Microsurgical hypospadias repair

Successful primary hypospadias repair depends on careful execution of surgical principles, particularly during the urethroplasty portion of the procedure. These principles include careful tissue handling, development of well-vascularized flaps, and avoidance of placing sutures in the uroepithelial surface. Despite meticulous repair, the complication rate requiring secondary surgery is 15 to 30 percent. For the past year, the authors have utilized an operating room microscope, microsurgical instruments, a specially designed microsuture, and a Biooclusive dressing to decrease the postoperative morbidity and subsequent complications requiring secondary surgery (6.5 percent). A comparison of 50 hypospadias patients on whom no microsurgical repair had been used was made with 62 patients on whom microsurgical techniques were employed. The nonmicrosurgical group had 17 complications, 12 of which required reoperation (24 percent). The microsurgical group had 8 complications, 4 of which required reoperation (6.5 percent).     

A Rapid Method of Obtaining Epidermal Peels in Plants by Treatment with Cupric Sulphate and Hydrochloric Acid

In a microsurgical study currently in progress, the need arose to prepare accurately oriented longitudinal sections of artery 1 mm in external diameter. The specimens were rats' common femoral arteries which had been subjected to division and microsuture anastomosis. We wanted to see the extent of any narrowing at the site of the microsurgical repair or histological changes on either side of the repair, and to relate such changes to possible causes of surgical trauma.     

A Method for Accurately Orienting Microsurgical Blood Vessel Specimens for Longitudinal Sectioning

In a microsurgical study currently in progress, the need arose to prepare accurately oriented longitudinal sections of artery 1 mm in external diameter. The specimens were rats' common femoral arteries which had been subjected to division and microsuture anastomosis. We wanted to see the extent of any narrowing at the site of the microsurgical repair or histological changes on either side of the repair, and to relate such changes to possible causes of surgical trauma.     

Functional evaluation of the effectiveness of experimental reconstructive and plastic operations on the uterine horns

In the given experimental model, tubo-plastic surgery was done with the use of locally produced fibrin glue (Fk-1), and commercial product "Beriplast" (FRG). In the control group microsurgical technique was adopted in the uterine horn of rats. It was proved that Fibrin glue reduces the frequency of post operative adhesion formation in the area of anastomosis and the chance of pregnancy is increased in comparison to microsurgical techniques.     

Microsurgery in the clinic and laboratory

Laboratory animals have been crucial to the development of modern microsurgical techniques which are now routinely used in many clinical departments worldwide. In return, microsurgical techniques are important in biomedical research as they allow many surgical procedures to be performed on rodents instead of dogs, pigs or primates. This has obvious advantages in terms of low cost, the use statistically valid numbers for comparison and the availability of genetically defined animals which are more likely to give valid answers to immunological questions. Microsurgical reconstruction is important in plastic, orthopaedic, urogenital, vascular and peripheral nerve surgery in man and it is likely that it will become part of every surgeon's training in the near future. In this review, the instrumentation essential to any microsurgical enterprise and the sutures available are described. Basic microsurgical techniques for end-to-end and end-to-side anastomosis of small vessels and for joining peripheral nerves, oviducts and other tubular structures are given in outline. Techniques for transplanting kidney, heart, heart and lung, liver, spleen, pancreas, small bowel, stomach, testicle, ovary and whole joint are only outlined but key references are given. Finally, some of the clinical indications for microsurgical reconstruction are reviewed.     

Bipolar anastomosis technique with removable instruments: an easy, fast, and reliable technique for vascular anastomosis

The interrupted suture technique is most commonly used for microsurgical vascular anastomosis. For several reasons (e.g., exposure of suture material to blood, time needed), many attempts have been made to find other solutions. This article describes a new means of performing a microsurgical vascular anastomosis. The aim of this study was to show the feasibility and possible advantages of this new technique. The basic components at work here are a modified cuff and electrically generated heat used to unite the vessel walls. In this way, both endothelial layers are adapted without manipulating the inside of the vessel or leaving behind foreign matter. Various energy/coagulation time settings were used to perform arterial anastomoses (n = 42) in an isogeneic abdominal aorta interposition model in the rat. The quality of anastomosis was evaluated at days 1, 10, 21, and 120. Immediately after the welding process all anastomoses (n = 42) were patent. No stenosis was found at any observation time. Anastomosis time ranged from 3 to 18 minutes (average, 11 minutes). This new technique permits a vascular anastomosis to be performed easily and reliably with a high patency rate. With this technique, the authors are convinced that a skilled surgeon can create a high-quality anastomosis in a fraction of the time needed to sew an anastomosis.     

