Morphology and survival of cryopreserved-thawed ovarian tissues after heterotopic autotransplantation

Objective： The aim of our study was to observe the survival and morphological changes of thawed ovarian tis- sues after heterotopic transplantation. Methods： Twenty SPF-SD female rats （5-6 weeks old） were equally randomized into the control group and experimental group. In control group, the freshly isolated ovaries were fixed in formalin. In experimental group, the freshly isolated ovaries were vitrified immediately and cut into thin slices. After stored in liquid nitrogen for 21 days, the tissues of experimental group were rapidly thawed and transplanted into back muscles of rats for 2 or 4 weeks, respectively. After that, all rats in experimental group were sacrificed and the ovarian tissues were collected and fixed in 4% formaldehyde solution. Then the ovarian tissues were stained with HE and observed under the light confocal microscope. Re- suits： With the naked eyes, there was no specific alteration except the size reduction with color changing. Under microscopy, we found normal cortex and medulla in the ovary, and the primordial follicles and follicles in various stages were observed in the cortex. The normal oocytes in ovarian tissues of experimental group were significant decreased than in the control group. Conclusion： The ovarian tissues survive well in experimental group and there is no significant difference in the proportion of follicles between different times （2 and 4 weeks） after grafting. Our results suggest that thawed ovarian tissues could survive after heterotopic transplantation into back muscles of rat models and maintain their morphology and function.

Effects of heterotopic autotransplantation of cryopreserved-thawed ovarian tissues on sex hormone level

Objective: The aim of our study was to measure and compare the serum hormone level of transplant group with blank control and castrated control groups after heterotopic autotransplantation of cryopreserved-thawed ovarian tissues into back muscles. Methods: A total of 40 SPF-SD female rats(5–6 week-old) were randomly divided into three groups: blank control group(group A), castration control group(group B) and transplant group(group C). Ovaries were removed by surgical procedure, then after cryopreservation and thawing procedures the ovarian tissues were implanted into the back muscles of mice in group C. After 4 weeks of ovarian tissues transplantation, all rats blood sampling were measured for E2, LH and FSH hormone levels by ELISA. Results: E2 level was significantly higher in group C and group A than group B [(38.98 ± 5.66) pg/mL,(8.14 ± 3.24) pg/mL; P < 0.05) and [(36.30 ± 6.90) pg/mL,(8.14 ± 3.24) pg/mL; P < 0.05)]. However, E2 level in group C and group A had no significant difference. FSH level in group B, group A and group C was(18.87 ± 2.54) nmol/L,(7.77 ± 0.87) nmol/L and(9.39 ± 2.12) nmol/L respectively. FSH level increased significantly in group B compared with group A, and the difference had statistical significance(P < 0.05). FSH level was slightly increased in group C compared with group A, and the difference was not statistically significant(P > 0.05), but compared with group B, FSH level was significantly reduced and being statistically significant(P < 0.05). Conclusion: Autotransplantation of cryopreserved-thawed ovarian tissue into back muscles can sustain follicular development and re-establish endogenous hormone production by restoring the factors such as angiogenesis and innervations at the graft site.

Effect of acoustic pulses and EHF radiation on multipotent marrow stromal cells in tissue engineering constructs

In this paper,we studied the effects of physical factors,such as,acoustic pulses of laser-induced hydrodynamics(ALIH)and extremely-high frequencies(EHF)radiation,on the formation of heterotopic bone marrow organs.A suspension of precipitated bone marrow cells from CBA mice were exposed to ALIH pulses and EHF radiation separately and in their com bination tissue engineering constructs,presenting gelatin sponges 2 by 2 by 2 mm in size containing 10^(7)nucleated bone marrow cells,were exposed to physical factors and were implanted under the renal capsules of syngeneic mice.The newly formed hematopoietic organs were examined in three and five months later after treatment.The five months old transplants were bigger in size than the three months old transplants.The number of hematopoietic cells in the rest of the groups increased during this period by a factor from 3 to 10,the increase being as high as 20-fold in the:ALH+EHF group.Maximal concentration of multipotent stromal cells(MSCs)was in the EHF+ALIH,and minimal concentration was in the ALIH+EHF.The accumulation rate of bone capsule weight was highest for the transplants of EHF+ALIH and ALIH-sponge groups during the first three months.These data showed that the combined impact of the EHF+ALIH on MSCs is the most effective for the formation of bone marrow transplantation.

An experimental study on autologous transplantation of fresh ovarian tissue in sheep

Objective:To investigate current autologous transplantation methods and sites of ovarian tissues in sheep.Methods:Sheep ovaries were resected.Ovarian cortices were sliced and transplanted orthotopically into the ovarian mesangial latum and heterotopically into the greater omentum and under groin skin.The grafts were removed two months after transplantation and examined to evaluate the survival of follicles(hematoxylin-eosin staining)and help determining feasible graft sites and transplantation methods.Results:Graft nodules were found in the transplanted sites.HE staining of the grafts showed that multiple primordial follicles were able to survive in the grafts on both sides of the ovarian mesangial latum,the right side of the greater omentum,and the left inguinal subcutaneous tissue.Secondary or cystic follicles were found in almost all of the grafts.Conclusion:The ovarian mesangial latum,the greater omentum and the inguinal subcutaneous tissue can be used as autologous transplantation sites,where sheep ovarian tissue can survive and the follicles grow and develop in good condition.

Rand reconstruction using heterotopic replantation of amputated index and little fingers

In cases of severe segmental injury across the hand and wrist, but one or other fingers are still in peak condition, the fingers can be selected for replantation at the forearm bones to restore pinch function. Here we reported an unusual case with a severe crush-avulsion amputated injury to the right hand caused by a machine accident. We conducted hand reconstruction using heterotopic replantation of the amputated index and little fingers.During 19 months follow-up, the bone union healed well with satisfactory outcome. The interphalangeal and metacarpophalangeal joint of the fingers after the heterotopic replantation had a good holding activity. This is a worthwhile procedure and the patient is satisfied with the result. The major disadvantage of this method is the poor appearance of the reconstructed fingers.