Tail Vein Of Rats Research Articles

Mesenchymal Stem Cells (MSC) Prevented the Progression of Renovascular Hypertension, Improved Renal Function and Architecture

Renovascular hypertension induced by 2 Kidney-1 Clip (2K-1C) is a renin-angiotensin-system (RAS)-dependent model, leading to renal vascular rarefaction and renal failure. RAS inhibitors are not able to reduce arterial pressure (AP) and/or preserve the renal function, and thus, alternative therapies are needed. Three weeks after left renal artery occlusion, fluorescently tagged mesenchymal stem cells (MSC) (2×105 cells/animal) were injected weekly into the tail vein in 2K-1C hypertensive rats. Flow cytometry showed labeled MSC in the cortex and medulla of the clipped kidney. MSC prevented a further increase in the AP, significantly reduced proteinuria and decreased sympathetic hyperactivity in 2K-1C rats. Renal function parameters were unchanged, except for an increase in urinary volume observed in 2K-1C rats, which was not corrected by MSC. The treatment improved the morphology and decreased the fibrotic areas in the clipped kidney and also significantly reduced renal vascular rarefaction typical of 2K-1C model. Expression levels of IL-1β, TNF-α angiotensinogen, ACE, and Ang II receptor AT1 were elevated, whereas AT2 levels were decreased in the medulla of the clipped kidney. MSC normalized these expression levels. In conclusion, MSC therapy in the 2K-1C model (i) prevented the progressive increase of AP, (ii) improved renal morphology and microvascular rarefaction, (iii) reduced fibrosis, proteinuria and inflammatory cytokines, (iv) suppressed the intrarenal RAS, iv) decreased sympathetic hyperactivity in anesthetized animals and v) MSC were detected at the CNS suggesting that the cells crossed the blood-brain barrier. This therapy may be a promising strategy to treat renovascular hypertension and its renal consequences in the near future.

Inhibition of Wnt/β‐Catenin Signaling Promotes Engraftment of Mesenchymal Stem Cells to Repair Lung Injury

We sought to explore the treatment effects and the repair mechanisms of bone marrow derived mesenchymal stem cells (MSCs) during HCl-induced acute lung injury (ALI). MSCs were delivered through the tail veins of rats 24 h after intranasal instillation of HCl. The results showed that MSCs did not ameliorate the histopathologic changes of ALI and pulmonary fibrosis. We found that the activated Wnt/β-catenin signaling may regulate the differentiation of MSCs and is associated with lung fibroblasts activation, pulmonary fibrosis and tissue repair process in ALI rats. Immunofluorescence and histology analysis indicated that activated canonical Wnt/β-catenin signaling induced most MSCs to differentiate into myofibroblasts or fibroblasts in vivo. However, inhibition of Wnt/β-catenin signaling by Dickkopf-1 (DKK1) promotes epithelial differentiation of MSCs induced by native alveolar epithelial cells which are beneficial to repair the injured lung epithelium. Inhibition of Wnt/β-catenin signaling after MSCs transplantation ameliorated pulmonary fibrosis and improved pulmonary function which attenuated the lung injury. In vitro study, activation of the Wnt/β-catenin signaling stimulated MSCs to express myofibroblasts markers, which was attenuated by DKK1. Furthermore, Wnt3α activated Wnt/β-catenin signaling in lung fibroblasts to enhance the expression of collagen I, vimentin and α-smooth muscle actin, but DKK1 attenuated these proteins expression. These findings demonstrated that canonical Wnt/β-catenin signaling plays a key role in regulating differentiation of MSCs in vivo or in vitro and the pathogenesis of fibrotic diseases. Our study suggested that inhibition of abnormal activated Wnt/β-catenin signaling would promote MSCs epithelial differentiation to repair lung injury and reduce pulmonary fibrosis.

