Endothelial Progenitor Cells Group Research Articles

Dickkopf-1 Treatment Stimulates Hematopoietic Regenerative Function in Infused Endothelial Progenitor Cells.

Acute high-dose radiation injury damages the bone marrow hematopoietic stem and progenitor cell compartment. This damage compromises the functional ability of the bone marrow to produce mature blood cells and results in an increased risk of death due to hematopoietic complications. Past work has shown that the bone marrow endothelium provides critical cues, which promote hematopoietic stem cell regeneration after injury. Additionally, transfusion of endothelial cells after radiation injury has been shown to promote recovery of both the bone marrow vasculature and hematopoietic systems. In this work, we examined the regenerative capacity of intravenous infusion of umbilical cord-blood derived endothelial progenitor cells (EPCs) since this is a cell source which is easy to obtain, expand and cryopreserve. We show that pre-treatment with the Wnt-antagonist Dickkopf1 (Dkk1) augments EPC regenerative function in an allogeneic mouse transplant model. Here, hematopoietic recovery was assessed in Balb/c mice after 5 Gy total-body irradiation and transplantation with C57/BL6-derived EPCs either with or without Dkk1 pre-treatment. The Dkk1-treated EPC group had significantly faster recovery of peripheral white blood cells, total bone marrow cellularity, bone marrow progenitors and BM endothelial cells compared to EPC treatment alone or saline controls. Importantly, after an LD50/30 dose of 8 Gy in the Balb/c mouse, Dkk1-treated EPCs were able to rescue 100% of irradiated mice versus 80% in the EPC control group and only 33% in the saline-treated group. To understand how Dkk1 induces regenerative function in the EPCs, we screened for pro-regenerative factors secreted by the EPC in response to Dkk1. Dkk1-treated EPCs were observed to secrete high levels of the anti-fibrotic protein follistatin as well as several proteins known to promote regeneration including EGF, VEGF and G-CSF. This work demonstrates the potential for Dkk1-treated EPCs as a rescue therapeutic for victims of acute radiation injury.

Clinical Assessment of Intravenous Endothelial Progenitor Cell Transplantation in Dogs

Endothelial progenitor cells (EPCs) have been applied for cell therapy because of their roles in angiogenesis and neovascularization in ischemic tissue. However, adverse responses caused by EPC therapy have not been fully investigated. In this study, a human peripheral blood sample was collected from a healthy donor and peripheral blood mononuclear cells were separated using Ficoll-Hypaque. There were four experimental groups: 10 ml saline infusion group (injection rate; 3 ml/min), 10 ml saline bolus group (injection rate; 60 ml/min), 10 ml EPCs infusion group (2 x 105 cells/ml, injection rate; 3 ml/min), 10 ml EPCs bolus group (2 × 105 cells/ml, injection rate; 60 ml/min). Clinical assessment included physical examination and laboratory examination for intravenous human EPC transplantation in dogs. The results revealed no remarkable findings in vital signs among the dogs used. In blood analysis, platelet counts in saline infusion groups were significantly higher than in the EPC groups within normal ranges, and no significant differences were observed except K+, Cl– and blood urea nitrogen/urea. In ELISA assay, no significant difference was observed in serum tumor necrosis factor alpha. The serum concentration of vascular endothelial growth factor was significantly higher in EPC groups than in saline groups, and interleukin 10 was significantly up-regulated in the EPC infusion group compared with other groups. In conclusion, we demonstrated that no clinical abnormalities were detected after intravenous transplantation of human EPCs in dogs. The transplanted xenogenic EPCs might be involved in anti-inflammatory and angiogenic functions in dogs.

17β-estradiol promotes recovery after myocardial infarction by enhancing homing and angiogenic capacity of bone marrow-derived endothelial progenitor cells through ERα-SDF-1/CXCR4 crosstalking.

17β-estradiol (E2) has been shown to mediate endothelial progenitor cells (EPCs) to repair infarcted myocardium. Both estrogen receptor α (ERα) and stromal derived factor-1 (SDF-1)/CXCR4 signaling pathways may play a critical role in regulating homing and angiogenesis of EPCs in this process. However, the interaction between ERα and SDF-1/CXCR4 signaling pathways remains unclear. In response to E2, the expression of SDF-1 and CXCR4 in EPCs from ovariectomized BALB/C mice was obviously up-regulated, in addition, the migration and tube formation of EPCs in vitro were also significantly enhanced. However, ERα antagonist (MMP) and CXCR4 inhibitor (AMD3100) significantly decreased the migration and tube length of EPCs, even if mediated by E2. The combined treatment of MMP and AMD3100 exerted more inhibitory effects on migration and tube formation of EPCs induced by E2. In in vivo studies, ovariectomized mice were induced acute myocardial infarction (AMI), and divided into four groups (n = 6): non-preconditioned EPCs (3 × 106) group, E2-preconditioned EPCs group, MMP + AMD3100 preconditioned EPCs group, and EPCs pretreated with E2 + MMP + AMD3100 group. E2 group displayed a greater number of homing EPCs, increased capillary density in infarcted myocardium, decreased left ventricular (LV) fibrosis. Nevertheless, these effects of E2 were almost completely blocked by the combined treatment of MMP and AMD3100. E2 can produce cardiovascular protective effects in AMI setting by enhancing homing and angiogenic capacity of EPCs through ERα and CXCR4 signaling pathways, which means that ERα and CXCR4 pathways are effective targets for the development of treatment strategies for AMI.

