The effect of detergents on the basement membrane complex of a biologic scaffold material.
The effect of detergents on the basement membrane complex of a biologic scaffold material.
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DOI：
10.1016/j.actbio.2013.09.006
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发表时间：
2014-01
影响因子：
9.7
通讯作者：
Badylak, S. F.
中科院分区：
文献类型：
--
作者：
Faulk, D. M.;Carruthers, C. A.;Warner, H. J.;Kramer, C. R.;Reing, J. E.;Zhang, L.;D'Amore, A.;Badylak, S. F.
基底膜复合物（BMC）是细胞外基质（ECM）的重要组成部分，支持并促进细胞的生长。本研究探讨了组织脱细胞过程中常用的四种洗涤剂及其对BMC的影响。将猪膀胱的BMC置于3% Triton-X 100、8 mM 3-[（3-胆酰胺丙基）二甲基铵]-1-丙磺酸盐（CHAPS）、4%脱氧胆酸钠或1%十二烷基硫酸钠（SDS）中24小时。通过免疫标记、扫描电子显微镜（SEM）和纤维网络的二次谐波发生（SHG）成像评估每个处理组的BMC结构。通过定量dsDNA、糖胺聚糖（GAG）和胶原蛋白含量来评估组合物。结果显示，与用3%Triton X-100或4%脱氧胆酸钠处理的样品相比，用1%SDS和8 mM CHAPS处理的样品内的胶原纤维变性，并且ECM含有较少的GAG。将人微血管内皮细胞（HMEC）接种到每个BMC上并培养7天。通过整合素β-1的免疫标记、SEM成像和细胞浸润、表型和融合的半定量评估来研究细胞-ECM相互作用。与在用4%脱氧胆酸钠、8 mM CHAPS和1% SDS处理的BMC上培养的HMEC相比，在用3% Triton X-100处理的BMC上培养的HMEC更汇合，并且具有正常表型。8 mM CHAPS和1% SDS均损伤BMC，以至于接种的HMEC能够浸润受损的基底膜下组织，显示融合减少和非典型表型。用于组织脱细胞化的去污剂的选择可对所得生物支架的BMC的完整性具有显著影响。
The basement membrane complex (BMC) is a critical component of the extracellular matrix (ECM) that supports and facilitates the growth of cells. This study investigates four detergents commonly used in the process of tissue decellularization and their effect upon the BMC. The BMC of porcine urinary bladder was subjected to either 3% Triton-X 100, 8 mM 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 4% sodium deoxycholate, or 1% sodium dodecyl sulfate (SDS) for 24 hours. The BMC structure for each treatment group was assessed by immunolabeling, scanning electron microscopy (SEM), and second harmonic generation (SHG) imaging of the fiber network. The composition was assessed by quantification of dsDNA, glycosaminoglycans (GAGs), and collagen content. Results showed that collagen fibers within samples treated with 1% SDS and 8 mM CHAPS were denatured and the ECM contained less GAGs compared to samples treated with 3% Triton X-100 or 4% sodium deoxycholate. Human microvascular endothelial cells (HMECs) were seeded onto each BMC and cultured for 7 days. Cell-ECM interactions were investigated by immunolabeling for integrin β-1, SEM imaging, and semi-quantitative assessment of cellular infiltration, phenotype, and confluence. HMECs cultured on a BMC treated with 3% Triton X-100 were more confluent and had a normal phenotype compared to HMECs cultured on a BMC treated with 4% sodium deoxycholate, 8 mM CHAPS, and 1% SDS. Both 8 mM CHAPS and 1% SDS damaged the BMC to the extent that seeded HMECs were able to infiltrate the damaged sub-basement membrane tissue, showed decreased confluence, and an atypical phenotype. The choice of detergents used for tissue decellularization can have a marked effect upon the integrity of the BMC of the resultant bioscaffold.
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影响因子：
14
作者：
D'Amore A;Stella JA;Wagner WR;Sacks MS
通讯作者：
Sacks MS
影响因子：
14
作者：
Brown, Bryan N.;Barnes, Christopher A.;Kasick, Rena T.;Michel, Roger;Gilbert, Thomas W.;Beer-Stolz, Donna;Castner, David G.;Ratner, BuddyD.;Badylak, Stephen F.
通讯作者：
Badylak, Stephen F.
影响因子：
82.9
作者：
Ott, Harald C.;Clippinger, Ben;Vacanti, Joseph P.
通讯作者：
Vacanti, Joseph P.
影响因子：
4
作者：
Arenas-Herrera, J. E.;Ko, I. K.;Yoo, J. J.
通讯作者：
Yoo, J. J.
影响因子：
--
作者：
Kelly DJ;Rosen AB;Schuldt AJ;Kochupura PV;Doronin SV;Potapova IA;Azeloglu EU;Badylak SF;Brink PR;Cohen IS;Gaudette GR
通讯作者：
Gaudette GR