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Morphology of rabbit oviduct after reanostomosis with fibrin glue
295
Inr. J. Gynaecol. Obstet., 1984,22: 295-301 International Federation of Gynaecology & Obstetrics
MORPHOLOGY OF RABBIT OVIDUCT AFTER REANOSTOMOSIS WITH FIBRIN GLUE
R. SPERNOL, P. RISS, K. SCHINDLERa, A. BECK and G. BREITENECKER II. Universittits-Frauenklinik, Spitalgasse 23, A-1090 Wein and aKIinik fiir Geburtshilfe, Gynikologie und Andrologie, Veten’tirmedizinische Universittit, Linke Bahngasse I I, A-I 030 Wien (Austrti) (Received January 12th, 1984) (Accepted February 27th, 1984)
Abstract Spernol R, Riss P, Schindler K, Beck A, Breitenecker G (II, Universitiits-Frauenklinik, Spitalgasse 23, A-l 090 Wien and Klinik fur Geburtshilfe, Gyndkologie uhd Andrologie, Universitat, Linke Veterinarmedizinische Bahngasse I I, A-l 030 Wien, Austria). Morphology of rabbit oviduct after reanastomosis with fibrin glue). Int J Gynaecol Obstet 22: 29.5-301,1984 We examined the site of experimental reanastomosis with fibrin glue in rabbit oviducts 3 months after the operation with light and scanning electron microscopy. Local tissue reaction was minimal, no fibrin clots were seen and inflammatory reactions were almost absent. On scanning electron microscopy ciliated cells were reduced in number. Morphologic changes at the site of anastomosis after the use of fibrin glue appear comparable to the alterations seen after the use of microsutures.
tube and on the prevention of scarring and of postoperative adhesions [ 81. Microsurgical suture of the anastomosis is a well-established technique. However, in the search for simpler and less traumatic procedures the use of fibrin glue, which has been used successfully in vas deferens [4] . in nerve transplants [3 I as well as in general surgery [9], has been suggested. Several authors have used fibrin glue for reconstruction of the oviduct in animals and reported satisfactory patency rates [ 2,.5,7 I. An important consideration is the morphological result after reanastomosis. Though the microscopic chtnges after surgical reconstruction are well documented [ 1 I, little is known on light and electron microscopic alterations in the fallopian tube after the use of fibrin glue. We therefore performed experimental reanastomosis of rabbit oviducts with fibrin glue and investigated tissue reactions by means of light and scanning electron microscopy.
Materials and methods Keywords: phology;
Rabbit oviduct; Histology; Mor-
Anastomosis;
Fibrin glue.
Introduction Microsurgical reanastomosis of the fallopian tube has gained increased importance in the last few years. The postoperative results depend on the morphology of the fallopian 0020-7292/84/$03.00 0 1984 International Federation of Gynaecology & Obstetrics Published and Printed in Ireland
The study was performed on 16 silver rabbits. After midline laparotomy the concomitant vessels of one oviduct were coagulated using bipolar microsurgical forceps and an operation microscope. After sharp transsection of the oviduct 1 cm distally from the tubo-cornual junction a nylon splint was inserted into both parts of the tube. The sectional planes were approximated and 1 Inr J Glwaecol Obstet 22
296
Spernol et al.
drop of the fibrinogen glue was applied by means of a mixing syringe. The 2 ends of the oviduct were kept in tight contact until drying of the glue after 2-3 min. The contralateral tube was occluded by means of tantal hemoclips. The splint remained in place for up to 12 h. Twenty rabbits could be covered 3 weeks after the operation, neither gonadotropins nor insemination were used. Six rabbits became pregnant, 2 of them twice, and delivered. Three months after the operation the oviducts were removed and the site of the anastomosis placed in formaldehyde or glutaraldehyde, respectively. For histological examination paraffin sections were prepared and stained with hematoxylin-eosin (HE) and ironhematoxylin, fuchsin, phosphormolybdenorange and light green according to Goldner. The samples for scanning electron microscopy were cut longitudinally and vaporized with gold.
