Effect of low-molecular-weight heparin on the commitment of bone marrow cells to liver sinusoidal endothelial cells in CCl(4)-induced liver injury.

BACKGROUND/AIMS: Recently liver regeneration by bone marrow transplantation has been proposed as an alternative source of functional liver cells. We investigate commitment of bone marrow cells (BMCs) to liver regeneration and the effect of dalteparin sodium (DS) on regeneration of the damaged liver caused by carbon tetrachloride (CCl(4)) administration in the mice. METHODS: Liver injury was produced in 8-week-old mice by treating with CCl(4) for 4 weeks. Thereafter, mice received a lethal dose of irradiation (10Gy) to whole body, followed by injection of 1x10(7) green fluorescent protein (GFP)-positive BMCs via the tail vein. DS (50IU/kg, intraperitoneally) was administered daily for 28 consecutive days starting at 1 day post-BMC transplantation. Lineage marker analysis of GFP-positive liver cells was performed immunostaining with a CD31 antibody. RESULT: Four weeks after BMC transplantation, GFP-positive cells in the CCl(4)-damaged liver could be detected in the lobule displaying a meshwork architecture extending from the periportal to pericentral regions, a pattern simulating sinusoidal lining. This localization of GFP-positive cells suggested that these cells were closely associated with sinusoidal endothelial cells. By staining the GFP-positive cells for CD31, it was confirmed that the majority of the GFP-positive cells are also positive for CD31. The GFP(+)CD31(+) cells were barely detected in the control group (1.0+/-1.2 per field). In marked contrast, a numerous number of GFP(+)CD31(+) cells were detected in the liver section obtained from the CCl(4)-induced liver damage group (3.8+/-1.3 per field, P<0.05 versus control). The number of GFP(+)CD31(+) cells in CCl(4) plus DS-treated group was further increased to 8.3+/-1.3 per field (P<0.05 versus CCl(4)-induced liver damage group). CONCLUSION: The majority of GFP-positive BMCs was committed to sinusoidal endothelial cells. DS promoted BMC differentiation into sinusoidal endothelial cells in the CCl(4)-damaged liver.     

Early in vivo experience with tissue-engineered heart valve leaflets from autologous bone marrow-derived cells

Background Currently used heart valve prostheses are associated with anticoagulation complications or limited durability. The advancement of stem cell study and tissue-engineered heart valve research may offer a relatively ideal solution to these problems. Methods Bone marrow was aspirated from sternum of lamb goats to isolate BMCs. Cells were identified by flow cytometry and its capacity of differentiation. Cellular viability was assessed with Rhdomine 123 staining. 1 × 107 cells were seeded on a patch of ...     

A single portion of blueberry (Vaccinium corymbosum L) improves protection against DNA damage but not vascular function in healthy male volunteers

It has been suggested that anthocyanin-rich foods may exert antioxidant effects and improve vascular function as demonstrated mainly in vitro and in the animal model. Blueberries are rich sources of anthocyanins and we hypothesized that their intake could improve cell protection against oxidative stress and affect endothelial function in humans. The aim of the study was to investigate the effect of one portion (300 g) of blueberries on selected markers of oxidative stress and antioxidant protection (endogenous and oxidatively induced DNA damage) and of vascular function (changes in peripheral arterial tone and plasma nitric oxide levels) in male subjects. In a randomized cross-over design, separated by a wash out period ten young volunteers received one portion of blueberries ground by blender or one portion of a control jelly. Before and after consumption (at 1, 2, and 24 hours), blood samples were collected and used to evaluate anthocyanin absorption (through mass spectrometry), endogenous and H₂O₂-induced DNA damage in blood mononuclear cells (through the comet assay), and plasma nitric oxide concentrations (through a fluorometric assay). Peripheral arterial function was assessed by means of Endo-PAT 2000. Blueberries significantly reduced (P < .01) H₂O₂-induced DNA damage (−18%) 1 hour after blueberry consumption compared to control. No significant differences were observed for endogenous DNA damage, peripheral arterial function and nitric oxide levels after blueberry intake. In conclusion, one portion of blueberries seems sufficient to improve cell antioxidant defense against DNA damage, but further studies are necessary to understand their role on vascular function.     

