Fujio Sato

Identification of human placenta-derived mesenchymal stem cells involved in re-endothelialization†

Human placenta is an attractive source of mesenchymal stem cells (MSC) for regenerative medicine. The cell surface markers expressed on MSC have been proposed as useful tools for the isolation of MSC from other cell populations. However, the correlation between the expression of MSC markers and the ability to support tissue regeneration in vivo has not been well examined. Here, we established several MSC lines from human placenta and examined the expression of their cell surface markers and their ability to differentiate toward mesenchymal cell lineages. We found that the expression of CD349/frizzled‐9, a receptor for Wnt ligands, was positive in placenta‐derived MSC. So, we isolated CD349‐negative and ‐positive fractions from an MSC line and examined how successfully cell engraftment repaired fractured bone and recovered blood flow in ischemic regions using mouse models. CD349‐negative and ‐positive cells displayed a similar expression pattern of cell surface markers and facilitated the repair of fractured bone in transplantation experiments in mice. Interestingly, CD349‐negative, but not CD349‐positive cells, showed significant effects on recovering blood flow following vascular occlusion. We found that induction of PDGFβ and bFGF mRNAs by hypoxia was greater in CD349‐negative cells than in CD349‐positive cells while the expression of VEGF was not significantly different in CD349‐negative and CD349‐positive cells. These findings suggest the possibility that CD349 could be utilized as a specialized marker for MSC isolation for re‐endothelialization. J. Cell. Physiol. 226: 224–235, 2010.

The minimum coronary artery diameter in which coronary spasm can be identified by synchrotron radiation coronary angiography

BACKGROUND Coronary vasospasm is defined as a temporary, intense narrowing of the coronary conduit artery. It brings about ischemic chest pain and becomes one of the causes of myocardial infarction. Coronary spasms are divided into two categories. One is the coronary spasm of the conduit artery and the other is the coronary microvascular spasm. Although coronary spasms are diagnosed with the images of coronary angiography, microvascular spasms cannot be diagnosed because of the limitations of conventional angiographic systems. However, synchrotron radiation coronary angiography (SRCA) can identify coronary arteries down to 100 microm in diameter in the beating heart and 50 microm in arrested heart. AIM The purpose of this study was to confirm whether microvascular spasms could be identified or not using SRCA, and then down that size identification was possible. METHODS The Langendorff perfusion system with isolated rat hearts was employed. Krebs-Henseleit solution (KH solution) was used as a perfusate. 10mM of 4-aminopyridine (4-AP: a voltage-gated potassium channel blocker; spasm inducer) was added to the KH solution and maintained for 5 min. SRCA was performed at pre-, during and 10 min after cessation of the KH solution with 4-AP. Coronary spasms were defined as a temporal 75% reduction of coronary arterial diameter. RESULTS AND CONCLUSION Multiple sizes of coronary arteries showed coronary spasms. The minimum stenosed coronary artery size was 100 microm. Since coronary microvascular spasms are seen in the arterioles (50-400 microm), coronary microvascular spasms may be diagnosed with the use of synchrotron radiation coronary angiography.

Endoscopic harvesting of the radial artery as a coronary artery bypass graft

With an endoscopic approach we harvested the radial artery as a coronary artery bypass graft, using two transverse 2-cm incisions in the forearm. There were no complications in the forearm and hand. The short-term results of the radial artery graft were good.

The Role of CCL5 in the Ability of Adipose Tissue-Derived Mesenchymal Stem Cells to Support Repair of Ischemic Regions

Mesenchymal stem cells (MSC) are multipotent and possess high proliferative activity, and thus are thought to be a reliable cell source for cell therapies. Here, we isolated MSC from adult tissues--bone marrow (BM-MSC), dental tissue (DT-MSC), and adipose tissue (AT-MSC)--to compare how autotransplantation of these MSC effectively supports the repair of bone fracture and ischemic tissue. An analysis by in vitro differentiation assays showed no significant difference among these MSC. The degree of calcification at the joint region of bone fracture was higher in mice transplanted with AT-MSC than in mice transplanted with BM-MSC or DT-MSC. To compare the abilities of MSC, characterize how those MSC affect the repair of ischemic tissue, vascular occlusion was performed by ligation of the femoral artery and vein. Of note, the blood flow in the ischemic region rapidly increased in mice injected with AT-MSC, as contrasted with mice injected with BM- or DT-MSC. The number of CD45- and F4/80-positive cells at the femoral region was higher in AT-MSC recipients than in recipients of BM-MSC or DT-MSC. We evaluated the mRNA expression of angiogenic and migration factors in MSC and found the expression of CCL5 mRNA was higher in AT-MSC than in BM-MSC or DT-MSC. Transplantation of AT-MSC with impaired expression of CCL5 clearly showed a significant delay in the recovery of blood flow compared with the control. These findings have fundamental implications for the modulation of AT-MSC in the repair of vasculature and bone fracture.