Results Light microscopy
Figures l-3 show light microscopic findings 3 months after the anastomosis. There were stout bands and fibrosis in the muscular layer. The mucosal folds were slightly flattened with partly uneven scarring. In the mesosalpinx foreign body giant cells were found (Fig. 3). Intraluminal fibrin deposits were not seen, inflammatory reactions in the mucosa or in the wall of the tube were minimal. Electronmicroscopy
Scanning electron microscopy revealed interrupted and lowered folds of the mucosa. Some detritus was seen near the site of the anastomosis. Though some undifferentiated peg cells were observed in proximity of the anastomosis the number of cilia was only slightly reduced and the cilia themselves appeared normal (Figs. 4-6).
Fig. 1. Rabbit oviduct 3 months after reanastomosis with fibrin glue, GoldneMaining, magnification 25 : 1. he gular folds of the mucosa, dispersed glands and fibrotic muscular layer at the site of the anastomosis. Int J Gynaecol Obstet 22
Reanustomosis with fibrin glue
Fig. 2. HE staining, magnification anastomosis.
Fig. 3.
63 : 1. Moderate lymphocytic
infiltration
297
and stout bands of muscularis 2It the site of the
HE staining, magnification 100: 1. Foreign body giant cells in the rabbit mesosalpinx after reanastomosis with flkin glue. Int J Gynaecol Obstet 22
298
Spernol et al.
Fig. 4. Scanning electron microscopy: magnification 150 : 1. (a) Normal mucosa of the rabbit oviduct; (b) site of the anastomosis 3 months after reanastomosis with fibrin glue: interrupted and slightly lowered mucosal folds with some intraluminal detritus.
Discussion The good patency and pregnancy rates which we and others observed after experimental reanastomosis of the rabbit oviduct are encouraging [6], Morphologic studies of the oviduct permit evaluation of local tissue reaction and seem to explain the good ZntJ Gynaecol Obstet 22
clinical results. We were able to show that 3 months after the operation very little morphologic changes persisted at the site of reanastomosis. One of the most important factors influencing ovum transport are the cilia. While Bernhard-Huth et al. [ 11 reported that cilia were absent over sutures on scanning electron
Reanastomosis with fibrin glue
299
Fig. 5. Scanning electron microscopy: magnification 4800 : 1. (a) Normal mucosa of the rabbit oviduct; (b) site of the anastomosis: undifferentiated cells and ciliated cells, which are reduced in number.
microscopy, we did not observe an area near the anastomosis where cilia were absent though their number seemed to be reduced (Fig. 5). Both light and electron microscopy showed that the area of morphologic changes around the anastomosis was extremely small, an important point in view of the fact that the epithelial lining of the fallopian tube is critical for adequate ovum transport.
A possible source of complications could be the persistence of fibrin in or around the oviduct. We did not see any fibrin clots attached to the site of anastomosis, nor did we observe intraluminal fibrin on histologic examination. Scheidel et al. [7] saw intraluminal fibrin clots up to 7 weeks after the operation. The fibrinolytic activity of the fallopian tube has not yet been adequately Int J Gynaecol Obstet 22
300
Spew01 et al.
Fig. 6. Scanning electron microscopy: Magnification 14000 : 1. (a) Normal mucosa of the rabbit oviduct; (b) site of the anastomosis: the ciliated cells are reduced in number but the cilia appear normal.
determined; unpublished results suggest that the oviduct has a high fibrinolytic activity, possibly explaining the complete absorption of fibrin from the site of operation. In summary tissue reaction after use of fibrin glue is comparable to the changes seen after the use of microsutures. It appears Int J Gynaecol Obstet 22
possible that by changing the composition of the fibrin glue a further reduction in tissue reaction can be achieved. Though the numbers in our study are too small to allow definite conclusions, the good healing and minimal tissue reaction after the use of fibrin glue appears encouraging.
