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 lacrimal gland acinar cells in albino rats following prolonged use of phenylephrine decongestant eye drops

Tears, secreted by the acini of lacrimal gland, keep the corneal epithelium moist, thas preserving the most refractive mechanism of the eye. The over use of over-the-counter decongestant ophthalmic solutions is nowadays a public practice. The aim of the present work was to study the possible histological changes in the acinar cells of the lacrimal gland in albino rats after prolonged use of phenylephrine decongestant eye drops. 24 adult female albino rats were divided equally into 2 experimental groups, a control and a treated group that was subjected to repeated daily instillation of 2-3drops of phenylephrine 2.5% ophthalmic solution twice daily for 3 months. The intraorbital lacrimal glands of both groups were processed for light microscopic examination of H and E-stained paraffin sections, and toluidine blue-stained semithjn sections. Electron microscopic examination was also done. lacrimal gland acini of the control group revealed tall pyramidal secretory cells bordering a central lumen and contained large numbers of lightly- stained secretory granules. In the treated group, various histological changes were manifested in different acini. Manifestations of enhanced secretion with enormous number of secretory granules, extruding from short cuboidal acinar cells, into the widely distended lumen were depicted. Sustained stimulation of phenylephrine to the adrenergic receptors on the acinar cell membrane is suggested to enhance exocytosis. Repeated episodes of stimulation, by phenylephrine, produced manifestations of exhaustions and depleted activity in acinar cells. These manifestations were represented by wide areas of granule-free cytoplasm with dense shrunken nuclei. This might be attributed to the relatively limited energy and protein synthetic reserve of the lacrimal acinar cells. Long-standing application of phenylephrine eye drops resulted in multiforcal histological alterations of lacrimal gland acinar cells, which may contribute to dry eye state by depleting the functional reserve of the lacrimal gland. More attention should he paid during the use of such drugs. Further work is needed to investigate the possible reversibility of lacrimal gland histological changes after stoppage of the drug

Effect of dietary arginine on the jejunal mucosa in indomethacin-induced intestinal injury: light and scanning electron microscopic study in male mice

The non-steroidal anti-inflammatory drug [NSAID] indomethacin causes, via its adverse effects, damage to the gastrointestinal mucosa of humans and experimental animals. The indomethacin-induced intestinal injury in mice jejunum is considered an experimental model of Crohn's disease, as one of the inflammatory bowel diseases [IBD's]. The semi-essential amino acid arginine, which is a precursor of nitric oxide, is proposed to promote gastrointestinal mucosal integrity. The present work aimed to study the possible protective role of dietary supplementation with arginine in ameliorating indomethacin-induced mucosal injury of mice jejunum. The present study was carried out on forty adult male mice which were divided into 4 equal groups; group I [negative control group] which received no treatment, group II [positive control]; mice received dietary L-arginine alone in a daily dose of 300 mg/Kg, group III [indomethacin group]; mice of this group received 2 consecutive subcutaneous injections of indomethacin in a dose of 7.5 mg/kg, 24 hours apart and group IV [arginine group]; in which arginine supplementation was provided 2 days before the administration of indomethacin, maintained during the administration and continued 3 days later till the end of the experiment. Mice of all groups were sacrificed by the end of the 7[th] day and specimens from the jejunum were dissected and processed for light and scanning electron microscopic examinations. Indomethacin-treated group exhibited jejunal mucosal injury. Light microscopic examination of this group, using H and E and toluidine blue- stained semithin sections showed distorted villi and sloughing of some of their apical parts with extrusion of many degenerated cells, intermingled with excess mucous. Some enterocytes appeared degenerated with loss of their regular arrangement, while the goblet cells appeared distended with excess mucous secretion. Increased cellular infiltration and edema of the villous core and in the lamina propria between the glands were noticed. Increased mucous secretion was also demonstrated by combined alcian blue-periodic acid Schiff reaction. Scanning electron microscope revealed alteration in the architecture of many villi which appeared short, blunt or with denuded surface and occasionally covered with membrane-like structure. The group of mice received arginine [group IV] revealed restoration of mucosal integrity in the form of regular villi with intact epithelial coverings including enterocytes and goblet cells. Some villi appeared shorter, while others showed partially denuded apical surface. Arginine evoked a remarkable cellular infiltration. Lymphocytes and macrophages were among the infiltrating cells and their roles were suggested to be vital in the healing process. Dietary L-arginine provided satisfactory protection against indomethacin-induced mucosal injury in mice, most probably via its role as a nitric oxide donor. So supplementation with dietary arginine is recommended

Histological evaluation of local verapamil hydrochloride injection in the prevention of keloid recurrence after excision

Keloids occur as the result of an exaggerated wound healing of skin following various types of injury. In addition to presenting a cosmetic concern, they are extremely difficult to treat. The aim of the present work was to assess histologically the efficacy of verapamil hydrochloride [a calcium channel blocker] local injection on prevention of recurrence of surgically excised keloid. 40 keloid subjects divided into two equal groups: Group I; subjected to only surgical excision of their keloids. Group II; subjected to intradermal verapamil injection for 2 months after surgical excision of keloid. Skin biopsies from the healed wound of both experimental groups were taken after 3 months [subgroups Ib, IIb] and 6 months [subgroups Ib, IIb] postoperatively. Tissue specimens were also collected from the excised keloids. Informed consents were taken from subjects of both groups including all steps of the study. The obtained specimens were further processed for light microscopic study by haematoxylin and eosin [H and E] and trichrome stains. Semithin sections and electron microscopic examination were also done. Examination of the healed skin after keloid surgical excision [group Ia. Ib], showed prominent neutrophils and mast cells. Large spindle- shaped fibroblasts with dilated cisternae of endoplasmic reticulum were frequently encountered among coarse collagen bundles denoting accelerated rate of collagen production. This appearance may point to local histological recurrence of keloid. On the other hand, verapamil- injected skin after keloid excision [verapamil- injected group IIa, IIb] revealed little collagen in the dermal interstitium forming thin dispersed fibrils. Ultrastructuraly, the dermal fibroblasts of this group exhibited oval to rounded outlines and showed few narrow cisternae of endoplasmic reticulum. The verapamil- induced cell rounding is related to the calcium- dependent change in the cytoskeleton of fibroblasts. This appearance is associated with decreased synthesis and secretion of the extracellular matrix. Verapamil injection provided a satisfactory tool for prevention of recurrence of surgically excised keloid

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