Quantitative stain-free and continuous multimodal monitoring of wound healing in vitro with digital holographic microscopy.
Quantitative stain-free and continuous multimodal monitoring of wound healing in vitro with digital holographic microscopy.
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通过数字全息显微镜在体外对伤口愈合的无定量染色和连续多模式监测。
DOI：
10.1371/journal.pone.0107317
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Kemper B
中科院分区：
文献类型：
--
作者：
Bettenworth D;Lenz P;Krausewitz P;Brückner M;Ketelhut S;Domagk D;Kemper B
上皮伤口愈合受损在多种情况下具有显着的病理生理学意义，包括胃肠道溃疡、吻合口瘘和静脉或糖尿病皮肤溃疡。加速伤口闭合的有前景的候选药物通常在体外伤口测定中进行评估。然而，染色程序和不连续监测是阻碍伤口分析准确评估的主要缺点。因此，我们研究了数字全息显微镜（DHM），以适当监测体外伤口愈合，其次，提供有关形态和功能细胞变化以及细胞因子刺激后运动变化的多模式定量信息。在刺激表皮生长因子 (EGF) 和抑制丝裂霉素 c 的情况下，对伤口愈合试验中 Caco-2 细胞的增殖和迁移所反映的伤口闭合进行了 40 小时的延时系列研究和评估。因此，每三十分钟连续记录一次数字全息图。通过定量数字全息相位显微镜的数据分析形态变化，包括细胞厚度、干质量和组织密度。与对照细胞相比，用 EGF 或丝裂霉素 c 刺激 Caco-2 细胞会在伤口愈合过程中导致显着的形态变化。总之，DHM 可以对体外伤口愈合进行准确、无污染和连续的多模式定量监测，并且可能是一种有前途的伤口愈合评估新技术。
Impaired epithelial wound healing has significant pathophysiological implications in several conditions including gastrointestinal ulcers, anastomotic leakage and venous or diabetic skin ulcers. Promising drug candidates for accelerating wound closure are commonly evaluated in in vitro wound assays. However, staining procedures and discontinuous monitoring are major drawbacks hampering accurate assessment of wound assays. We therefore investigated digital holographic microscopy (DHM) to appropriately monitor wound healing in vitro and secondly, to provide multimodal quantitative information on morphological and functional cell alterations as well as on motility changes upon cytokine stimulation. Wound closure as reflected by proliferation and migration of Caco-2 cells in wound healing assays was studied and assessed in time-lapse series for 40 h in the presence of stimulating epidermal growth factor (EGF) and inhibiting mitomycin c. Therefore, digital holograms were recorded continuously every thirty minutes. Morphological changes including cell thickness, dry mass and tissue density were analyzed by data from quantitative digital holographic phase microscopy. Stimulation of Caco-2 cells with EGF or mitomycin c resulted in significant morphological changes during wound healing compared to control cells. In conclusion, DHM allows accurate, stain-free and continuous multimodal quantitative monitoring of wound healing in vitro and could be a promising new technique for assessment of wound healing.
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影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
--
作者：
CROSS, HS;QUARONI, A
通讯作者：
QUARONI, A
影响因子：
5.6
作者：
Karrasch, T;Steinbrecher, KA;Jobin, C
通讯作者：
Jobin, C
影响因子：
4.5
作者：
Curl, CL;Harris, T;Delbridge, LMD
通讯作者：
Delbridge, LMD
影响因子：
3.1
作者：
Greve, B.;Sheikh-Mounessi, F.;Eich, H. T.
通讯作者：
Eich, H. T.