Autologous Endothelial Progenitor Cells Transplantation for Acute Ischemic Stroke: A 4-Year Follow-Up Study.

Transplantation of endothelial progenitor cells (EPCs) is a proven safe and effective method for treatment of cerebral ischemia in animal experiments. However, safety and efficacy need to be determined in clinical trials. We performed a two‐center, randomized, placebo‐controlled phase I/IIa trial with blinded outcome assessment on 18 patients with acute cerebral infarct within the middle cerebral artery territory, and followed for up to 4 years. Autologous ex vivo expanded EPCs were injected intravenously in the EPC group, and patients who received saline or autologous bone marrow stromal cells served as control groups. Mortality of any cause, adverse events, and new‐onset comorbidities were monitored. Changes in neurological deficits were assessed at different time points. We found no toxicity events or infusional or allergic reactions in any treated group. Three patients in the placebo group died during the 4‐year follow‐up. We found that the EPC group had fewer serious adverse events compared with the placebo‐controlled group, although there were no statistical differences in mortality among the three groups. Furthermore, there was no significant difference in neurological or functional improvement observed among the three groups, except for the Scandinavia Stroke Scale score at 3 months between the EPC group and placebo‐controlled group. Autologous transplantation of EPCs appears to improve long‐term safety in acute cerebral infarct patients, supporting the feasibility of this novel method for treatment of ischemic stroke (ClinicalTrials.gov: NCT01468064). Stem Cells Translational Medicine 2019;8:14–21

KAI1/CD82 Genetically Engineered Endothelial Progenitor Cells Inhibit Metastasis of Human Nasopharyngeal Carcinoma in a Mouse Model.

BackgroundEndothelial progenitor cells (EPCs) are regarded as promising targeted vectors for delivering therapeutic genes or agents in cancer therapy. The purpose of this study was to investigate the role of intravenously administered KAI1/CD82 genetically transduced EPCs in the tumorigenesis and metastasis of nasopharyngeal carcinoma (NPC).Material/MethodsEPCs were isolated from human umbilical cord blood, expanded in culture, and stably transduced with lentiviral vectors expressing KAI1/CD82. The KAI1/CD82 EPCs were injected intravenously into nude mice bearing human NPC xenografts. Tumor growth and the incidence of liver and lung metastases were observed. Expression of KAI1/CD82 was determined by immunofluorescent staining.ResultsThe NPC model was successfully established. Tumor growth was not suppressed when mice were injected with KAI1/CD82 EPCs (KAI1/CD82 EPCs group) compared with when non-transduced EPCs was present (EPCs group) or the control (1.485±0.234, 1.388±0.204, and 1.487±0.223g, respectively; P>0.05). However, the incidence of lung metastasis was significantly reduced in the KAI1/CD82+ EPCs group compared with the EPCs group and the control group (10%, 55% and 45%, respectively; P=0.005), and there was a significant decrease in the number of metastatic foci on the lung surface (17.50±3.54, 34.27±5.35, and 38.44±9.63 respectively; P=0.007). Moreover, KAI1/CD82 was expressed in lung metastatic foci of the KAI1/CD82 EPCs group, but not in the EPCs group and control group.ConclusionsEPCs can be used as a delivery vehicle for suppressor genes KAI1/CD82 to NPC, and the migration of KAI1/CD82 genetically engineered EPCs can inhibit NPC lung metastasis in a mouse model.

