Osteoblast Response to Rest Periods During Bioreactor Culture of Collagen–Glycosaminoglycan Scaffolds

Plunkett, Niamh A.; Partap, Sonia; O’Brien, Fergal J.

doi:10.1089/ten.tea.2009.0345

Cited by 44 publications

(37 citation statements)

References 33 publications

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“…Additionally, the procedures involved in the preparation of biocompatible scaffolds, such as blending, freeze-drying, and dehydrothermal cross-linking, may alter the conformation of proteins present, reducing potential integrin-ligand interactions. Experimental evidence corroborating this can been seen from published studies; Jaasma and O'Brien, 74 Partap et al, 75 and Plunkett et al 76 all experienced 40%-50% cell loss at shear forces two to four orders of magnitude (8Â10 À4 to 2Â10 À2 Pa) below the lowest seen in 2D experiments for a 49 h time frame.…”

Section: 71supporting

confidence: 65%

Influence of Shear Stress in Perfusion Bioreactor Cultures for the Development of Three-Dimensional Bone Tissue Constructs: A Review

McCoy

O’Brien

2010

Tissue Engineering Part B: Reviews

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184

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Section: 71supporting

confidence: 65%

Influence of Shear Stress in Perfusion Bioreactor Cultures for the Development of Three-Dimensional Bone Tissue Constructs: A Review

McCoy

O’Brien

2010

Tissue Engineering Part B: Reviews

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184

202

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“…Cells have non-uniform attachment lengths or orientations to the direction of flow, with larger cells and orientations more normal to the direction of flow resulting in greater levels of cell deformation. Computational modelling has shown that bridged cells experience up to 500 fold greater levels of cytoskeletal deformation than flat cells (Jungreuthmayer et al, 2009b) under flow conditions that have been shown to promote osteogenic differentiation, but cause cell detachment from the scaffold (Alvarez-Barreto and Cartmell, S. et al, 2003;Partap et al, 2009;Plunkett et al, 2010). We hypothesised the greater levels of cell deformation experienced by bridging cells is the primary contributing factor to cell detachment observed in cell-seeded scaffolds undergoing flow perfusion.…”

Section: Discussionmentioning

confidence: 97%

“…Therefore, in-vitro bone tissue cultivation strategies have sought to enhance the osteogenic differentiation of cell-seeded constructs by employing mechanical stimulation via flow-perfusion (Bancroft et al, 2002;Cartmell, S. et al, 2003;Keogh et al, 2011;Sikavitsas et al, 2003;Sikavitsas et al, 2005;Vance et al, 2005). We have successfully employed an in-house designed perfusion bioreactor to examine the effects of mechanical stimulation on gene expression levels of key osteogenic markers for MC3T3 (pre-osteoblasts) cell-seeded collagen-GAG (CG) scaffolds subjected to a range of fluid flow regimes and for exposure times of 1h to 14 days Partap et al, 2009;Plunkett et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

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Influence of flow rate and scaffold pore size on cell behavior during mechanical stimulation in a flow perfusion bioreactor

McCoy

Jungreuthmayer

O’Brien

2012

Biotech & Bioengineering

104

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FJ. Influence of flow rate and scaffold pore size on cell behavior during mechanical stimulation in a flow perfusion bioreactor. Biotechnology and Bioengineering. 2012;109(6):1583-1594.-Use LicenceAttribution-Non-Commercial-ShareAlike 1.0 You are free:• to copy, distribute, display, and perform the work.• to make derivative works. Under the following conditions:• Attribution -You must give the original author credit.• Non-Commercial -You may not use this work for commercial purposes.• AbstractMechanically stimulating cell-seeded scaffolds by flow-perfusion is one approach utilised for developing clinically applicable bone graft substitutes. A key challenge is determining the magnitude of stimuli to apply that enhances cell differentiation but minimises cell detachment from the scaffold. In this study we employed a combined computational modelling and experimental approach to examine how the scaffold mean pore size influences cell attachment morphology and subsequently impacts upon cell deformation and detachment when subjected to fluid-flow. Cell detachment from osteoblast-seeded collagen-GAG scaffolds was evaluated experimentally across a range of scaffold pore sizes subjected to different flow-rates and exposure times in a perfusion bioreactor. Cell detachment was found to be proportional to flow-rate and inversely proportional to pore size. Using this data, a theoretical model was derived that accurately predicted cell detachment as a function of mean shear stress, mean pore size and time. Computational modelling of cell deformation in response to fluid flow showed the percentage of cells exceeding a critical threshold of deformation correlated with cell detachment experimentally and the majority of these cells were of a bridging morphology (cells stretched across pores). These findings will help researchers optimise the mean pore size of scaffolds and perfusion bioreactor operating conditions to manage cell detachment when mechanically simulating cells via flow perfusion.

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“…We agree with the reviewer that the application of shear stress might have beneficial effect on vasculogenesis. Ongoing research in our laboratory is using a flow perfusion bioreactor to provide a biophysical environment to enhance osteogenesis and vasculogenesis (see for example Plunkett et al, 2010). However, we feel that these types of experiments are beyond the scope of the current www.ecmjournal.org GP Duffy et al Pre-vascularisation of collagen-GAG scaffold study and the inclusion of any bioreactor data would detract from the focus of the current paper.…”

Section: Discussion With Reviewersmentioning

confidence: 98%

Towards in vitro vascularisation of collagen-GAG scaffolds

Duffy¹,

McFadden²,

Byrne³

et al. 2011

eCM

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Collagen-glycosaminoglycan scaffolds that have been used clinically for skin regeneration have also shown significant promise for other applications in tissue engineering. However, regeneration of thicker tissues with the aid of implanted biomaterials is likely to depend on, or be accelerated by, the ability to establish rapid vascularisation of the implant. The present study aims to establish a nascent vascular network in vitro within a CG scaffold as a first step towards that goal. Mesenchymal stem cells (MSCs) were chosen as primary vasculogenic candidate cells and a culture medium that promoted maximal network formation on Matrigel by these cells was selected. MSCs seeded in the CG scaffold formed networks of cord-like structures after one to two weeks in the presence of the vasculogenic medium; similar structures were formed by aortic endothelial cells (ECs) cultured for comparison. Gene expression analysis suggested that the MSCs began to adopt an endothelial phenotype, with RNA for PECAM and VCAM rising while that for α-smooth muscle actin fell. However there was no increase in Tie-2 and vWF expression. Addition of smooth muscle cells (SMCs) as a potential perivascular stabilising component did not have a noticeable effect on MSC-derived networks, although it enhanced EC-derived structures.

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Osteoblast Response to Rest Periods During Bioreactor Culture of Collagen–Glycosaminoglycan Scaffolds

Cited by 44 publications

References 33 publications

Influence of Shear Stress in Perfusion Bioreactor Cultures for the Development of Three-Dimensional Bone Tissue Constructs: A Review

Influence of Shear Stress in Perfusion Bioreactor Cultures for the Development of Three-Dimensional Bone Tissue Constructs: A Review

Influence of flow rate and scaffold pore size on cell behavior during mechanical stimulation in a flow perfusion bioreactor

Towards in vitro vascularisation of collagen-GAG scaffolds

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