Endogenous TGF-beta 1 and TGF-beta 2 are not essential for epithelialization and neovascularization in the hairless mouse ear wound model.

BACKGROUND AND AIMS: TGF-beta stimulates neovascularization and epithelialization in healing wounds, yet relatively little is known about the mechanisms involved. Using the hairless mouse ear wound model, we studied the effects endogenous TGF-beta 1 and TGF-beta 2 have on epithelialization and neovascularization following the application of neutralizing antibodies. MATERIAL AND METHODS: Forty-three adult male hairless mice had an excisional wound made on the dorsum of each ear. Using vital microscopy, epithelialization and neovascularization were measured every third day until completion. TGF-beta 1 and TGF-beta 2 antibody, control-IgG, or phosphate buffered saline (PBS) were applied to the wounds. RESULTS AND CONCLUSIONS: Excisional wounds treated with anti-TGF-beta 1 and anti-TGF-beta 2, IgGcontrol IgG, and PBS epithelialized in 11.2 +/- 0.5 days (N = 22), 10.9 +/- 0.6 days (N = 17), and 10.6 +/- 0.6 days (N = 15), respectively and neovascularized in 27.9 +/- 0.5 days (N = 17), 27.1 +/- 0.8 days (N = 14), and 26.1 +/- 0.8 days (N = 10), respectively (mean +/- SEM). There were no significant differences in time to epithelialization and neovascularization between the three groups. Furthermore, there were no differences in the average time course of epithelialization and neovascularization between the three groups throughout the healing process. We conclude that endogenous TGF-beta 1 and TGF-beta 2 are not essential for epithelialization and neovascularization in the hairless mouse ear wound model.

Ketanserin accelerates wound epithelialization and neovascularization.

We investigated the acute effects of topical ketanserin, a 5-HT(2) (serotonin) receptor blocker, on wound epithelialization and vascularization with the use of the hairless mouse ear model. Varying concentrations of Ketanserin (0%, 0.2%, 2.0%, 20% weight/volume) were administered to standardized full-thickness skin wounds on the dorsum of the hairless mouse ear immediately after surgery and daily thereafter. With the use of video microscopy and computer-assisted planimetry, vascularization and epithelialization were traced every third day until the wounds were fully healed. Arteriole diameters at selected sites near the skin wound were measured before wound creation and after wounding. It was concluded that topically administered ketanserin significantly accelerates both the vascular (p < 0.001 at 2% and 20% concentrations) and epithelial (p < 0.001 at 20% concentration) rates of wound healing in full-thickness nonpathologic skin wounds. Vasodilation of terminal arterioles was not a major response to Ketanserin. Faster epithelialization was possibly due to direct effect of ketanserin on epithelial cells.

Biofeedback treatment of children and adults with idiopathic detrusor instability.

In adults and children suffering from urge, urge incontinence, frequent voidings, and nocturia, urodynamic examination often confirms the diagnosis of detrusor instability. We assessed the outcome of biofeedback therapy in 15 children aged 6-12 years and 7 adults aged 20-52 years, all with cystometrically proven detrusor instability. The detrusor pressure was visually conveyed to the patient during repeated bladder fillings. The patient was instructed to interrupt detrusor pressure increments by tensing the pelvic floor musculature. None of the children were found completely cured; however, 9 showed a marked decrease in either the number of the extent of symptoms. Two children showed moderate improvement, while 4 remained unaffected by the treatment. One adult was completely cured, 2 improved moderately, and 4 showed no improvement. None of these patients were converted to stable cystometry. Upon termination of the BF treatment a follow-up study was conducted for patients exhibiting pronounced or moderate improvement. The beneficial effect was maintained except in one case.

Direct visualization and measurements of wound neovascularization: application in microsurgery research.

