Samaa Kamar

BACKGROUND: Cryptorchidism, one or both hidden testes, is the most common abnormality of male sexual development. Subfertility or infertility is associated with both unilateral and bilateral cryptorchidism. In this study, we investigate the possible ameliorative effect of curcumin (Cur) on the induced-unilateral cryptorchidism testicular injury in both cryptorchid (Cryp) and non-cryptorchid (non-Cryp) scrotal testes through histological, immunohistochemical and morphometrics.

MATERIALS AND METHODS: Forty adult male albino rats were divided into: control group, Cur control group, Cryp group, and Cryp+Cur group. The rat model was surgically established by fixing the left testis in the abdomen. The treated groups were subjected to surgically induced-unilateral cryptorchidism on the left side then were given Cur (80 mg/kg) orally, for 20 days. Histological analysis using haematoxylin and eosin and periodic acid Schiff's reaction was done. Immunohistochemistry was performed for proliferating cell nuclear antigen (PCNA); to estimate the proliferation in the germinal epithelium, and vimentin; to evaluate Sertoli cells. The results were confirmed by statistical evaluation of the spermatogenic epithelium height, the seminiferous tubules diameter, the basement membrane thickness, the number of PCNA immunostained cells and the area per cent of vimentin immunostaining.

RESULTS: Distorted seminiferous tubules, substantial degeneration of the germinal epithelium, thickening of the basement membrane with a significant decrease in PCNA and vimentin immunostaining were observed in Cryp group; mainly in the cryptorchid testis. These structural changes were significantly reversed in Crypt+Cur group.

CONCLUSIONS: Curcumin proved to be an important and effective medical line for protecting against the unfavourable sequels of cryptorchidism in a rat model.

BACKGROUND: Oligospermia is one of the common causative factors of male infertility. Some medical and hormonal therapy for male infertility is typically with unsatisfactory outcome. Stem cell therapy has become a new therapeutic strategy for restoring function in addition to inducing proliferation and differentiation of malfunctioning germ cells. This work aims at investigating the potential ability of BM-MSCs to repair the spermatogenic arrest in oligospermic rat model.

METHODS: In this work, a rat model of oligospermia was induced using two intraperitoneal injections of busulfan (15 mg/kg) with two weeks interval. Rats were divided into (i) donor group [source of the bone marrow mesenchymal stem cells (BM-MSCs) that were labelled and transfected with green fluorescent protein (GFP)] and (ii) experimental groups that were subdivided into: GpI (control), GpII (spermatogenic arrest model), GpIII (untreated rats), and GpIV (BM-MSCs treated rats). Estimation of the testicular weight, sperm count and motility % were performed. Histological and immunohistochemical staining for inducible nitric oxide synthase (iNOS) and caspase-3 (Cas-3) were conducted. Besides, the level of the testicular malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α) and testicular testosterone were estimated by ELISA.

RESULTS: Oligospermic rats illustrated hypospermatogenesis of the seminiferous tubule with spermatocyte and spermatid arrest, focal thickening of the basement membrane and significant increase in germ cells apoptosis and testicular oxidative stress. Compared with the control, MDA and TNF-α were markedly elevated with marked suppression of the testicular testosterone. Intra-testicular injection of BM-MSCs substantially ameliorated these changes and effectively improved the sperm count and motility %.

CONCLUSIONS: BM-MSCs improved the induced-spermatogenic arrest in the rat model mainly through anti-apoptotic and antioxidant paracrine effects.

