly during the first 7 days and then increased slowly thereafter. There was a fourfold increase in the amount of nitrotyrosine in the 7 day obstructed groups when compared to sham controls and the levels remain elevated at 14 and 28 days of obstruction. Contractile dysfunction in response to FS (8 and 32 Hz) was noted as early as 1 day after obstruction and increased progressively over the study period. The decrease in response to carbachol and KCl was significant only after 3 days of obstruction and the progressive increase in dysfunction was slower than with FS. CONCLUSIONS: PBOO is accompanied by an increase in nitrotyrosine, a marker of free radical damage. Simultaneously there was a progressive decrease in contractility of detrusor smooth muscles (DSMs). Nitrotyrosine may be usable as a marker of free radical damage and reperfusion injury.

L-NAME, a nitric oxide synthase inhibitor, diminishes oxidative damage in urinary bladder partial outlet obstruction.

Partial bladder outlet obstruction (PBOO) results in cellular damage due to ischemia and reperfusion injury. Our study seeks to establish how early this damage can occur and the role that nitric oxide may play in its pathophysiology. Surgical PBOO (1, 3, and 7 days) were performed on male New Zealand White rabbits. Half of the animals were premedicated for 3 days with N(G)-nitro-l-arginine methyl ester(l-NAME), an inhibitor of nitric oxide synthase before obstruction. Bladder weight increased with duration of PBOO but was significantly lower at 3 and 7 days in animals treated with l-NAME compared with their untreated counterparts. Contractile function decreased progressively with PBOO duration. At 1 day postobstruction, bladder contractility was significantly lower in the l-NAME rabbits than in the untreated rabbits. At 3 and 7 days, contractility of the l-NAME bladders was equal or higher than the untreated bladders. The level of hypoxia at 1 day after obstruction was significantly higher in the l-NAME-treated animals than in the untreated controls but equal at 3 and 7 days obstruction. Increased nitrotyrosine was seen by Western blot in all obstructed animals. However, the amount was significantly less in the l-NAME-treated animals at 3 and especially at 7 days. Nerve density decreased progressively after obstruction; however, it decreased to a significantly lesser degree in the l-NAME-treated bladders than in the untreated groups. These results suggest that l-NAME pretreatment enhanced ischemic damage at 1 day after obstruction but protected the bladder from nitric oxide-generated free radical damage at the later time periods by inhibiting the generation of nitrotyrosine.

Protective effects of grape suspension on in vivo ischaemia/reperfusion of the rabbit bladder.

OBJECTIVE: To investigate the potential protective effect of a grape suspension in a rabbit model of in vivo bilateral ischaemia/reperfusion (I/R), which is a causal factor in obstructive bladder dysfunction. MATERIALS AND METHODS: Six groups of four New Zealand White rabbits were treated by twice-daily gavage with aqueous grape suspension (groups 1-3) or sugar-water vehicle (groups 4-6) for 3 weeks. Groups 1 and 4 then received bilateral ischaemia for 2 h, and groups 2 and 5 received bilateral ischaemia for 2 h and reperfusion (recovery) for 1 week. Groups 3 and 6 were controls (sham-operated). The effects on cystometry, in vitro contractile responses, and morphology were evaluated. RESULTS: Ischaemia resulted in significant reductions in the contractile responses to all forms of stimulation in vehicle-fed rabbits, whereas there were no reductions in grape-fed rabbits. Contractile responses were significantly reduced in both I/R groups, but significantly more in vehicle-fed than in grape-fed rabbits. Immunohistochemical studies showed less hypoxia in the bladders of grape-fed rabbits than in vehicle-fed rabbits for both ischaemia-only and I/R groups. CONCLUSIONS: Feeding rabbits with grape suspension provided significant protection against the hypoxic effects of bilateral ischaemia and I/R.

Nocturnal enuresis in older adults.

BACKGROUND: Nocturnal enuresis is uncommon in older adults. The paucity of literature about this problem prompts us to review our cases to determine the management strategy. METHODS: Six older adults, including 2 females and 4 males, were evaluated for refractory nocturnal enuresis. Only 2 of them had minor daytime urge symptom. Most of them had failed in the treatment using anticholinergics and/or alpha-adrenergic blocker. Evaluation consisted of detailed medical history, voiding diary, and urodynamic studies. Clinical follow-up persisted for 12 months. We define nocturnal polyuria as nighttime urine amount being more than 35% of total daily urine amount. Bladder outlet obstruction in men was diagnosed based on the definition described by International Continence Society. RESULTS: The average age was 71 years (range 61-84). The average duration of the symptom was 3.1 months (range 0.5-6). Two patients had bladder outlet obstruction. Four patients used hypnotics for insomnia, which might result in difficult awakening on bladder distension. Nocturnal polyuria was found in 3 patients. Most patients had multiple factors contributing to their nocturnal enuresis except 1, who was found to have an enlarged prostate with chronic bladder distension. Specific treatments were given based on the causes for each patient. Hypnotics were discontinued for a certain meanwhile in some patients. Nocturnal polyuria was managed with afternoon diuretic or bedtime desmopressin. Bedtime anticholinergic agent was used in patients with detrusor overactivity. The patient with enlarged prostate and urinary retention was managed with indwelling catheter followed by elective transurethral prostatectomy. All patients were dry in the night following the treatment. CONCLUSIONS: Nocturnal enuresis in older adult is usually multi-factorial. Hypnotic usage and nocturnal polyuria are frequently overlooked. Detailed investigation is necessary to identify the causes. Tailored treatment may achieve satisfactory results.