Amelioration of cognitive ability in senescence-accelerated mouse prone 8 (SAMP8) by intra-bone marrow-bone marrow transplantation

Bone marrow cells (BMCs) can increase the number of activated microglias, which play a central role in the inflammatory response in Alzheimer's disease (AD). Senescence-accelerated mouse (SAM) prone 8 (SAMP8) are widely used in various experiments because of cognitive deficits observed with age. In the present study, 4-month-old SAMP8 were reconstituted with BMCs of C57BL/6 mice by intra-bone marrow-bone marrow transplantation (IBM-BMT), which can reconstitute both donor-derived hemopoietic stem cells and mesenchymal stem cells. Three months after IBM-BMT, the impairment of spatial memory in SAMP8 was found to be ameliorated after analyzing the results of the water maze test. Although IL-1β, IL-6 and iNOS increased and TGF-β decreased in 7 M SAMP8, IL-1β, IL-6 and iNOS decreased while TGF-β increased after IBM-BMT by RT-PCR. Moreover, oxidative stress-related heme oxygenase-1 (HO-1) increased in 7 M SAMP8, but significantly decreased after IBM-BMT. In conclusion, this is the first report suggesting that the impaired cognitive ability of SAMP8 is ameliorated by IBM-BMT. It seems likely that decreases in IL-1β, IL-6, iNOS and HO-1 are a result of the development of donor-derived BMCs.     

Clinical Trial Updates and Hotline Sessions presented at the Scientific Session 2007 of the American heart association

This article provides information and commentaries on trials which were presented at Clinical Trial Updates and Hotline Sessions presented at the Scientific Sessions 2007 of the American Heart Association in Orlando, Florida. The comprehensive summaries have been generated from the oral presentations and the webcasts of the American Heart Association. Most reports have not been published as full papers and therefore have to be considered as preliminary data, as the analysis may change in the final publications. The following papers are discussed: TRITON TIMI-38, EVA-AMI, BRIEF-PCI, RACE, MASS Stent, HF-ART, STITCH, CORONA, ILLUMINATE, CORE-64, OAT Substudy, AFCHF, MASCOT, RETHINQ, MASTER I, POISE, COUMA-GEN, HIJ-CREATE, PROVIDENCE I, CAUSMIC, IC-BMC, IC/IM BMCs.     

Human heart failure: Is cell therapy a valid option?

The concept of the heart as a terminally differentiated organ incapable of replacing damaged myocytes has been at the center of cardiovascular research and therapeutic development for the past 50 years. The progressive decline in myocyte number with aging and the formation of scarred tissue following myocardial infarction have been interpreted as irrefutable proofs of the post-mitotic characteristics of the adult heart. However, emerging evidence supports a more dynamic view of the myocardium in which cell death and cell restoration are vital components of the remodeling process that governs organ homeostasis, aging and disease. The identification of dividing myocytes throughout the life span of the organisms and the recognition that undifferentiated primitive cells regulate myocyte turnover and tissue regeneration indicate that the heart is a self-renewing organ controlled by a compartment of resident stem cells. Moreover, exogenous progenitors of bone marrow origin transdifferentiate and acquire the cardiomyocyte and vascular lineages. This new reality constitutes the foundation of the numerous cell-based clinical trials that have been conducted in the last decade for the treatment of ischemic and non-ischemic cardiomyopathies.     

TiO2纳米管负载BMP-2指节肽对骨髓间充质干细胞增殖与分化的影响

目的 检测负载骨形态发生蛋白2(bone morphogenetic protein 2, BMP-2)指节肽的二氧化钛(TiO2)纳米管复合结构对骨髓间充质干细胞增殖与成骨分化的影响.方法 通过阳极氧化法在钛片表面制备TiO2纳米管阵列.全骨髓差速贴壁法分离培养Wistar大鼠骨髓间充质干细胞.实验组负载BMP-2指节肽TiO2纳米管;对照组只采用TiO2纳米管.2组纳米管钛片接种间充质干细胞培养,测定细胞增殖、碱性磷酸酶活性、骨钙素(osteocalcin,OCN)及骨桥蛋白(osteopontin,OPN)RNA表达,并进行统计学分析.结果 2组细胞培养1、3、5天,细胞增殖实验组>对照组(P<0.05);2组细胞培养7、10、15天,碱性磷酸酶活性实验组>对照组(P<0.01);2组细胞培养21天后,骨钙素和骨桥蛋白的差异倍数(fold change)显示为实验组>对照组.结论 TiO2纳米管负载BMP-2指节肽对骨髓间充质干细胞增殖和早期成骨分化的促进作用强于单纯TiO2纳米管结构.     

