CD44 mediates hyaluronan to promote the differentiation of human amniotic mesenchymal stem cells into chondrocytes.

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2023-03-01 DOI:10.1007/s10529-022-03322-2

Yan Xu, Ai-Tong Wang, Jian-Hui Xiao

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引用次数: 2

Abstract

Objectives: CD44 is the major receptor for hyaluronan (HA), but its effect on HA-induced differentiation of human amnion mesenchymal stem cells into chondrocytes is unclear. This study aimed to investigate the effects and mechanisms of CD44 in HA-induced chondrogenesis.

Methods: Immunocytochemistry and toluidine blue staining were used to assess the secretion of type II collagen and aggrecan, respectively. qRT-PCR and western blotting were performed to evaluate the expression of key genes and proteins.

Results: The expression of aggrecan and type II collagen was downregulated after using the anti-CD44 antibody (A3D8). The transcriptional levels of chondrocytes‑associated genes SRY‑box transcription factor 9, aggrecan, and collagen type II alpha 1 chain were also decreased. Thus, CD44 may mediate HA-induced differentiation of hAMSCs into chondrocytes. Further investigation indicated that expression of phosphorylated (p)‑Erk1/2 and p‑Smad2 decreased following CD44 inhibition. The changes in the expression of p-Erk1/2 and p-Smad2 were consistent after using the ERK1/2 inhibitor (U0126) and agonist (EGF), respectively. After administering the p-Smad2 inhibitor, the expression levels of p-ERK1/2 and p-Smad2 appeared downregulated. The results showed crosstalk between Erk1/2 and Smad2. Moreover, inhibition of p-Erk1/2 and p-Smad2 significantly reduced the accumulation of aggrecan and type II collagen.

Conclusion: These data indicate that CD44 mediates HA-induced differentiation of hAMSCs into chondrocytes by regulating Erk1/2 and Smad2 signaling.

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CD44介导透明质酸促进人羊膜间充质干细胞向软骨细胞分化。

目的:CD44是透明质酸(HA)的主要受体，但其在HA诱导的人羊膜间充质干细胞向软骨细胞分化中的作用尚不清楚。本研究旨在探讨CD44在ha诱导的软骨形成中的作用及其机制。方法:采用免疫细胞化学法和甲苯胺蓝染色法分别检测ⅱ型胶原和聚集蛋白的分泌情况。采用qRT-PCR和western blotting检测关键基因和蛋白的表达情况。结果:使用抗cd44抗体(A3D8)后，聚集蛋白和II型胶原的表达下调。软骨细胞相关基因SRY - box转录因子9、聚集蛋白和II型胶原α 1链的转录水平也下降。因此，CD44可能介导ha诱导的hAMSCs向软骨细胞的分化。进一步的研究表明，磷酸化(p)‑Erk1/2和p‑Smad2的表达在CD44抑制后下降。使用ERK1/2抑制剂(U0126)和激动剂(EGF)后，p-Erk1/2和p-Smad2的表达变化一致。给予p-Smad2抑制剂后，p-ERK1/2和p-Smad2的表达水平出现下调。结果表明，Erk1/2与Smad2之间存在串扰。此外，抑制p-Erk1/2和p-Smad2可显著减少聚集蛋白和II型胶原的积累。结论:这些数据表明CD44通过调节Erk1/2和Smad2信号通路介导ha诱导的hAMSCs向软骨细胞的分化。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.