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Biomedical subjects

B Ludwikowski

Publications and source records attributed to B Ludwikowski.

7 recordsLinked to original sources

Clinical anatomy of the pelvic floor.

The study presented here comparing cross-sectional anatomy of the fetal and the adult pelvic connective tissue with the results of modern imaging techniques and actual surgical techniques shows that the classical concepts concerning the subdivision of the pelvic connective tissue and muscles need to be revised. According to clinical requirements, the subdivision of the pelvic cavity into anterior, posterior, and middle compartments is feasible. Predominating connecting tissue structures within the different compartments are: Paravisceral fat pad within the anterior compartment (Fig. 17, I), rectal adventitia or perirectal tissue within the posterior compartment (Fig. 17, II), and uterosacral ligaments within the middle compartment. The nerve-vessel guiding plate can be found in all of these compartments; it starts within the posterior compartment and it ends within the anterior one. It constitutes the morphological border between the anterior and posterior compartments in the male. This border is supplied by the uterosacral ligaments in the female. Whereas in gross anatomy no further border is discernable between anterior and posterior or middle compartment, the rectal fascia (hardly visible in embalmed cadavers) demarcates the rectal adventitia and is one of the most important pelvic structures for the surgeon. In principle, the outlined subdivision of the pelvic connective tissue is identical in the male and in the female; facts that become clear from early human life and that are already established during this period (Fig. 18). The uterus is interposed between the bladder and rectum and subdivides the pelvic peritoneum into two pouches thus establishing the only real difference between male and female pelvic cavity. The preferential direction of the pelvic connective tissue fibers is not changed by the interposition of the uterovaginal complex. The pelvic floor muscles are composed of the portions of the levator ani muscle, the muscles of the cavernous organs and the deep transverse perineal muscle in the male. The latter does not exist in the female. We have clearly shown that the different muscles can already be found in early human life and that they are never intermingled with the muscular walls of the pelvic organs. The levator ani muscle of the female, however, is intermingled with connective tissue long before the female sexual hormones exert influence. We have also shown that the distinct sexual differences within the pelvic floor muscles as well as within the sphincter muscles can already be found in early human life. Both the external urethral and the external anal sphincter muscles are not completely circular. The external anal sphincter is intimately connected with the internal sphincter as well as with the longitudinal muscle. Whereas the innervation and function of the urethral sphincter muscles are mostly clear, cloacal development, innervation, and function of all parts of anal sphincter complex are not completely clarified. As to the support of the pelvic viscera, we believe that intact pelvic floor muscles, an undisturbed topography of the pelvic organs, and an undisturbed perineum are of more importance than the so-called pelvic ligaments. Our hypothesis points to the fact that the support of pelvic viscera is multistructural. Thus in pelvic surgery, a lot of techniques have to be revised with the aim to preserve or to reconstruct all the structures mentioned. This is a multidisciplinary task that can only be solved by cooperation of morphologists, urologists, gynecologists, and coloproctologic surgeons or by creating a multidisciplinary pelvic floor specialist.

Blood Vessels↗

Total urogenital sinus mobilization: a modified perineal approach for feminizing genitoplasty and urogenital sinus repair.

PURPOSE: We report a modification of the total urogenital sinus mobilization technique adapted to the repair of isolated persistent urogenital sinus and masculinized external female genitalia. MATERIALS AND METHODS: The records of 6 girls undergoing total urogenital sinus mobilization were reviewed. Diagnosis was the adrenogenital syndrome in 4 girls, and persistent urogenital sinus and true hermaphroditism in 1 girl each. Mean patient age at the time of surgery was 5.9 years (range 4.5 months to 19.5 years). The surgical technique was modified by using the perineal approach since all patients had normal anorectal anatomy. A posterior perineal skin flap was used to widen the vaginal introitus. The wall of the mobilized urogenital sinus was opened and used to create a mucous lined vestibule. In those cases of the adrenogenital syndrome reduction clitoroplasty was performed at the same time. Postoperative results were assessed subjectively by observation of the vulvar appearance and objectively by determining the location of the urethral meatus and caliber of the vagina. RESULTS: Mean followup was 3.7 months (range 1 to 9). There were no postoperative complications. Most patients were discharged home within 3 days of surgery. All patients have a satisfactory cosmetic appearance. The urethral meatus was situated in the vestibule and easily accessible. There were no changes in voiding habits postoperatively in those patients who were toilet trained before surgery. Vaginal calibration was performed in 4 patients and mean vaginal caliber was 10.5 Hegar (range 6 to 14). CONCLUSIONS: The repair of persistent urogenital sinus less than 3 cm. long can be accomplished with total urogenital sinus mobilization through the perineal approach. The technique can be combined with reduction clitoroplasty for the surgical management of girls with masculinized external genitalia.

Adolescent↗

The development of the external urethral sphincter in humans.

