Cell shape and spreading of stromal (mesenchymal) stem cells cultured on fibronectin coated gold and hydroxyapatite surfaces

Dolatshahi-Pirouz, Alireza; Jensen, Tonny; Kolind, Kristian; Bünger, Cody; Kassem, Moustapha; Foss, Morten; Besenbacher, Flemming

doi:10.1016/j.colsurfb.2010.12.004

Cited by 43 publications

(37 citation statements)

References 48 publications

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“…All STF nanoto- 44 pographies investigated in the absence of adsorbed FN were found to significantly enhance cell adhesion 45 relative to flat substrates; and the addition of FN to STFs was found to have cell-dependent effects on 46 enhancing cell-material interactions. Furthermore, the amount of FN adsorbed to the STFs did not corre- 47 late with comparative enhancements of cell-material interactions, suggesting that nanotopography pre- 48 dominantly contributes to the biocompatibility of homogenous nanocolumnar surfaces. This is the first 49 study to correlate precisely defined nanostructured features with protein distribution and cell-nanoma- 50 terial interactions.…”

mentioning

confidence: 84%

Use of precisely sculptured thin film (STF) substrates with generalized ellipsometry to determine spatial distribution of adsorbed fibronectin to nanostructured columnar topographies and effect on cell adhesion

et al. 2015

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Sculptured thin film (STF) substrates consist of nanocolumns with precise orientation, intercolumnar spacing, and optical anisotropy, which can be used as model biomaterial substrates to study the effect of homogenous nanotopogrophies on the three-dimensional distribution of adsorbed proteins. Generalized ellipsometry was used to discriminate between the distributions of adsorbed FN either on top of or within the intercolumnar void spaces of STFs, afforded by the optical properties of these precisely crafted substrates. Generalized ellipsometry indicated that STFs with vertical nanocolumns enhanced total FN adsorption two-fold relative to flat control substrates and the FN adsorption studies demonstrate different STF characteristics influence the degree of FN immobilization both on top and within intercolumnar spaces, with increasing spacing and surface area enhancing total protein adsorption. Mouse fibroblasts or mouse mesenchymal stem cells were subsequently cultured on STFs, to investigate the effect of highly ordered and defined nanotopographies on cell adhesion, spreading, and proliferation. All STF nanotopographies investigated in the absence of adsorbed FN were found to significantly enhance cell adhesion relative to flat substrates; and the addition of FN to STFs was found to have cell-dependent effects on enhancing cell-material interactions. Furthermore, the amount of FN adsorbed to the STFs did not correlate with comparative enhancements of cell-material interactions, suggesting that nanotopography predominantly contributes to the biocompatibility of homogenous nanocolumnar surfaces. This is the first study to correlate precisely defined nanostructured features with protein distribution and cell-nanomaterial interactions. STFs demonstrate immense potential as biomaterial surfaces for applications in tissue engineering, drug delivery, and biosensing.

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mentioning

confidence: 84%

Use of precisely sculptured thin film (STF) substrates with generalized ellipsometry to determine spatial distribution of adsorbed fibronectin to nanostructured columnar topographies and effect on cell adhesion

et al. 2015

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“…26 Dolatshahi-Pirouz et al demonstrated the importance of fibronectin-coated gold and hydroxyapatite surfaces for shape and spreading of MSCs. 40 Garcia et al demonstrated that conformation of the adsorbed protein (fibronectin) was influenced by surface topography, because its specific RGD binding sequence can be either freely available or hidden. 41 Fewer MSCs were attached to the alumina substrate with 100 nm pores due to less deposited protein, a hidden RGD binding sequence, and a larger pore-to-pore distance on a surface with larger pores.…”

Section: Figurementioning

confidence: 99%

In vitro proliferation and osteogenic differentiation of mesenchymal stem cells on nanoporous alumina

