Investigation of the influence of UV irradiation on collagen thin films by AFM imaging

Stylianou, Andreas; Yova, Dido; Alexandratou, Eleni

doi:10.1016/j.msec.2014.09.006

Cited by 39 publications

(31 citation statements)

References 78 publications

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“…To that regard, we generated collagen I gels containing 0.5, 1.0 or 3.0 mg/ml collagen I. In order to characterize the generated gels in terms of structure and stiffness, we employed AFM, which can be used for imaging and characterization of the mechanical properties of collagen samples without destroying the fibrillar structure of collagen [32, 33]. As shown in Figure 1A, collagen gels consisted of fibers with 3D random orientations, confirming that the formed gels mimic collagen-rich tissues.…”

Section: Resultsmentioning

confidence: 99%

Identification of Ras suppressor-1 (RSU-1) as a potential breast cancer metastasis biomarker using a three-dimensionalin vitroapproach

et al. 2017

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Breast cancer (BC) is the most common malignant disease in women, with most patients dying from metastasis to distant organs, making discovery of novel metastasis biomarkers and therapeutic targets imperative. Extracellular matrix (ECM)-related adhesion proteins as well as tumor matrix stiffness are important determinants for metastasis. As traditional two-dimensional culture does not take into account ECM stiffness, we employed 3-dimensional collagen I gels of increasing concentration and stiffness to embed BC cells of different invasiveness (MCF-7, MDA-MB-231 and MDA-MB-231-LM2) or tumor spheroids. We tested the expression of cell-ECM adhesion proteins and found that Ras Suppressor-1 (RSU-1) is significantly upregulated in increased stiffness conditions. Interestingly, RSU-1 siRNA-mediated silencing inhibited Urokinase Plasminogen Activator, and metalloproteinase-13, whereas tumor spheroids formed from RSU-1-depleted cells lost their invasive capacity in all cell lines and stiffness conditions. Kaplan-Meier survival plot analysis corroborated our findings showing that high RSU-1 expression is associated with poor prognosis for distant metastasis-free and remission-free survival in BC patients. Taken together, our results indicate the important role of RSU-1 in BC metastasis and set the foundations for its validation as potential BC metastasis marker.

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Section: Resultsmentioning

confidence: 99%

Identification of Ras suppressor-1 (RSU-1) as a potential breast cancer metastasis biomarker using a three-dimensionalin vitroapproach

et al. 2017

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“…Our experimental approach involved the formation of cell spheroids embedded in 3D gels made by purified type I collagen, with tunable stiffness. The structure and elastic properties of gels containing 0.5, 1.0 or 3.0 mg ml 21 collagen were analysed using AFM, which does not destroy the gel's microstructure [29,[42][43][44][45]. Figure 5a depicts the microstructure of the gels consisted of fibres with random orientations, similar to the ECM structure of collagen-rich tumours [46].…”

Section: Matrix Stiffening Inhibits Cell Spheroid Invasion In Three-dmentioning

confidence: 99%

Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts

Stylianou

Gkretsi

Louca

et al. 2019

J. R. Soc. Interface.

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In many solid tumours a desmoplastic reaction takes place, which results in tumour tissue stiffening due to the extensive production of extracellular matrix (ECM) proteins, such as collagen, by stromal cells, mainly fibroblasts (FBs) and cancer-associated fibroblasts (CAFs). In this study, we investigated the effect of collagen stiffness on pancreatic FBs and CAFs, particularly on specific cytoskeleton properties and gene expression involved in tumour invasion. We found that cells become stiffer when they are cultured on stiff substrates and express higher levels of alpha-smooth muscle actin (α-SMA). Also, it was confirmed that on stiff substrates, CAFs are softer than FBs, while on soft substrates they have comparable Young's moduli. Furthermore, the number of spread FBs and CAFs was higher in stiffer substrates, which was also confirmed by Ras-related C3 botulinum toxin substrate 1 ( RAC1 ) mRNA expression, which mediates cell spreading. Although stress fibres in FBs become more oriented on stiff substrates, CAFs have oriented stress fibres regardless of substrate stiffness. Subsequently, we demonstrated that cells' invasion has a differential response to stiffness, which was associated with regulation of Ras homologue family member ( RhoA ) and Rho-associated, coiled-coil containing protein kinase 1 ( ROCK-1 ) mRNA expression. Overall, our results demonstrate that collagen stiffness modulates FBs and CAFs cytoskeleton remodelling and alters their invasion properties.

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“…The use of AFM can significantly help in the research in this direction and especially on issues related to surface properties, which are very important in biomaterials. It has been demonstrated that the characterization of collagen thin films with AFM allowed the exploration of the effects of UV irradiation (254 nm) on the optical properties of collagen (fluorescence, absorption), its topography, roughness, and the influence on cell culturing [94,95]. It must be noticed that surface roughness is a crucial factor in biomedical applications and plays a significant role in cellsurface crosstalk [96,97] as alterations in roughness change the surface that is available for cell adhesion and growth [98].…”

Section: Collagen Thin Films For Studying Collagen-optical Radiation mentioning

confidence: 99%

“…It must be noticed that surface roughness is a crucial factor in biomedical applications and plays a significant role in cellsurface crosstalk [96,97] as alterations in roughness change the surface that is available for cell adhesion and growth [98]. It has been shown that for UV irradiation of the order of magnitude as those used for sterilization/crosslinking purposes, photodegradation occurs and AFM imaging demonstrates modifications in collagen surface roughness [95]. Also, it was demonstrated that fibrils retained their characteristic band and structure D-periodicity (see Figure 4).…”

Section: Collagen Thin Films For Studying Collagen-optical Radiation mentioning

confidence: 99%

Atomic Force Microscopy for Collagen-Based Nanobiomaterials

Stylianou

2017

Journal of Nanomaterials

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Novel nanobiomaterials are increasingly gaining ground in bioengineering research. Among the numerous biomaterials, collagen-nanobiomaterials, such as collagen thin films, are of great interest since they present a wide range of applications in the fields of biomaterials, tissue engineering, and biomedicine. Collagen type I is the most abundant protein within extracellular matrix and, due to its unique characteristics, is widely used as biomaterial. A thorough characterization of the structure and properties of nanomaterials can be achieved by Atomic Force Microscopy (AFM). AFM is a very powerful tool which can be used to obtain qualitative or quantitative information without destroying the collagen fibrillar structure. This mini review covers issues related to the use of AFM for studying the structure and mechanical properties of collagen-based nanobiomaterials, collagen-substrate interactions during the formation of collagen thin films, collagen-cells interactions, and the collagen-optical radiation interactions.

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Investigation of the influence of UV irradiation on collagen thin films by AFM imaging

Cited by 39 publications

References 78 publications

Identification of Ras suppressor-1 (RSU-1) as a potential breast cancer metastasis biomarker using a three-dimensionalin vitroapproach

Identification of Ras suppressor-1 (RSU-1) as a potential breast cancer metastasis biomarker using a three-dimensionalin vitroapproach

Collagen content and extracellular matrix cause cytoskeletal remodelling in pancreatic fibroblasts

Atomic Force Microscopy for Collagen-Based Nanobiomaterials

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