Hala Gabr Metwally

Stem cell therapy as a novel therapeutic intervention for resistant cases of alopecia areata and androgenetic alopecia

Abstract Background: Management of alopecia areata (AA) and androgenetic alopecia (AGA) is often challenging as patients may be resistant to currently available modalities of treatment. The use of stem cells may be a novel option for resistant cases. Objective: To evaluate the safety and efficacy of the use of autologous bone marrow derived mononuclear cells (including stem cells) as compared to follicular stems cells for the management of resistant cases of AA and AGA. Methods: This study included 40 patients (20 AA patients and 20 AGA patients), all patients were treated with a single session of intradermal injection of autologous stem cells (SCs) therapy. They were divided into four groups according to the applied modality [either autologous bone marrow derived mononuclear cells (bone marrow mononuclear cells [BMMCs] or autologous follicular stem cells [FSC]). Results: Six months after stem cell therapy (SCT) injection, there was a significant improvement, confirmed by immunostaining and digital dermoscopy. The mean improvement in all groups was “very good”. There was no significant difference between both methods in either type of alopecia. No serious adverse events were reported. Conclusion: Autologous BMMCs and FSC seem to be a safe tolerable and effective treatment for the management of both resistant AA and AGA.

Comparative Study between Intravenous and Intraperitoneal Stem Cell Therapy in Amiodarone Induced Lung Injury in Rat.

Background and Objectives: The fibrosing form of lung injury (occupational, environmental, infective or drug induced) is associated with significant morbidity and mortality. Amiodarone (AM), often prescribed for control of arrhythmias is considered a potential cause. No effective treatment was confirmed, except lung transplantation. Intravenous (IV) stem cell therapy may produce pulmonary emboli or infarctions. Despite being commonly used in clinical practice, the intraperitoneal (IP.) route has been rarely used for cell delivery. The present study aimed at investigating and comparing the possible effect of IP stem cell therapy (SCT) on pulmonary toxicity versus the intravenous route in a rat model of amiodarone induced lung damage. Methods and Results: 36 adult male albino rats were divided into 4 groups. Rats of AM group were given 30 mg/kg daily orally for 4 weeks. Rats of IV SCT group were injected with stem cells in the tail vein. Rats of IP SCT group received IP cell therapy. Histological, histochemical, immunohistochemical and morphometric studies were performed. Obstructed bronchioles, overdistended alveoli, reduced type I pneumocytes, increased thickness of alveolar septa and vessels wall besides increased area% of collagen fibers regressed in response to IV and IP SCT. The improvement was more obvious in IV group. The area% of Prussion blue +ve and CD105 +ve cells was significantly higher in IV group. Conclusions: Cord blood MSC therapy proved definite amelioration of lung injury ending in fibrosis. The effect of IP SCT was slightly inferior to that of IV SCT, which may be overwhelmed by repeated IP injection.

The effect of stem cell therapy versus melatonin on the changes induced by busulfan in the testes of adult rat: histological and immunohistochemical studies

Introduction Chemotherapy is associated with significant gonadal damage. Melatonin has been found to be a potent free radical scavenger and antioxidant. Stem cells have the capacity to generate multiple distinct cell lineages. Aim of the work The present study aimed to compare the effect of stem cell therapy and melatonin in the amelioration of harmful changes induced by busulfan in rat testes. Materials and methods Forty-three adult male rats were used in the present work. Animals were divided into the following groups: group Iwas the control group, the rats in group II received a single dose of intraperitoneal busulfan (20 mg/kg), the rats in group III rats received 0.5 ml stem cells upon an initial dose of busulfan, the animals in group IV received a single dose of intraperitoneal melatonin (10 mg/kg) for 5 days, and the rats in group V received melatonin for 5 days upon an initial dose of busulfan. The testes were stained with H&E, prussian blue, and immunohistochemical stains for the detection of caspase-3-positive and proliferating cell nuclear antigen (PCNA)-positive cells. Digital image analysis was used to determine the mean number of active caspase-3-positive and PCNA-positive cells. The results were compared statistically. Results Busulfan-treated animals showed atrophy and irregularity of the tubules. Marked depletion of germ cells was supported by an increase in the mean number of caspase-3-positive cells in the semineferous tubules. In addition, decreased proliferative capacity was observed in the present study and confirmed by adecrease in the mean number of PCNA-positive cells. Sections of the stem cell-treated group (group III) showed incomplete or partial regeneration ofgerm cells with an absence of elongated and rounded spermatids. Sections ofthe melatonin-treated group (group V) showed almost complete regeneration of germ cells with the appearance of rounded and elongated spermatids. Conclusion: It could be concluded that melatonin was more effective than stem cells in ameliorating busulfan-induced testicular damage.