The mechanism of bone marrow-derived endothelial progenitor cells in the treatment of peripheral neuropathy of diabetic rats

Objective To explore the mechanism of bone marrow-derived endothelial progenitor cells(EPCs)in the treatment of peripheral neuropathy of diabetic rats. Methods Rats with diabetic peripheral neuropathy(DPN)were induced by streptozotocin(60 mg/kg). Male SD rats were divided into normal control group(NC group), DPN group, DPN+ saline group(DPN+ S group), and DPN+ EPCs group. Sciatic nerve motor nerve conduction velocity(MNCV)was measured. The expressions of NF-κB and myelin basic protein(MBP)in sciatic nerve were detected by immunohistochemistry. Results Compared with DPN group, the expression of NF-κB was reduced in the sciatic nerve of DPN+ EPCs group, while the expression of NF-κB was increased in the sciatic nerve of DPN+ S group. There was no statistical difference in the expression of MBP between DPN and DPN+ EPCs groups. Compared with DPN+ S group, the expression of MBP was higher in DPN+ EPCs group. Conclusion Transplantation of EPCs inhibits the expression of NF-κB and increases the expression of MBP, which might be conducive to the repairs of nerve injury. (Chin J Endocrinol Metab, 2018, 34: 312-317) Key words: Endothelial progenitor cells; Diabetic rats; Diabetic peripheral neuropathy

Effect of Moxibustion on Delayed Memory and Expression of Hippocampal Nestin and Doublecortin Proteins in Dementia Rats

To observe the effect of "Huayu Tongluo"(Blood-stasis Dispersing and Meridian-collateral Dredging) moxibustion on the delayed memory and expression of Nestin and Doublecortin (DCX) proteins in the hippocampus in vascular dementia (VD) rats in the view of neurogenesis produced by intracerebral transplantation of neural stem cells (NSCs) and endothelial progenitor cells (EPCs). Healthy male Wistar rats were randomized into control group, VD model group,NSCs＋EPCs group and NSCs＋EPCs moxibustion group. The VD model was established by using a modified 2-vessels occlusion method, and neurogenesis was produced by transplantation of NSCs＋EPCs (2×106cell/10 µL) into the lateral ventricle for rats of the NSCs＋EPCs groups 3 days after successful VD-modeling. Moxibustion was applied to "Dazhui" (GV 14), "Baihui" (GV 20) and "Shenting" (GV 24) once daily for 21 days with an interval of one day between every two 7 days. The Morris Water Maze was used to test the rat's delayed memory ability before and 24 h after the treatment. The expression of Nestin and DCX proteins in the hippocampus tissues was detected using double-labeled immunofluorescence technique. Following modeling, Morris Water Maze tests showed that the average escape latency of location navigation task was significantly prolonged in VD rats(P<0.008)and the times of target platform crossing (spatial probing task) within 120 s were remarkably reduced in VD rats (P<0.008). Compared with pre-treatment in the same one group, the escape latency of NSCs＋EPCs and NSCs＋EPCs moxibustion groups were considerably reduced (P<0.05), and the average times of target platform crossing of the NSCs＋EPCs moxibustion group were markedly increased(P<0.05). The effect of NSCs＋EPCs moxibustion was evidently superior to that of simple NSCs＋EPCs in shortening the escape latency (P<0.008). The expression levels of Nestin protein were significantly higher in the NSCs＋EPCs moxibustion group after 1 and 3 period treatment than those in the NSCs＋EPCs group (P<0.05)．. Moxibustion intervention is able to improve the delayed memory in VD rats, which may be related to its effect in up-regulating the expression of hippocampal Nestin and DCX proteins within 15 days via accelerating neurogenesis.

Effect of co-culture human endothelial progenitor cells with porcine oocytes during maturation and subsequent embryo development of parthenotes in vitro.

Human endothelial progenitor cells (EPCs) have been applied to regenerative medicine for their roles in angiogenesis as well as neovascularization, and these angiogenetic functions have beneficial effects on maturation of ovarian follicles. However, little information is available on whether EPCs on culture systems affect oocyte maturation and subsequent embryo development. Therefore, the objective of this study was to investigate the effect of EPC co-culture on porcine oocytes during in vitro maturation (IVM) and subsequent embryo development, and to examine gene expression in cumulus cells, oocytes and blastocysts. The effect of co-culture using EPC on porcine oocyte IVM was investigated. Oocytes were activated using electrical stimulation and embryo developmental competence was estimated. The expression of the genes related to cumulus expansion, oocyte maturation, embryo development, and apoptosis were analyzed. In result, there was a significantly increased maturation rate in EPC group compared with control (p < 0.05). Also, oocytes co-cultured with EPCs exhibited significantly improved blastocyst formation rates (p < 0.05). The expression of mRNAs associated with cumulus expansion and apoptosis in cumulus cells was significantly up-regulated in EPC group. Also, markedly increased levels of GDF9, BMP15, and BCL2 were observed in oocytes from the EPC group. Blastocysts in the co-culture group showed significantly higher SOX2, OCT4, and NANOG levels. In conclusion, co-culturing porcine oocytes with EPCs improves their maturation by regulating genes involved in cumulus cell expansion, oocyte maturation, and apoptosis. Moreover, EPC co-culture during IVM enhanced embryo development as shown by increased blastocyst formation rate and pluripotency-related gene expression.

Endothelial progenitor cell miR-126 promotes homing of endothelial progenitor cells within arterial thrombus in patients with cerebral infarction and its molecular mechanism.