Tubo-ovarian transplantation in the treatment of sterility. Experimental research]

Microsurgical transposition of fallopian tube and ovary has the potential of being an efficient therapeutic treatment in patients with tubal sterility. The Authors present their experience of microsurgical adnexal transplantation in rabbit by two different techniques: the first procedure by microvascular anastomosis of the ovarian vessels, the second one without vascular pedicle. Function is evaluated at various time after grafting by: exploratory laparotomy on day 30 to establish whether circulation to the grafts was still maintained; macroscopic and microscopic examination of ovaries and fallopian tubes. The microvascular techniques prove highly reliable in terms of immediate vascular patency rate but it is disappointing that 50% of the autografts has failed with blocked vessels by day 30. Perhaps this is due to the difficult techniques in anastomosing the ovarian vessels of small caliber. In spite of these outcomes the vascularized autografts were viable and functional after transplantation in contrast with the non-vascularized tubo-ovarian grafts which all failed. This experience encourages to believe that the microsurgical technique could be employed for homograft transplantation in woman with extensive ovarian and tubal damages.     

Significance of venous anastomosis in fingertip replantation

Adequate venous outflow is the most important factor for successful fingertip replantation. The authors have attempted venous anastomosis in all cases of fingertip replantation to overcome postoperative congestion. In this article, the significance of venous repair for fingertip replantation is described from the authors' results of 64 complete fingertip amputations in 55 consecutive patients, which were replanted from January of 1996 to June of 2001. The overall survival rate was 86 percent. Of the 44 replantations in zone I, 37 survived, and the success rate was 84 percent. Of the 20 replantations in zone II, 18 survived, and the success rate was 90 percent. Venous anastomosis was attempted in all cases, but it was possible in 39 zone I and in all zone II replantations. For arterial repair, vein grafts were necessary in 17 of the 44 zone I and in one of the 20 zone II replantations; for venous repair, they were necessary in six zone I replantations and one zone II replantation. Postoperative vascular complications occurred in 15 replantations. There were five cases of arterial thrombosis and 10 cases of venous congestion. Venous congestion occurred in nine zone I and one zone II replantations. In five of these 10 replantations, venous anastomosis was not possible. In another five replantations, venous outflow was established at the time of surgery, but occlusion occurred subsequently. Except for the five failures resulting from arterial thrombosis, successful venous repair was possible in 49 of 59 replantations (83 percent). Despite the demand for skillful microsurgical technique and longer operation time, the authors' results using venous anastomosis in successful fingertip replantations are encouraging. By performing venous anastomosis, external bleeding can be avoided and a higher survival rate can be achieved. Venous anastomosis for fingertip replantation is a reliable and worthwhile procedure.     

A triangular cutting arteriotomy scissors facilitating end-to-side anastomoses.

A new microsurgical triangular cutting scissors facilitating end-to-side anastomoses has been developed. The instrument removes oval pieces of tissue from the vascular wall with a single cut. This has several advantages over conventional double cuts with straight microscissors. Preferential use of end-to-side anastomoses has been advocated by several authors, one of the main reasons for this being preservation of the distal blood flow in the artery used in the recipient bed. It also has been reported that vascular occlusions are less frequent with this type of coupling. Another important indication for end-to-side anastomoses is a significant discrepancy in vessel size. One of the more demanding tasks in making an end-to-side anastomosis is the performance of a perfect arteriotomy. With the aim of facilitating the arteriotomy in end-to-side anastomoses, a new microsurgical instrument has been developed.     

