| Part one Experimental study of IVD sterilization in male dogsObjectiveTaken the male Beagle dog as experimental animal, to detect sperm density, concentration of glycosidase, copper ions and zinc ions and histological morphology changes of vas deferens, epididymis and testis, to judge the contraceptive efficacy, safety and reverilibity of this IVD, provding basis for the animal and clincial experiments of next step.Material and methodsAccording to stochastic, comparable principles, 20 healthy, adult and male Beagle dogs were randomly allocated to four sterilization group-sham operation group, vasectomy group, IVD group and reverilibity group( n=5 in each group). Semen samples(4-8ml) were obtained from each dog by using hand-stimulating method before operation and then were tested by Computer-aided analysis system to calculate its sperm density and living rate. Sham operation, IVD implantation operation and vasectomy were conducted to four groups, then semen semples were collected by hand-stimulating method in postoperation 1, 3, 6, 12 month and all semen samples were tested by CASA. Seminal plasma was frozen in -20℃for the detection ofа-glucosidase, copper ions and zinc ions. In 12th postoperative month, the tissues of testis, epididymis and vas deferens of sham-operation group, vasectomy group and IVD group were obtained and slides were made to be observed in optical and electron microscope. Apoptosis in testis, epididymis and vas deferens was detected by TUNEL method. In 12th postoperative month, IVDs in dogs of reverilibity group were taken out and in postoperation 1 month, semen samples were collected by hand-stimulation method. The parameters of semen samples were tested by CASA examination.ResultThe quantity and quality of seminal fluid of each group collected in preoperation is normal. Surgery processes is smooth. After operation, all dogs do not have complications such as pain, infection. In postoperation 1, 3, 6 and 12 month, dogs in IVD group and vasectomy group were all azoospermia and the rate of sucessful contraception was 100%. The parameters of semen samples of sham-operation group were not great different from those in preoperation. The operation to take out IVD of reverilibity group was smooth and the semen parameters of each dog become normal within post-reverilibity 1 month. The concentration of glycosidase, copper ions and zinc ions of sham-operation group, IVD group, reverilibity group were similar to those detected in preoperation and there were no significant statistics difference between preoperation and postoperation; But the statistics difference of glycosidase, copper ions and zinc ions of vasectomy group in postoperation was significant compared with preoperation. In observation of optical and electron microscope, the tissues of testis, epididymis and vas deferens of sham-operation group, IVD group were damaged more slighter than vasectomy group. Spermatogenic cells of seminiferous tubules in testis of sham-operation group, IVD group were normal and arranged regularly; Epididmal duct and its lumen were not expanded; the vas deferens was expanded slightly or not at all. The Spermatogenic cells in vasectomy group were arranged irregularly; quantity of cells were reduced; The mesenchymal was loose. Moreover, the Apoptosis cells in sham-operatiom and vasectomy group were also less serious than vasectomy group.ConclusionsThe experiment showed the rate of contraceptive success was 100% and was equal between IVD group and vasectomy group. The Filtering Nano-copper Complex/polymer Composites Intra-Vas Device have good biocompatibility and safety and do not result in adverse impact on partial and whole body tissues or organs. The results of reverilibity operation indicate it can achieve easily the patency of vas deferens after IVD was taken out. Moreover, operative procedure is simple and there are no postoperative complications. Part two Experimental study of IVD sterilization in male rabbitsObjectiveTaken New Zealand Big Eared White Rabbit as experimental animal, to judge IVD's efficacy, safety and reversibility by observing the result of the breeding experiments of male and female rabbits and histological morpology changes of testis, epididymis and vas deferens, therefore providing theoretical basis for animal and clinical experiment of next step.Material and methodsAccording to the stochastic, comparable principles, 40 healthy, adult, male New Zealand Big Eared White Rabbit were randomly allocated to four groups: sham- operation group, vasectomy group, IVD group and reverilibity group( n=10 in each group). Seminal fluid were obtained from each rabbit by artificial-vagina method before operation and then were observed in optical microscope. Meanwhile, 40 healthy, adult and female New Zealand Big Eared White Rabbit were aslo randomly allocated to four groups: sham-operation group, vasectomy group, IVD group and reveribility group( n=10 in each group), which were prepared for mating experiment. Sham operation, vasectomy and IVD implantation were conducted to sham-operation group, vasectomy group and IVD group. After operation, the breeding experiment between male and female New Zealand Big Eared White Rabbit was conducted and the sucessful rate of contraception was observed and recorded. In postoperation 12 month, the IVD of reverilibity group had been taken out from lumen of vas deferens and breeding experiment with ten female New Zealand Big Eared White Rabbit was performed again and the pregnant rate of female rabbits was recorded. In postoperation 12 month, the tissues of testis, epididymis and vas deferens of sham-operation group, vasectomy group and IVD group were collected and then detected in optical microscope to observe histological morphology changes. Apoptosis of testis, epididymis and vas deferens in sham-operation group, vasectomy group and IVD group was conducted by TUNEL method.ResultBefore operation, semen samples were obtained by artificial-vagina method, The density and living rate of sperm of each male rabbits was normal. The processes of IVD-inplantation and vasectomy were smooth. After breeding experiment, all female rabbits in IVD, reverilibity and vasectomy group were not pregnant and contraception rate of them was zero. In contrast, 4 female rabbits in sham-operation group were pregnant and successful rate was 80%. The operation processes to take out IVD in male rabbit of reverilibity group were smooth and there were no postoperative complications such as hematoma, wound infection. The postoperative male rabbits were mated with female rabbits again. After 15 days, 3 female rabbits were conceived and the pregnant rate of female rabbits were 60%. Histological morphology examination of testis, epididymis and vas deferens under optical microscope showed sham-operation group and IVD group had suffered much less impairment than vasectomy group. The testis was normal, its spermatogenic cell of all level was ranged neatly on wall of siminiferous tubule; The tubule of epimidmis and vas deferens were expanded slightly. The tissues in vasectomy group were demaged seriously, the quantity of spermatogenic cell was reduced and arranged irregularly; The tubule of epimidmis and vas deferens were expanded obviously; Cilia was fell off. Apoptosis in sham-operation group and IVD group was aslo much slighter than vasectomy group. ConclusionsThe Filtering Nano-copper Complex/polymer Composites IVD has significantly contraceptive effect within postoperation 1 year and the sterilzation efficacy equal to vasectomy, the sucessful contraception rate of them all achieved 100%. Moreover, the operation can be easily grasped and the hemorrage of wound is less. Postoperative complications of IVD and reverilibity group are happened fewer than vasectomy group. This IVD aslo have showed good security and biocompatibility. The histological morphology of testis, epididymis and vas deferens of rabbits in IVD group have comfirmed this IVD has not adverse effect on partial and whole body organization. Moreover, process of reverilibity operation is a simple, less trauma with good patency rate of vas lumen.
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