The present study was designed to investigate the effect of microRNA-126 (miR-126) on the migration and homing of endothelial progenitor cells (EPCs) within arterial thrombus of cerebral infarction patients. EPCs from rat bone marrow were isolated, and miR-126 overexpressed EPCs were constructed by lentiviral transfection. Then, the middle cerebral artery occlusion (MCAO) model was established by the method of thread ligation. Successfully established model rats were randomly divided into miR-126 overexpression EPC group, miR-126 wild type EPC group, and normal saline group. One day after the infarction, the miR-126 overexpression EPCs, miR-126 wild type EPCs, and normal saline, were injected into the lateral ventricle of the corresponding groups. Also, the transplanted cells were tracked by cell dye SPDiIC18. The expression of tight junction proteins ZO-1 and Claudin-5 in brain tissue was detected by Western blotting. Transplanted cells were detected in the cerebral infarction area 3 days after transplantation by cell dye SP-DiIC18. The number of homing EPCs in miR-126 overexpression group was significantly higher than that of miR-126 wild type EPC group (p < 0.05). Also, the protein expression of ZO-1 and Claudin-5 in the miR- 126 overexpression EPC group was significantly higher compared with that of the miR-126 wild type EPC group and the normal saline group. miR-126 overexpression EPCs, which were transplanted in the lateral ventricle, can home to the cerebral infarction areas via increasing increase.

Local transplantation of human thrombomodulin gene-modified endothelial progenitor cells to prevent restenosis after angioplasty

Objective To investigate whether restenosis can be inhibited by human thrombomodulin (hTM) gene-modified endothelial progenitor cells (EPCs) after angioplasty. Methods EPCs derived from rabbit periphery blood were modified with hTM gene using lentiviral vector and were incubated with Fe2O3-arginine. Thirty-six New Zealand rabbits were randomly divided into 3 groups. Angioplastywas underwent by Fogarty ballooncatheter via catheterization of right common carotid artery, and then hTM-EPCs, EPCs or equal volume of normal saline were transplanted to the injury sites with blood flow blocked respectively. Histology and Western blotting were performed on 1th week for 4 rabbits from each group to evaluate cell adherence. Magnetic resonance (MR) was performed on 4th and 8th week after transplantation to evaluate restenosis. All of the rest rabbits were sacrificed at the end of 8th week after injury and the injured vessels were harvested for hematoxylin and eosin (HE), elastic tissue and proliferating cell nuclear antigen (PCNA) staining to observe the vascular wall structure and degree of intimal hyperplasia and smooth muscle proliferation. Results Prussian blue staining showed iron-labeled cells distributed in the injured artery intima. Western blotting indicated that injured vascular transplanted with hTM-EPCs overexpressed thrombomodulin than other two groups. MR showed that the stenotic degree of the lumen in hTM-EPCs group was less serious than other two groups. The histopathological study on 8th week showed that the neointimal/media ratio (N/M) in hTM-EPCs group, EPCs group and control group was 0.348±0.040, 0.704±0.060 and 1.294±0.051, respectively (P=0.000); the ratio of restenosis in each group was (11.4±2.3)%, (24.3±4.8)% and (35.2±9.4)%, respectively (P=0.005); and the PCNA positive cell index in each group was (12.3±3.9)%, (23.4±4.2)% and (49.3±6.8)%, respectively (P=0.000). Conclusion The local transplantation of hTM-EPCs can prevent restenosis through Fogarty balloon after angioplasty in rabbit model. Key words: Thrombomodulin; Endothelial progenitor cells; Balloon injury; Restenosis

The Therapeutic Value of Bone Marrow-Derived Endothelial Progenitor Cell Transplantation after Intracerebral Hemorrhage in Rats.

To study the effect of endothelial progenitor cell (EPC) treatment on intracerebral hemorrhage (ICH) in rats and elucidate possible mechanisms. The rats were randomly divided into three groups: (1) EPC group: ICH + EPC, (2) phosphate-buffered saline group: ICH + PBS, and (3) sham group. EPCs were transplanted intravenously 6 h after ICH. Modified neurological severity score was used to evaluate neurological function. Blood-brain barrier (BBB) integrity was evaluated. Dead cells, inflammatory cytokines, and neuroprotective cytokines were assessed to investigate possible mechanisms. The animals in the EPC group showed significant improvement in neurological function at 48 h, 72 h, and 7 days after ICH, compared with those in the PBS group. EPC transplantation significantly reduced brain edema and the number of dead cells in the hematoma boundary areas. The intensity of Evans Blue was decreased, and expression levels of zonula occluden-1 and claudin-5 were increased in the EPC group. Proinflammatory cytokines, including interferon-γ, IL-6, and TNF-α, were decreased, whereas anti-inflammatory cytokines, including transforming growth factor-β1 and IL-10, were increased in the EPC group. In addition, expression levels of brain-derived neurotrophic factor, vascular endothelial growth factor, and neurotrophic growth factor were increased following transplantation of EPCs. EPC transplantation could improve neurological function of ICH rats. The protective effect may be mediated by promotion of neuroprotective cytokine secretion, restoration of the BBB, reduction of cell death, and the decrease in inflammation.

