Surface nanoscale imaging of collagen thin films by Atomic Force Microscopy

Stylianou, Andreas; Yova, Dido

doi:10.1016/j.msec.2013.03.029

Cited by 58 publications

(49 citation statements)

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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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“…26 Because of this, high spin speeds were used in this report to form films with closely packed fibrils with a random orientation. The orientation of the fibers is the result of a number of different parameters, such as the collagensubstrate interactions, collagen adsorption, friction forces, solvent and solution evaporation rate, and centrifugal forces.…”

Section: Discussionmentioning

confidence: 99%

“…The stock solution had a significantly higher concentration than the critical concentration for collagen fibrillogenesis in bulk solution. 26 26 …”

Section: Preparation Of the Stock Solutionsmentioning

confidence: 99%

“…During adsorption, collagen molecules were both adsorbed and diffused, while fibrils were formed and grown on the substrate 23,26,27 because of the collagen-mica interactions. Part of the 8-mg∕mL stock collagen solution (50 μL) was flushed on the substrate, freshly cleaved mica discs (V1, 9.5 diameter, 71856-01 Electron Microscopy Science, Hatfield, PA), and spin coated for 40 s at 6000 rpm (WS-400B-6NPP/LITE Laurell Technologies spin coater, North Wales, PA), in order to form randomly oriented thin films.…”

Section: Collagen Thin Film Formationmentioning

confidence: 99%

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Nanotopography of collagen thin films in correlation with fibroblast response

2013

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Collagen thin films consisting of randomly oriented and oriented collagen fibrils/ fibers are fabricated by hydrodynamic flow and spin coating, and then they are characterized by atomic force microscopy (AFM). Fibroblasts are cultured on these films in order to correlate their morphology and alignment, which are assessed with fluorescence and AFM imaging with different film characteristics. The results showed that the formed films could be used as substrates for culturing cells. Furthermore, cells reacted to film nanocharacteristics and especially to the orientation of fibrils/fibers. The investigation of the influence that the substrate nanotopography has on cells will help to elucidate the mechanisms of cell-biomaterial interactions, and will enable the design of intelligent coatings for implants and tissue engineering purposes.

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“…Фèбрèëëярíàя ñтрóктóрà кîëëàгåíà I è II тèпîâ ñ îт÷åтëèâîй пîпåрå÷íîй èñ÷åр÷åííîñтью (шàг 67 íм) õîрîшî дèàгíîñтèрóåтñя àтîмíî-ñèëîâîй мè-крîñкîпèåй дàжå â ñóщåñтâåííî дåфîрмèрîâàííыõ îñàждåíèåм è âыñыõàíèåм прåпàрàтàõ [8,25]. Этî пîçâîëяåт прåдпîëîжèть, ÷тî кîëëàгåí кîíîдîíтî-âыõ эëåмåíтîâ íå прèíàдëåжèт к грóппå фèбрèëëяр-íыõ, т. å. к I, II, III, V, XI, XXIV è XXVII тèпàм [11,27].…”

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Structure of the organic matter of conodont elements: atomic force microscopy data

Журавлев¹

2017

Vestnik IG KomiSC

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Èзó÷åíèå дåìèíåðàëèзîвàííых в 1Í-ñîëяíîй кèñëîòå фðàíñкèх кîíîдîíòîвых эëåìåíòîв хîðîшåй ñîхðàííîñòè (òåìпåðàòóðà кàòàãåíåзà ìåíåå 60 °С) ñ пîìîщью àòîìíî-ñèëîвîй ìèкðîñкîпèè пîкàзàëî пðèñóòñòвèå ñòðóкòóð пðåдпîëîжèòåëьíî èз íåфèбðèë-ëяðíîãî кîëëàãåíà. Сðàвíåíèå ñ îпóбëèкîвàííыìè дàííыìè пî жåëàòèíàì пîкàзывàåò íèзкóю вåðîяòíîñòь òîãî, ÷òî эòè ñòðóкòó-ðы îòðàжàюò ðåзóëьòàò дåíàòóðàöèè фèбðèëëяðíîãî кîëëàãåíà. В ñî÷åòàíèè ñ ìàòåðèàëàìè пðåдшåñòвóющèх èññëåдîвàíèй пîëó-÷åííыå ðåзóëьòàòы дàюò вîзìîжíîñòь пðåдпîëîжèòь фîðìèðîвàíèå îðãàíè÷åñкîй ìàòðèöы кîíîдîíòîвых эëåìåíòîв èз кîëëàãå-íà IV òèпà, à ñàìèх эëåìåíòîв -зà ñ÷åò ìèíåðàëèзàöèè íå ñîåдèíèòåëьíîй (кàк кîñòíыå òкàíè è дåíòèí зóбîв пîзвîíî÷íых), à эпè-òåëèàëьíîй òкàíè.Ключевые слова: кîíîäîíты, îðгàíè÷åскîå вåщåствî, àтîìíî-сèлîвàя ìèкðîскîпèя.struCture of the organiC matter of Conodont elements: atomiC forCe miCrosCopy data A. V. ZhuravlevInstitute of Geology Komi SC UrB RAS, Syktyvkar micropalaeontology@gmail.comThe mineral composition of conodont elements is known in details; however their organic matter is studied in a less degree. The previous investigations had demonstrated that organic matter, consisting less than 4 % of a conodont element, was composed of collagenlike protein (Kemp, , 2000Zhuravlev & Shevchuk, 2017). This work is aimed at characterizing of the structure of organic matter of the lamellar tissue of the Frasnian conodont element. The studied conodont element (CAI = 1, temperature of catagenesis <60 °C) was demineralized in 1N HCl, dried, and investigated with AFM (Ntegra Prima, NT-MDT, CKP «Geonauka», Syktyvkar). The organic matter of conodont element possesses globular, not fibrillary, nanostructured. The globules of 40-60 nm in size are ordered into linear (0.3-1.5 um wide) and isometric (200-260 nm) structures. Absence of any residues of fibrillar nanostructures, characteristic for collagen I and II, suggests either denaturation of collagen, or presence of non-fibrillar collagen (e.g. IV type of collagen). The observed nanostructures differ significantly from those of gelatins, thus denaturation of collagen is of low probability. The supposition, that collagen of conodont elements is non-fibrillar, is more probable. The conodonts demonstrate extremely specific biomineralization based on non-fibrillar collagen probably of epithelial origin. Thus vertebrate affinities of conodonts are not proved by study of the organic component of conodont elements.

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Surface nanoscale imaging of collagen thin films by Atomic Force Microscopy

Cited by 58 publications

References 67 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

Nanotopography of collagen thin films in correlation with fibroblast response

Structure of the organic matter of conodont elements: atomic force microscopy data

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