Modeling the Maturation of the Vocal Fold Lamina Propria Using a Bioorthogonally Tunable Hydrogel Platform.
Modeling the Maturation of the Vocal Fold Lamina Propria Using a Bioorthogonally Tunable Hydrogel Platform.
复制标题
使用生物正交可调谐水凝胶平台模拟声带固有层的成熟。
DOI：
10.1002/adhm.202301701
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发表时间：
2023
影响因子：
10
通讯作者：
Jia,Xinqiao
中科院分区：
文献类型：
--
作者：
Zou,Xiaoyu;Zhang,He;Benson,JamieM;Gao,Hanyuan;Burris,DavidL;Fox,JosephM;Jia,Xinqiao
为了建立声带固有层（LP）的工程化模型，采用四嗪与应变烯烃连接，将间充质干细胞（MSC）封装在基于透明质酸（HA）的水凝胶中。为了模拟LP成熟过程中的基质硬化，使用用凶猛的亲二烯体反式环辛烯（TCO）改性的HA对软细胞结构进行扩散控制界面生物正交交联。将培养物在MSC生长培养基中维持14天以提供均匀柔软的新生LP（nLP）模型、细胞包封后零天（sLP 0）或7天（sLP 7）的均匀硬化的构建体以及具有硬顶层和软底层的成熟LP模型（mLP）。安装额外的HA交叉链路限制了小区扩展。与nLP对照相比，sLP 7条件上调纤维基质蛋白（Col I、DCN和FN EDA）、经典成纤维细胞标志物（TNC、FAP和FSP 1）和基质重塑酶（MMP 2、TIMP 1和HAS 3）的表达。第7天硬化也上调分解代谢活动，增强ECM周转，并促进YAP表达。总体而言，原位延迟基质硬化促进了从MSC的成纤维细胞转化，并增强了雅普调节的机械传感。
Toward the goal of establishing an engineered model of the vocal fold lamina propria (LP), mesenchymal stem cells (MSCs) are encapsulated in hyaluronic acid (HA)‐based hydrogels employing tetrazine ligation with strained alkenes. To mimic matrix stiffening during LP maturation, diffusion‐controlled interfacial bioorthogonal crosslinking is carried out on the soft cellular construct using HA modified with a ferocious dienophile,trans‐cyclooctene (TCO). Cultures are maintained in MSC growth media for 14 days to afford a model of a newborn LP that is homogeneously soft (nLP), a homogeneously stiffened construct zero (sLP0) or 7 days (sLP7) post cell encapsulation, and a mature LP model (mLP) with a stiff top layer and a soft bottom layer. Installation of additional HA crosslinks restricts cell spreading. Compared to the nLP controls, sLP7 conditions upregulate the expression of fibrous matrix proteins (Col I, DCN, andFN EDA), classic fibroblastic markers (TNC, FAP, andFSP1), and matrix remodeling enzymes (MMP2, TIMP1, andHAS3). Day 7 stiffening also upregulates the catabolic activities, enhances ECM turnover, and promotesYAPexpression. Overall, in situ delayed matrix stiffening promotes a fibroblast transition from MSCs and enhances YAP‐regulated mechanosensing.