Bio-distribution study of Reolysin® (pelareorep) through a single intravenous infusion in Sprague-Dawley rats

Numerous pre-clinical and clinical studies on reovirus have generated valuable information which supports the use of this orphan virus as an investigational drug for cancer treatment. Reolysin® (pelareorep) is a clinical formulation of the human Reovirus Type 3 Dearing strain. The clinical safety and efficacy of Reolysin® in humans is being tested on an assortment of cancer indications as a mono and/or combination therapy. Reovirus has many inherent characteristics that make it a potential candidate for virotherapy, including: the rapid and natural spread through the haematogenous route, the ability to overcome immunological barriers thereby reaching tumor sites, and being replication-competent. The purpose of this study was to elucidate the bio-distribution pattern of Reolysin® in healthy Sprague-Dawley rats. Following a single 15-min intravenous infusion via the tail vein in Sprague-Dawley rats, the levels of virus genome were determined in 16 organs/tissues by RT-qPCR (Reverse Transcriptase- Quantitative Polymerase Chain Reaction) over a 336 h (Day 15) incubation regime. Consistent with previous studies, maximal reovirus RNA levels were observed in the spleen; indicating its involvement in viral uptake and clearance, followed by heart, ovaries, tail (infusion site), liver and lungs. All the organs/tissues demonstrated unquantifiable levels of reovirus genome at the end of incubation, suggesting substantial to complete viral clearance. Several studies in the last decade have described the use of reovirus for treating ovarian cancers. An increase of reovirus genome in ovaries at 24 h post infection was noted. The results will aid in the design of additional exploratory clinical trials for Reolysin®.

The effect of carvedilol and nebivolol on contrast nephropathy

Aim: This study examined biochemical, histopathological and antioxidant effects of the carvedilol and nebivolol on contrast induced nephropathy (CIN). Methods: Forty male Wistar-albino rats were divided into six groups. Control group (C, n=6), contrast medium group (CM, n=6), carvedilol group (CV, n=7), carvedilol+contrast group (CVCM, n=7), nebivolol group (N, n=7), nebivolol+contrast group (NCM, n=7). At the beginning of the study, blood samples were taken from tail veins of rats. All rats were taken to a 72 hour dehydration period. For three days, 2mg/kg carvedilol which was diluted in 1% DMSO was administered to the rats in group 3 and 4 and 1mg/kg nebivolol was administered to the rats in group 5 and 6 with orogastric lavage. Also 2 ml/kg 1% DMSO consisting 0.9% NaCl solution was administered with orogastric lavage to C and CM groups. In the third day of dehydration, 6 ml/kg diatrizoate was intravenously administered via tail vein to the groups CM, N and NCM. 6 ml/kg 0.9%NaCl solution was intravenously administered via tail vein to C, CV and N groups. Rats were sacrificed, and right kidney was kept to measure total antioxidant status (TAS), superoxide dismutase (SOD) and malondialdehyde (MDA) levels. Results: Histopathologically changes were significantly higher in CM group (Table 1). Although it was not statistically significant, in the CM group creatinine levels were increased. Despite exposure to contrast medium, CVCM and NCM groups did not differ in terms of histopathological findings from control group. In CV group, interstitial inflammation and other histopathological findings were remarkably lower compared to the C group. MDA levels were decreased and TAS levels were significantly increased in CVCM and NCM compared to the CM group. Although not statistically significant, levels of SOD increased in both group compared to CM group. View this table: Table 1 Conclusion: This study demonstrated the protective role of carvedilol and nebivolol against CIN. Antioxidant properties of these drugs may have an effect in this.

Study on a Single-Dose Toxicity Test of D-Amino Acid Oxidase (DAAO) Extracts Injected into the Tail Vein of Rats.

Objective:This study was performed to analyze the single-dose toxicity of D-amino acid oxidase (DAAO) extracts.Methods:All experiments were conducted at the Korea Testing & Research Institute (KTR), an institution authorized to perform non-clinical studies, under the regulations of Good Laboratory Practice (GLP). Sprague-Dawley rats were chosen for the pilot study. Doses of DAAO extracts, 0.1 to 0.3 cc, were administered to the experimental group, and the same doses of normal saline solution were administered to the control group. This study was conducted under the approval of the Institutional Animal Ethics Committee.Results:In all 4 groups, no deaths occurred, and the LD50 of DAAO extracts administered by IV was over 0.3 ml/kg.No significant changes in the weight between the control group and the experimental group were observed.To check for abnormalities in organs and tissues, we used microscopy to examine representative histological sections of each specified organ, the results showed no significant differences in any organs or tissues.Conclusion:The above findings suggest that treatment with D-amino acid oxidase extracts is relatively safe. Further studies on this subject should be conducted to yield more concrete evidence.