Prospective intraindividual comparison of unenhanced magnetic resonance imaging vs contrast-enhanced computed tomography for the planning of endovascular abdominal aortic aneurysm repair

OBJECTIVE This study clarified whether unenhanced magnetic resonance imaging (MRI) is an alternative to contrast-enhanced computed tomography (CT) for aortoiliac arterial measurement before endovascular abdominal aortic aneurysm repair (EVAR). METHODS The institutional review board approved this prospective study. Twenty patients being considered for EVAR underwent MRI using a steady-state free-precession sequence in a 1.5-T system and contrast-enhanced CT within 4 weeks of each other. Two independent observers reviewed MRI and CT in random order using vessel analysis software and measured seven diameters, four lengths, and the angle of the aortoiliac arteries. The intermodality, interobserver, and intraobserver agreements were assessed for each measurement by intraclass correlation coefficients (ICCs) and the Altman-Bland method. Additionally, the observers independently recorded the number of bilateral renal arteries, decided EVAR suitability, and selected the main endograft on each modality. RESULTS Intermodality ICCs for observers A and B showed ranges of 0.83 to 0.99 and 0.70 to 0.98; interobserver ICCs for MRI and CT showed ranges of 0.73 to 0.99 and 0.65 to 0.99; and intraobserver ICCs for MRI and CT showed ranges of 0.59 to 0.99 and 0.59 to 0.99. In intermodality, interobserver, and intraobserver comparisons, mean differences in diameters were included within the range -1 to +1 mm, excluding three of seven diameters on CT in interobserver comparison and one of seven on CT in intraobserver comparison. Mean differences in lengths were included within the range -5 to +5 mm, excluding one of four lengths in observer B in intermodality comparison and one of four on MRI and CT in interobserver comparison. All mean differences in angles were included within the range -5° to +5°. Both observers detected all 40 bilateral main renal arteries on MRI and CT. Of the 13 accessory renal arteries, observers A and B detected four (31%) and nine (69%), respectively, on MRI; in contrast, both observers detected 11 (85%) on CT. The observers independently determined that the same seven patients were suitable for EVAR on MRI and CT. Of the seven selected main endografts, seven and six diameters and five and six lengths agreed exactly between MRI and CT for observers A and B, respectively. CONCLUSIONS Although contrast-enhanced CT remains the gold standard for preoperative EVAR planning, unenhanced MRI with steady-state free-precession sequence can be an alternative modality for patients with contraindications for CT, such as renal impairment, because the intermodality agreement for preoperative measurements is as good as interobserver and intraobserver agreement.

Increased Expression of EGR-1 in Diabetic Human Adipose Tissue-Derived Mesenchymal Stem Cells Reduces Their Wound Healing Capacity.

The prevalence of type 2 diabetes mellitus (T2DM), which leads to diabetic complications, has been increasing worldwide. The possible applications of T2DM-derived stem cells in cell therapy are limited because their characteristics are still not fully understood. In this study, we characterized adipose tissue-derived mesenchymal stem cells (AT-MSCs) from diabetic patients (dAT-MSCs) and found that insulin receptor substrate-1 (IRS-1) was highly phosphorylated at serine 636/639 in dAT-MSCs. Moreover, we found that early growth response factor-1 (EGR-1) and its target genes of PTEN and GGPS1 were highly expressed in dAT-MSCs in comparison to healthy donor-derived AT-MSCs (nAT-MSCs). We observed impaired wound healing after the injection of dAT-MSCs in the ischemic flap mouse model. The expressions of EGR-1 and its target genes were diminished by small hairpin RNA-targeted EGR-1 (shEGR-1) and treatment with a mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) inhibitor (PD98059). Importantly, dAT-MSCs with shEGR-1 were able to restore the wound healing ability in the mouse model. Interestingly, under hypoxic conditions, hypoxia-inducible factor-1α (HIF-1α) can bind to the EGR-1 promoter in dAT-MSCs, but not in nAT-MSCs. Together, these results demonstrate that the expression of EGR-1 was upregulated in dAT-MSCs through two pathways: the main regulatory pathway is the MAPK/ERK pathway, the other is mediated by HIF-1α through direct transcriptional activation at the promoter region of the EGR1 gene. Our study suggests that dAT-MSCs may contribute to microvascular damage and delay wound healing through the overexpression of EGR-1. Interrupting the expression of EGR-1 in dAT-MSCs may be a useful treatment for chronic wounds in diabetic patients.

