Micro-CT evaluation of high pressure-decellularized cardiovascular tissues transplanted in rat subcutaneous accelerated-calcification model
Micro-CT evaluation of high pressure-decellularized cardiovascular tissues transplanted in rat subcutaneous accelerated-calcification model
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DOI：
10.1007/s10047-014-0808-7
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发表时间：
2015-06-01
影响因子：
1.3
通讯作者：
Yamaoka, Tetsuji
中科院分区：
文献类型：
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作者：
Mahara, Atsushi;Sago, Mitsuru;Yamaoka, Tetsuji
我们已经成功地通过去除DNA和磷脂来减少猪异体系统中脱细胞主动脉或瓣膜的钙化，但其进一步减少是可取的。本研究在大鼠皮下移植模型（即钙化模型）中对脱细胞组织的钙化进行了评价。采用高静水压力（high-hydrostatic pressure， HHP）方法制备脱细胞样品，植入不同的洗涤介质，在微计算机断层扫描下监测钙化1个月和3个月。采用原子吸收光谱法定量测定钙沉积量。一种细胞培养基表现出很好的细胞去除能力，但在1个月时导致严重的钙化，令人惊讶的是，随着洗涤时间的延长，钙沉积增加。通过去除高DNA酶浓度的DNA片段可以抑制这种钙化。另一方面，即使在低DNase浓度下，用生理盐水洗涤2周后，钙化也大大减少。这些结果表明，洗涤介质中的离子种类和残留的dna酶共同影响体内钙化的趋势，这使我们有可能减少非细胞心脏组织的钙化。
We have succeeded in reducing the calcification of acellular aortas or valves in porcine allogeneic system by removing the DNA and phospholipids, but its further reduction is desirable. Here, the calcification of the acellular tissue was evaluated in rat subcutaneous transplantation model which is known as calcification model. Acellular samples prepared by high-hydrostatic pressure (HHP) protocols with different washing media were implanted and the calcification was monitored under micro-computed tomography for 1 and 3 months. The amount of the calcium deposition was quantitatively evaluated by atomic absorption spectroscopy. A cell culture medium showed very good cell removal ability but led to severe calcification at 1 month, and surprisingly the calcium deposition increased as the washing period increased. This calcification was suppressed by removing the DNA fraction with high DNase concentration. On the other hand, the calcification was greatly reduced when washed with saline even at low DNase concentration after 2 weeks washing. These results suggest that the ion species in the washing medium and the residual DNase cooperatively affect the tendency of in vivo calcification, which led us to the possibility of reduced calcification of acellular cardiac tissues.