The Therapeutic Potential of Amniotic Fluid-Derived Stem Cells on Busulfan-Induced Azoospermia in Adult Rats

BACKGROUND@#Busulfan is an alkylating chemotherapeutic agent that is routinely prescribed for leukemic patients to induce myelo-ablation. However, it also results in azoospermia and infertility in cancer survivors. This research was constructed to explore the possible therapeutic role of amniotic fluid-derived stem cells (AFSCs) in improving busulfaninduced azoospermia in adult rats. @*METHODS@#Forty two adult male albino rats were randomized into: (1) control group, (2) azoospermia group, (3) spontaneous recovery group, and (4) AFSCs-treated group, in which AFSCs were transplanted through their injection into the testicular efferent ducts. The assessment included a histo-pathological examination of the seminiferous tubules by the light and transmission electron microscopes. Additionally, the confocal laser scanning microscope was used for confirmation of homing of the implanted cells. Moreover, we conducted an immuno-fluorescence study for detection of the proliferating cell nuclear antigen (PCNA) in the spermatogenic cells, epididymal sperm count, and a histo-morphometric study. @*RESULTS@#AFSCs successfully homed over the basement membrane of the injured seminiferous tubules. They greatly attenuated busulfan-induced degenerative and oxidative changes. They also caused a re-expression of PCNA in the germ cells, leading to resumption of spermatogenesis and re-appearance of spermatozoa. @*CONCLUSION@#AFSCs could be a promising treatment modality for male infertility induced by chemotherapy, as they possess prominent regenerative, anti-apoptotic, and anti-inflammatory potentials.

The Therapeutic Potential of Amniotic Fluid-Derived Stem Cells on Busulfan-Induced Azoospermia in Adult Rats

BACKGROUND@#Busulfan is an alkylating chemotherapeutic agent that is routinely prescribed for leukemic patients to induce myelo-ablation. However, it also results in azoospermia and infertility in cancer survivors. This research was constructed to explore the possible therapeutic role of amniotic fluid-derived stem cells (AFSCs) in improving busulfaninduced azoospermia in adult rats. @*METHODS@#Forty two adult male albino rats were randomized into: (1) control group, (2) azoospermia group, (3) spontaneous recovery group, and (4) AFSCs-treated group, in which AFSCs were transplanted through their injection into the testicular efferent ducts. The assessment included a histo-pathological examination of the seminiferous tubules by the light and transmission electron microscopes. Additionally, the confocal laser scanning microscope was used for confirmation of homing of the implanted cells. Moreover, we conducted an immuno-fluorescence study for detection of the proliferating cell nuclear antigen (PCNA) in the spermatogenic cells, epididymal sperm count, and a histo-morphometric study. @*RESULTS@#AFSCs successfully homed over the basement membrane of the injured seminiferous tubules. They greatly attenuated busulfan-induced degenerative and oxidative changes. They also caused a re-expression of PCNA in the germ cells, leading to resumption of spermatogenesis and re-appearance of spermatozoa. @*CONCLUSION@#AFSCs could be a promising treatment modality for male infertility induced by chemotherapy, as they possess prominent regenerative, anti-apoptotic, and anti-inflammatory potentials.

Histological study of the effect of amiodarone on the follicular cells of the thyroid gland of male albino rats