Serum-free isolation and culture system to enhance the proliferation and bone regeneration of adipose tissue-derived mesenchymal stem cells

Cell therapy using human mesenchymal stem cells (MSCs) is an attractive approach for many refractory diseases. Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are considered as a favorable tool due to its abundance in the body, easy proliferation, and high cytokine production potency. In order to avoid the risks associated with the use of fetal bovine serum (FBS) in culture that includes batch variations and contamination with pathogens, development of serum-free culture system has been initiated. We have formulated a completely serum-free culture medium (SFM) that could be used not only for the expansion of AT-MSCs but also for initial isolation. We demonstrate that the AT-MSCs isolated and cultured in serum-free medium (AT-MSCs/SFM) possess high proliferation capacity and differentiation potency to osteoblast, adipocyte, and chondrocyte lineages in vitro. In in vivo bone fraction model analysis, AT-MSCs/SFM showed higher bone repair potency and quality of the regenerated bone than the cells cultured in serum-containing medium (AT-MSCs/SCM). This was attributed to the (i) presence of translated cells in the bone, as evidenced by in vivo imaging of the illuminated translated cells and (ii) high level of expression and induction capacity of AT-MSCs/SFM for cytokine BMP2, CCL2, and CCL5. Taken together, we report a new serum-free culture system for AT-MSCs that is suitable for cell therapy.

An approach for acute disruption of large arteries in patients with advanced cervical cancer: endoluminal balloon occlusion technique.

OBJECTIVE To test the feasibility of an intraluminal balloon occlusion technique for the control of sudden arterial disruption from cervical branches in patients with advanced malignancy. BACKGROUND A sudden disruption of large cervical arteries is a devastating complication of advanced cervical malignancy and local infection. METHODS Three patients with this complication underwent endoluminal balloon occlusion before surgical management. A double-lumen balloon catheter was introduced from the femoral artery to the bleeding point. Angiographic examination of the intracranial cross-filling could be performed by the injection of contrast medium from the opposite carotid artery, and consciousness levels could be directly confirmed under unilateral carotid occlusion. RESULTS Resection of the carotid or innominate artery was safely performed in all the patients. No neurologic complications occurred. Bleeding did not recur during follow-up (range 5-32 months). CONCLUSIONS This experience, although limited, suggests that surgical intervention to control cervical arterial bleeding with intraluminal balloon occlusion prevents excessive bleeding, decreases the risk of damage to the central nervous system, and improves the outcome in these critically ill patients.

A gas sampling calorimeter using conductive plastic tubes (II)

Abstract A prototype module for the electromagnetic shower calorimeter has been constructed and its characteristics have been measured. It consists of 38 layers of planar proportional chambers interleaved with 3 mm thick lead plates. The proportional chamber is made of conductive plastic tubes as a dc cathode and pickup electrodes as an rf cathode. The pickup electrodes are copper-clad G-10 panels etched with finely segmented patterns which, when assembled, form complete conical towers. The module was tested using high energy electron and hadron beams of energy range from 25 GeV to 150 GeV. Energy resolution is 24% E( GeV ) , and the position resolution is 1.5 mm or better at and above 50 GeV. The lateral and longitudinal distributions of the energy deposit are studied and they are applied to π e discrimination.

Surgical removal of infected pacemaker leads without cardiopulmonary bypass after failed extraction using the Excimer Laser Sheath Extraction System

With the growing number of cardiac pacemakers and internal cardioverter defibrillator implantations, problems with endocardial lead infection have been increasing. The newly developed Excimer Laser Sheath Lead Extraction System has been recognized as being highly useful for removing chronic infected leads. However, serious bleeding complications are a concern when this system is used. Here we report our experience with a 67-year-old man who was diagnosed with pacemaker endocarditis. Initially, lead removal was attempted using the Excimer Laser Sheath Extraction System, though this was abandoned because of severe adhesion of the leads and the junction of the supra vena cava (SVC) with the right atrium. Surgical removal of the leads was performed without using cardiopulmonary bypass and the leads were removed without any complications. During surgery, we found there was a silent perforation of the innominate vein brought about by the Excimer Laser Sheath System. Also, the junction of the SVC with the right atrium was thought to be an area potentially at high risk of perforation, because of a lack of surrounding tissue. It is our opinion that those who carry out procedures with the Excimer Laser Sheath System should understand the potential risk of perforation based on cardiac anatomy and should be prepared for lethal bleeding complications. Also, for emergent situations, we believe that close backup by a cardiovascular surgical team should be considered essential for performing the Excimer Laser Sheath Lead Extraction safely.