Reanastomosis with fibrin glue
Acknowledgment We are grateful to H. Hbrantner, Institute of Micromorphology and Electron Microscopy (Head: Prof. Dr. L. Stockinger), University of Vienna, Medical School, for the electron microscopic evaluations. References Bernhardt-Huth D, Frantzen Ch, Schlisser HW: Morphology of rabbit oviduct after microsurgical techniques for reanastomosis of the isthmus or ampulla. Arch Gynecol230: 251, 1981. lntraphuvasak J, Stofft E, Osada H, Friedberg V: Tuben End-zu-End-Anastomose mit Fibrinkleber. Abstrakt 17, 44. Tagung der Deutschen Gesellschaft fiir Gynlkologie und Geburtshilfe, Miinchen 13.9-17.9.1982. Kudema H: Klinische Anwendung der Klebung von Nervenanastomosen mit Fibrinogen Fortschr KieferGesichtschir 21: 135, 1976. Pfliiger H, Lunglmayr G, Breitenecker G: Vaso-Vasoanastomose unter Verwendung von Frbrinogenkonzentraten: Versuche an Kaninchen. Wien Klin Wochenschr 88; 800,1976.
301
Osada H, Noda M, Fujli TK, Tsubata K, Takagi S: Basic studies on the use of a fibrinogen-adhesive system during fallopian tubal transplants. Abstract 396, XIth World Congress on Fertility and Sterility, June 26-July 1, 1983, Dublin, Ireland. Riss P, Spernol R, Beck A, Schindler K: Reanastomose der Kaninchentube mit Fibrinogenkonzentrat. Wien Klln Wochenschr 00: 000,1984. Scheidel PH, Wallwiener DR, Wiedemann RA, HEPP HK: Experimental anastomosis of the rabbit fallopian tube using fibrin glue. Fertil Steril38: 471,1982. Winston RML: Microsurgery of the fallopian tube: from fantasy to reality. Fertil Steril34: 521, 1980. Wolner E: Die Fibrinklebung in der Chirurgie, Wien Klin Wochenschr 94: 181,1982.
Address for reprints: Dr. R. Spemol LKH Klagenfurt St. Veiterstrapse 47 A-9020 Klagenfurt Austria
int J Gynaecol Obstet 22
Inr. J. Gynaecol. Obstet., 1984,22: 295-301 International Federation of Gynaecology & Obstetrics
MORPHOLOGY OF RABBIT OVIDUCT AFTER REANOSTOMOSIS WITH FIBRIN GLUE
R. SPERNOL, P. RISS, K. SCHINDLERa, A. BECK and G. BREITENECKER II. Universittits-Frauenklinik, Spitalgasse 23, A-1090 Wein and aKIinik fiir Geburtshilfe, Gynikologie und Andrologie, Veten’tirmedizinische Universittit, Linke Bahngasse I I, A-I 030 Wien (Austrti) (Received January 12th, 1984) (Accepted February 27th, 1984)
Abstract Spernol R, Riss P, Schindler K, Beck A, Breitenecker G (II, Universitiits-Frauenklinik, Spitalgasse 23, A-l 090 Wien and Klinik fur Geburtshilfe, Gyndkologie uhd Andrologie, Universitat, Linke Veterinarmedizinische Bahngasse I I, A-l 030 Wien, Austria). Morphology of rabbit oviduct after reanastomosis with fibrin glue). Int J Gynaecol Obstet 22: 29.5-301,1984 We examined the site of experimental reanastomosis with fibrin glue in rabbit oviducts 3 months after the operation with light and scanning electron microscopy. Local tissue reaction was minimal, no fibrin clots were seen and inflammatory reactions were almost absent. On scanning electron microscopy ciliated cells were reduced in number. Morphologic changes at the site of anastomosis after the use of fibrin glue appear comparable to the alterations seen after the use of microsutures.
tube and on the prevention of scarring and of postoperative adhesions [ 81. Microsurgical suture of the anastomosis is a well-established technique. However, in the search for simpler and less traumatic procedures the use of fibrin glue, which has been used successfully in vas deferens [4] . in nerve transplants [3 I as well as in general surgery [9], has been suggested. Several authors have used fibrin glue for reconstruction of the oviduct in animals and reported satisfactory patency rates [ 2,.5,7 I. An important consideration is the morphological result after reanastomosis. Though the microscopic chtnges after surgical reconstruction are well documented [ 1 I, little is known on light and electron microscopic alterations in the fallopian tube after the use of fibrin glue. We therefore performed experimental reanastomosis of rabbit oviducts with fibrin glue and investigated tissue reactions by means of light and scanning electron microscopy.
