Yoshiaki Takeshita

Angiogenesis and Vasculogenesis Are Impaired in the Precocious-Aging klotho Mouse

Background—The effects of aging on angiogenesis (vascular sprouting) and vasculogenesis (endothelial precursor cell [EPC] incorporation into vessels) are not well known. We examined whether ischemia-induced angiogenesis/vasculogenesis is altered in klotho (kl) mutant mice, an animal model of typical aging. Methods and Results—After unilateral hindlimb ischemia, laser Doppler blood-flow (LDBF) analysis revealed a decreased ischemic-normal LDBF ratio in kl mice. Tissue capillary density was also suppressed in kl mice (+/+>+/kl>kl/kl). Aortic-ring culture assay showed impaired angiogenesis in kl/kl mice, accompanied by reduced endothelium-derived nitric oxide release. Moreover, the rate of transplanted homologous bone marrow cells incorporated into capillaries in ischemic tissues (vasculogenesis) was lower in kl/kl mice than in wild-type (+/+) mice, which was associated with a decrease in the number of c-Kit+CD31+ EPC-like mononuclear cells in bone marrow and in peripheral blood. Finally, the 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor cerivastatin restored the impaired neovascularization in kl/kl mice, accompanied by an increase in c-Kit+CD31+ cells in bone marrow and peripheral blood, and enhanced angiogenesis in the aortic-ring culture. Conclusions—Angiogenesis and vasculogenesis are impaired in kl mutant mice, a model of typical aging. Moreover, the age-associated impairment of neovascularization might be a new target of statin therapy.

Autologous transplantation of bone marrow mononuclear cells improved ischemic peripheral neuropathy in humans.

To the Editor: Ischemic peripheral neuropathy is a term used to describe the neurological deficits of peripheral nerves in patients with peripheral artery obstructive disease ([1][1]). Although ischemic peripheral neuropathy is a major complication of critical limb ischemia resulting in impaired

Clinical Survey of Cell Therapy in Japan

Background: Therapeutic angiogenesis by using cells is being performed in Japan. However, it is unknown in how many centers and how these therapies are performed. The efficacy and side effects are also unknown. Thus, we conducted the survey by mailing questionnaire within Japan. Methods and results: Two surveys were performed in 2003. The first survey unveiled that cell therapy was performed in 32 facilities until October 2003. The second survey unveiled the followings. (1) The total number of performed cases was 221. 153 patients (69.2%) had arterio-sclerosis obliterance (ASO), 56 patients (25.3%) thromboangitis obliterans (TAO, Burger’s disease), and 12 patients (5.4%) other conditions. (2) The sources of cells were bone marrow-mononuclear cells (61.5%), peripheral-mononuclear cells (9.5%), and peripheral CD34+ cells (22.1%). A few patients (6.7%) were treated with a cytokine only (granulocyte-colony stimulating factor: G-CSF). (3) Inclusion criteria were the same for most facilities, such as patients with PAOD, especially with critical limb ischemia with rest pain, non-healing ischemic ulcers and non-candidates for non-surgical or surgical revascularization. All facilities excluded patients with histories of malignant disorder during the past 5 years, proliferative diabetic retinopathy, pregnancy, proliferative blood disease, uncontrolled ischemic heart disease, rheumatic arthritis, or psychiatric disease. (4) Subjective improvement was observed in 138 of 199 patients (69%). Objective improvements for ABI, TcO2 or angiographic findings were observed in 98 of 182 patients (53.8%). (6) Three of 221 patients (1.4%) died after cell therapy. One died from cerebro-vascular attack (thrombo-embolizm), and two died from acute myocardial infarction (AMI). Conclusion: Our clinical survey has shown that cell therapy is being performed in many medical centers in Japan. It seems to be safe and effective for patients with PAOD with no surgical options

Effect of Bone Marrow Transplantation in Patients with Critical Limb Ischemia

Background: Endothelial progenitor cells in the CD34+ stem-cell fraction of adult human peripheral blood take a part in postnatal neovascularization after mobilization from bone marrow. We investigated whether transplantation of bone marrow-mononuclear cells (BM-MNCs), including endothelial progenitor cells (EPCs), into ischemic limbs rescues ischemia in patients with peripheral artery occulusive disease (PAOD). Methods and Results: 22 patients with severe PAOD (atherosclerosis obliterance and Thromboangiitis obliterance / Burger’s disease) were enrolled. After collecting MNCs from the autologous bone marrow using the density gradient centrifugation method, we transplanted them into the ischemic limb. Four weeks after transplantation, pain quantified by the Visual Analog Scale was significantly improved. Skin ulcers healed in 6 of 10 patients and the size was reduced in the rest of them. Ankle brachial blood pressure ratio index (ABI) increased (0.58 ±0.2 to 0.69±0.26 at 4weeks). Furthermore, most of patients revealed angiographical improvements. No adverse event was observed.