Amiodarone is a potent antiarrhythmic drug used for treatment of many types of cardiac arrhythmias. Its structural formula contains a high dose of iodine with a considerable potential to cause thyroid gland dysfunction. The aim of the present work was to study the possible histological changes in the follicular cells of the thyroid gland in male albino rats after long-term administration of amiodarone. Reversibility of these changes was also investigated. The study was confirmed by hormonal assay, to evaluate the thyroid gland endocrine function. The present study was carried out on 40 adult male albino rats, which were divided into 3 study groups; group I, considered as control, group II [the amiodarone- treated group] received amiodarone orally in a daily dose of 30 mg/kg for 12weeks and group III [the withdrawal group] that received the same dose of amiodarone for 12 weeks and were sacrificed 6 weeks after withdrawal of the drug. Rats were sacrificed after ether anesthesia and blood samples were subjected to hormonal assay of TSH, T[4] and T[3] levels. The thyroid gland tissue samples were removed and prepared for light microscopic examination of H and E stain and toluidine blue- stained semithin sections. Electron microscopic examination was also done. Histological examination of thyroid gland in amiodarone treated- rats [group II] revealed different light microscopic and ultrastructural changes. Some follicular cells showed signs of degeneration in the form of irregular dilation of the rough endoplasmic reticulum, vacuolation of the cytoplasm and increased number of lysosomes with irregular nuclei. These changes might be due to direct cytotoxic effect of the high iodine-containing amiodarone on the follicular cells. Other follicles showed signs of hyperactivity manifested by vacuolated colloid with scalloped appearance of its edges and papillary projections of the follicular cells with epithelial stratification. The manifestations of hyperactivity appeared to be compensatory to the direct effect of the drug. Alterations of thyroid function in group II, was further confirmed by results of hormonal assay, which revealed significant increase in serum levels of both TSH and T[3] with significant decrease in the level of T[3]. In group HI, after withdrawal of the drug, many follicles exhibited histological reversibility to almost normal pattern. Manifestations of sustained activity were still depicted in few follicles with focal areas of stratification and dilation of the rough endoplasmic reticulum. Hormonal assay showed that TSH and T[3] returned to nearly normal values while T[4] remained higher than the control level. From the present study, it could be concluded that prolonged amiodarone administration caused manifest histological and biochemical changes in the follicular cells of the thyroid gland. The obtained structural and hormonal changes were almost reversible after stoppage of the drug administration. Baseline and repeated levels of thyroid hormones should be monitored in every patient on amiodarone therapy

Protective role of antioxidant vitamins [C and E] on DI-2-ethylhexyl phthalate induced testicular toxicity in rats. histological and ultrastructural studies

Di-2-ethylhexyl phthalate [DEHP] is one of the most common synthetic environmental pollutants to which nearly everyone is exposed. It is used as a plasticizer that imparts flexibility, durability and softness to rigid polyvinyl chloride products. The aim of the present work was to study histologically the effects of DEHP on the testis and the role of antioxidant vitamins in prevention of such effects. The present study was carried out on 55 adult male albino rats. The animals were divided into three main groups. Group I served as control group. Group II, received oral daily dose of] gm/kg b.w. of DEHP for 4 weeks. Group III was further subdivided into three subgroups; IIIa, IIIb, IIIc. They received the same dose of DEHP for the same period simultaneously with vitamin C [100 mg/kg b.w.], vitamin E [20 mg/kg b.w.] and vitamin C and E respectively. At the end of the experiments, specimens were taken from the testes of all animals and were subjected to histological and ultrastructural studies Animals of group Ii revealed moderate degenerative changes in most of the somniferous tubules of the testis. Few tubules showed severe atrophy of the germinal epithelium. These tubules appeared lined only with spermatogonia and Sertoli cells. There was a decrease in the number of spermatozoa. Electron microscopic examination revealed considerable vacuolation of the cytoplasm of Sertoli cells. They exhibited dilatation of the smooth endoplasmic reticulum and bizzar shaped dense mitochondria. Degenerative changes were also seen in Leydig cells. Simultaneous administration of antioxidant vitamins with DEHP [group Hi] ameliorated the toxic effects of DEHP on the somniferous tubules of rats. This protective effect was more evident in animals that received both vitamins C and E together with DEHP. The use of vitamin E provided better protection than vitamin C. Conclusion: It could be concluded that DEHP has an injurious effect on the testis of the exposed animals. This effect could be prevented by regular use of antioxidant vitamins

Histological and biochemical study on the toxic effects of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin [TCDD] on the follicular cells of thyroid gland of rats