Neovascularization or angiogenesis is an essential yet poorly understood component of the healing process. In wound healing research, there is a lack of models enabling quantitative and continuous measurements of wound neovascularization. The hairless mouse ear wound model permits quantitative measurements of wound epithelialization and neovascularization continuously throughout the healing process. On the ears of male homozygous (hr/hr) hairless mice, standardized circular full thickness dermal wounds are produced; then, using vital microscopy, these two processes are directly viewed and measured at day 0 and every third day thereafter until these are complete. This model system and its application to clinically relevant situations are reviewed.

Urodynamic biofeedback training for children with bladder-sphincter dyscoordination during voiding.

Neurologically normal children with recurrent urinary tract infections (UTIs), night- and daytime wetting, and urge and painful voiding may have staccato voiding due to pelvic floor contractions. The immediate effect of non-invasive urodynamic biofeedback (BF) therapy was assessed using a historical follow-up study in 31 children aged 5-15 years suffering from urodynamically proven overactive urethra during voiding. A long-term follow-up study was performed to investigate whether improvement was maintained. Twenty-four children (77.5%) benefited from the treatment. Of these 16 (51.5%) were cured, while 8 (26%) had a pronounced reduction in their symptoms. Although the flow was normalized in 17 (55%) and nearly normalized in 7 (22.5%), there was no significant correlation between subjective and objective criteria of improvement. Similarly, no relationship was found between the initial urodynamic characteristics and the treatment outcome. During a mean follow-up time of 4 years (range: 1-7.5 years) two of the initially cured patients relapsed. They were recurred with a refresher course. Three had had a single or a few episodes of cystitis in the course of several years. Of the patients with pronounced reduction in their symptoms, three relapsed. A refresher course was attempted in two patients; one was successful. It can thus be concluded that BF is an effective way of treating this disturbance and the beneficial effect is to a wide degree maintained.

The microcirculation of myocutaneous island flaps in pigs studied with radioactive blood volume tracers and microspheres of different sizes.

In order to further improve the understanding of hemodynamic changes in the immediate postoperative phase after elevation of myocutaneous flaps, regional blood flow and arteriovenous (A-V) shunting were measured in rectus abdominis island flaps in 8 pigs. Radioactive microspheres of two sizes (15 and 50 micron) were used. Approximately half (53.4 +/- 6 percent) of the 15-micron microspheres and one-fourth (24.1 +/- 6 percent) of the 50-micron microspheres entering the flap appeared in the venous outflow. Compared with the control area, A-V shunting was significantly increased in muscle and substantially more pronounced in skin. Nutritional blood flow, total blood flow, and vascular volume were increased in muscle and unchanged in skin and subcutis. The lowest tissue hematocrit of 7 +/- 1 percent was found in skin as compared with a central hematocrit of 35 +/- 2 percent. Tissue hematocrit in flap muscle was decreased to 17 +/- 2 percent when compared with control muscle (22 +/- 3 percent), and the mean transit time for blood was correspondingly decreased. Thus vasodilation provided increased perfusion through muscular capillaries and through A-V shunts. Shunting of 15-micron microspheres appeared to take place not only in skin, but also in subcutis and muscle, which challenges the widespread belief that A-V shunting does not occur in muscle.

Fuel metabolism in a pig myocutaneous island flap model.

Unilateral denervated myocutaneous island flaps based on the superior epigastric vessels were raised in 24 pigs and the metabolic changes during the first 6 postoperative hours were monitored. Secondary to flap elevation, decreased arteriovenous (A-V) differences in oxygen, glucose, and alanine levels were observed, indicating the opening of A-V shunts and increased arterialization of the venous blood. Venous outflow increased during the first 3 hours, but the A-V differences in all metabolites were constant over the entire 6-hour observation period. Exchange of intermediary metabolites therefore increased within the first 3 hours, after which a steady state was established. The main flap fuels seemed to be fatty acids, muscle proteins, and glycogen, whereas blood-borne carbohydrates and ketone bodies played only a minor role as energy sources. Anaerobic metabolism was increased secondary to flap elevation from 2 to 6 percent as compared with preelevation values. No changes were found in concentrations of plasma catecholamines, which were constantly high. An average weight gain of 3 to 4 percent per hour was equally distributed to skin, subcutis, panniculus carnosus, and muscle. Thus the flap seemed to adapt to the new perfusion pattern within a few hours by a slightly increased anaerobic metabolism, but still with an oxidative metabolism of more than 90 percent.