The Therapeutic Effects of Bone Marrow-Derived Mesenchymal Stem Cells and Simvastatin in a Rat Model of Liver Fibrosis

Liver fibrosis is the excessive accumulation of extracellular matrix (ECM) proteins including collagen that occurs in most types of chronic liver diseases. Studies concerning the capacity of mesenchymal stem cells (MSCs) and simvasatain (SIMV) to repair fibrotic tissues through reducing inflammation, collagen deposition, are still controversial. This study aimed to investigate the therapeutic efficacy of bone marrow (BM)-derived MSCs and SIMV on carbon tetrachloride (CCl4)-induced liver fibrosis in rats. Rats were divided into: normal, CCl4, CCl4/MSCs, CCl4/SIMV, CCl4/MSCs/SIMV, and SIMV groups. BM-derived MSCs were detected by RT-PCR of CD29 and were then infused into the tail vein of female rats that received CCl4 injection to induce liver fibrosis. Sex-determining region Y (SRY) gene on Y-chromosome gene was assessed by PCR to confirm homing of the male stem cells in liver tissue of the female recipients. Serum liver function tests, liver procollagens I and III, tissue inhibitors of metalloproteinase-1 (TIMP-1), endoglin, matrix metalloproteinase-1 (MMP-1) gene expressions, transforming growth factor-beta (TGF-β1) immunostaining, and histopathologicl examination were performed. MSCs and SIMV decreased liver procollagens I and III, TIMP-1 and endoglin gene expressions, TGF-β1 immunostaining, and serum liver function tests compared with the CCl4 group. MMP-1 expression was increased in the CCl4/MSCs group. Histopathological examination as well as fibrosis score supports the biochemical and molecular findings. It can be concluded that MSCs and SIMV were effective in the treatment of hepatic CCl4-induced fibrosis-rat model. Treatment with MSCs was superior to SIMV. This antifibrotic effect can be attributed to their effect on the MMPs/TIMPs balance which is central in fibrogenesis.

1. The anticonvulsant effects of n-3 PUFA: Animal studies

The omega-3 polyunsaturated acids (n-3 PUFA) show great promise as a therapy for epileptic seizures. There has been considerable confusion, however, in the animal data related to the n-3 PUFAs’ anticonvulsant effects. Our own recent work has clarified this confusion, and led to a simple, reproducible animal preparation in which to study the n-3 PUFAs’ mechanism(s) of action. We first attempted to replicate the studies of Yehuda et al. (1994), who had reported dramatic seizure suppression after 21 days of 40 mg/kg injections of the “SR 3” formula i.p in rats. In our replication, we found no effect at all at this dose (Taha et al., 2006). We therefore tested the higher dose of 200 mg/kg of the SR-3 mixture. At 200 mg/kg, we found a significant increase in PTZ seizure latency, although no change in severity (Taha et al., 2009). We subsequently tried acute administration of a single dose in rats, switching from SR 3 to docosahexaenoic acid (DHA). Injecting subcutaneously (s.c.), we found a significant elevation in PTZ-seizure latency following after a single injection (Taha et al., 2010). Our dose–response curves in this study, however, were “inverted U” shaped. In parallel studies, we tested dietary (p.o.) administration. Working in rats with chronically implanted electrodes, we found that dietary PUFAs elevated thresholds in both the cortex and the amygdala of rats. These effects, however, took several months to occur (Taha et al., in preparation). To simplify the kinetics, we have moved to intravenous (i.v.) injections of DHA (tail vein in rats). Our i.v. experiments have now shown anticonvulsant effects 5 min after injection and plasma concentrations that rise rapidly, plateau at 5 min, and drop to baseline a minute after infusion (Trepanier et al., in preparation). We have therefore finally evolved a preparation in which anticonvulsant effects and plasma concentrations parallel the known kinetics of DHA. This preparation will now allow a meaningful study of DHA’s mechanisms of anticonvulsant action.