Epidemiology and pathogenesis of cardiovascular diseases (CVD) and osteoporosis are strikingly overlapping. This study presents matrix metalloproteinase-9 (MMP-9), as a simple molecular link more consistently associated with the pathophysiology of both osteoporosis and CVD risk factors. 40 adult female rats were randomly distributed into 4 groups [control sham-operated, untreated osteoporosis, carvedilol-treated osteoporosis and alendronate-treated osteoporosis]. After 8 weeks, blood samples were collected to estimate Lipid profile (Total cholesterol, HDL, Triglycerides), inflammatory markers (IL-6, TNF alpha, CRP and NO), and Bone turnover markers (BTM) (Alkaline phosphatase, osteocalcin and pyridinoline). The tibias were dissected to estimate MMP-9 and NF-kB gene expression, OPG, RANKL levels and for histological examination. Induction of osteoporosis resulted in a significant elevation in BTM, inflammatory markers and dyslipidemia. MMP-9 was significantly elevated and positively correlated with BTM, inflammation and dyslipidemia markers. Carvedilol and alendronate exerted a bone preservative role and attenuated dyslipidaemia and inflammation in accordance with their respective effect on MMP-9.

INTRODUCTION: . Worldwide, nanoparticles especially gold-nanoparticles (Au-NPs) are widely used in medicine, cancer treatment and cosmetic industry. They are easily conjugated with different biomedical and biological agents and effortlessly absorbed with few side effects. The pars distalis of the pituitary gland is considered as the maestro of the endocrine peripheral glands since it secrets trophic hormones that controls their functions. 5-10% of the non-granular pars distalis cells are folliculo-stellate cells (FSCs) that support the granular cells' functions. The aim of the study was to explore the histological and the biochemical effects of repeated exposure to Au-NPs on the pars distalis in adult male albino rats with highlighting the impact on FSCs.

MATERIAL AND METHODS: . Thirty-six adult male albino rats were divided equally into control group and Au-NPs group (received 40 μg/kg/day of 11 ± 2 nm spherical Au-NPs orally for 2 weeks). Then, they were euthanized after 2 weeks and deposition of Au-NPs in pars distalis was investigated. Biochemical investigations and histological studies including hematoxylin and eosin staining, periodic acid Schiff's reaction, immunohistochemistry (IHC) for S-100, connexin 43 (Cx43) and Cytochrome-C (Cyt-C) as well as electron microscopic and morphometric studies were carried out.

RESULTS: . The Au-NPs group demonstrated structural disorganization in the pars distalis, inflammation, congestion and increased extracellular PAS-positive colloid deposition due to the accumulation of Au-NPs. A significant increase in the immunoreactivity of S-100, Cx43 and Cyt-C, along with a significant increase in TNF-α, MDA, and bFGF content in the pituitary homogenates, was noted as compared to the control group. Ultrastructurally, degenerative changes were observed in the secretory cells. FSCs showed proliferation and increased phagocytic activity.

CONCLUSIONS: . Repetitive exposure of adult male albino rats to Au-NPs prompted the accumulation of these nanoparticles in the pars distalis that was accompanied by cellular degeneration and dysfunction of the secretory cell and proliferation of FSCs. Thus, monitoring of the pars distalis hormonal levels might be useful for early detection of some hazardous effects possibly associated with the use of gold-nanoparticles.

Diet pattern is an emerging risk factor for renal disease. The mechanism by which high-fat high fructose (western) diet mediates renal injury is not yet fully understood. The objective of the present study was to investigate the relationship between endoplasmic reticulum (ER) stress and autophagy in the development of renal impairment and aggravation of the inflammatory response. Eighty male rats were randomly divided into four groups as follows: a standard diet-fed (ConD), a high-fat high fructose diet fed (HFHF-V), ConD fed and orally supplemented with vitamin E (ConD-E), and HFHF fed and orally supplemented vitamin E (HFHF-E). After 12 weeks, either lipopolysaccharide (LPS) or saline was injected. We found that upregulation of endoplasmic reticulum stress-related proteins rendered the cells susceptible to injury induced by dysbiosis and microbiota-derived metabolites. A downregulation of autophagy and upregulation of caspase-12 resulted in the loss of intestinal integrity and renal tubular injury. Maintained ER stress also increased the inflammatory response to LPS. In contrast, vitamin E effectively ameliorated ER stress and promoted autophagy to protect intestinal and renal tissues. Our results provide insight into the influences of sustained ER stress activation and autophagy inhibition on the development of renal injury, which may contribute also to the enhanced inflammatory response.

