Ichihara Yuki, Shinoka Toshiharu, Matsumura Goki, Ikada Yoshito, Yamazaki Kenji
Department of Cardiovascular Surgery, Tokyo Women's Medical University, Shinjuku-ku, Tokyo, Japan.
Department of Surgery, Ohio State University, The Heart Center, Nationwide Children's Hospital, Columbus, OH, USA
Interact Cardiovasc Thorac Surg. 2015 Jun;20(6):768-76. doi: 10.1093/icvts/ivv017. Epub 2015 Feb 26.
Ideal alternatives for replacing native arteries, which have biocompatibility such as growth potential, anti-thrombogenesis and durability, have yet to be discovered. We previously demonstrated the utility of tissue-engineered vascular autografts; however, the use of these autografts is limited to low-pressure conditions. The aim of this study was to create the tissue-engineered arterial patch (TEAP) that could be used in high-pressure systems, and to evaluate the maturation in this regenerative tissue.
We developed a new biodegradable polymer scaffold, which is composed of a co-polymer of epsilon-caprolactone and lactide acid [P(CL/LA)] and a poly-L-lactide acid (PLLA). To obtain mechanical strength, we modified PLLA that is degraded by hydrolysis for 1-2 years in contrast to polyglycolic acid in our low-pressure study previously. We implanted an oval-shaped patch (30 × 15 mm) of this polymer without cell seeding into the descending aorta of 12 dogs, and followed the animals for 1, 3 and 6 months (n = 4 in each group). The cell proliferation in the patch was evaluated with histological and immunohistochemical methods. Additionally, the expression of vascular endothelial growth factor (VEGF) and smooth muscle myosin heavy chain (smMHC) mRNA in the patches was determined with reverse transcriptase-polymerase chain reaction.
Macroscopically, there was no incidence of rupture or aneurysmal formation on the patch. The luminal surface of the TEAP was covered with a single layer of endothelial cells stained with vWF immunohistochemically at 1 month after implantation. αSMA-positive cells that indicated smooth muscle cells and collagen fibres were observed in the patch and they increased over time. The VEGF mRNA expression in the patch at 1 month was significantly higher than that of native arterial tissue (1 month; 0.124 ± 0.017 ng/µl, native; 0.009 ± 0.003 ng/µl, P < 0.05). The smMHC mRNA expression gradually increased, and reached ∼ 60% of that of the native artery at 6 months (6 months: 0.351 ± 0.028 ng/µl, native: 0.540 ± 0.027 ng/µl).
We demonstrated the maturation of endothelial and smooth muscle cells in TEAP, suggesting that this biodegradable polymer scaffold could be used as an alternative vascular material even in high-pressure systems.
具有生物相容性(如生长潜力、抗血栓形成能力和耐久性)的理想天然动脉替代物尚未被发现。我们之前已证明组织工程化血管自体移植物的实用性;然而,这些自体移植物的使用仅限于低压条件。本研究的目的是创建可用于高压系统的组织工程化动脉补片(TEAP),并评估该再生组织的成熟情况。
我们开发了一种新型可生物降解聚合物支架,其由ε-己内酯和乳酸的共聚物[P(CL/LA)]以及聚-L-乳酸(PLLA)组成。为了获得机械强度,与我们之前在低压研究中使用的聚乙醇酸相比,我们对PLLA进行了改性,使其通过水解在1 - 2年内降解。我们将这种未接种细胞的聚合物椭圆形补片(30×15毫米)植入12只犬的降主动脉,并对动物进行1、3和6个月的随访(每组n = 4)。用组织学和免疫组织化学方法评估补片中的细胞增殖情况。此外,用逆转录聚合酶链反应测定补片中血管内皮生长因子(VEGF)和平滑肌肌球蛋白重链(smMHC)mRNA的表达。
宏观上,补片未发生破裂或动脉瘤形成。植入后1个月,TEAP的管腔表面免疫组织化学染色显示被单层内皮细胞覆盖,vWF呈阳性。在补片中观察到指示平滑肌细胞的αSMA阳性细胞和胶原纤维,且它们随时间增加。补片中1个月时VEGF mRNA表达显著高于天然动脉组织(1个月;0.124±0.017 ng/µl,天然;0.009±0.003 ng/µl,P < 0.05)。smMHC mRNA表达逐渐增加,在6个月时达到天然动脉的约60%(6个月:0.351±0.028 ng/µl,天然:0.540±0.027 ng/µl)。
我们证明了TEAP中内皮细胞和平滑肌细胞的成熟,表明这种可生物降解聚合物支架即使在高压系统中也可作为替代血管材料使用。