Up-regulation of production of TGF-beta and IL-4 and down-regulation of IL-6 by apoptotic human bronchial epithelial cells

Human bronchial epithelial cells secrete cytokines that play a role in immune responses in the lung. However, the roles of these cytokines in regulating epithelial repair following acute lung injury are largely unknown. Responses to injury include hyperplasia of epithelial cells and squamous metaplasia. The resolution stage is characterized by discontinuation of hyperplasia. Apoptosis is considered to be the most efficient mechanism of removal of unwanted cells without causing inflammation. The presence of TGF-beta1 increases apoptosis, induces squamous metaplasia and inhibits proliferation of airway epithelial cells. Interleukin-4 increases the ability of macrophages to phagocytose epithelial cells and produce inflammatory cytokines. The purpose of this study was to investigate the hypothesis that apoptotic lung epithelial cells produce cytokines, which could act in an autocrine manner to control hyperplasia and induce squamous differentiation following acute lung injury. A bronchial epithelial cell line (16 HBE) was used as an in vitro model, to study the production of TGF-beta, IL-4 and IL-6 by lung epithelial cells undergoing apoptosis. Apoptotic and live cells were sorted on the basis of bright and negative staining with FITC-conjugated Annexin V, respectively. Intracellular IL-6, TGF-beta and IL-4 was measured using flow cytometric techniques. Electron microscopy, immunohistochemistry and ELISA were used as supportive techniques. Apoptotic cells produced significantly more TGF-beta and IL-4 (but less IL-6) than viable cells. Increased production of TGF-beta and IL-4 by epithelial cells undergoing apoptosis may contribute to the inhibition of proliferation, squamous metaplasia, and reduction of the inflammatory response in acute lung injury.     

Present status and new perspectives in laser welding of vascular tissues

The laser welding of biological tissues is a particular use of lasers in surgery. The technique has been proposed since the 1970s for surgical applications, such as repairing blood vessels, nerves, tendons, bronchial fistulae, skin and ocular tissues. In vascular surgery, two procedures have been tested and optimized in animal models, both ex vivo and in vivo, in order to design different approaches for blood vessels anastomoses and for the repair of vascular lesions: the laser-assisted vascular anastomosis (LAVA) and the laser-assisted vessel repair (LAVR). Sealing tissues by laser may overcome the problems related to the use of conventional closuring methods that are generally associated with various degrees of vascular wall damage that can ultimately predispose to vessel thrombosis and occlusion. In fact, the use of a laser welding technique provides several advantages such as simplification of the surgical procedure, reduction of the operative time, suppression of bleeding, and may guarantee an optimal healing process of vascular structures, very similar to restitutio ad integrum. Despite the numerous preclinical studies performed by several research groups, the clinical applications of laser-assisted anastomosis or vessel repair are still far off. Substantial breakthrough in the laser welding of biological tissues may come from the advent of nanotechnologies. Herein we describe the present status and the future perspectives in laser welding of vascular structures.     

New methods of X-ray surgical diagnosis and treatment of varicocele in children and adolescents

The authors studied 520 patients with left-sided varicocele by left-sided phleborenotesticulography of Doppler ultrasonography, tensiometry of the left renotesticular venous system, blood gas composition, hormonal parameters, and etc. They present rationale for and evidence illustrations of aortomesenteric compression of the left renal vein, left-sided phleborenal hypertension prior to and after surgery for endovascular occlusion of the left testicular vein, before and following intervenous proximal testiculoiliac venous bypass surgery. The data of andrological dysfunction in patients with varicosis of the pampiniform plexus and spermatic cord (varicocele) are presented. An algorithm of examination and treatment of this cohort of patients, by using both miniinvasive techniques (for X-ray endovascular occlusion of the temporal veins under local anesthesia) and high tech operations using microsurgical techniques (testiculoiliac venous anastomosis), is outlined.     