BACKGROUND: Cardiovascular diseases (CVD) represent one of the major sequelae of obesity. On the other hand, the relationship between bone diseases and obesity remains unclear. An increasing number of biological and epidemiological studies suggest the presence of a link between atherosclerosis and osteoporosis, however, the precise molecular pathways underlying this close association remain poorly understood. The present work thus aimed to study Matrix Metalloproteinase 9 (MMP-9), as a proposed link between atherosclerosis and osteoporosis in high fat diet fed rats.

METHODS AND FINDINGS: 40 rats were randomly divided into 4 groups: control, untreated atherosclerosis group, atherosclerotic rats treated with carvedilol (10mg/kg/d) and atherosclerotic rats treated with alendronate sodium (10mg/kg/d). After 8 weeks, blood samples were collected for estimation of Lipid profile (Total cholesterol, HDL, TGs), inflammatory markers (IL-6, TNF-α, CRP and NO) and Bone turnover markers (BTMs) (Alkaline phosphatase, osteocalcin and pyridinoline). Rats were then euthanized and the aortas and tibias were dissected for histological examination and estimation of MMP-9, N-terminal propeptide of type I procollagen (PINP), C-terminal telopeptide of type I collagen (CTX) and NF-kB expression. Induction of atherosclerosis via high fat diet and chronic stress induced a significant increase in BTMs, inflammatory markers and resulted in a state of dyslipidaemia. MMP-9 has also shown to be significantly increased in the untreated atherosclerosis rats and showed a significant correlation with all measured parameters. Interestingly, Carvedilol and bisphosphonate had almost equal effects restoring the measured parameters back to normal, partially or completely.

CONCLUSION: MMP-9 is a pivotal molecule that impact the atherogenic environment of the vessel wall. A strong cross talk exists between MMP-9, cytokine production and macrophage function. It also plays an important regulatory role in osteoclastogenesis. So, it may be a key molecule in charge for coupling CVD and bone diseases in high fat diet fed rats. Therefore, we suggest MMP-9 as a worthy molecule to be targeted pharmacologically in order to control both conditions simultaneously. Further studies are needed to support, to invest and to translate this hypothesis into clinical studies and guidelines.

Bone marrow-derived mesenchymal stem cells (BM-MSCs) have demonstrated potential in treating diabetic cardiomyopathy. However, patients with diabetes are on multiple drugs and there is a lack of understanding of how transplanted stem cells would respond in presence of such drugs. Metformin is an AMP kinase (AMPK) activator, the widest used antidiabetic drug. In this study, we investigated the effect of metformin on the efficacy of stem cell therapy in a diabetic cardiomyopathy animal model using streptozotocin (STZ) in male Wistar rats. To comprehend the effect of metformin on the efficacy of BM-MSCs, we transplanted BM-MSCs (1 million cells/rat) with or without metformin. Our data demonstrate that transplantation of BM-MSCs prevented cardiac fibrosis and promoted angiogenesis in diabetic hearts. However, metformin supplementation downregulated BM-MSC-mediated cardioprotection. Interestingly, both BM-MSCs and metformin treatment individually improved cardiac function with no synergistic effect of metformin supplementation along with BM-MSCs. Investigating the mechanisms of loss of efficacy of BM-MSCs in the presence of metformin, we found that metformin treatment impairs homing of implanted BM-MSCs in the heart and leads to poor survival of transplanted cells. Furthermore, our data demonstrate that metformin-mediated activation of AMPK is responsible for poor homing and survival of BM-MSCs in the diabetic heart. Hence, the current study confirms that a conflict arises between metformin and BM-MSCs for treating diabetic cardiomyopathy. Approximately 10% of the world population is diabetic to which metformin is prescribed very commonly. Hence, future cell replacement therapies in combination with AMPK inhibitors may be more effective for patients with diabetes. Metformin treatment reduces the efficacy of mesenchymal stem cell therapy for cardiac repair during diabetic cardiomyopathy. Stem cell therapy in diabetics may be more effective in combination with AMPK inhibitors.

