Regulation of Human Mesenchymal Stem Cell Osteogenesis by Specific Surface Density of Fibronectin: a Gradient Study

Faia-Torres, Ana Bela; Goren, Tolga; Ihalainen, Teemu O.; Guimond-Lischer, Stefanie; Charnley, Mirren; Rottmar, Markus; Maniura‐Weber, Katharina; Spencer, Nicholas D.; Reis, Rui L.; Textor, Marcus; Neves, Nuno M.

doi:10.1021/am506951c

Cited by 42 publications

(31 citation statements)

References 40 publications

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“…Specifically, the matrix produced by chondrocytes has a lower fibronectin content and consists of type II collagen, which is not found in fibroblast matrix (Supplemental Figure 8) [46]. The osteo-inductive potential observed in ESF scaffolds could be attributed to the fibronectin enriched decellularized matrix from fibroblasts, given the established role of fibronectin in osteogenesis within hMSCs [47, 48]. Recent studies show that rat bone marrow mesenchymal stromal cells were more osteogenic when cultured on fibroblast derived decellularized matrices as compared to chondrocyte derived decellularized matrices [43].…”

Section: Discussionmentioning

confidence: 99%

Profiling stem cell states in three-dimensional biomaterial niches using high content image informatics

et al. 2016

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A predictive framework for the evolution of stem cell biology in 3-D is currently lacking. In this study we propose deep image informatics of the nuclear biology of stem cells to elucidate how 3-D biomaterials steer stem cell lineage phenotypes. The approach is based on high content imaging informatics to capture minute variations in the 3-D spatial organization of splicing factor SC-35 in the nucleoplasm as a marker to classify emergent cell phenotypes of human mesenchymal stem cells (hMSCs). The cells were cultured in varied 3-D culture systems including hydrogels, electrospun mats and salt leached scaffolds. The approach encompasses high resolution 3-D imaging of SC-35 domains and high content image analysis (HCIA) to compute quantitative 3-D nuclear metrics for SC-35 organization in single cells in concert with machine learning approaches to construct a predictive cell-state classification model. Our findings indicate that hMSCs cultured in collagen hydrogels and induced to differentiate into osteogenic or adipogenic lineages could be classified into the three lineages (stem, adipogenic, osteogenic) with ≥ 80% precision and sensitivity, within 72 hours. Using this framework, the augmentation of osteogenesis by scaffold design exerted by porogen leached scaffolds was also profiled within 72 hours with ~80% high sensitivity. Furthermore, by employing 3-D SC-35 organizational metrics, differential osteogenesis induced by novel electrospun fibrous polymer mats incorporating decellularized matrix could also be elucidated and predictably modeled at just 3 days with high precision. We demonstrate that 3-D SC-35 organizational metrics can be applied to model the stem cell state in 3-D scaffolds. We propose that this methodology can robustly discern minute changes in stem cell states within complex 3-D architectures and map single cell biological readouts that are critical to assessing population level cell heterogeneity.

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Section: Discussionmentioning

confidence: 99%

Profiling stem cell states in three-dimensional biomaterial niches using high content image informatics

et al. 2016

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“…Prior to the cell culture, the PLA-based samples (the scaffolds and sheets) were coated by incubating them in 25 ug/ml fibronectin in phosphate-buffered saline (PBS, BE17-515 F, BioWhittaker ® , Lonza) solution for 48 h at + 37 ºC. The used fibronectin was isolated from human plasma as described earlier 15 . Following the coating, 100 000 cells were applied to each PLA scaffold in a volume of 100 µl.…”