Therapeutic Effect of Adipose Derived Stem Cells versus Atorvastatin on Amiodarone Induced Lung Injury in Male Rat.

Background and Objectives Amiodarone (AM), a class 3 antiarrhythmic drug, has been associated with variety of adverse effects, the most serious of which is pulmonary toxicity. Ator (A) is a statin, known for their immunomodulatory and anti-inflammatory activities. Recent studies provide evidence of potential therapeutic effect of statins on lung injury. Adipose derived stem cells (ADSCs) have shown great promise in the repair of various tissues. The present study aimed at investigating and comparing the possible therapeutic effect of A and ADSCs on AM induced lung injury in albino rats. Methods and Results 34 adult male albino rats were divided into 5 groups: control group (Gp I), A group (Gp II) received 10 mg/kg of A orally 6 days (d)/week (w) for 4 weeks (ws), AM group (Gp III) received 30 mg/kg of AM orally 6 d/w for 4 ws, AM&A group (Gp IV) received AM for 4ws then A for other 4 ws and AM&SCs group (Gp V) received AM for 4 ws then injected with 0.5 ml ADSCs on 2 successive days intravenously (IV). Histological, histochemical, immunohistochemical and morphometric studies were performed. Group III displayed bronchiolitis obliterans, thickened interalveolar septa (IAS) and thickened vascular wall which were proven morphometrically. Increased area% of collagen fibers and apoptotic changes were recorded. All findings regressed on A administration and ADSCs therapy. Conclusion Ator proved a definite ameliorating effect on the degenerative, inflammatory, apoptotic and fibrotic changes induced by AM. ADSCs administration denoted more remarkable therapeutic effect compared to A.

In-vitro differentiation of adipose-derived stem cells into melanocytic lineage: a possible new alternative treatment for vitiligo

Introduction Adipose tissue has been shown to contain adipose-derived stem cells (ADSCs), which constitute a very convenient source of stem cells because it can be obtained easily in large quantities. Vitiligo is a pigmentation disorder caused by loss of skin melanocytes and can be treated by grafting autologous melanocytes. However, it is difficult to obtain sufficient skin for autologous cell transplantation. If ADSCs can differentiate into melanocytes, adipose tissue could be used as an alternative source of melanocytes in vitiligo treatment. Aim The aim of this study was to evaluate the potential differentiation of ADSCs into melanocyte precursor cells. Materials and methods Human subcutaneous adipose tissue was obtained. Isolation and culture of ADSCs was done. After 1 week, differentiation of ADSCs into melanocytic lineage was initiated using basic medium M254 supplemented with Human Melanocyte Growth Supplement, which was replaced every other day for 3 weeks. C-kit immunocytochemistry and Melan-A/MART-1 immunofluorescence were performed to assess the differentiation of ADSCs into melanocytic lineage. Functional characterization of melanocytic differentiation was validated by detecting tyrosinase activity assay after 1, 2, and 3 weeks of differentiation. Data obtained from quantitative morphometric study and tyrosinase activity assay were statistically analyzed. Results After 1 week of culture in the differentiation medium, ADSCs began to exhibit melanocytic morphology with multiple cytoplasmic processes, which became more apparent by the end of the third week. Melanocytic differentiation was confirmed by positive C-kit and Melan-A/MART-1 immunophenotyping as well as by positive tyrosinase activity, which increased significantly by the end of week 3 after differentiation. Conclusion ADSCs could be differentiated into morphological and functional melanocytic lineages, which might be useful as an alternative treatment for vitiligo.

Histological comparative study between endogenous and therapeutic stem cells in adriamycin-induced renal cortical toxicity in male rats