Magnetic Resonance Guided High-Intensity Focused Ultrasound Mediated Hyperthermia Improves the Intratumoral Distribution of Temperature-Sensitive Liposomal Doxorubicin

The aim of this study was to investigate the intratumoral distribution of a temperature-sensitive liposomal carrier and its encapsulated compounds, doxorubicin, and a magnetic resonance (MR) imaging contrast agent after high-intensity focused ultrasound (HIFU)-mediated hyperthermia-induced local drug release. (111)In-labeled temperature-sensitive liposomes encapsulating doxorubicin and [Gd(HPDO3A) (H(2)O)] were injected intravenously in the tail vein of rats (n = 12) bearing a subcutaneous rhabdomyosarcoma tumor on the hind leg. Immediately after the injection, local tumor hyperthermia (2 × 15 minutes) was applied using a clinical 3 T MR-HIFU system. Release of [Gd(HPDO3A)(H(2)O)] was studied in vivo by measuring the longitudinal relaxation rate R(1) with MR imaging. The presence of the liposomal carriers and the intratumoral distribution of doxorubicin were imaged ex vivo with autoradiography and fluorescence microscopy, respectively, for 2 different time points after injection (90 minutes and 48 hours). In hyperthermia-treated tumors, radiolabeled liposomes were distributed more homogeneously across the tumor than in the control tumors (coefficient of variation(hyp, 90 min) = 0.7 ± 0.2; coefficient of variation(cntrl, 90 min) = 1.1 ± 0.2). At 48 hours after injection, the liposomal accumulation in the tumor was enhanced in the hyperthermia group in comparison with the controls. A change in R(1) was observed in the HIFU-treated tumors, suggesting release of the contrast agent. Fluorescence images showed perivascular doxorubicin in control tumors, whereas in the HIFU-treated tumors, the delivered drug was spread over a much larger area and also taken up by tumor cells at a larger distance from blood vessels. Treatment with HIFU hyperthermia not only improved the immediate drug delivery, bioavailability, and intratumoral distribution but also enhanced liposomal accumulation over time. The sum of these effects may have a significant contribution to the therapeutic outcome.

Effects of triterpenic acid from Prunella vulgaris L. on glycemia and pancreas in rat model of streptozotozin diabetes

Background The effects of triterpenic acid from Prunella vulgaris L. (TAP) on diabetes and its mechanism are uncertain. The aim of this study was to investigate the effects of TAP on antihyperglycemic, antioxidant, and pancreas-protective in streptozotozin (STZ)-diabetic rats. Methods The diabetic model was produced by injection of 60 mg/kg STZ. Blood was drawn from the tail vein of rats after 72 hours. Rats with blood glucose ≥16.7 mmol/L were considered diabetic. Diabetic rats were randomly divided into four groups: (1) Diabetes rat (STZ), (2) Diabetic rats treated with 50 mg/kg of triterpenic acid from Prunella vulgaris L (STZ+TAP50), (3) Diabetic rats treated with 100 mg/kg TAP (STZ+TAP100), and (4) Diabetic rats treated with 200 mg/kg TAP (STZ+TAP200). Normal rats (n=10) acted as the control group (NC). TAP was administered by the intragastric route once each day for six weeks. Body weight and the concentration of blood glucose (BG) were measured after three and six weeks. Fructosamine (FMN), malondialdehyde (MDA), and nitric oxide (NO), and the activities of nitric oxide synthase (NOS) and superoxide dismutase (SOD) in serum were determined after six weeks using commercially available kits following the manufacturer's instructions. Pathologic changes in pancreatic β-cells were also investigated by microscopic examination after hematoxylin-eosin (HE) staining. The level of SOD mRNA in pancreatic β-cells was measured by polymerase chain reaction (PCR). Results The levels of BG, FMN, NO, and MDA and the activities of NOS in serum in the four diabetes groups were significantly increased compared with the control group (P <0.01). The activity of SOD in serum and the body weight was significantly decreased compared with the control group (P <0.01). After administration of TAP to diabetic rats for six weeks, the body weight and the levels of BG, FMN, MDA, NO and the activity of NOS in serum decreased significantly compared with the STZ group in a dose-dependent manner. The activity of SOD in serum and body weight increased significantly compared with the STZ group in a dose-dependent manner. In addition, diabetic rats showed a significant decrease in SOD mRNA expression in pancreatic β cells. However, these changes were reversed by TAP. Histopathological examination also showed the protective effect of TAP on pancreatic β cells. Conclusions Triterpenic acid from Prunella vulgaris L. has an anti-diabetic effect, by controlling blood glucose and antioxidants, and has a protective effect on the pancreas.