OBJECTIVE: To assess the hypothesis that during fetal development, the external urethral sphincter changes from a concentric sphincter of undifferentiated muscle fibres to a transient ring of striated muscle which regresses caudo-cranially in the posterior urethra during the first year of life, when the sphincter assumes its omega-shaped configuration. MATERIALS AND METHODS: The anatomy and development of the external urinary sphincter was assessed in human males and females during fetal life. Plastic-embedded sections (transverse, sagittal and frontal planes; 300-700 microm) of the pelvis of 31 females and 31 males (9 weeks of gestation to newborn) were stained with azure II/methylene blue/basic fuchsin and viewed at x 4-80. The sections of interest were taken from the bladder neck to the perineum. The sections of the membranous urethra were reconstructed three-dimensionally using a computer program. RESULTS: In both male and female an omega-shaped external sphincter was apparent in all specimens at > 10 weeks of gestation. In the early fetal period (ninth week), there was undifferentiated mesenchyme; in this period the mesenchyme was more dense in the anterior part and loose in the posterior part of the urethra. In females, there was a close connection between the urethra and the anterior wall of the vagina. CONCLUSION: The omega-shaped configuration of the external urethral sphincter was recognisable from 10 weeks of gestation in both sexes. There was no suggestion of a change from a cylindrical to an omega-shaped sphincter in the fetal period to birth. Also, a transient 'tail' posterior to the sphincter was not apparent. The rectovesical septum was well developed in neonates. There is no reason to assume that the development of the septum leads to an apoptosis of muscle cells in the posterior part of the external sphincter in males after birth. The anatomical development of the external sphincter does not explain transient outlet obstruction during fetal life. The function of the muscle may change during development because of neuronal maturation.

Embryonic and Fetal Development↗

The long-term culture of porcine urothelial cells and induction of urothelial stratification.

OBJECTIVE: To assess porcine urothelial cell cultures and the in vitro induction of urothelial stratification in long-term cultures, to study their morphological, functional and genetic behaviour, and thus provide potential autologous urothelium for tissue-engineered substitutes for demucosalized gastric or colonic tissue. MATERIALS AND METHODS: Primary cultures of porcine urothelium were established and the cells passaged thereafter. Cell specificity was confirmed by cytokeratin analysis, cell membrane stability assessed using lactate dehydrogenase leakage, cell de-differentiation by gamma-glutamyl transferase activity and genomic stability by karyotype investigations. Histology and scanning electron microscopy were performed to study the cultured cells and the stratified constructs. Furthermore, collagen matrices were tested as cell scaffolds. RESULTS: The cells were cultured for 180 days; 10 subcultures were established during this period. Stratification was induced in a culture flask and on a collagen matrix. Cytokeratins 7, 8, 17 and 18 were expressed in all cultures, and cell membranes were stable, with no evident de-differentiation. The cultures were stable in their genotype and no chromosomal aberrations were found. The histology and immunohistochemistry of the stratified porcine constructs, and cell membrane stability and cell de-differentiation, were compared with those in the human system. CONCLUSION: Pig and human urothelial cells can be cultured over a long period with no signs of senescence. Urothelial stratification can be induced in vitro. The collagen matrix seems to be an excellent scaffold, allowing cell adherence and growth.

Animals↗

Total urogenital sinus mobilization: expanded applications.

OBJECTIVE: To report further applications of total urogenital sinus mobilization, earlier described as an easier method to correct a cloaca. PATIENTS AND METHODS: Seven children (six girls and one boy, mean age 4 years, range 3 months to 10.5 years) underwent surgery and were followed for a mean of 1 year; their diagnoses included persistent cloaca and congenital adrenal hyperplasia (CAH) in two each, and a urogenital sinus (UGS), bladder exstrophy and penile agenesis in one each. The UGS is approached through a posterior sagittal incision and dissected circumferentially to the retropubic space, allowing the UGS to descend. It is then excised and separate openings of the vagina and urethra created. This technique is applicable to a UGS of </=3 cm. RESULTS: In all patients, separate openings for the urethra and vagina were created. In three patients urinary continence was preserved after surgery. The patient with bladder exstrophy remains incontinent. The remaining patients are too young to assess (not yet toilet-trained). CONCLUSION: This technique simplifies the surgical correction of UGS malformation; we confirm its usefulness in cases of persistent cloaca. It is also valuable in patients with CAH, primary UGS and in selected patients with bladder exstrophy and penile agenesis. When the UGS is not associated with a cloaca, the procedure can be performed perineally. Despite circumferential mobilization of the UGS, urinary continence is preserved.

Adrenal Hyperplasia, Congenital↗

Expansion and long-term culture of differentiated normal rat urothelial cells in vitro.

The objective of this study is to establish a reliable cell culture system for the long-term culture of rat urothelial cells (RUC), in which the cells multiply in vitro and form stratified polarized urothelium. Urothelial cells were harvested by the enzymatic digestion of the urothelium exposed by the eversion of resected rat bladders. Primary cultures were initiated in keratinocyte serum-free medium (KSFM) for selective proliferation of urothelial cells. Subsequently, the cells were propagated in a mixture of conditioned medium (CM) derived from Swiss 3T3 cell culture supernatant and KSFM (CM-KSFM). Mean population doubling time was 13.8 +/- 0.9 h. RUC were successfully maintained for 18 passages over a period of 4-5 mo. Detailed investigations of culture conditions showed that CM-KSFM yielded a differentiated multilayer structure. The stratified urothelial sheets measuring 4 x 6 cm2 could be formed and then detached using dispase. Cytokeratin pattern in both the cultured urothelial monolayer and engineered stratified layers was similar to those seen in vivo, as assessed with monoclonal antibody against cytokeratin 17. Ultrastructural morphology showed microvilli, basal cell layer, and desmosomes between adjacent cells in the stratified urothelium.

3T3 Cells↗