Cholestasis caused by slowing or blockage of bile flow is a serious liver disease that can lead to liver fibrosis and cirrhosis. The link between transforming growth factor beta 1 (TGFβ1), Smad family member 3 (Smad3), and microRNA 21 (miR21) in bile duct ligation (BDL)-induced liver fibrosis in the presence and absence of the anti-inflammatory and antioxidant compound, resveratrol (RSV), has not been previously studied. Therefore, we tested whether RSV can protect against BDL-induced liver fibrosis associated with the inhibition of the TGFβ1-Smad3-miR21 axis and profibrogenic and hepatic injury biomarkers. The model group of rats had their bile duct ligated (BDL) for 3 weeks before being killed, whereas, the BDL-treated rats were separated into three groups that received 10, 20, and 30 mg/kg RSV daily until the end of the experiment. Using light microscopy and ultrasound examinations, we documented in the BDL group, the development of hepatic injury and fibrosis as demonstrated by hepatocytes necrosis, bile duct hyperplasia, collagen deposition, enlarged liver with increased echogenicity, irregular nodular border and dilated common bile duct, which were more effectively inhibited by the highest used RSV dosage. In addition, RSV significantly (p ≤ 0.0027) inhibited BDL-induced hepatic TGFβ1, Smad3, miR21, the profibrogenic biomarker tissue inhibitor of metalloproteinases-1 (TIMP-1), malondialdehyde (MDA), interleukin-17a (IL-17a), and blood levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin. These findings show that RSV at 30 mg/kg substantially protects against BDL-induced liver injuries, which is associated with the inhibition of TGFβ1-Smad3-miR21 axis, and biomarkers of profibrogenesis, oxidative stress, and inflammation.

Synchronized uterine receptivity with the time of implantation is crucial for pregnancy continuity. Vitamin D (VD) deficiency has been linked to the failure of implantation. Therefore, we tested the link between the Homeobox transcription factor-10/immunophilin FK506-binding protein 52 (HOXA-10/FKBP52) axis and the uterine receptivity in VD-deficient rats. The effect of VD supplementation at different doses was also investigated. Forty-eight pregnant rats were divided into six groups (eight/group); normal control rats fed with standard chow (control), control rats supplemented with VD (equivalent dose of 400 IU/day) (control-D400). VD-deficient group (DEF) and the three VD deficiency groups with VD supplementation were equivalent to 400, 4,000, and 10,000 IU/day (DEF-D400, DEF-D4000, and DEF-D10000, respectively). The expression levels of HOXA-10/FKBP52, progesterone level, and histological evaluation of decidualization using osteopontin (OSN) and progesterone receptor (PGR) were estimated. An assessment of the uterine contractility was conducted for all rats. This study showed the downregulation of HOXA-10/FKBP52 together with increased amplitude and frequency of the uterine contractility in the DEF group compared to control. VD dose-dependent supplementation restored progesterone/receptor competency, upregulated the expressional response of HOXA-10 and its downstream FKBP52, and improved uterine receptivity and endometrial decidualization at the time of implantation that was documented by increased area% of OSN and the number of implantation beads.