β-Amyloid peptide (1–40) in the brain reaches the nasal cavity via a non-blood pathway

We examined the distribution patterns of human β-amyloid (1–40) peptide labeled with iodine 125 (125I-Aβ40) after injections into the cerebral ventricle or tail vein of rats. In rats receiving an intravenous injection, the radioactive concentration of 125I-Aβ40 in the nasal area was similar to other extracranial organs. In contrast, the caudal part of the nasal area in rats receiving an intracerebroventricular injection displayed a high level of 125I-Aβ40 radioactivity. These results suggest that Aβ reaches the nasal cavity from the brain via a non-blood pathway.

Effects of Magnetically Labeled Exogenous Endothelial Progenitor Cells on Cerebral Blood Perfusion and Microvasculature Alterations after Traumatic Brain Injury in Rat Model

Increasing evidence suggests that endothelial progenitor cells (EPCs), a subgroup of bone marrow hematopoietic stromal cells, play a critical role in neovascularization and tissue repair. To explore the effect of exogenous EPCs on the cerebral blood perfusion and microvessels in the injured region in rat model with traumatic brain injury (TBI). EPCs were collected from the spleens of healthy Sprague-Dawley rats. Fifty-four Sprague-Dawley rats were randomly divided into six groups. The controlled cortical impact TBI was performed. Spleen-derived exogenous EPCs labeled with super-paramagnetic iron oxide (SPIO) (SPIO-EPCs) were transplanted into the blood by tail vein of rats at 6 and 12 h after TBI, respectively. Magnetic resonance imaging (MRI) and computed tomography perfusion imaging were performed at various time points. Microvascular density was determined by immunohistochemistry. In SPIO-EPCs group, patchlike hypointensities were detected in the injured region at 24 h after transplantation, and the range of hypointensities tended to expand gradually over time on MRI, which was confirmed by Prussian blue staining. Computed tomography perfusion imaging parameters were gradually developed from hyperperfusion to normal, while, microvascular density was gradually increased during 72 to 168 h after injury. The values of these indices in SPIO-EPCs group were significantly lower than those in SPIO-alone group at the same time point, but no significant differences were found in different time groups. The intravenously transplanted EPCs diminish the brain injury through restoring cerebral blood perfusion and increasing the cerebral microvasculature in the injured region in rat model with TBI.

The role of NK1 receptor antagonist L‐703, 606 in the experimental traumatic brain injury

Background and purposeL‐703, 606 is a NK1 receptor antagonist. the purpose of the research is to explore the the role of L‐703, 606 in the experimental traumatic brain injury£TBI£©.Methods45 male Wistar rats were randomly divided into control group, L‐703, 606 intervention group (group L) ,TBI trauma group (group T). L‐703, 606 was injected into the tail vein of rats in group L after TBI. HE staining were used to observe the pathological changes in each group. The gene and protein expression of substance P(SP), calcitonin gene related peptide (CGRP) and nuclear transcription factor‐kappa B (NF‐¦ÊB) were detected by immunohistochemical staining and RT‐PCR.ResultsThe gene and protein expression of SP, CGRP and NF‐¦ÊB in rat cerebral cortical lesions of group L and group T was increased compared with that of control group. Above indicators of group L was significantly reduced compared with that of group T. Only mild congestion, a small amount of inflammatory cell infiltration showed in group L. (P <0.05)ConclusionL‐703, 606 has a protective effect on the brain injury in rats. The possible mechanism maybe reducing the inflammatory reaction induced by SP and CGRP which released from cerebral cortical lesion.