Song¹,

Ju²,

Song³

et al. 2013

IJN

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Cell adhesion, migration, and proliferation are significantly affected by the surface topography of the substrates on which the cells are cultured. Alumina is one of the most popular implant materials used in orthopedics, but few data are available concerning the cellular responses of mesenchymal stem cells (MSCs) grown on nanoporous structures. MSCs were cultured on smooth alumina substrates and nanoporous alumina substrates to investigate the interaction between surface topographies of nanoporous alumina and cellular behavior. Nanoporous alumina substrates with pore sizes of 20 nm and 100 nm were used to evaluate the effect of pore size on MSCs as measured by proliferation, morphology, expression of integrin β1, and osteogenic differentiation. An MTT assay was used to measure cell viability of MSCs on different substrates, and determined that cell viability decreased with increasing pore size. Scanning electron microscopy was used to investigate the effect of pore size on cell morphology. Extremely elongated cells and prominent cell membrane protrusions were observed in cells cultured on alumina with the larger pore size. The expression of integrin β1 was enhanced in MSCs cultured on porous alumina, revealing that porous alumina substrates were more favorable for cell growth than smooth alumina substrates. Higher levels of osteoblastic differentiation markers such as alkaline phosphatase, osteocalcin, and mineralization were detected in cells cultured on alumina with 100 nm pores compared with cells cultured on alumina with either 20 nm pores or smooth alumina. This work demonstrates that cellular behavior is affected by variation in pore size, providing new insight into the potential application of this novel biocompatible material for the developing field of tissue engineering.

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“…7 Protocol I-cultured cells exhibited reduced cell size, 15 and cultured mesenchymal cells exhibited stellate, thinner, and sparse cellular aspects, and decreased cell clusters and numbers, and mineralized nodule formations were not observed over the time. All cells cultured using Protocol I conditions did not reach confluence at day 28, and exhibited large gaps between colonies.…”

Section: Figmentioning

confidence: 99%

“…In contrast, cells cultured using Protocol II, III, and IV conditions exhibited significantly increased cell proliferation between 7th and 28th day, consistent with published reports. 10,15 The importance of supplementing cell culture media with human blood products was also analyzed, to determine its potential for cellular differentiation and mineralized tissue formation. In this study, Protocols III and IV exhibited the highest cell proliferation between the 7th and 28th day, similar to that obtained using Protocol II, with no significant changes in cell size, but rather the formation of brownish mineralized nodules suggestive of cell differentiation.…”

Section: Figmentioning

confidence: 99%

Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells

Pisciolaro

Duailibi

Novo

et al. 2015

Tissue Engineering Part A

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One of the goals in using cells for tissue engineering (TE) and cell therapy consists of optimizing the medium for cell culture. The present study compares three different blood product supplements for improved cell proliferation and protection against DNA damage in cultured human dental pulp stem cells for tooth TE applications. Human cells from dental pulp were first characterized as adult stem cells (ectomesenchymal mixed origin) by flow cytometry. Next, four different cell culture conditions were tested: I, supplement-free; II, supplemented with fetal bovine serum; III, allogeneic human serum; and IV, autologous human serum. Cultured cells were then characterized for cell proliferation, mineralized nodule formation, and colony-forming units (CFU) capability. After 28 days in culture, the comet assay was performed to assess possible damage in cellular DNA. Our results revealed that Protocol IV achieved higher cell proliferation than Protocol I ( p = 0.0112). Protocols II and III resulted in higher cell proliferation than Protocol I, but no statistical differences were found relative to Protocol IV. The comet assay revealed less cell damage in cells cultured using Protocol IV as compared to Protocols II and III. The damage percentage observed on Protocol II was significantly higher than all other protocols. CFUs capability was highest using Protocol IV ( p = 0.0018) and III, respectively, and the highest degree of mineralization was observed using Protocol IV as compared to Protocols II and III. Protocol IV resulted in significantly improved cell proliferation, and no cell damage was observed. These results demonstrate that human blood product supplements can be used as feasible supplements for culturing adult human dental stem cells.

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Cell shape and spreading of stromal (mesenchymal) stem cells cultured on fibronectin coated gold and hydroxyapatite surfaces

Cited by 43 publications

References 48 publications

Use of precisely sculptured thin film (STF) substrates with generalized ellipsometry to determine spatial distribution of adsorbed fibronectin to nanostructured columnar topographies and effect on cell adhesion

Use of precisely sculptured thin film (STF) substrates with generalized ellipsometry to determine spatial distribution of adsorbed fibronectin to nanostructured columnar topographies and effect on cell adhesion

In vitro proliferation and osteogenic differentiation of mesenchymal stem cells on nanoporous alumina

Tooth Tissue Engineering: The Importance of Blood Products as a Supplement in Tissue Culture Medium for Human Pulp Dental Stem Cells

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