Effects of allogeneic hematopoietic stem cell transplantation in combination with infusion of endothelial progenitor cells on bone marrow inflammatory injury

目的探索异基因造血干细胞移植联合内皮祖细胞（EPC）输注对骨髓炎性损伤的影响。方法6~8周龄BALB/c（H-2Kd）小鼠经致死剂量全身照射（TBI）预处理后建立异基因骨髓移植模型（BMT组），并在BMT基础上同时输注体外培养的EPC，建立骨髓联合EPC移植模型（EPC组）。移植后第7、14、21、28天分别获取各组小鼠骨髓细胞，流式细胞术检测EPC培养比率及两组嵌合率变化，通过骨髓细胞和外周血细胞计数比较两组骨髓造血情况，Western blot法检测骨髓巨噬细胞表面特异性标志CD68表达水平；实时定量PCR及Western blot法分析骨髓炎性复合体nlrp1、nlrp6、caspase-1 mRNA及蛋白表达水平。结果移植后第7天BMT组嵌合率为（83.69±1.26）%，明显低于EPC组的（91.65±2.77）%（P<0.05）。在移植后第10、15、20天，EPC组小鼠外周血中的WBC、网织红细胞、PLT均高于BMT组（P值均<0.05），在第7、21天EPC组骨髓中原始红细胞较BMT组高（P值均<0.05）。HE染色显示，移植后第7、14、21天，EPC组骨髓微环境恢复较BMT组快。Western blot检测显示EPC组骨髓CD68蛋白表达水平较BMT组低。EPC组nlrp1、nlrp6、casepase-1 mRNA及蛋白表达水平在移植后各时间点接近正常表达水平，与BMT组相比，差异均有统计学意义（P值均<0.05）。结论异基因造血干细胞移植联合输注EPC可减少炎性细胞的浸润及减少炎性复合体的活化，促进骨髓造血重建。

Transplantation of Amniotic Scaffold-Seeded Mesenchymal Stem Cells and/or Endothelial Progenitor Cells From Bone Marrow to Efficiently Repair 3-cm Circumferential Urethral Defect in Model Dogs.

The treatment options for patients with a urethral defect are limited by the availability of autologous tissues. We hypothesized that transplantation of decellularized human amniotic scaffolds (dHAS) seeded with allogeneic bone marrow mesenchymal cells (BMSCs) and/or endothelial progenitor cells (EPCs) may serve as a promising repair strategy for long segment of circumferential urethral defect. To verify the hypothesis, with urinary catheterization, a 3-cm segment of whole urethra in 25 male mongrel dogs was excised and replaced by dHAS seeded with allogeneic BMSCs and/or EPCs. Postoperative observation and ascending urethrogram found that dHAS+BMSCs+EPCs and dHAS+EPCs groups demonstrated unhindered urination and capacious urethral caliber, which were similar to the normal group, while urethrostenosis was revealed in dHAS+BMSCs, dHAS, and sham-operated groups, with the shortest narrow section in dHAS+BMSCs group and the longest in sham-operated group. Urethral anatomy check and histological analyses showed that new urethral mucosa composed of stratified columnar epithelium completely covered on the inner surface of the graft site in dHAS+BMSCs+EPCs and dHAS+EPCs groups, but the middle epithelium was thin in dHAS+EPCs group, while incompletely covered in dHAS+BMSCs, dHAS, and sham-operated groups, and there were monolayer epithelial cells at the urethrostenosis in dHAS+BMSCs and dHAS groups. In addition, abundant new vessel and blood sinus showed at submucosa in dHAS+BMSCs+EPCs and dHAS+EPCs groups, instead of the scar tissue of collagen deposition and structural distortion at the urethrostenosis in dHAS+BMSCs, dHAS, and sham-operated groups. This study demonstrates that dHAS seeded with BMSCs+EPCs or EPCs can successfully repair a 3-cm circumferential urethral defect in model dogs, but the former works best. This technology may provide some references for human clinical trials on long segment of circumferential urethral defect repair.