Background Renal toxicity resulting from kidney injury poses an important problem to public health. Exogenous and endogenous stem cells (SCs) derived from various tissues hold great promise for regeneration. Aim The aim of this study was to assess and compare the effect of both endogenous and therapeutic SCs in rat model of adriamycin (ADR)-induced renal cortical toxicity. Materials and methods Twenty-four adult male albino rats were divided into three groups: the control group, the ADR group, and the SC therapy group. The ADR group received single intraperitoneal injection of 5 mg/kg ADR; this group was subdivided into subgroups IIa and IIb, and the rats were sacrificed at 2 and 4 weeks following the expected confirmed renal cortical injury, respectively. The SC therapy group received ADR and was injected with 0.5 ml of human umbilical cord blood SCs in the tail vein. The SC therapy group was subdivided into subgroups IIIa and IIIb that were sacrificed at 2 and 4 weeks following SC therapy, respectively. Kidney sections were stained with H&E, Prussian blue stain, and CD105 immunostaining. Morphometric study and statistical analysis were performed. Results A significant decrease in the mean glomerular area was found in subgroup IIb compared with control and other subgroups. A significant increase in the mean area of vacuolated cells with dark nuclei was found in subgroups IIa and IIb compared with other subgroups. A significant increase in the mean area of CD105+ cells was detected in subgroup IIIa and IIIb compared with other subgroups. Conclusion Endogenous SCs need to be activated by exogenous administration of SCs for more rapid improvement to attain normal function as early as possible.

Comparative Histological Study on the Therapeutic Effect of Green Tea and Stem Cells in Alzheimer's Disease Complicating Experimentally Induced Diabetes.

Background and Objectives Alzheimer’s disease (AD) is a devastating neurodegenerative disorder. Increasing evidence implicates diabetes mellitus (DM) as a risk factor for AD. Green tea (GT) has several beneficial effects attributed to its anti-oxidant phenolic compounds. Adipose tissue is a rich source of adipose-derived mesenchymal stem cells (ADSCs). This study was designed to evaluate and compare the possible therapeutic effect of green tea extract (GTE) and ADSCs on AD complicating induced DM in male rat. Methods 31 adult male albino rats were divided into 5 groups. Group I (Control), Group II received GTE, 50 mg/kg daily orally for 4 weeks, Group III received a single intraperitoneal injection of Streptozotocin (STZ), 50 mg/kg, Group IV: received STZ followed by GTE and Group V: received STZ followed by human ADSCs (hADSCs) intravenously. Results Multiple acidophilic masses, deformed neurons, Congo red +ve masses and Caspase 3 +ve neurons were seen in group III, became few in group IV and occasional in group V. Multiple Prussian blue +ve cells were detected in group V. Some CD44 +ve cells were noticed in group III, became multiple in groups IV and V. The mean area of neurons exhibiting acidophilic cytoplasm, mean area of amyloid plaques and mean area % of Caspase 3 +ve cells indicated a significant increase in group III. The mean area % of CD44 +ve cells recorded a significant increase in group IV. Conclusions hADSCs exerted a more marked therapeutic effect on the neurodegenerative changes complicating DM and corresponding to AD.

Effect of stem cell therapy on amiodarone-induced liver injury in albino rats

IntroductionSustained liver injury causes the development of fibrosis. For patients with end-stage fibrosis (cirrhosis) with subsequent portal hypertension, liver failure, and hepatocellular cancer, liver transplantation is the only effective method of treatment. However, it is associated with several complications and side effects. Aim of the workThe present study aimed to determine the possible effect of human cord blood mesenchymal stem cell therapy on liver injury using amiodarone as a model of induced liver damage in albino rats. Materials and methodsTwenty-three adult male albino rats were divided into three groups: group I (control) included six rats that were given 0.5ml Tween 80 orally for 2 weeks, group II included 10 rats that were given 5.4mg of amiodarone orally for 2 weeks, and group III included seven rats that were injected with stem cells in the tail vein following confirmation of liver damage and held for 4 weeks before sacrifice. Liver specimens were processed. Sections were subjected to the following stains: H&E, Massons trichrome, and Prussian blue. Immunohistochemical studies were carried out for CD44 and human APF. Digital image analysis was used to determine the area % of collagen fibers and the optical density of &agr;-fetoprotein-positive cells. The results were compared statistically. ResultsIn group II, congested dilated blood sinusoids were observed. Some hepatocytes showed dark nuclei. Some hepatocytes appeared with dark nuclei and a strong acidophilic cytoplasm; others were ballooned. Mallory bodies were observed. Some portal areas showed intense mononuclear cellular infiltration. Extensive collagen fibers existed around some central veins and portal tracts and increased area % of collagen fibers was observed. Most histological findings were improved in group III. In addition, multiple-positive &agr;-fetoprotein immunostained cells were detected and proved morphometrically by their increased mean optical density in comparison with group II. ConclusionIt can be concluded that cord blood mesenchymal stem cell therapy induces amelioration in morphological changes associated with amiodarone-induced liver injury, provided therapy is initiated early in the development of the injury.