Deferoxamine preconditioning potentiates mesenchymal stem cell homing in vitro and in streptozotocin-diabetic rats

Objective: Today, cell therapy is considered a promising alternative in treatment of several diseases such as type 1 diabetes. Loss of transplanted stem cell and more importantly scarcity in the number of cells reaching to target tissue is a major obstacle in cell therapy. There is evidences showing that deferoxamine (DFO), an iron chelator, increases the mobilization and homing of progenitor cells through increasing the stability of hypoxia-inducible factor 1α (HIF-1α) protein. In this study, the effect of DFO on some factors involved in homing of bone marrow-derived mesenchymal stem cell was investigated, and the other objectives of this research were to determine whether DFO is able to increase migration and subsequent homing of mesenchymal stem cell (MSCs) both in vitro and in vivo in streptozotocin-diabetic rats.Research design and methods: MSCs were treated by DFO in minimal essential medium α (αMEM) for 24 h. The expression and localization of HIF-1α were evaluated by western blotting and immunocytochemistry. The expression of C-X-C chemokine receptor type 4 (CXCR-4) and chemokine receptor 2 (CCR2) were assessed by western blotting and RT-PCR. The activity of matrix metalloproteinases (MMP) -2 and -9 were measured by gelatin zymography. Finally, in vitro migration of MSCs toward different concentrations of stromal cell-derived factor and monocyte chemotactic protein-1 were also evaluated. To demonstrate the homing of MSCs in vivo, DFO-treated chloromethyl-benzamidodialkylcarbocyanine-labeled MSCs were injected into the tail vein of rats, and the number of stained MSCs reaching to the pancreas were determined after 24 h.Results: In DFO-treated MSCs, expression of HIF-1α (p < 0.001), CXCR4 (p < 0.001), CCR2 (p < 0.001), and the activity of MMP-2 (p < 0.01) and MMP-9 (p < 0.05) were significantly increased compared to control groups. Elevation of HIF-1α, upregulation of CXCR4/CCR2 and higher activity of MMP-2/MMP-9 in DFO-treated MSCs were reversed by 2-methoxyestradiol (2-ME; 5 μmol), a HIF-1α inhibitor. The in vitro migrations as well as in vivo homing of DFO-treated MSCs were also significantly higher than control groups (p < 0.05).Conclusions: Preconditioning of MSCs by DFO prior to transplantation could increase homing of MSCs through affecting some chemokine receptors as well as proteases involved and eventually improving the efficacy of cell therapy.

Enhancement of recombinant adeno-associated virus mediated transgene expression by targeted echo-contrast agent

Ultrasound-targeted microbubble destruction (UTMD) has been recently developed for destroying bubbles carrying drugs or genes, thereby permitting local release of these target molecules. We investigated whether SonoVue®, a new contrast agent that contains phospholipid-stabilized microbubbles filled with sulfur hexafluoride vapor, is effective at delivering a recombinant adeno-associated viral (rAAV) vector to the rat heart by UTMD. Serotype-2 (rAAV2) marked with green fluorescent protein (GFP) as a reporter gene was attached to the surface of sulfur hexafluoride-filled microbubbles. Microbubbles were infused into the tail vein of rats with or without simultaneous echocardiography. Additional controls included ultrasound microbubbles that did not contain virus, virus alone, and virus plus ultrasound. One group underwent echocardiographic destruction of microbubbles followed by rAAV2-GFP infusion. Rats were killed after 4 weeks and examined for GFP expression. Green fluorescence was detected in all groups that received the rAAV2-GFP vector, indicating expression of the rAAV2 transgene; however, GFP expression in the UTMD group was significantly higher than that in control groups. We conclude that ultrasound-mediated destruction mediated by SonoVue is a promising method for delivery of rAAV2 to the heart in vivo.

The Effect of Bone Marrow Transplantation on Oocyte-Granulosa Cell Interaction and Follicular Development of Cisplatin-Induced Ovarian Failure in Rat

