Hydrogel-Supported, Engineered Model of Vocal Fold Epithelium.
Hydrogel-Supported, Engineered Model of Vocal Fold Epithelium.
复制标题
水凝胶支撑的声带上皮工程模型。
DOI：
10.1021/acsbiomaterials.0c01741
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发表时间：
2021-09-13
影响因子：
5.8
通讯作者：
Jia X
中科院分区：
文献类型：
--
作者：
Ravikrishnan A;Fowler EW;Stuffer AJ;Jia X
目前迫切需要建立一个工程模型的声带上皮，可用于获得了解其在声带健康和疾病的作用，并促进新的治疗方案的发展。为了实现这一目标，从健康猪喉分离原代声带上皮细胞（VFEC），并在第3代内使用。培养扩增的VFEC表达基底上上皮标记物、细胞角蛋白13和细胞间连接蛋白、闭合蛋白、E-钙粘蛋白和闭合小带-1。为了建立工程化模型，将VFEC在透明质酸衍生的合成基底膜上培养，所述基底膜展示纤连蛋白衍生的整合素结合肽（RGDSP）和/或层粘连蛋白111衍生的多配体结合肽AG 73（RKRLQVQLSIRT）。我们的研究结果表明，基质硬度和成分协同调节VFECs的粘附，增殖和分层。在具有生理刚度（弹性剪切模量，G' = 1828 Pa）的水凝胶上培养的细胞采用具有紧密细胞-细胞接触的鹅卵石形态，而在较软基质（G' = 41 Pa）上培养的细胞为具有广泛细胞内应力纤维的纺锤形。具有增殖的基底细胞和额外的（1-2）基底上层的复层上皮的发育需要RGDSP和AG 73肽信号两者的存在。在VFEC培养物中补充波形蛋白阳性的原代猪声带成纤维细胞产生的细胞因子导致建立4-5个不同的细胞层。工程化声带上皮在形态上类似于天然组织，表达细胞角蛋白13、粘蛋白1和紧密/粘附连接标记物，并分泌基底膜蛋白IV型胶原和层粘连蛋白5。总的来说，我们的研究结果表明，刚度匹配，细胞-基质接合和旁分泌信号合作有助于VFEC的分层。工程化的上皮细胞可以作为一种通用的工具，用于声带健康和疾病的遗传和分子机制的调查。
There is a critical need for the establishment of an engineered model of the vocal fold epithelium that can be used to gain understanding of its role in vocal fold health and disease and facilitate the development of new treatment options. Towards this goal, primary vocal fold epithelial cells (VFECs) were isolated from healthy porcine larynxes and used within passage 3. Culture expanded VFECs expressed the suprabasal epithelial marker, cytokeratin 13 and intercellular junctional proteins, occludin, E-cadherin and zonula occludens-1. To establish the engineered model, VFECs were cultured on hyaluronic acid-derived synthetic basement membrane displaying fibronectin-derived integrin-binding peptide (RGDSP) and/or laminin 111-derived syndecan-binding peptide AG73 (RKRLQVQLSIRT). Our results show that matrix stiffness and composition cooperatively regulate the adhesion, proliferation and stratification of VFECs. Cells cultured on hydrogels with physiological stiffness (elastic shear modulus, G’ = 1828 Pa) adopted a cobblestone morphology with close cell-cell contacts, while those on softer matrices (G’ = 41 Pa) were spindle-shaped with extensive intracellular stress fibers. The development of stratified epithelium with proliferating basal cells and additional (1–2) suprabasal layers requires the presence of both RGDSP and AG73 peptide signals. Supplementation of cytokines produced by vimentin positive primary porcine vocal fold fibroblasts in the VFEC culture led to the establishment of 4–5 distinct cell layers. The engineered vocal fold epithelium resembled native tissue morphologically, expressed cytokeratin 13, mucin 1 and tight/adherens junction markers and secreted basement membrane proteins collagen IV and laminin 5. Collectively, our results demonstrate that stiffness matching, cell-matrix engagement and paracrine signaling cooperatively contribute to the stratification of VFECs. The engineered epithelium can be used as a versatile tool for investigations of genetic and molecular mechanisms in vocal fold health and disease.
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影响因子：
2.6
作者：
Leydon, Ciara;Imaizumi, Mitsuyoshi;Fried, Marvin P.
通讯作者：
Fried, Marvin P.
DOI：
10.1155/2012/248759
发表时间：
2012
期刊：
Journal of signal transduction
影响因子：
--
作者：
Benoit YD;Groulx JF;Gagné D;Beaulieu JF
通讯作者：
Beaulieu JF
影响因子：
9.7
作者：
Dicker, Kevin T.;Gurski, Lisa A.;Pradhan-Bhatt, Swati;Witt, Robert L.;Farach-Carson, Mary C.;Jia, Xinqiao
通讯作者：
Jia, Xinqiao
影响因子：
4.8
作者：
Hoffman, MP;Engbring, JA;Kleinman, HK
通讯作者：
Kleinman, HK
影响因子：
13.7
作者：
Koizumi, N;Inatomi, T;Kinoshita, S
通讯作者：
Kinoshita, S