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Atomic Force Microscopy-Based Nanoscopy of Chondrogenically Differentiating Human Adipose-Derived Stem Cells: Nanostructure and Integrin ?1 Expression.

ABSTRACT: Integrin ?1 is known to be involved in differentiation, migration, proliferation, wound repair, tissue development, and organogenesis. In order to analyze the binding probability between integrin ?1 ligand and cluster of differentiation 29 (CD29) receptors, atomic force microscopy (AFM) was used to detect native integrin ?1-coupled receptors on the surface of human adipose-derived stem cells (hADSc). The binding probability of integrin ?1 ligand-receptor interaction was probed by integrin ?1-functionalized tips on hADSc during early chondrogenic differentiation at the two-dimensional cell culture level. Cell morphology and ultrastructure of hADSc were measured by AFM, which demonstrated that long spindled cells became polygonal cells with decreased length/width ratios and increased roughness during chondrogenic induction. The binding of integrin ?1 ligand and CD29 receptors was detected by ?1-functionalized tips for living hADSc. A total of 1200 curves were recorded at 0, 6, and 12 days of chondrogenic induction. Average rupture forces were, respectively, 61.8?±?22.2 pN, 60?±?20.2 pN, and 67.2?±?22.0 pN. Rupture events were 19.58?±?1.74%, 28.03?±?2.05%, and 33.4?±?1.89%, respectively, which demonstrated that binding probability was increased between integrin ?1 ligand and receptors on the surface of hADSc during chondrogenic induction. Integrin ?1 and the ?-catenin/SOX signaling pathway were correlated during chondrogenic differentiation. The results of this investigation imply that AFM offers kinetic and visual insight into the changes in integrin ?1 ligand-CD29 receptor binding on hADSc during chondrogenesis. Changes in cellular morphology, membrane ultrastructure, and the probability of ligand-transmembrane receptor binding were demonstrated to be useful markers for evaluation of the chondrogenic differentiation process.

SUBMITTER: Yang J

PROVIDER: S-EPMC6199198 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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Atomic Force Microscopy-Based Nanoscopy of Chondrogenically Differentiating Human Adipose-Derived Stem Cells: Nanostructure and Integrin β1 Expression.

Yang Jie J He Ming-Tang MT Huang Xun X Wang Qiu-Shi QS Pi Jiang J Wang Hua-Jun HJ Rahhal Ali Hasan AH Luo Si-Min SM Zha Zhen-Gang ZG

Nanoscale research letters 20181023 1

Integrin β1 is known to be involved in differentiation, migration, proliferation, wound repair, tissue development, and organogenesis. In order to analyze the binding probability between integrin β1 ligand and cluster of differentiation 29 (CD29) receptors, atomic force microscopy (AFM) was used to detect native integrin β1-coupled receptors on the surface of human adipose-derived stem cells (hADSc). The binding probability of integrin β1 ligand-receptor interaction was probed by integrin β1-fun ...[more]

PMID: 30353236

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Project description:CD44 ligand-receptor interactions are known to be involved in regulating cell migration and tumor cell metastasis. High expression levels of CD44 correlate with a poor prognosis of melanoma patients. In order to understand not only the mechanistic basis for dacarbazine (DTIC)-based melanoma treatment but also the reason for the poor prognosis of melanoma patients treated with DTIC, dynamic force spectroscopy was used to structurally map single native CD44-coupled receptors on the surface of melanoma cells. The effect of DTIC treatment was quantified by the dynamic binding strength and the ligand-binding free-energy landscape. The results demonstrated no obvious effect of DTIC on the unbinding force between CD44 ligand and its receptor, even when the CD44 nanodomains were reduced significantly. However, DTIC did perturb the kinetic and thermodynamic interactions of the CD44 ligand-receptor, with a resultant greater dissociation rate, lower affinity, lower binding free energy, and a narrower energy valley for the free-energy landscape. For cells treated with 25 and 75 μg/mL DTIC for 24 hours, the dissociation constant for CD44 increased 9- and 70-fold, respectively. The CD44 ligand binding free energy decreased from 9.94 for untreated cells to 8.65 and 7.39 kcal/mol for DTIC-treated cells, which indicated that the CD44 ligand-receptor complexes on DTIC-treated melanoma cells were less stable than on untreated cells. However, affinity remained in the micromolar range, rather than the millimolar range associated with nonaffinity ligands. Hence, the CD44 receptor could still be activated, resulting in intracellular signaling that could trigger a cellular response. These results demonstrate DTIC perturbs, but not completely inhibits, the binding of CD44 ligand to membrane receptors, suggesting a basis for the poor prognosis associated with DTIC treatment of melanoma. Overall, atomic force microscopy-based nanoscopic methods offer thermodynamic and kinetic insight into the effect of DTIC on the CD44 ligand-binding process.

Dataset Information

Atomic Force Microscopy-Based Nanoscopy of Chondrogenically Differentiating Human Adipose-Derived Stem Cells: Nanostructure and Integrin ?1 Expression.

Publications

Atomic Force Microscopy-Based Nanoscopy of Chondrogenically Differentiating Human Adipose-Derived Stem Cells: Nanostructure and Integrin β1 Expression.

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