2,3,7,8, TCDD and related substances are ubiquitous environmental pollutants causing a wide variety of pathological alterations. They are capable of altering endocrine homeostasis. This work was carried out to study the histological and biochemical changes of acute and chronic administration of TCDD on the follicular cells of thyroid gland of rats. Thirty six male Sprague-Dawley rats were used in this study. They were divided into three groups. Group I was given corn oil and served as a control group. Group II was given 10 micro g/Kg TCDD as an acute single oral dose. Group III was further subdivided into subgroup IIIa and subgroup IIIb that were given 0.125 micro g /Kg/ day TCDD for 4 weeks and 16 weeks respectively. At the end of the experiment, animals were sacrificed and thyroid glands were removed and examined by light and electron microscope [EM]. Biochemical estimation of serum thyroxin [T[4]], triiodothyronine [T[3]] and thyroid stimulating hormone [TSH] were also done to all animals. Histologically, Thyroid gland of group II showed beginning of follicular hypertrophy and hyperplasia in some follicles. These follicles were lined by high cuboidal epithelium with appearance of follicles lined by more than one layer of cells. Dilated rough endoplasmic reticulum [rER], well developed Golgi apparatus and numerous lysosomes were evident. Microvilli of some follicular cells were short and blunted. Subgroup IIIa and IIIb showed signs of hyperactivity affecting most of the follicles. These changes were more pronounced in animals of subgroup IIIb that received TCDD for 16 weeks. Most of the follicles were small and irregular. Many of them were lined by tall columnar epithelium with appearance of papillary projections in their lumens that appeared empty or with sparse scalloped colloid. EM examination revealed marked dilatation of rough endoplasmic reticulum and prominent well developed Golgi complexes with accumulation of numerous lysosomes and membrane bound colloid droplets. Some follicles of this group revealed degenerative changes due to direct toxic effect of TCDD. These histological changes were accompanied by a significant decrease in serum level of T[4] and a significant increase in TSH level. These biochemical changes were more pronounced in animals that received TCDD for 16 weeks. TCDD had deleterious effect on thyroid gland which is time and dose dependent

harmful effect of chronic hypervitaminosis D[3] on the skin and cardiac muscle of albino rat. A histological study

Is to investigate the effect of hypervitaminosis D[3] on the skin and cardiac muscle of albino rats. Eighteen adult male albino rats were used in this study. Rats were divided into two main groups. Group I [6 rats] served as a control group. Group II. [12 rats] to which vitamin D[3] were administered in a daily dose of 50,000 IU/kg body weight and subdivided into two subgroups. [Subgroup IIa] 6 rats received the preparation for two weeks and [Subgroup IIb]: 6 rats received the preparation for eight weeks. At the end of experimental periods, specimens were taken from the skin covering the back and the left ventricles of the heart of each animal. Animals of subgroup IIa showed epidermal affection as focal areas of decreased number of epidermal layers except for thickened stratum corneum, widening of the intercellular spaces between keratinocytes, loosened desmosomal junctions and microvillous transformation of the plasma membrane. These changes were marked and diffuse in animals of subgroup IIb. Layers beneath the stratum corneum were only one or two layers. Keratinocytes showed affection of the nuclei. The cytoplasm showed increased cytokeratin filaments and keratohyaline granules. Stratum corneum appeared thickened with wide spaces in between its layers and failure to form intact wavy bundles. Increased deposition of collagen bundles were noticeable in the dermis of most animals of this subgroup. The cardiac muscle of subgroup IIa showed damage of muscle fibers with widening of the spaces in between and interstitial odema, Breaking of continuity of myofibrils at the site of I band, partial degradation of Z-line material in many sarcomeres, thinning of myofibrils and mitochondrial swelling. The sarcolemma appeared lifted away from the underlying sarcomeres. Rats of subgroup IIb presented aggravation of these changes with severe cardiac muscle damage. Some cardiac myocytes appeared thinned with dark shrunken nuclei, others revealed dissolution with either karyolytic or pyknotic nuclei. Dehiscence of the intercalated disc with widened disc space, contraction bands with subsequent shrinkage of the sarcomere were also observed. Many myofibrils were disorganized, distorted and fragmented. The use of excessive doses of vitamin D resulted in severe damage to the skin and cardiac muscle. The use of vitamins should be restricted for individuals with documented deficiency or those at risk, especially fat-soluble vitamins because of their cummulative effect in the body