Hydrogen Sulfide-Preconditioning of Human Endothelial Progenitor Cells Transplantation Improves Re-Endothelialization in Nude Mice with Carotid Artery Injury

Background/Aims: The aim of present study was to test the hypothesis that preconditioning with sodium hydrosulfide (NaHS) could enhance the capacity of migration, adhesion and proliferation of endothelial progenitor cells (EPCs) in vitro, and also could improve the efficacy of EPCs transplantation for re-endothelialization in nude mice with carotid artery injury. The paper further addressed the underlying mechanisms. Methods: EPCs were isolated from peripheral blood mononuclear cells of healthy male volunteers and the markers of EPCs were analyzed by flow cytometry. Thereafter, different concentrations of NaHS (25, 50, 100, 200 and 500 uM) were used for preconditioning EPCs. In vitro and in vivo migration, adhesion and proliferation as well as nitric oxide (NO) production of EPCs were evaluated. Carotid artery injury model was produced in nude mice and thereafter, NaHS-preconditioned EPCs were transplanted in order to evaluate their capacity of re-endothelialization. Results: Cellular immuno-staining showed that isolated cells expressed the key markers of EPCs. In vitro, EPCs proliferation rates and NO production were gradually increased in a NaHS-concentration dependent manner, while these benefits were blocked at a concentration of 500 uM NaHS. Similarly, the migration and adhesion rates of EPCs were also increased the most prominently at a concentration of 200 µM NaHS. In vivo, compared to the control group, treatment with NaHS-preconditioned EPCs significantly enhanced the capacity of re-endothelialization of EPCs. Fluorescent microscope revealed that there were more EPCs homing to the injury vessels in the NaHS-preconditioned EPCs group than the non-preconditioned group. With the administration of AMPK or eNOS inhibitors respectively, the above benefits of NaHS-preconditioning were abrogated. Conclusion: These results suggested that NaHS-preconditioning enhanced the biological function and re-endothelialization of EPCs through the AMPK/eNOS signaling pathway.

105 Does Neoadjuvant Androgen Deprivation Therapy Impact Erectile Function Recovery Post-Radical Prostatectomy?

INTRODUCTION AND OBJECTIVES: Erectile dysfunction (ED) is one of the most common complications of diabetes mellitus (DM). The efficacy of treatments for diabetes mellitus-induced erectile dysfunction (DMED) is quite poor, and stem cell therapy is emerging as a useful method. This study evaluated the possibility and mechanisms of DMED treatment with endothelial progenitor cells (EPCs) genetically modified to express human telomerase reverse transcriptase (hTERT). METHODS: Rat EPCs were isolated and transfected with hTERT (EPCs-hTERT). Proliferation capacity, intracellular reactive oxygen species, paracrine character and resistance to oxidative stress of EPCs-hTERT were assessed in vitro. Diabetes was induced via an intraperitoneal injection of streptozotocin. Twenty-four male DMED rats were subjected to four treatments: DMED (DMED group), EPCs (EPCs group), EPCs transfected with control lentivirus (EPC-control group) and EPCs-hTERT (EPCs-hTERT group). A group of health rats were used as a normal control group. Intracavernosal pressure (ICP) was measured using electrical stimulation for each group after 2 weeks of EPC injections. Masson’s trichrome staining was performed to determine collagen content in penile tissues. The degree of apoptosis was assessed using western blotting analysis for terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) signaling pathway was detected. RESULTS: EPCs-hTERT secreted more growth factors than EPCs and EPCs-control. EPC resistance to oxidative stress and proliferation was dramatically improved by hTERT transfection. The ICP/ mean arterial pressure (MAP) ratio induced by electrical stimulation was markedly increased in the EPCs-hTERT group compared with the EPCs and EPCs-control group. Immunofluorescence demonstrated increased cell survival in penile tissues after EPCs-hTERT implantation. The smooth muscle/collagen ratio was markedly increased in the EPCshTERT group. TGF-b1 and p-Smad expression decreased significantly in DMED rats after EPCs-hTERT treatment. The degree of apoptosis in penile tissues of the EPCs-hTERT group was considerably reduced, as demonstrated by the decreased Bax/Bcl-2 ratio. NOS expression and NO and cGMP concentrations increased significantly in the EPCshTERT group. CONCLUSIONS: The enhanced paracrine effect, resistance to oxidative stress and EPCs-hTERT proliferation contributed to the improvement of erectile function in DMED rats.

Effect of endothelial progenitor cells derived from human umbilical cord blood on oxygen-induced retinopathy in mice by intravitreal transplantation.

