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Synthetic PAMPS gel induces chondrogenic differentiation of ATDC5 cells via a novel protein reservoir function.
Journal of Biomedical Materials Research Part A ( IF 3.9 ) Pub Date : 2020-06-04 , DOI: 10.1002/jbm.a.37028
Shingo Semba 1 , Nobuto Kitamura 1, 2 , Masumi Tsuda 2, 3, 4 , Keiko Goto 1 , Sadamu Kurono 5 , Yoshihiro Ohmiya 6 , Takayuki Kurokawa 2, 7 , Jian Ping Gong 2, 4, 7 , Kazunori Yasuda 1, 2 , Shinya Tanaka 2, 3, 4
Affiliation  

We previously demonstrated that a synthetic negatively charged poly(2‐acrylamido‐2‐methylpropanesulfonic acid) (PAMPS) gel induced chondrogenic differentiation of ATDC5 cells. In this study, we clarified the underlying molecular mechanism, in particular, focusing on the events that occurred at the interface between the gel and the cells. Gene expression profiling revealed that the expression of extracellular components was enhanced in the ATDC5 cells that were cultured on the PAMPS gel, suggesting that extracellular proteins secreted from the ATDC5 cells might be adsorbed in the PAMPS gel, thereby contributing to the induction of chondrogenic differentiation. Therefore, we created “Treated‐PAMPS gel,” which adsorbed various proteins secreted from the cultured ATDC5 cells during 7 days. Proteomic analysis identified 27 proteins, including extracellular matrix proteins such as Types I, III, and V collagens and thrombospondin (THBS) in the Treated‐PAMPS gel. The Treated‐PAMPS gel preferentially induced expression of chondrogenic markers, namely, aggrecan and Type II collagen, in the ATDC5 cells compared with the untreated PAMPS gel. Addition of recombinant THBS1 to the ATDC5 cells significantly enhanced the PAMPS‐induced chondrogenic differentiation, whereas knockdown of THBS1 completely abolished this response. In conclusion, we demonstrated that the PAMPS gel has the potential to induce chondrogenic differentiation through novel reservoir functions, and the adsorbed THBS plays a significant role in the induction.

中文翻译:

合成 PAMPS 凝胶通过新的蛋白质库功能诱导 ATDC5 细胞的软骨分化。

我们之前证明了合成的带负电荷的聚(2-丙烯酰胺-2-甲基丙磺酸)(PAMPS)凝胶诱导了 ATDC5 细胞的软骨分化。在这项研究中,我们阐明了潜在的分子机制,特别是关注凝胶和细胞界面处发生的事件。基因表达谱显示,在 PAMPS 凝胶上培养的 ATDC5 细胞中细胞外成分的表达增强,表明 ATDC5 细胞分泌的细胞外蛋白可能被 PAMPS 凝胶吸附,从而有助于诱导软骨分化。因此,我们创建了“处理过的 PAMPS 凝胶”,它在 7 天内吸附了培养的 ATDC5 细胞分泌的各种蛋白质。蛋白质组学分析鉴定了 27 种蛋白质,包括细胞外基质蛋白,例如 I、III 和 V 型胶原蛋白以及治疗 PAMPS 凝胶中的血小板反应蛋白 (THBS)。与未经处理的 PAMPS 凝胶相比,经处理的 PAMPS 凝胶优先诱导 ATDC5 细胞中软骨形成标记物(即蛋白聚糖和 II 型胶原蛋白)的表达。向 ATDC5 细胞中添加重组 THBS1 显着增强了 PAMPS 诱导的软骨形成分化,而敲除 THBS1 则完全消除了这种反应。总之,我们证明了 PAMPS 凝胶具有通过新的储库功能诱导软骨形成分化的潜力,并且吸附的 THBS 在诱导中起重要作用。与未经处理的 PAMPS 凝胶相比,经处理的 PAMPS 凝胶优先诱导 ATDC5 细胞中软骨形成标记物(即蛋白聚糖和 II 型胶原蛋白)的表达。向 ATDC5 细胞中添加重组 THBS1 显着增强了 PAMPS 诱导的软骨形成分化,而敲除 THBS1 则完全消除了这种反应。总之,我们证明了 PAMPS 凝胶具有通过新的储库功能诱导软骨形成分化的潜力,并且吸附的 THBS 在诱导中起重要作用。与未经处理的 PAMPS 凝胶相比,经处理的 PAMPS 凝胶优先诱导 ATDC5 细胞中软骨形成标记物(即蛋白聚糖和 II 型胶原蛋白)的表达。向 ATDC5 细胞中添加重组 THBS1 显着增强了 PAMPS 诱导的软骨形成分化,而敲除 THBS1 则完全消除了这种反应。总之,我们证明了 PAMPS 凝胶具有通过新的储库功能诱导软骨形成分化的潜力,并且吸附的 THBS 在诱导中起重要作用。而敲除 THBS1 则完全消除了这种反应。总之,我们证明了 PAMPS 凝胶具有通过新的储库功能诱导软骨形成分化的潜力,并且吸附的 THBS 在诱导中起重要作用。而敲除 THBS1 则完全消除了这种反应。总之,我们证明了 PAMPS 凝胶具有通过新的储库功能诱导软骨形成分化的潜力,并且吸附的 THBS 在诱导中起重要作用。
更新日期:2020-06-04
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