A 5-fluorouracil (5-FU) is used for cancer treatment despite its cytotoxic sequelae on healthy cells, especially the rapid proliferating ones. Intestinal mucositis is one of the most frequent chemotherapeutic debilitating sequelae. Rhubarb (Rh), an ancient herb, is known for its curing effect on gastrointestinal complications. This study aims to detect the role of aquaporin-4 (AQP-4), tumour necrosis factor-α (TNF-α), nuclear factor-kappa B (NF-κB), and matrix metalloproteinase-9 (MMP-9) in 5-FU-induced ileal histological and biochemical changes and the potential therapeutic effect of Rh water extract on these changes in rats. A 45 rats were divided into 3 groups; control, 5-FU (single intraperitoneal injection of 150 mg/kg/rat) and Rh-treated (oral 20 mg/kg/day/rat for 8 days). The change in animals' weight, incidence of diarrhoea and AQP-4 and TNF-α values in ileal homogenates were measured. Ileal sections were subjected to hematoxylin and eosin stain, periodic acid Schiff (PAS)-reaction and MMP-9, NF-κB and AQP-4 immunohistochemical staining. A 5-FU group revealed marked ileal mucosal damage associated with a significant decrease in the mean body weight, AQP-4 level and area percent of PAS and AQP-4 positive reaction. Significant increase in the mean incidence of diarrhoea, TNF-α value and area percent of MMP-9 and NF-κB was detected. These changes were significantly corrected with Rh administration. A 5-FU resulted in severe ileal mucositis through TNF-α, NF-κB, MMP-9, and AQP-4 disturbances. Rh treatment was highly effective in preventing such mucositis.

Gastric ulcer is one of the most serious diseases. Nebivolol (Neb), a β1-blocker, exhibits vasodilator and anti-oxidative properties, simvastatin (Sim) antihyperlipidemic drug, exhibits anti-oxidative, anti-inflammatory properties and promote endogenous nitric oxide (NO) production. The aim of this study was to evaluate the gastroprotective effects of Neb and Sim against cold restraint stress (CRS)-induced gastric ulcer in rats. Rats were restrained, and maintained at 4°C for 3 hours. Animals were divided into six groups; control group, CRS group, and four treatment groups received ranitidine (Ran), Neb, Sim and both Neb and Sim. Treatments were given orally on a daily basis for 7 days prior to CRS. The gastroprotective effects of Neb and Sim were assessed biochemically by measuring variations in prostaglandins E2, NO, reduced glutathione and malondialdehyde, and functionally by estimating force of contractions of isolated rat fundus in the studied groups in response to acetylecholine stimulation and morphologically using hematoxylin and eosin staining, periodic acid Schiff's reaction and immunohistochemistry for cyclooxygenase 2 in gastric mucosa. CRS caused significant ulcerogenic effect. Alternatively, pretreatment with Ran, Neb, and Sim significantly corrected biochemical findings, pharmacological and histological studies.

We sought to determine whether TDZD-8, the inhibitor of the glycogen synthase kinase-3β (GSK3β), can protect the synovial membrane of the knee joint against injuries induced by collagen type II immunization (CIA) possibly via the downregulation of synovial leukocyte infiltration, endoplasmic reticulum stress (ERS), and autophagy. The model group of rats (CIA) were immunized over a period of 3 weeks with collagen type II, whereas the treated group of rats (CIA + TDZD-8) were treated with TDZD-8 (1 mg/kg) for 21 days after the completion of the immunization regimen. All rats were then killed at week 6. Harvested synovial tissues were prepared for immunohistochemistry staining, and synovial homogenates were assayed for biomarkers of ERS, autophagy, apoptosis, and cell survival and proliferation. In addition, blood samples were assayed for biomarkers of arthritis. Synovial tissue images showed that CIA enhanced leukocyte recruitment as demonstrated by an increased CD45+ (leukocyte common antigen) immunostaining, which was markedly decreased by TDZD-8. TDZD-8 also significantly (P < .05) inhibited collagen-induced autophagy biomarkers Beclin-1 and LC3II, the ERS biomarkers GRP-78, IRE1-α, XBPIs, and eIF2a, and the survival protein Bcl-2. Whereas, the collagen-induced proliferative biomarkers Akt and mTOR were not inhibited by TDZD-8, and CIA inhibited the apoptotic proteins CHOP and cleaved caspase-3, which were augmented by TDZD-8. We further demonstrated a significant (P < .05) correlation between autoantibodies generated during the course of arthritis and biomarkers of ERS and autophagy. We conclude that TDZD-8 inhibits CIA and decreases synovial leukocyte infiltration, ERS, and autophagy, which is independent of Akt/mTOR signalling.