Introduction: Chemotherapy has cytotoxic effect that induces follicular damage and abnormal folliculogenesisleads to ovarian failure. Two crucial growth factors in abnormal folliculogenesis, Growth Differentiation Factor-9 (GDF-9) and Kit-Ligand, will be disrupted and affect follicular development. In this study, we evaluate whether bone marrow transplantation (BMT) has a role on oocyte-granulosa cell interaction by analyzing GDF-9 and Kit-Ligand expressions and also follicular development by analyzing primordial, primary, secondary and graafian follicles of cisplatin-induced ovarian failure in rat. Material and method: Forty eight rats were divided into three groups: control, cisplatin and cisplatin+BMT. Ovarian failure was induced by administration of intraperitoneal cisplatin 5 mg/kg body weight for 1 week. BMT 2 × 107 cells were injected through rat tail vein after cisplatin administration. Bone marrow was isolated from rat femur and characterized by CD44(+), CD45(-), CD105(+). Immunohistochemistry examinations for ovarian GDF-9, Kit-Ligand and follicle development evaluation were performed after 2 weeks of BMT injection. Results: The expressions of Kit-Ligand among three groups by ANOVA were significant different (p=0.00), whereas by Post Hoc: cisplatin group lower than control group (p=0.00); cisplatin+BMT group higher than cisplatin group (p=0.00); and no significant different between control group and cisplatin+BMT group (p=0.955). The expressions of GDF-9 by Kruskal Wallis showed significant different (p=0.00) among three groups whereas cisplatin+BMT group higher than cisplatin group and control group. In cisplatin+BMT group the number of primordial, primary, secondary and graafian follicles were higher than those in cisplatin group; but were lower than those in control group (p=0.000). Positive Paul Karl Horan (PKH) labeling was seen in cisplatin+BMT group only. Conclusion: In cisplatin-induced ovarian failure in rat, bone marrow transplantation may improve oocytegranulosa cell interaction and follicular development.

Serial MR Imaging of Magnetically Labeled Humen Umbilical Vein Endothelial Cells in Acute Renal Failure Rat Model

Purpose : To evaluate the usefulness of in vivo magnetic resonance (MR) imaging for tracking intravenously injected superparamagnetic iron oxide (SPIO)-labeled human umbilical vein endothelial cells (HUVECs) in an acute renal failure (ARF) rat model. Materials and Methods: HUVECs were labeled with SPIO and poly-L-lysine (PLL) complex. Relaxation rates at 1.5-T MR, cell viability, and labeling stability were assessed. HUVECs were injected into the tail vein of ARF rats (labeled cells in 10 rats, unlabeled cells in 2 rats). Follow-up serial T2*-weighted gradient-echo MR imaging was performed at 1, 3, 5 and 7 days after injection, and the MR findings were compared with histologic findings. Results: There was an average of 98.4±2.4% Prussian blue stain-positive cells after labeling with SPIOPLL complex. Relaxation rates (R2*) of all cultured HUVECs at day 3 and 5 were not markedly decreased compared with that at day 1. The stability of SPIO in HUVECs was maintained during the proliferation of HUVECs in culture media. In the presence of left unilateral renal artery ischemia, T2*-weighted MR imaging performed 1 day after the intravenous injection of labeled HUVECs revealed a significant signal intensity (SI) loss exclusively in the left renal outer medulla regions, but not in the right kidney. The MR imaging findings at days 3, 5 and 7 after intravenous injection of HUVECs showed a SI loss in the outer medulla regions of the ischemically injured kidney, but the SI progressively recovered with time and the right kidney did not have a significant change in SI in the same period. Upon histologic analysis, the SI loss on MR images was correspondent to the presence of Prussian blue stained cells, primarily in the renal outer medulla. Conclusion: MR imaging appears to be useful for in vivo monitoring of intravenously injected SPIO-labeled HUVECs in an ischemically injured rat kidney. Index words : Magnetic resonance imaging (MRI)∙Superparamagnetic iron oxide (SPIO) Human umbilical vein endothelial cells (HUVECs)∙Cell tracking

American Pancreatic Association

American Pancreatic Association

Hypoxic preconditioning enhances the therapeutic potential of the secretome from cultured human mesenchymal stem cells in experimental traumatic brain injury