Section: Methodsmentioning

confidence: 99%

A tube-source X-ray microtomography approach for quantitative 3D microscopy of optically challenging cell-cultured samples

et al. 2020

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Development and study of cell-cultured constructs, such as tissue-engineering scaffolds or organ-on-a-chip platforms require a comprehensive, representative view on the cells inside the used materials. However, common characteristics of biomedical materials, for example, in porous, fibrous, rough-surfaced, and composite materials, can severely disturb low-energy imaging. In order to image and quantify cell structures in optically challenging samples, we combined labeling, 3D X-ray imaging, and in silico processing into a methodological pipeline. Cell-structure images were acquired by a tube-source X-ray microtomography device and compared to optical references for assessing the visual and quantitative accuracy. The spatial coverage of the X-ray imaging was demonstrated by investigating stem-cell nuclei inside clinically relevant-sized tissue-engineering scaffolds (5x13 mm) that were difficult to examine with the optical methods. Our results highlight the potential of the readily available X-ray microtomography devices that can be used to thoroughly study relative large cell-cultured samples with microscopic 3D accuracy.

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“…Combined with anionically charged polymers such as PLGA and polyvinylpyrrolidone-I, these collagenbased matrices promote osteogenic differentiation of MSCs, increase fibronectin binding onto the scaffold surface, and enhance COL I synthesis as well as mineralization of the cultured MSCs in vitro. [47][48][49] β1 integrin immunofluorescence analysis…”

Section: Cell Proliferation Analysismentioning

confidence: 99%

The preosteoblast response of electrospinning PLGA/PCL nanofibers: effects of biomimetic architecture and collagen I

Qian¹,

Chen²,

Yang³

et al. 2016

IJN

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Constructing biomimetic structure and incorporating bioactive molecules is an effective strategy to achieve a more favorable cell response. To explore the effect of electrospinning (ES) nanofibrous architecture and collagen I (COL I)-incorporated modification on tuning osteoblast response, a resorbable membrane composed of poly(lactic-co-glycolic acid)/poly(caprolactone) (PLGA/PCL; 7:3 w/w) was developed via ES. COL I was blended into PLGA/PCL solution to prepare composite ES membrane. Notably, relatively better cell response was delivered by the bioactive ES-based membrane which was fabricated by modification of 3,4-dihydroxyphenylalanine and COL I. After investigation by field emission scanning electron microscopy, Fourier transform infrared spectroscopy, contact angle measurement, and mechanical test, polyporous three-dimensional nanofibrous structure with low tensile force and the successful integration of COL I was obtained by the ES method. Compared with traditional PLGA/PCL membrane, the surface hydrophilicity of collagen-incorporated membranes was largely enhanced. The behavior of mouse preosteoblast MC3T3-E1 cell infiltration and proliferation on membranes was studied at 24 and 48 hours. The negative control was fabricated by solvent casting. Evaluation of cell adhesion and morphology demonstrated that all the ES membranes were more favorable for promoting the cell adhesion and spreading than the casting membrane. Cell Counting Kit-8 assays revealed that biomimetic architecture, surface topography, and bioactive properties of membranes were favorable for cell growth. Analysis of β1 integrin expression level by immunofluorescence indicated that such biomimetic architecture, especially COL I-grafted surface, plays a key role in cell adhesion and proliferation. The real-time polymerase chain reaction suggested that both surface topography and bioactive properties could facilitate the cell adhesion. The combined effect of biomimetic architecture with enhanced surface activity by 3,4-dihydroxyphenylalanine-assisted modification and COL I incorporation of PLGA/PCL electrospun membranes could successfully fill osteogenic defects and allow for better cell proliferation and differentiation.

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Regulation of Human Mesenchymal Stem Cell Osteogenesis by Specific Surface Density of Fibronectin: a Gradient Study

Cited by 42 publications

References 40 publications

Profiling stem cell states in three-dimensional biomaterial niches using high content image informatics

Profiling stem cell states in three-dimensional biomaterial niches using high content image informatics

A tube-source X-ray microtomography approach for quantitative 3D microscopy of optically challenging cell-cultured samples

The preosteoblast response of electrospinning PLGA/PCL nanofibers: effects of biomimetic architecture and collagen I

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