Osteoarthritis (OA) secondary to diabetes affects millions of people worldwide and can lead to disability. The protective effect of metformin pretreatment against alterations to the articular cartilage ultrastructure induced by type 2 diabetes mellitus (T2DM) associated with the inhibition of oxidative stress and inflammation has not been investigated before. Therefore, we induced T2DM in rats (the model group) using high carbohydrate and fat diet and a single injection of streptozotocin (50 mg/kg body weight). The protective group of rats started metformin (200 mg/kg body weight) treatment 14 days before diabetic induction and continued on metformin until the end of the experiment at week 12. Harvested tissues obtained from knee joints were prepared for staining with hematoxylin and eosin (H&E), safranin o staining, and electron microscopy. Histology images showed that OA was developed in the T2DM rats as demonstrated by a substantial damage to the articular cartilage and profound chondrocyte and territorial matrix ultrastructural alterations, which were partially protected by metformin. In addition, metformin significantly ( < .05) reduced hyperglycemia, glycated hemoglobin (HbA1 c), malondialdehyde (MDA), high sensitivity C-reactive protein (hs-CRP), and interleukin-6 blood levels induced by diabetes. Furthermore, a significant (p ≤ 0.015) correlation between either OA cartilage grade score or the thickness of the articular cartilage and the blood levels of HbA1 c, hs-CRP, MDA, superoxide dismutase (SOD) were observed. These findings demonstrate effective protection of the articular cartilage by metformin against damage induced secondary to T2DM in rats, possibly due to the inhibition of hyperglycemia and biomarkers of oxidative stress and inflammation.

BACKGROUND: The potential protective effects of resveratrol (RES) on the modulation of hepatic biomarkers of apoptosis and survival, p53-Bax axis, and B-cell lymphoma 2 (Bcl-2) in an animal model of paracetamol-induced acute liver injury have not been investigated before.

METHODS: The model group of rats received a single dose of paracetamol (2 g/kg, orally), whereas the protective group of rats were pretreated for 7 days with RES (30 mg/kg, i.p.) before they were given a single dose of paracetamol. All rats were then sacrificed 24-h post paracetamol ingestion.

RESULTS: Histology images showed that paracetamol overdose induced acute liver injury, which was substantially protected by RES. Paracetamol significantly (p < 0.05) modulated p53, apoptosis regulator Bax, Bcl-2, tumor necrosis factor-alpha, interleukin-6, inducible nitric oxide synthase, malondialdehyde, superoxide dismutase, glutathione peroxidase, alanine aminotransferase, and aspartate aminotransferase, which were significantly protected by RES. We further demonstrated a significant (p< 0.01) correlation between either p53 or Bcl-2 scoring and the levels of inflammatory, nitrosative stress, and liver injury biomarkers.

CONCLUSION: We demonstrate a substantial protection by RES pretreatment against paracetamol-induced modulation of p53-Bax axis, Bcl-2, and other acute liver injury biomarkers in rats.

We investigated whether the anti-inflammatory and antioxidant agent, resveratrol can inhibit type 2 diabetes mellitus (T2DM)-induced osteoarthritis (OA) in rats and whether it is associated with the suppression of glycaemia, dyslipidemia and inflammatory and oxidative stress biomarkers. T2DM was induced by streptozotocin (50 mg/kg body weight) and high carbohydrate and fat diet (HCFD). The protective group was put on resveratrol (30 mg/kg) 14 days prior to the induction of diabetes and continued on resveratrol and HCFD until being sacrificed at week 12. Diabetic rats showed a substantial damage to the knee joints and loss of proteoglycans from the articular cartilage, which were effectively but not completly protected by resveratrol. Resveratrol also significantly ( ≤ .0029) reduced diabetic up-regulation of HbA1c, hyperlipidaemia, inflammation and oxidative stress. Resveratrol protects against T2DM-induced OA associated with the inhibition of glycated haemoglobin, dyslipidemia, and biomarkers of oxidative stress and inflammation.