The use of lasers in end-to-end anastomosis of the uterine horn in the rat. Histological considerations

The Authors have verified the utility of the Laser CO2 instrument for termino-terminal anastomosis of the uterine horns of the rat. In this study Laser tecnique is compared with the same traditional microsurgical operation. No significant difference was seen in the number of pregnancies nor fetuses born. On the contrary, hystological observation has shown lower reactivity in the group treated with Laser technique 10 days. In this group it was also noted that there was less formation of postoperative adhesions, and, during the operation, there was easier dissection of the tissues accompanied by lower blood loss and necrotic debris.     

Use of prostheses made of ePTFE in anastomoses of the rat uterine horn

ePTFE prostheses were used in the microsurgical anastomosis of the rat uterine horn. Handling characteristics of this synthetic material during the performance of anastomosis were tested. Patency rates, infections and local tissue reaction were evaluated after thirty and sixty days. Connective tissue infiltration and incomplete endothelialization by normal endometrial elements were observed on the luminal surface of the prosthesis.     

Microvascular anastomosis: vascular cuff technique

A technique is described for reinforcing and sealing a microvascular anastomosis by using a detached vessel segment as a cuff around the anastomosis. This may allow fewer microvascular sutures to be used. This would allow a simpler, less traumatic, and more rapid repair and may allow "more vessels to be repaired by more people."     

Rat model of pulmonary arteriovenous malformations after right superior cavopulmonary anastomosis

We developed a rat model of pulmonary arteriovenous malformations after cavopulmonary anastomosis. We sought to determine whether this model reproduces the angiographic and histologic features seen in the human condition. Eight Sprague-Dawley rats underwent a right superior cavopulmonary anastomosis with the use of microsurgical techniques. Between 2 and 13 mo, pulmonary angiography was performed, the animals were euthanized, and the lungs were removed. Microscopic sections of the lung were stained with an endothelial-specific antibody (von Willebrand factor). Microvessel density was determined by counting vessels staining positively for von Willebrand factor, and the shunted and nonshunted (control) lungs were compared for each animal. Pulmonary angiography revealed time-dependent development of arteriovenous malformations. Microvessel density demonstrated a time-dependent increase in the shunted lung compared with the control lung (simple linear regression of the ratio of the microvessel density of the shunted lung divided by the microvessel density of the control lung on time; R(2) = 0.79, P = 0.003). This animal model reproduces the same angiographic and microscopic features of pulmonary arteriovenous malformations that develop in humans after cavopulmonary anastomosis. This appears to be a valid model that may be used to further study etiologic mechanisms for this phenomenon.     

Arterial repair after microvascular anastomosis. Scanning and transmission electron microscopy study

In order to study the morphological aspects of endothelial regeneration and vascular wall reaction after microvascular anastomosis, rat femoral arteries were sectioned and successively sutured (end-to-end anastomosis) with microsurgical techniques. Control arteries and anastomosed vessels (recovered after 1, 4, 7, 14, 21, 30, 60, 120, 180 and 360 days) were studied by means of scanning (SEM) and transmission electron microscopy (TEM). The reendothelialization phenomena started after 7 days and were mainly evident at 21 days. Areas of subendothelial connective tissue with fibrin deposition remained exposed to the blood stream up to 21-30 days. Thrombus formations or post-anastomotic stenosis have been occasionally observed. Regenerating endothelium showed evident morphological differences from the control. These changes mainly consisted of shortened cell length, absence of pinocytotic vesicles, presence of cytoplasmic prolongations, and microvillous proliferations. The arterial wall showed subintimal thickening. The anastomotic site appeared completely covered by new endothelium after 30-60 days. Subintimal vascular wall changes (thickening of the media) as well as slight alterations of endothelial cells (shortened length, reduced number of pinocytotic vesicles) were evident in 60-day vessels. Lumen reduction, due to the protruding of endothelial-covered sutures, was occasionally observed in 60- to 120-day arteries. Endothelial cell morphology normalized after 60-120 days. However, thickening of the media and occasional lumen reduction were observed also after 180-360 days. Although the endothelial regeneration phenomena were clearly evident after 2 weeks, nevertheless the reestablishment of arterial wall took longer time.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pneumonia Due to Mycoplasma in Gnotobiotic Mice IV. Localization and Identification of Mycoplasma pulmonis in the Bronchi of Infected Gnotobiotic Mice by Immunofluorescence and by Light Microscopy

