Study On Effects Of The Spermatogenesis Of NOA Model Treated With Testicular Sustained-release FSH And SCF

ObjectiveTo investigate the effect FSH and SCF testicular sustained-release on spermatogenesis of NOA model of Wistar rat.Materials and methods1. Determine the concentration of FSH/SCF in normal Wistar rat testis and the dosage of FSH/SCF(1)6rats were gained randomly, bilateral testis were exsected, peeled albuginea,polished into tissue homogenate and centrifuged, then the concentration of FSH and SCF of the liquid supernatant was tested with chemiluminescence and enzyme-linked immunosorbent assay, respectively.(2) The recipe of the sustained-release was estimated according to the testicular volume, FSH/SCF concentration in the testis and the results of the preliminary work. The sustained-release was made by the multiple emulsion-solvent evaporation. The using dosage of FSH/SCF microspheres attained the5/2times concentration of the normal tissue was worked out according to the release in vitro and the drug-loading capacity.36rats were taken randomly and divided into two groups:the drug group (18rats) and blank group (18rats). One lateral testis was injected with FSH microspheres and other lateral with SCF microspheres in the drug group. Bilateral testis of the blank group was injected with commensuration of the blank microspheres. Testises of6rats of two groups were resected after injection3-day,10-day and15-day, respectively. The injection site was resected firstly for evaluating the lesion by HE stain slice. The remained testis peeled the albuginea was cut into small chop, digested with protease, polished into tissue homogenate, then centrifuged, FSH/SCF of the liquid supernate was tested with chemiluminescence and enzyme-linked immunosorbent assay, respectively. The drug concentration-time curves was drawed.2. Assessment of effect FSH/SCF sustained-release on spermatogenesis of NO A The NOA model of Wistar rat was made by using busulfan15mg-kg"1single intraperitoneal injection.90rats were injected. Sampling randomly6rat of them were undergone the testis biopsy for evaluating whether the model was success or not. After the model success was affirmed,36rats were divided into3groups (12rats in each), randomly, namely A, B and C group. The superior and inferior polar of testis of A group was injected underneath of albuginea with FSH/SCF microspheres, respectively. The dosage of FSH/SCF was5times/2times of concentration of the normal. Repeat injection interval15days was3with equal dose. B group was injected in muscle of thigh with equal dose as A group FSH/SCF microspheres. Repeat injection interval15days was3with equal dose,also. The testis of C group were injected with commensuration of the blank microsphere, the dosage and time of given the drug was similar to A group. After using drug19days,38days and57days, HE stain slice of the testis and epididymis of every group was made, the number of the spermatogonia, spermatocytes, spermatids and sperm in every vertical section of contorted25compliant senimiferous tubules of each group was counted, and the number of sperm in every vertical section of the epididymis tubules of each group was counted as well.The results was analyzed with SPSS17.0, the statistically significant differences was provided when P<0.01.Results1. The testicular FSH and SCF of Wistar rat was0.31±0.06mIU/gå'Œ0.02±0.003pg/g. The testicular volume was1.72±0.34ml. The recipe of FSH sustained-release:PVA1gplus100ml water, PLGA2g plus8ml CH2C12and FSH70IU. The FSH microspheres loading capacity was4IU/mg. The recipe of SFC sustained-release:SCF20ng, other ingredient was ditto. The release duration of the new FSH/SCF microspheres was15days approximately of the release in vitro. Average daily release quantity of FSH/SCF microspheres26mIU/g and0.14pg/g, respectively. The dosage of FSH/SCF with5times/2times of concentration of the normal, release sustainedly for15days, was lmg FSH microspheres and1.6mg SCF microspheres, respectively. After injection lmg FSH and1.6mg SCF in3,10and15days, the concentration of FSH/SCF in the testis was1.21±0.03mIU/g/0.05±0.008pg/g,1.17±0.52mIU/g/0.05±0.003pg/g and0.84±0.27mIU/g/0.03±0.007pg/g, respectively. These results showed effect on sustained-release of FSH/SCF. Some extent of inflammatory cell infiltration and mild fibrillation in inject site could be observed.2. A group of testicular spermatogenic function recovery was better than B and C group, B group was better than C group. After fifty-seventh days, the spermatogonia, spermatocytes, spermatids and spermatozoa of A group was73.51±21.73,75.75±19.45and109.47±38.57, B group was45.81±16.03,41.76±21.43and37.27±16.69, and C group was35.37±9.64,17.73±16.58and25.28±10.68, A group was compared with B group, P<0.01, A and B group was compared with C group, P<0.01. The sperm counting of epididymal of A, B and C group was51.09±75.94,14.73±29.01,5.57±17.13, respectively. Pairwise comparison, P<0.01.ConclusionOur study suggested that testicular injection and intramuscular injection of FSH and SCF microsphere might promote to spermatogenesis recovery of NOA model and testicular injection seem better than intramuscular injection. A further study was worthy.