INTRODUCTION: Asherman syndrome (AS) is a symptomatic intrauterine adhesion caused by endometrial basal layer fibrosis as a result of either uterine cavity surgery or infection leading to many complications. There is a concern to repair the injured tissues by using bone marrow mesenchymal stem cells (BM-MSCs). We aimed in this study to develop an animal model of AS and evaluate the anti-inflammatory and anti-fibrotic effects of BM-MSCs in this model through histological, immunohistochemical, and morphometric studies.

MATERIAL AND METHODS: Forty-two adult female adult albino rats were divided into (i) donor group composed of 2 rats used for isolation and propagation of BM-MSCs, and (ii) experimental groups: 40 rats equally divided into 4 groups: GpI (control), GpII (AS model), GpIII (BM-MSCs-treated AS rats), GpIV (untreated AS rats). Histological staining and immunohistochemical (IHC) detection of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), and nuclear factor-kappa beta (NF-kB) were performed. The results were evaluated by morphometric and statistical analysis.

RESULTS: Significant endometrial thinning, fibrosis, and degeneration of the endometrial epithelium with a significant decrease in PCNA and VEGF immunoexpression and a significant increase in NF-kB immunoexpression were detected in GpII and GpIV groups. These changes were substantially reversed in BM-MSCs-treated animals.

CONCLUSIONS: BM-MSCs treatment resulted in substantial improvement of intrauterine adhesion in the rat model of Asherman syndrome.

The multifetal reduction (MFR) procedure is usually reserved for high-order multiple pregnancies, and aspirated tissues are typically discarded. In this study, cells obtained from MFR tissue (termed multifetal reduction embryonic cells (MFR-ECs)), were characterized by genotypic and phenotypic analyses and tested by injection under the kidney capsule of nude mice. MFR-ECs were highly proliferative in culture and showed a normal karyotype by microarray CGH. Immunohistochemical analysis at day zero showed positive focal staining for desmin, S-100 protein, synaptophysin and chromogranin. Histology examination showed a mixture of cells from the three germ layers at different stages of differentiation. Markers of these stages included important developmental transcription factors, such as beta three-tubulin (ectoderm), paired box 6 (ectoderm) and alpha-smooth muscle actin (mesoderm). Quantitative polymerase chain reaction (qPCR) showed down-regulation of the mRNAs of cancer-related genes such as TP53. transplantation in nude mice showed a typical hyaline cartilage plate and no teratoma formation. Thus, MFR-ECs represent a rich, unique source for studying stem cell development, embryogenesis and cell differentiation.

Management of diabetic wounds remains a major challenge in the medical field, mostly due to incompetent outcomes of treatments. Curcumin has been documented as anti-inflammatory, antioxidant, antimicrobial and antineoplastic agent in addition to wound healing activities. However, its poor aqueous solubility and impaired skin permeation handicap its topical pharmaceutical usage. Hydrogel loaded curcumin nanoparticle (Cur-NP/HG) could overcome this pitfall and enable extended topical delivery of curcumin. Rat model of diabetes mellitus (DM) type I was induced using single injection of 70mg/kg streptozotocin (STZ) followed by full thickness skin wound. Rats were divided into 4 groups. GpI: control non-diabetic, GpII: diabetic non-treated, GpIII: diabetic treated with topical curcumin hydrogel (Cur/HG) and GpIV: diabetic treated with topical Cur-NP/HG. Histological assessment of epidermal regeneration, dermo-epidermal junction, leukocyte infiltration and collagen deposition, in addition to immunohistochemical staining for vascular endothelial growth factor (VEGF) and aquaporin-3 (AQP3) were performed. Diabetic rat possessed impaired wound closure, persistence of inflammation and decreased collagen deposition as compared to non-diabetic control. Application of Cur/HG induced partial improvement of the healing process in diabetic rats. Cur-NP/HG treatment provoked obvious improvement of the healing process with complete re-epithelization, intact dermo-epidermal junction, reorganization of the dermis with significantly increased collagen deposition and VEGF and AQP3 expression. These results illustrated that Cur-NP/HG have effectively improved the healing process in diabetic skin wound with substantial differences in the wound healing kinetics compared to wounds that received Cur/HG.