Materials and methods Keywords: phology;
Rabbit oviduct; Histology; Mor-
Anastomosis;
Fibrin glue.
Introduction Microsurgical reanastomosis of the fallopian tube has gained increased importance in the last few years. The postoperative results depend on the morphology of the fallopian 0020-7292/84/$03.00 0 1984 International Federation of Gynaecology & Obstetrics Published and Printed in Ireland
The study was performed on 16 silver rabbits. After midline laparotomy the concomitant vessels of one oviduct were coagulated using bipolar microsurgical forceps and an operation microscope. After sharp transsection of the oviduct 1 cm distally from the tubo-cornual junction a nylon splint was inserted into both parts of the tube. The sectional planes were approximated and 1 Inr J Glwaecol Obstet 22
296
Spernol et al.
drop of the fibrinogen glue was applied by means of a mixing syringe. The 2 ends of the oviduct were kept in tight contact until drying of the glue after 2-3 min. The contralateral tube was occluded by means of tantal hemoclips. The splint remained in place for up to 12 h. Twenty rabbits could be covered 3 weeks after the operation, neither gonadotropins nor insemination were used. Six rabbits became pregnant, 2 of them twice, and delivered. Three months after the operation the oviducts were removed and the site of the anastomosis placed in formaldehyde or glutaraldehyde, respectively. For histological examination paraffin sections were prepared and stained with hematoxylin-eosin (HE) and ironhematoxylin, fuchsin, phosphormolybdenorange and light green according to Goldner. The samples for scanning electron microscopy were cut longitudinally and vaporized with gold.
Results Light microscopy
Figures l-3 show light microscopic findings 3 months after the anastomosis. There were stout bands and fibrosis in the muscular layer. The mucosal folds were slightly flattened with partly uneven scarring. In the mesosalpinx foreign body giant cells were found (Fig. 3). Intraluminal fibrin deposits were not seen, inflammatory reactions in the mucosa or in the wall of the tube were minimal. Electronmicroscopy
Scanning electron microscopy revealed interrupted and lowered folds of the mucosa. Some detritus was seen near the site of the anastomosis. Though some undifferentiated peg cells were observed in proximity of the anastomosis the number of cilia was only slightly reduced and the cilia themselves appeared normal (Figs. 4-6).
Fig. 1. Rabbit oviduct 3 months after reanastomosis with fibrin glue, GoldneMaining, magnification 25 : 1. he gular folds of the mucosa, dispersed glands and fibrotic muscular layer at the site of the anastomosis. Int J Gynaecol Obstet 22
Reanustomosis with fibrin glue
Fig. 2. HE staining, magnification anastomosis.
Fig. 3.
63 : 1. Moderate lymphocytic
infiltration
297
and stout bands of muscularis 2It the site of the
HE staining, magnification 100: 1. Foreign body giant cells in the rabbit mesosalpinx after reanastomosis with flkin glue. Int J Gynaecol Obstet 22
298
Spernol et al.
Fig. 4. Scanning electron microscopy: magnification 150 : 1. (a) Normal mucosa of the rabbit oviduct; (b) site of the anastomosis 3 months after reanastomosis with fibrin glue: interrupted and slightly lowered mucosal folds with some intraluminal detritus.