To investigate the effect of endothelial progenitor cells (EPCs) labeled by carboxy fluorescein diacetate succinimidyl ester (CFSE) on murine oxygen-induced retinopathy (OIR) by intravitreal transplantation. After isolated from human umbilical cord blood mononuclear cells, EPCs were cultivated and then labeled with CFSE in vitro. C57BL/6J mice were placed to 75% hyperoxia chamber from P7 to P12 to establish OIR model. At P12, OIR mice were intravitreally injected with 1 µL suspension contained 2×105 EPCs (EPCs group) or isometric phosphate buffered saline (PBS group). The contralateral eye of each mice received no injection (OIR group). Evans blue angiography and frozen section were examined to track the labeled cells in OIR group at P15 and P19. Using retina paraffin sections and adenosinediphos phatase staining at P12 and P19, the effect of EPCs on OIR mice was evaluated quantitatively and qualitatively. The retinas from EPCs group with less non-perfusion area and fewer peripheral tufts were observed at P19, comparing with that from PBS or OIR group. The retinopathy in EPCs group receded earlier with less non-ganglion cells and neovascular nuclei, together with relatively regular distribution. The counts of the neovascular nuclei at P19 were reduced by 44% or 45%, compared with those of OIR group or PBS group respectively. Three days after EPCs injection, a large number of EPCs appeared in the vitreous cavity and adhered to the retinal surface. While at one week, the cells gathered between the internal plexiform layer and the inner limiting membrane, and some EPCs appeared in retinal vessels. EPCs transplantation can participate in the reparative procedure of the neovascularization in OIR.

Autologous endothelial progenitor cells improve allograft survival in porcine lung transplantation with prolonged ischemia.

As endothelial progenitor cells (EPCs) attenuated acute lung injury (ALI) in rabbit model, we hypothesized that autologous EPCs preserved lung graft function during the acute reperfusion period of lung transplantation and tested the therapeutic potential of EPCs in a porcine model of lung transplantation with prolonged graft ischemia. Day-7 EPCs isolated from the recipient subjects or plain culture media were administered into the left pulmonary artery immediately before restoration of pulmonary blood flow in a porcine lung allotransplantation model, with the transplantation surgeons blinded to the content of injection. Hemodynamics and arterial blood gas were recorded, and the right pulmonary artery was occluded 30 min after reperfusion to evaluate the lung graft function. The lung grafts were sectioned for histological examination at the end of experiments. The total ischemic time for lung graft was approximately 14 h. All animals receiving plain medium died within 40 min after reperfusion, but 3 out of 5 (60%) piglets receiving EPCs survived up to 4 h after diversion of the entire cardiac output into the lung graft (P<0.01). The donor body weight, recipient body weight, cold ischemic time, and time for anastomosis were comparable between the EPC and control group (P=0.989, 0.822, 0.843, and 0.452, respectively). The mean aortic pressure decreased, and the cardiac output and mean pulmonary artery pressure elevated after right pulmonary artery occlusion. All these parameters were gradually compensated in the EPC group but decompensated in the control group. Better preservation of gas exchange function, reduced thrombi formation in the terminal pulmonary arterioles, and attenuated interstitial hemorrhage of the lung graft were observed in the EPC group. We concluded autologous EPCs significantly enhanced the function of lung allograft and improved survival in a porcine model of lung transplantation with prolonged ischemia.

Treatment of diabetes mellitus-induced erectile dysfunction using endothelial progenitor cells genetically modified with human telomerase reverse transcriptase.

The efficacy of treatments for diabetes mellitus-induced erectile dysfunction (DMED) is quite poor, and stem cell therapy is emerging as a useful method. In this study, we used endothelial progenitor cells (EPCs) overexpressing human telomerase reverse transcriptase (hTERT) for the treatment of DMED. Rat EPCs were transfected with hTERT (EPCs-hTERT). EPCs-hTERT secreted more growth factors and demonstrated enhanced proliferation and resistance to oxidative stress. Twenty-four male DMED rats were subjected to four treatments: DMED (DMED group), EPCs (EPCs group), EPCs transduced with control lentivirus (EPC-control group) and EPCs-hTERT (EPCs-hTERT group). A group of healthy rats were used as the normal control group. The erectile function in the EPCs-hTERT group was markedly increased compared with the EPCs and EPCs-control groups. The EPCs-hTERT group exhibited more growth factors, smooth muscle content and retained stem cells in penile tissues. The degree of apoptosis and collagen/smooth muscle ratio in penile tissues of the EPCs-hTERT group was considerably reduced. Endothelial nitric oxide synthase (eNOS) expression increased significantly in the EPCs-hTERT group. Taken together, these data showed that the enhanced paracrine effect, resistance to oxidative stress and proliferation of EPCs-hTERT may contribute to the improvements of erectile function in DMED rats.

