Mechanical stress potentiates the differentiation of periodontal ligament stem cells into keratocytes.
Mechanical stress potentiates the differentiation of periodontal ligament stem cells into keratocytes.
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DOI：
10.1136/bjophthalmol-2017-311150
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发表时间：
2018-04
期刊：
影响因子：
--
通讯作者：
Danielson P
中科院分区：
文献类型：
--
作者：
Chen J;Zhang W;Backman LJ;Kelk P;Danielson P
探讨角膜静态机械牵张对人牙周膜干细胞（PDLSCs）向角膜基质细胞分化的影响，以及机械牵张与诱导液的协同作用。将PDLSC在Flexcell张力系统中暴露于3%静态圆顶形机械应变3天和7天。角膜细胞表型由角膜细胞标志物的基因表达决定。将角质细胞分化（诱导）培养基引入Flexcell系统中，连续或间歇地与机械刺激组合。通过角膜基质细胞标志物基因和蛋白表达的检测，评价力学和诱导液对角膜基质细胞分化的协同作用。最后，通过将PDLSC接种在胶原膜上并诱导角膜细胞分化来组装多层细胞片。评估细胞片层的透明度，并通过免疫荧光染色评价天然人角膜基质的典型标志物。圆顶形机械刺激促进PDLSC向角膜基质细胞分化，如通过ALDH 3A1、CD 34、LUM、COL I和COL V的上调所示。机械刺激后整合素的表达也上调，包括整合素α 1、α 2、β 1和非肌肉肌球蛋白II B。力学和诱导培养基对角膜基质细胞分化有协同作用。在机械力和诱导培养基的共同作用下，细胞片层进行组装。细胞片层透明，呈多层结构，表达角膜基质的典型标志物。圆顶形机械刺激促进PDLSCs向角膜基质细胞分化，并与诱导培养基有协同作用。类似于天然人角膜基质的多层细胞片显示出未来临床应用的潜力。
To explore the role of corneal-shaped static mechanical strain on the differentiation of human periodontal ligament stem cells (PDLSCs) into keratocytes and the possible synergistic effects of mechanics and inducing medium. PDLSCs were exposed to 3% static dome-shaped mechanical strain in a Flexcell Tension System for 3 days and 7 days. Keratocyte phenotype was determined by gene expression of keratocyte markers. Keratocyte differentiation (inducing) medium was introduced in the Flexcell system, either continuously or intermittently combined with mechanical stimulation. The synergistic effects of mechanics and inducing medium on keratocyte differentiation was evaluated by gene and protein expression of keratocyte markers. Finally, a multilamellar cell sheet was assembled by seeding PDLSCs on a collagen membrane and inducing keratocyte differentiation. The transparency of the cell sheet was assessed, and typical markers of native human corneal stroma were evaluated by immunofluorescence staining. Dome-shaped mechanical stimulation promoted PDLSCs to differentiate into keratocytes, as shown by the upregulation of ALDH3A1, CD34, LUM, COL I and COL V. The expression of integrins were also upregulated after mechanical stimulation, including integrin alpha 1, alpha 2, beta 1 and non-muscle myosin II B. A synergistic effect of mechanics and inducing medium was found on keratocyte differentiation. The cell sheets were assembled under the treatment of mechanics and inducing medium simultaneously. The cell sheets were transparent, multilamellar and expressed typical markers of corneal stroma. Dome-shaped mechanical stimulation promotes differentiation of PDLSCs into keratocytes and has synergistic effects with inducing medium. Multilamellar cell sheets that resemble native human corneal stroma show potential for future clinical applications.
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