Bone-marrow-derived human MSCs (mesenchymal stem cells) support repair when administered to animals with TBI (traumatic brain injury) in large part through secreted trophic factors. We directly tested the ability of the culture medium (or secretome) collected from human MSCs under normoxic or hypoxic conditions to protect neurons in a rat model of TBI. Concentrated conditioned medium from cultured human MSCs or control medium was infused through the tail vein of rats subjected to TBI. We have demonstrated that MSCs cultured in hypoxia were superior to those cultured in normoxia in inducing expression of both HGF (hepatocyte growth factor) and VEGF (vascular endothelial growth factor) in the cultured medium. We showed further that rats treated with the secretome from both normoxic- and hypoxic-preconditioned MSCs performed significantly better than the controls in both motor and cognitive functional test. Subsequent post-mortem evaluation of brain damage at the 4-day time point confirmed that both normoxic- and hypoxic-preconditioned MSC secretome-treated rats had significantly greater numbers of newly forming neurons, but significantly less than the controls in brain damaged volume and apoptosis. The TBI rats treated with hypoxic-preconditioned MSC secretome performed significantly better in both motor and cognitive function tests and neurogenesis, and had significantly less brain damage than the TBI rats treated with the normoxic-preconditioned MSC secretome. Collectively, these findings suggest that MSCs secrete bioactive factors, including HGF and VEGF, that stimulate neurogenesis and improve outcomes of TBI in a rat model. Hypoxic preconditioning enhances the secretion of these bioactive factors from the MSCs and the therapeutic potential of the cultured MSC secretome in experimental TBI.

In-vivo pharmacokinetics, tissue distribution and anti-tumour effect of hydroxycamptothecin delivered in oil-in-water submicron emulsions

The aim of this study was to investigate the pharmacokinetics, tissue distribution and anti-tumour effect of hydroxycamptothecin submicron emulsions (HCPT-SEs). HCPT-SEs or HCPT injection (HCPT-I) was administered intravenously into the tail vein of rats or S180 tumour-bearing mice. HCPT-SEs increased the plasma concentration of HCPT compared with HCPT-I at all time points. The AUC(0-∞) , elimination half-life and mean residence time of anionic submicron emulsions containing HCPT (HCPT-ASEs) and cationic submicron emulsions containing HCPT (HCPT-CSEs) were significantly greater than those of HCPT-I (P <0.01). Especially, a prolonged elimination half-life was found for HCPT-CSEs. HCPT-CSEs and HCPT-ASEs resulted in a 7.9-fold and 3.1-fold increase in AUC(0-6h) of tumour compared with HCPT-I, respectively. The targeting efficiency (T(e) ) of HCPT-ASEs and HCPT-CSEs indicated their selectivity to tumour and the T(e) of HCPT-CSEs was significantly higher than that of HCPT-ASEs (P<0.01). The anti-tumour effect studies showed that HCPT-SEs improved the therapeutic efficiency of HCPT compared with HCPT-I. The percentage of tumour growth suppression rate of mice treated with HCPT-CSEs (2.0 mg HCPT eq./kg) increased 2.1 fold compared with that of HCPT-I. Submicron emulsions can alter the pharmacokinetic characteristics and tissue distribution of HCPT, and enhance tumour targeting and anti-tumour activity.

Can Leptin-Derived Sequence-Modified Nanoparticles Be Suitable Tools for Brain Delivery?

In order to increase the knowledge on the use of nanoparticles (NPs) in brain targeting, this article describes the conjugation of the sequence 12-32 (g21) of leptin to poly-lactide-co-glycolide NPs. The capability of these modified NPs to reach the brain was evaluated in rats after intravenous administration. The g21 was linked on the surface of NPs labeled with tetramethylrhodamine by means of the Avidin-Biotin technology. The g21-labeled NPs were injected into the tail vein of rats and, after animal sacrifice, the brain localization was evaluated by confocal microscopy, fluorescence microscopy and electron microscopy. Studies to evaluate the biodistribution of the g21-modified NPs in comparison to the unmodified NPs were also carried out. Moreover, to confirm the absence of any anorectic effect of g21 linked on the surface of NPs, appropriate studies were used to assess the rats. After intravenous administration, the g21-modified NPs were able to cross the blood-brain barrier and to enter the brain parenchyma. The biodistribution studies of both unmodified and modified NPs pointed out an uptake at liver and spleen level, whereas only the g21-modified NPs showed brain localization. The food-intake experiments pointed out that the intravenous administration of g21 conjugated to the NP surface did not produce any anorectic effect in the rats. g21-modified NPs were able to cross the blood-brain barrier. These new modified NPs could be effectively considered as useful carrier systems for brain drug delivery.