Management of diabetic wounds remains a major challenge in the medical field, mostly due to incompetent outcomes of treatments. Curcumin has been documented as anti-inflammatory, antioxidant, antimicrobial and antineoplastic agent in addition to wound healing activities. However, its poor aqueous solubility and impaired skin permeation handicap its topical pharmaceutical usage. Hydrogel loaded curcumin nanoparticle (Cur-NP/HG) could overcome this pitfall and enable extended topical delivery of curcumin. Rat model of diabetes mellitus (DM) type I was induced using single injection of 70mg/kg streptozotocin (STZ) followed by full thickness skin wound. Rats were divided into 4 groups. GpI: control non-diabetic, GpII: diabetic non-treated, GpIII: diabetic treated with topical curcumin hydrogel (Cur/HG) and GpIV: diabetic treated with topical Cur-NP/HG. Histological assessment of epidermal regeneration, dermo-epidermal junction, leukocyte infiltration and collagen deposition, in addition to immunohistochemical staining for vascular endothelial growth factor (VEGF) and aquaporin-3 (AQP3) were performed. Diabetic rat possessed impaired wound closure, persistence of inflammation and decreased collagen deposition as compared to non-diabetic control. Application of Cur/HG induced partial improvement of the healing process in diabetic rats. Cur-NP/HG treatment provoked obvious improvement of the healing process with complete re-epithelization, intact dermo-epidermal junction, reorganization of the dermis with significantly increased collagen deposition and VEGF and AQP3 expression. These results illustrated that Cur-NP/HG have effectively improved the healing process in diabetic skin wound with substantial differences in the wound healing kinetics compared to wounds that received Cur/HG.

Wnt/β-catenin signaling pathway plays a crucial role in diabetic cardiomyopathy (DCM), thus we aimed at investigating the effect of one therapeutic approach with resveratrol (RSV) given systemically and combined treatment of RSV with mesenchymal stem cells (MSCs) that was either RSV-preconditioned or not on Wnt/β-catenin signaling pathway in streptozotocin-induced DCM, and to evaluate effects of RSV preconditioning on MSCs therapeutic potential. The rats were divided into control (C, n = 8), diabetic (DM, n = 8), diabetic treated with systemic RSV (DM-RSV, n = 8), diabetic treated with RSV and nonconditioned MSCs (DM-RSV-MSCs, n = 8), diabetic treated with RSV and RSV-incubated with MSCs (DM-RSV-MSCc, n = 8) and diabetic treated with RSV-conditioned MSCs (DM-MSCc, n = 8). Echocardiography (Echo) showed significant improvement of cardiac functions in all groups treated with RSV either systemic or added in culture media. Data of ejection fraction (EF%) of DM-RSV-MSCc (81.50; interquartile range [IQR], 80.00-83.00) was comparable to both DM-RSV-MSCs (77.50; IQR, 71.50-79.00), and DM-MSCc (71.50; IQR, 70.00-74.50). Histological examination of the left ventricles was performed for all groups. DM group revealed significant myocardial hypertrophy, apoptosis, interstitial fibrosis, and microvascular affection. All treated groups were associated, in variable degrees, with attenuation of cardiac hypertrophy and fibrosis, decreased area% for cardiac immunostaining of secreted frizzled-related protein (sFRP2) and Wnt/β-catenin and improvement of the microvasculature. In conclusion, MSCs pretreated with resveratrol for 7 days showed increased 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and combined use of RSV (systemically and in culture media) significantly could improve cardiac remodeling capacity of MSCs via attenuation of sFRP2-mediated fibrosis and the downstream Wnt/β-catenin pathway.