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基于氢氧化钠的非去污剂去细胞化溶液,用于大鼠肺。

Sodium hydroxide based non-detergent decellularizing solution for rat lung.

机构信息

a Department of Surgical Oncology , Nagasaki University Graduate School of Biomedical Sciences , 1-7-1 Sakamoto, Nagasaki City , Japan.

b Translational Research Center , Research Institute for Science & Technology, Tokyo University of Science , Chiba , Japan.

出版信息

Organogenesis. 2018;14(2):94-106. doi: 10.1080/15476278.2018.1462432. Epub 2018 Jun 11.

Abstract

Lung transplantation is the last option for the treatment of end stage chronic lung disorders. Because the shortage of donor lung organs represents the main hurdle, lung regeneration has been considered to overcome this hurdle. Recellularization of decellularized organ scaffold is a promising option for organ regeneration. Although detergents are ordinarily used for decellularization, other approaches are possible. Here we used high alkaline (pH12) sodium hydroxide (NaOH)-PBS solution without detergents for lung decellularization and compared the efficacy on DNA elimination and ECM preservation with detergent based decellularization solutions CHAPS and SDS. Immunohistochemical image analysis showed that cell components were removed by NaOH solution as well as other detergents. A Collagen and GAG assay showed that the collagen reduction of the NaOH group was comparable to that of the CHAPS and SDS groups. However, DNA reduction was more significant in the NaOH group than in other groups (p < 0.0001). The recellularization of HUVEC revealed cell attachment was not inferior to that of the SDS group. Ex vivo functional analysis showed 100% oxygen ventilation increased oxygen partial pressure as artificial hemoglobin vesicle-PBS solution passed through regenerated lungs in the SDS or NaOH group. It was concluded that the NaOH-PBS based decellularization solution was comparable to ordinal decellularizaton solutions and competitive in cost effectiveness and residues in the decellularized scaffold negligible, thus providing another potential option to detergent for future clinical usage.

摘要

肺移植是治疗终末期慢性肺部疾病的最后手段。由于供体肺器官的短缺是主要障碍,因此肺再生已被认为是克服这一障碍的方法。脱细胞器官支架的再细胞化是一种很有前途的器官再生方法。虽然通常使用去污剂进行脱细胞处理,但也有其他方法。在这里,我们使用高碱性(pH12)的氢氧化钠(NaOH)-PBS 溶液而不使用去污剂进行肺脱细胞处理,并将其与基于去污剂的脱细胞溶液 CHAPS 和 SDS 的 DNA 消除和 ECM 保存效果进行了比较。免疫组织化学图像分析表明,NaOH 溶液和其他去污剂均可去除细胞成分。胶原和 GAG 测定表明,NaOH 组的胶原减少与 CHAPS 和 SDS 组相当。然而,NaOH 组的 DNA 减少比其他组更显著(p < 0.0001)。HUVEC 的再细胞化表明,细胞附着并不逊于 SDS 组。体外功能分析表明,在 SDS 或 NaOH 组中,人工血红蛋白囊泡-PBS 溶液通过再生肺时,100%氧气通气会增加氧分压。因此,NaOH-PBS 基脱细胞溶液与常规脱细胞溶液相当,在成本效益和脱细胞支架中的残留物方面具有竞争力,因此为未来的临床应用提供了另一种替代去污剂的潜在选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/198a/6150056/233c5d7fcccd/kogg-14-02-1462432-g001.jpg

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