Biochemical indicators of implantation success of tissue-engineered oral mucosa.
Biochemical indicators of implantation success of tissue-engineered oral mucosa.
复制标题
组织工程口腔粘膜植入成功的生化指标。
DOI：
10.1177/0022034514554225
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发表时间：
2015
影响因子：
7.6
通讯作者：
Feinberg,SE
中科院分区：
文献类型：
--
作者：
Kuo,S;Zhou,Y;Kim,HM;Kato,H;Kim,RY;Bayar,GR;Marcelo,CL;Kennedy,RT;Feinberg,SE
在移植之前，实时（RT）测定体外组织工程构建体的健康状况对于预测植入组织工程移植体的成功至关重要。此外，美国食品和药物管理局要求在人体使用的组织工程设备发布之前，在RT中有特定的释放标准。原则上，评估细胞成分的活力和功能可以通过量化组织工程构建物的生长因子和趋化因子的分泌来实现。体外产生的口腔黏膜等效物（EVPOME）在43°C的热应力条件下制作24小时，以创建功能受损的EVPOME。我们使用微通道酶联免疫吸附法，通过测量血管内皮生长因子（VEGF）、白细胞介素-8 （IL-8）、人β-防御素1 （hBD-1）和金属蛋白酶组织抑制剂1和2 （TIMP-1和-2）在废培养基中的释放，来评估应激和非应激EVPOMEs的细胞成分、口腔角质形成细胞的功能。废培养基是在移植严重联合免疫缺陷小鼠的前一天收集的。移植EVPOMEs在移植后第7天的组织学将移植结果与相应EVPOMEs中IL-8、hBD-1、VEGF、TIMP-1和TIMP-2的分泌量相关联。我们的研究结果表明，与热应激evpome相比，对照组分泌的IL-8、hBD-1和TIMP-2水平明显更高。我们还发现VEGF和IL-8的分泌与植入evpome的血管计数直接相关。我们的结论是，测量这些因子的组成释放可以作为移植前健康组织工程evpome的无创预测指标。
Real-time (RT) determination of the health of in vitro tissue-engineered constructs prior to grafting is essential for prediction of success of the implanted tissue-engineered graft. In addition, the US Food and Drug Administration requires specific release criteria in RT prior to the release of tissue-engineered devices for human use. In principle, assessing the viability and functionality of the cellular component can be achieved by quantifying the secretion of growth factors and chemokines of tissue-engineered constructs. Ex vivo–produced oral mucosa equivalents (EVPOMEs) were fabricated under thermally stressed conditions at 43 °C for 24 h to create a functionally compromised EVPOME. We used microchannel enzyme-linked immunosorbent assay to evaluate the functionality of the cellular component, oral keratinocytes, of stressed and unstressed EVPOMEs by measuring the release of vascular endothelial growth factor (VEGF), interleukin-8 (IL-8), human β-defensin 1 (hBD-1), and tissue inhibitor of metalloproteinase 1 and 2 (TIMP-1 and -2) into the spent medium, which was collected on the same day prior to graft implantation into severe combined immunodeficiency mice. Implanted EVPOMEs’ histology on the seventh postimplantation day was used to correlate outcomes of grafting to secreted amounts of IL-8, hBD-1, VEGF, TIMP-1, and TIMP-2 from corresponding EVPOMEs. Our findings showed that significantly higher levels of IL-8, hBD-1, and TIMP-2 were secreted from controls than from thermally stressed EVPOMEs. We also found a direct correlation between secreted VEGF and IL-8 and blood vessel counts of implanted EVPOMEs. We concluded that measuring the constitutive release of these factors can be used as noninvasive predictors of healthy tissue-engineered EVPOMEs in RT, prior to their implantation.