Discussion The good patency and pregnancy rates which we and others observed after experimental reanastomosis of the rabbit oviduct are encouraging [6], Morphologic studies of the oviduct permit evaluation of local tissue reaction and seem to explain the good ZntJ Gynaecol Obstet 22
clinical results. We were able to show that 3 months after the operation very little morphologic changes persisted at the site of reanastomosis. One of the most important factors influencing ovum transport are the cilia. While Bernhard-Huth et al. [ 11 reported that cilia were absent over sutures on scanning electron
Reanastomosis with fibrin glue
299
Fig. 5. Scanning electron microscopy: magnification 4800 : 1. (a) Normal mucosa of the rabbit oviduct; (b) site of the anastomosis: undifferentiated cells and ciliated cells, which are reduced in number.
microscopy, we did not observe an area near the anastomosis where cilia were absent though their number seemed to be reduced (Fig. 5). Both light and electron microscopy showed that the area of morphologic changes around the anastomosis was extremely small, an important point in view of the fact that the epithelial lining of the fallopian tube is critical for adequate ovum transport.
A possible source of complications could be the persistence of fibrin in or around the oviduct. We did not see any fibrin clots attached to the site of anastomosis, nor did we observe intraluminal fibrin on histologic examination. Scheidel et al. [7] saw intraluminal fibrin clots up to 7 weeks after the operation. The fibrinolytic activity of the fallopian tube has not yet been adequately Int J Gynaecol Obstet 22
300
Spew01 et al.
Fig. 6. Scanning electron microscopy: Magnification 14000 : 1. (a) Normal mucosa of the rabbit oviduct; (b) site of the anastomosis: the ciliated cells are reduced in number but the cilia appear normal.
determined; unpublished results suggest that the oviduct has a high fibrinolytic activity, possibly explaining the complete absorption of fibrin from the site of operation. In summary tissue reaction after use of fibrin glue is comparable to the changes seen after the use of microsutures. It appears Int J Gynaecol Obstet 22
possible that by changing the composition of the fibrin glue a further reduction in tissue reaction can be achieved. Though the numbers in our study are too small to allow definite conclusions, the good healing and minimal tissue reaction after the use of fibrin glue appears encouraging.
Reanastomosis with fibrin glue
Acknowledgment We are grateful to H. Hbrantner, Institute of Micromorphology and Electron Microscopy (Head: Prof. Dr. L. Stockinger), University of Vienna, Medical School, for the electron microscopic evaluations. References Bernhardt-Huth D, Frantzen Ch, Schlisser HW: Morphology of rabbit oviduct after microsurgical techniques for reanastomosis of the isthmus or ampulla. Arch Gynecol230: 251, 1981. lntraphuvasak J, Stofft E, Osada H, Friedberg V: Tuben End-zu-End-Anastomose mit Fibrinkleber. Abstrakt 17, 44. Tagung der Deutschen Gesellschaft fiir Gynlkologie und Geburtshilfe, Miinchen 13.9-17.9.1982. Kudema H: Klinische Anwendung der Klebung von Nervenanastomosen mit Fibrinogen Fortschr KieferGesichtschir 21: 135, 1976. Pfliiger H, Lunglmayr G, Breitenecker G: Vaso-Vasoanastomose unter Verwendung von Frbrinogenkonzentraten: Versuche an Kaninchen. Wien Klin Wochenschr 88; 800,1976.
301
Osada H, Noda M, Fujli TK, Tsubata K, Takagi S: Basic studies on the use of a fibrinogen-adhesive system during fallopian tubal transplants. Abstract 396, XIth World Congress on Fertility and Sterility, June 26-July 1, 1983, Dublin, Ireland. Riss P, Spernol R, Beck A, Schindler K: Reanastomose der Kaninchentube mit Fibrinogenkonzentrat. Wien Klln Wochenschr 00: 000,1984. Scheidel PH, Wallwiener DR, Wiedemann RA, HEPP HK: Experimental anastomosis of the rabbit fallopian tube using fibrin glue. Fertil Steril38: 471,1982. Winston RML: Microsurgery of the fallopian tube: from fantasy to reality. Fertil Steril34: 521, 1980. Wolner E: Die Fibrinklebung in der Chirurgie, Wien Klin Wochenschr 94: 181,1982.
Address for reprints: Dr. R. Spemol LKH Klagenfurt St. Veiterstrapse 47 A-9020 Klagenfurt Austria
int J Gynaecol Obstet 22