The model of pneumonia produced by intranasal inoculation of Mycoplasma pulmonis in gnotobiotic mice provided the opportunity to study the localization and identification of the infecting organisms in the tissues by immunofluorescence techniques. Frozen sections of pneumonic mouse lung were fixed in acetone, layered with rabbit anti-M. pulmonis serum, washed, layered again with fluorescein-isothiocyanate-labeled goat anti-rabbit globulin, washed again, and examined by fluorescence microscopy. A bright line of fluorescence was seen at the bronchial epithelial surface, usually in a continuous layer. Occasional masses of fluorescence were seen in the polymorphonuclear leukocytic exudate in the bronchial lumen. Sections of tissues fixed in Helle's or 10% Formalin fixatives and stained with hematoxylin and eosin were reviewed by light microscopy and revealed a zone of blue-staining material composed of tiny coccoid bodies in the same locations at the bronchial epithelial surface as in the immunofluorescent preparations and in previously reported electron microscope studies.     

Inhibition of microsurgical thrombosis by the platelet glycoprotein IIb/IIIa antagonist SR121566A

Despite major improvements in tools and significant refinements of techniques, microsurgical anastomosis still carries a significant risk of failure due to microvascular thrombosis. The key to improving the success of microvascular surgery may lie in the pharmacologic control of thrombus formation. Central to pathologic arterial thrombosis are platelets. Glycoprotein IIb/IIIa is a highly abundant platelet surface receptor that plays a major role in platelet aggregation by binding platelets to each other through the coagulation factor fibrinogen. To explore the ability of antithrombotic agents to prevent microvascular thrombosis, a rabbit ear artery model was used in which a standardized arterial injury results in predictable thrombus formation. This model was used to examine whether SR121566A, a specific and potent glycoprotein IIb/IIIa inhibitor, can successfully prevent microsurgical thrombosis.Using a coded, double-blind experimental design, 20 rabbits (40 arteries) were assigned to four treatment groups: (1) saline injection (n = 10), (2) acetylsalicylic acid 10 mg/kg (n = 10), (3) heparin 0.5 mg/kg bolus with subsequent intermittent boluses of 0.25 mg/kg every 30 minutes (n = 10), and (4) SR121566A 2 mg/kg bolus (n = 10). After vessel damage and clamp release, arteries were assessed for patency at 5, 30, and 120 minutes by the Acland refill test. Coagulation assays, in vivo bleeding times, and ex vivo platelet aggregation studies were also conducted. Scanning electron microscopy was used to examine mural thrombus composition.A significant, fourfold increase in vessel patency following administration of SR121566A over saline control (80 percent versus 20 percent patency, respectively, at 35 minutes after reperfusion, p < 0.01) was noted. This was correlated with marked inhibition of ex vivo platelet aggregation. This antiplatelet treatment did not prolong coagulation assays (mean international normalized ratio: saline, 0.66 +/- 0.04; SR121566A, 0.64 +/- 0.03; mean thromboplastin time: saline, 19.63 +/- 0.67; SR121566A, 17.87 +/- 3.27) and bleeding times (mean bleeding time: saline, 42 +/- 4; SR121566A, 48 +/- 6). Scanning electron microscopy demonstrated extensive platelet and fibrin deposition in control vessel thrombi. In contrast, thrombi from SR121566A-treated vessels demonstrated predominance of fibrin with few platelets when examined under scanning electron microscopy.Administration of SR121566A was associated with a significant increase in vessel patency, without deleterious effects on coagulation assays or bleeding times. The increase in vessel patency was correlated with inhibition of platelet aggregation and decreased platelet deposition, as demonstrated by scanning electron microscopy. Glycoprotein IIb/IIIa antagonists represent a new class of anti-platelet agents that may be suited for inhibiting microsurgical thrombosis. This study supports further investigation into the use of these agents in microsurgery.