Takeda Koji, Fukumoto Shinya, Motoyama Koka, Morioka Tomoaki, Mori Katsuhito, Kageyama Ken, Sakai Yukimasa, Sato Hideki, Suzuki Masakazu, Koyama Hidenori, Shoji Tetsuo, Ishimura Eiji, Emoto Masanori, Furuzono Tsutomu, Nakajima Koichi, Inaba Masaaki
Department of Metabolism, Endocrinology, and Molecular Medicine, Osaka City University Graduate School of Medicine, Osaka, Japan.
Department of Premier Preventive Medicine, Osaka City University Graduate School of Medicine, Osaka, Japan.
Biochem Biophys Res Commun. 2014 Nov 7;454(1):119-24. doi: 10.1016/j.bbrc.2014.10.045. Epub 2014 Oct 18.
The clinical success of cell-based therapeutic angiogenesis has been limited in diabetic patients with critical limb ischemia. We previously reported that an injectable cell scaffold (ICS), which is a nano-scaled hydroxyapatite (HAp)-coated polymer microsphere, enhances therapeutic angiogenesis. Subsequently, we developed a modified ICS for clinical use, measuring 50 μm in diameter using poly(l-lactide-co-ε-caprolactone) as a biodegradable polymer, which achieved appropriately accelerated absorption in vivo. The aim of the present study was to evaluate the effectiveness of this practical ICS in diabetic hindlimb ischemia. Bone-marrow mononuclear cells (BMNCs) were intramuscularly injected, without or with a practical ICS, into the ischemic hindlimbs of mice (BMNCs or ICS+BMNCs group, respectively). Kaplan-Meier analysis demonstrated that the beneficial effects of BMNC transplantation for limb salvage after ischemic surgery were almost entirely abrogated in streptozotocin-induced diabetic mice. In contrast, injection of ICS+BMNCs revealed significant limb salvage in diabetic mice to a similar extent as in non-diabetic mice. The number of apoptotic transplanted BMNCs was 1.8-fold higher in diabetic mice 10 days after transplantation compared to non-diabetic mice, while that in the ICS+BMNCs group was markedly lower (8.3% of that in the BMNCs group) even in diabetic mice. The proangiogenic factors VEGF and FGF2, also known as antiapoptotic factors, mostly co-localized with transplanted GFP-positive BMNCs that were closely aggregated around the ICS in ischemic tissue. In conclusion, the practical ICS significantly augmented cell-based therapeutic angiogenesis even in diabetic animals, through local accumulation of proangiogenic factors and antiapoptotic effects in transplanted cells.
基于细胞的治疗性血管生成在患有严重肢体缺血的糖尿病患者中的临床成功率一直有限。我们之前报道过一种可注射细胞支架（ICS），它是一种纳米级羟基磷灰石（HAp）涂层的聚合物微球，可增强治疗性血管生成。随后，我们开发了一种用于临床的改良ICS，其直径为50μm，使用聚（L-丙交酯-共-ε-己内酯）作为可生物降解聚合物，在体内实现了适当加速吸收。本研究的目的是评估这种实用的ICS在糖尿病后肢缺血中的有效性。将骨髓单个核细胞（BMNCs）分别在不使用或使用实用ICS的情况下肌肉注射到小鼠的缺血后肢中（分别为BMNCs组或ICS + BMNCs组）。Kaplan-Meier分析表明，链脲佐菌素诱导的糖尿病小鼠中，缺血手术后BMNC移植对肢体挽救的有益作用几乎完全丧失。相比之下，注射ICS + BMNCs的糖尿病小鼠显示出与非糖尿病小鼠相似程度的显著肢体挽救。移植后10天，糖尿病小鼠中凋亡的移植BMNCs数量比非糖尿病小鼠高1.8倍，而即使在糖尿病小鼠中，ICS + BMNCs组中的凋亡BMNCs数量也明显更低（为BMNCs组的8.3%）。促血管生成因子VEGF和FGF2，也被称为抗凋亡因子，大多与移植的GFP阳性BMNCs共定位，这些细胞在缺血组织中紧密聚集在ICS周围。总之，实用的ICS即使在糖尿病动物中也能通过促血管生成因子的局部积累和对移植细胞的抗凋亡作用，显著增强基于细胞的治疗性血管生成。