Effects of chimerism on the mice heart transplanted survival with the bone marrow infusion

Aims

To evaluate the effects of chimerism on the mice heart transplanted survival with the bone marrow infusion.

Methods

Bone marrow cells (BMCs) were obtained from BALB/c mice. These BMCs were injected into the irradiated (2Gy-Co60) C57BL/6 mice through femoral vein. Then Group A mice were treated with Cyclosporine (1 mg/kg) for 21 days and Group B were not treated with Cyclosporine. Group C were treated as the control group without BMCs infusion. Group D were treated with Cyclosporine (1 mg/kg) for 21 days pre-hearttransplantation without BMCs infusion. After 21 days, the C57BL/6 mice received heart allografts from BALB/c. To determine the degree of chimerism in BMCs infusion recipients, peripheral blood were isolated on day 7, 14, 21. Allografts were harvested 10 days after heart transplantation for the histological analysis.

Results

(1) Chimerism detected in the peripheral blood of Group A mice on day 7 after BMCs infusion was 6.1 ± 2.5%, on day 14 was 15.4 ± 2.9% and on day 21 was 10.7 ± 2.6%. For the Group B mice on day 7 after BMCs infusion, the chimerism was 2.8 ± 1.1%, on day 14 was 11.2 ± 4.8% and on day 21 was 7.4 ± 3.7%. For the Groups C and D mice, no chimerism was observed. Group A mice had the tendency toward improved level of chimerism than Group B mice. (2) The survival time of Group A (n = 6) was 13.0 ± 1.4 days which was significantly longer than Group B (n = 6) with the survival time was 8.5 ± 1.3 days (p < 0.001), also longer than the mice in Groups C and D, the survival time of which were 10.0 ± 1.3 days (p = 0.008) and 9.4 ± 1.1 days (p = 0.004). There is no significant difference among Groups B, C, and D. (3) The HE staining showed the much more seriously heart rejection in Groups B, C and D than Group A.

Conclusions

The chimerism was found in the BMCs infusion groups. Without the CsA treatment combined with chimerism could not protect the transplanted heart. There was no obvious evidence showed that the chimerism alone could improve the survival time of cardiac allografts in mice.     

Anti-thrombosis effect of LRRFIP1 shRNA lentivirus in a mouse model of deep vein thrombosis

Introduction

Deep vein thrombosis (DVT) is one of the common complications of orthopedic surgery. Low molecular weight heparin (LMWH) is a usually used agent for DVT, but it would increase the risk of bleeding. LRRFIP1 has been shown to play an important role in the formation of thrombosis. Therefore, we investigated the effect of LRRFIP1 shRNA lentivirus on DVT in mice.

Materials and Methods

Lentiviral Vectors carrying LRRFIP1 shRNA were constructed and transfected into cultured mouse bone marrow cells (BMCs). Male ICR mice were irradiated with a single dose of 9.5 Gy and then were injected with different agents through the tail vein. Stasis venous thrombosis was induced by inferior vena cava (IVC) ligation. Mice were sacrificed on the 1st, 3rd and 7th day post operation and the thrombi were removed, blotted the excess blood on it with filter paper and immediately weighed. P-selectin and d-Dimer were determined by enzyme-linked immunosorbent assay (ELISA).

Results

LRRFIP1 shRNA significantly suppressed the expression of LRRFIP1 in the thrombi. In contrast, low molecular weight heparin (LMWH) and negative shRNA exhibited little effect on the expression of LRRFIP1. LRRFIP1 shRNA, LMWH and negative shRNA inhibited the thrombus formation in vivo significantly. The plasma P-selectin and d-Dimer levels were significantly increased after IVC ligation. LRRFIP1 shRNA significantly decreased the plasma P-selectin and d-Dimer levels. However, LMWH and negative shRNA showed little effects on the levels of plasma P-selectin and d-Dimer.

Conclusion

LRRFIP1 shRNA might represent a promising prevention strategy for DVT.