MP86-14 TREATMENT OF DIABETES MELLITUS-INDUCED ERECTILE DYSFUNCTION USING ENDOTHELIAL PROGENITOR CELLS GENETICALLY MODIFIED WITH HUMAN TELOMERASE REVERSE TRANSCRIPTASE

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology I1 Apr 2016MP86-14 TREATMENT OF DIABETES MELLITUS-INDUCED ERECTILE DYSFUNCTION USING ENDOTHELIAL PROGENITOR CELLS GENETICALLY MODIFIED WITH HUMAN TELOMERASE REVERSE TRANSCRIPTASE Yan Zhang, Tao Wang, Jun Yang, Rui Li, Zhi Chen, Shaogang Wang, Jihong Liu, and Zhangqun Ye Yan ZhangYan Zhang More articles by this author , Tao WangTao Wang More articles by this author , Jun YangJun Yang More articles by this author , Rui LiRui Li More articles by this author , Zhi ChenZhi Chen More articles by this author , Shaogang WangShaogang Wang More articles by this author , Jihong LiuJihong Liu More articles by this author , and Zhangqun YeZhangqun Ye More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2322AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Erectile dysfunction (ED) is one of the most common complications of diabetes mellitus (DM). The efficacy of treatments for diabetes mellitus-induced erectile dysfunction (DMED) is quite poor, and stem cell therapy is emerging as a useful method. This study evaluated the possibility and mechanisms of DMED treatment with endothelial progenitor cells (EPCs) genetically modified to express human telomerase reverse transcriptase (hTERT). METHODS Rat EPCs were isolated and transfected with hTERT (EPCs-hTERT). Proliferation capacity, intracellular reactive oxygen species, paracrine character and resistance to oxidative stress of EPCs-hTERT were assessed in vitro. Diabetes was induced via an intraperitoneal injection of streptozotocin. Twenty-four male DMED rats were subjected to four treatments: DMED (DMED group), EPCs (EPCs group), EPCs transfected with control lentivirus (EPC-control group) and EPCs-hTERT (EPCs-hTERT group). A group of health rats were used as a normal control group. Intracavernosal pressure (ICP) was measured using electrical stimulation for each group after 2 weeks of EPC injections. Masson's trichrome staining was performed to determine collagen content in penile tissues. The degree of apoptosis was assessed using western blotting analysis for terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) signaling pathway was detected. RESULTS EPCs-hTERT secreted more growth factors than EPCs and EPCs-control. EPC resistance to oxidative stress and proliferation was dramatically improved by hTERT transfection. The ICP/mean arterial pressure (MAP) ratio induced by electrical stimulation was markedly increased in the EPCs-hTERT group compared with the EPCs and EPCs-control group. Immunofluorescence demonstrated increased cell survival in penile tissues after EPCs-hTERT implantation. The smooth muscle/collagen ratio was markedly increased in the EPCs-hTERT group. TGF-β1 and p-Smad expression decreased significantly in DMED rats after EPCs-hTERT treatment. The degree of apoptosis in penile tissues of the EPCs-hTERT group was considerably reduced, as demonstrated by the decreased Bax/Bcl-2 ratio. NOS expression and NO and cGMP concentrations increased significantly in the EPCs-hTERT group. CONCLUSIONS The enhanced paracrine effect, resistance to oxidative stress and EPCs-hTERT proliferation contributed to the improvement of erectile function in DMED rats. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1111 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Yan Zhang More articles by this author Tao Wang More articles by this author Jun Yang More articles by this author Rui Li More articles by this author Zhi Chen More articles by this author Shaogang Wang More articles by this author Jihong Liu More articles by this author Zhangqun Ye More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

The use of endothelial progenitor cells combined with barrier membrane for the reconstruction of peri-implant osseous defects: an animal experimental study.

The aim of this research was to study the efficacy of endothelial progenitor cells (EPC) with ß tri-calcium phosphate (β-TCP) and barrier in the treatment of experimental peri-implantitis. Thirty implants were placed in five American foxhounds. Following osseointegration, ligature-induced peri-implantitis was initiated. EPC were isolated from peripheral blood. Two months later, open flap debridement was performed and implants' surface was decontaminated with 24% EDTA. The sites were grafted with β-TCP and collagen membrane (β-TCP group) or β-TCP loaded with EPC and covered with membrane (EPC group) or left un-grafted (OFD group). At 3 months, animals were killed and specimens sent for histomorphometric and micro-CT analysis. Bone height and %BIC served as primary and secondary outcomes. EPC group showed shorter distance to first BIC (3.29 ± 0.69 mm) compared to 4.2 ± 0.92 mm (β-TCP) and 3.82 ± 0.73 mm (OFD). Mean histological BIC was 2-3 folds higher in the EPC group (17.65 ± 3.3%) compared with OFD (7.55 ± 2.24%, p = 0.01) and β-TCP (5.68 ± 2.91%, p = 0.05). BIC greater than 25% was found only in the EPC group. The distance between the implant shoulder to 1st BIC was 2.51 ± 0.7 mm (lingual sites) compared to 3.64 ± 0.8 mm for buccal sites (p < 0.0001). β-TCP loaded with EPC and covered with a membrane-enhanced bone formation around previously contaminated dental implants in an experimental peri-implantitis canine model.