Protective trials against aflatoxin B[1] induced toxicity in albino rat hepatocytes. Histological and histochemical study

Is to investigate the effect of aflatoxin B[1] on hepatocytes and the possible protective effect of administration of selenium and high protein diet. The stud was carried out on 32 albino rats divided into 4 equal groups; a control group, a group receiving aflatoxin B[1] a group receiving selenium with aflatoxin and a fourth group receiving high protein diet with aflatoxin. Specimens were taken from the liver of all groups and subjected to histological, histochemical and ultrastructural studies. Animals receiving aflatoxin B[1] showed periportal hepatic degeneration in the form of ballooning of cells and vacuolation of the cytoplasm. Some cells revealed hepaloceliular necrosis. Portal tract showed bile duct proliferation. Fibrosis was occasionally seen in the portal tract. Ultrastructural results revealed irregularities in the nuclei with dilatation of the perinuclear cisternea. Some nuclei showed separation of the fibrillar and granular component of their nucleoli. The cytoplasm showed proliferation and dilatation of smooth and rough endoplasmic reticulum, degenerated mitochondria, decreased number of ribosomes and glycogen granules with the increase of lysosomes and appearance of fat droplets. Evident decrease in the DNA and RNA content were seen by methyl green pyronin stain. Administration of selenium showed evident improvement in the cellular structure and liver architecture. The electron microscopic study showed nearly normal nuclei and nucleoli. The cytoplasm showed more or less normal mitochondria, moderate number of ribosomes and glycogen granules Administration of high protein diet revealed moderate improvement. Administration of selenium antagonizes the deleterious effect induced by aflatoxin B[1] while high protein diet has less protective effect on the toxicity, of aflatoxin on the liver

Pulmonary changes that occur in the interalveolar septa of albino rats exposed to nickel and the possible protective effect of magnesium. A histological study

Is to investigate the effect of exposure to nickel chloride on the interalveolar septa of lung of albino rats and the possible protective effect of simultaneous administration of magnesium acetate. Thirty adult male albino rats were used in this study. They were divided into 3 groups. Group I included 6 rats. They were used as normal controls. Group II, included 12 rats that were exposed to nickel chloride 5 days/ week for 4 weeks. Group III, included 12 rats that were given magnesium acetate simultaneously with nickel chloride for 4 weeks. Specimens were taken from the lung of all animals and were subjected to histological and ultrastructural studies. Rats lung of group II revealed evident degenerative pulmonary changes in the interalveolar septa of the lung with its marked thickening and collapse of many alveoli. The interstitium showed marked cellular infiltration, focal areas of oedema and deposition of collagen fibers. Some blood vessels appeared congested with marked extravasation of blood. Focal areas showed homogeneous eosinophilic material within the alveolar lumen and in the interalveolar septa. Electron microscopic study revealed alteration of normal alveolar lining with predominance of pneumocyte type II which showed atypical vacuolar inclusion in some cells. Others appeared shrunken with blunting of its microvillous border. Some alveoli showed severe degenerative changes and its lining exhibit hyaline membrane formation. Endothelial cells were sloughed in some capillaries with deposition of fibrillar osmophilic material on the epithelial aspect of the basal lamina. Proliferation of active macrophages was also seen. These changes were not evident in group III except for some cellular infiltration, thickening of interalveolar septum and focal oedema. Exposure to nickel chloride has deleterious effect on the delicate lung tissue, however, simultaneous administration of magnesium with nickel ameliorated significantly the harmful effect of nickel