Tissue oxygen tension in myocutaneous flaps. Correlation with blood flow and blood gases.

The relationship between tissue oxygen tension (PO2) and blood flow, blood gas tensions, oxygen delivery, and oxygen consumption was investigated in myocutaneous island flaps in 24 pigs. The tissue PO2 was measured by the silicone rubber tonometer technique. Capillary blood flow was increased during the first 3-4 hours after implantation of tonometers and they must, therefore, be implanted 3-4 hours before use. After raising the flap, mean (SEM) total flap blood flow increased from 11.2 (0.8) ml/min to 16.0 (1.3) ml/min within the first three hours. Mean (SEM) subcutaneous tissue PO2 decreased from 9.5 (0.6) kPa to 5.2 (0.7) kPa over six hours after the procedure, whereas muscle PO2, blood oxygen tension, and arteriovenous oxygen difference were unchanged. Hence, both the supply to, and use of oxygen in, the flap increased. Muscular and subcutaneous oxygen tensions were about 3 and 1 kPa, respectively, lower than venous oxygen tension. Tissue PO2 did not correlate with blood flow, blood oxygen tensions, arteriovenous differences in oxygen tension, or with oxygen supply in a cross section of animals. Tissue PO2 seems to reflect accurately the balance between oxygen supply and consumption, which might be the most useful indicator for continuous flap monitoring.

Arteriovenous shunting and regional blood flow in myocutaneous island flaps: an experimental study in pigs.

In eight pigs, total blood flow, regional capillary blood flow distribution, and arteriovenous (AV) shunting were studied during the first 4 postoperative hours after elevation of a myocutaneous rectus abdominis island flap. Capillary blood flow and AV shunting were measured using radioactive microspheres before flap creation and 1 and 4 hours after surgery. Total blood flow, measured continuously as venous outflow, increased in the first postoperative hour (p less than 0.05). Elevation of the flap caused a slight decrease in skin capillary blood flow (p less than 0.05), whereas muscular capillary blood flow increased (p less than 0.01). AV shunting accounted for 50 percent of the total flap blood flow, whereas it was negligible in the abdominal wall prior to flap elevation. Thus stalk blood flow, skin appearance, and skin temperature may be poor indicators of nutritional capillary perfusion. However, the clinical and nutritional consequences of these findings remain to be established.

The influence of tissue tension on the microcirculation in myocutaneous flaps: an experimental study in dogs.

The effect of increased and decreased uniaxial tissue tension on capillary blood flow, plasma volume, edema formation, and vascular permeability in myocutaneous flaps was studied. A total of 64 equally sized arterialized myocutaneous flaps were outlined on the abdomen of 16 dogs. The four areas on each dog were randomized to either untouched control area, reattachment to normal tension, increased (one and one-half times the original length), or decreased (one-half times the original length). Capillary blood flow measured by the radioactive microsphere technique two hours after operation was decreased in the muscular part of flaps reattached at decreased and normal tension (p less than 0.05). The muscular blood flow was unchanged when flaps were reattached at increased tension and significantly higher than in flaps reattached at decreased tension (p less than 0.05). No differences were found in skin or subcutaneous blood flow. There were no differences in plasma volume, vascular permeability, or total water content between flaps attached under different tensions, whereas the plasma volume was increased in muscle in all three flap types (p less than 0.05) presumably secondary to venous stasis. The study implies that the decreased capillary blood flow seen in the muscular component of a myocutaneous flap in the immediate postoperative period may be counteracted by a somewhat increased reattachment tension.