Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds

Bjerre, Lea; Bünger, Cody; Kassem, Moustapha; Mygind, Tina

doi:10.1016/j.biomaterials.2008.03.003

Cited by 110 publications

(124 citation statements)

References 64 publications

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“…The material was selected because they originate from the cortical bone of cattle and, as such, its porosity size and porosity rate is similar to that of the human jaw. Moreover, the wide range of pore sizes and the rough surface provide for a good adhesion base for the cells [13].…”

Section: Discussionmentioning

confidence: 99%

“…Scaffolds were then divided into three groups. Group A and group B were placed into the fluid flow cell culture system with an initial flow rate of 0.3 ml/min on the first day to ensure good cell attachment; the flow rate was then subsequently increased to 1 ml/min for a total of 12 days [13]. The constructs in group B were subjected to 1 h fluid flow per day.…”

Section: Cell Seeding and Culturementioning

confidence: 99%

“…The use of spinner flasks subjects cells on the outer periphery of scaffolds to turbulent flow while the interior of the cells exhibit static molecular diffusion. Similar drawbacks exist in the use of a rotating wall bioreactor [6,[10][11][12][13]. As a result, scholars have focused on the study of fluid flow systems and have suggested that flow-derived shear stress may be the most significant and primary mediator in the mechanical stimulation [14].…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of different culture techniques of osteoblasts on 3D scaffolds

Wu¹,

Ban²,

Geng

et al. 2010

Open Life Sciences

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Bones adjust their structure to withstand the mechanical demands they experience. It is suggested that flow-derived shear stress may be the most significant and primary mediator of mechanical stimulation. In this study, we designed and fabricated a fluid flow cell culture system that can load shear stress onto cells cultured on 3D scaffolds. We evaluated the effect of different culture techniques, namely, (1) continuous perfusion fluid flow, (2) intermittent perfusion fluid flow, and (3) static condition, on the proliferation of osteoblasts seeded on partially deproteinized bones. The flow rate was set at 1 ml/min for all the cells cultured using flow perfusion and the experiment was conducted for 12 days. Scanning electron microscopy analysis indicated an increase in cell proliferation for scaffolds subjected to fluid shear stress. In addition, the long axes of these cells lengthened along the flowing fluid direction. Continuous perfusion significantly enhanced cell proliferation compared to either intermittent perfusion or static condition. All the results demonstrated that fluid shear stress is able to enhance the proliferation of cells and change the form of cells.© Versita Sp. z o.o.

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Section: Discussionmentioning

confidence: 99%

Section: Cell Seeding and Culturementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Evaluation of different culture techniques of osteoblasts on 3D scaffolds

Wu¹,

Ban²,

Geng

et al. 2010

Open Life Sciences

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“…A similar observation has been reported in previous studies in which localized cell aggregation and close contact with ECM enhanced hMSC calcium deposition. 12,33,34 However, for the PC T constructs with an even cell distribution from the PC, calcium depositions in both flow configurations were comparable and were higher than their PC P counterparts. For the PC T constructs, the short duration of the OI (e.g., 7 days) compared with what was used in other studies ( > 14 days) could be a contributing factor that resulted in the limited difference between the PC T -OI T and PC T -OI P constructs.…”

Section: Impact Of the Pc And The Flow Configuration On Hmsc Osteogenmentioning

confidence: 96%

Bioreactor Strategy in Bone Tissue Engineering: Pre-Culture and Osteogenic Differentiation Under Two Flow Configurations

Kim

2012

Tissue Engineering Part A

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Since robust osteogenic differentiation and mineralization are integral to the engineering of bone constructs, understanding the impact of the cellular microenvironments on human mesenchymal stem cell (hMSCs) osteogenic differentiation is crucial to optimize bioreactor strategy. Two perfusion flow conditions were utilized in order to understand the impact of the flow configuration on hMSC construct development during both preculture (PC) in growth media and its subsequent osteogenic induction (OI). The media in the in-house perfusion bioreactor was controlled to perfuse either around (termed parallel flow [PF]) the construct surfaces or penetrate through the construct (termed transverse flow [TF]) for 7 days of the PC followed by 7 days of the OI. The flow configuration during the PC not only changed growth kinetics but also influenced cell distribution and potency of osteogenic differentiation and mineralization during the subsequent OI. While shear stress resulted from the TF stimulated cell proliferation during PC, the convective removal of de novo extracellular matrix (ECM) proteins and growth factors (GFs) reduced cell proliferation on OI. In contrast, the effective retention of de novo ECM proteins and GFs in the PC constructs under the PF maintained cell proliferation under the OI but resulted in localized cell aggregations, which influenced their osteogenic differentiation. The results revealed the contrasting roles of the convective flow as a mechanical stimulus, the redistribution of the cells and macromolecules in 3D constructs, and their divergent impacts on cellular events, leading to bone construct formation. The results suggest that the modulation of the flow configuration in the perfusion bioreactor is an effective strategy that regulates the construct properties and maximizes the functional outcome.

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“…While millions of people are affected by these conditions, patients subjected to prolonged immobility or bed-rest (e.g., due to spinal cord injury, and nonunion), as well as (6,12). Moreover, more complex 3D systems have been used by tissue engineers to explore the role of mechanical forces on MSC adhesion and differentiation, as well as promoting the growth of both bone and cartilage (13)(14)(15)(16)(17)(18)(19). In vivo studies using mechanical stimulation, such as whole body vibration, have also demonstrated their effects on MSC proliferation and differentiation toward osteogenesis (20)(21)(22).…”

mentioning

confidence: 99%

Dynamic Fluid Flow Mechanical Stimulation Modulates Bone Marrow Mesenchymal Stem Cells

Yeh

Lien

et al. 2013

Bone Res.

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Osteoblasts are derived from mesenchymal stem cells (MSCs), which initiate and regulate bone formation. New strategies for osteoporosis treatments have aimed to control the fate of MSCs. While functional disuse decreases MSC growth and osteogenic potentials, mechanical signals enhance MSC quantity and bias their differentiation toward osteoblastogenesis. Through a non-invasive dynamic hydraulic stimulation (DHS), we have found that DHS can mitigate trabecular bone loss in a functional disuse model via rat hindlimb suspension (HLS). To further elucidate the downstream cellular effect of DHS and its potential mechanism underlying the bone quality enhancement, a longitudinal in vivo study was designed to evaluate the MSC populations in response to DHS over 3, 7, 14, and 21 days. Five-month old female Sprague Dawley rats were divided into three groups for each time point: age-matched control, HLS, and HLS+DHS. DHS was delivered to the right mid-tibiae with a daily "10 min on-5 min off-10 min on" loading regime for five days/week. At each sacrifice time point, bone marrow MSCs of the stimulated and control tibiae were isolated through specific cell surface markers and quantified by flow cytometry analysis. A strong time-dependent manner of bone marrow MSC induction was observed in response to DHS, which peaked on day 14. After 21 days, this effect of DHS was diminished. This study indicates that the MSC pool is positively influenced by the mechanical signals driven by DHS. Coinciding with our previous findings of mitigation of disuse bone loss, DHS induced changes in MSC number may bias the differentiation of the MSC population towards osteoblastogenesis, thereby promoting bone formation under disuse conditions. This study provides insights into the mechanism of time-sensitive MSC induction in response to mechanical loading, and for the optimal design of osteoporosis treatments. Keywords: bone adaptation; mechanical loading; noninvasive stimulation; osteoporosis; osteopenia Bone Research (2013) 1: 98-104. doi: 10.4248/BR201301007 IntroductionBone loss due to functional disuse osteopenia, classified as secondary osteoporosis ( 1), leads to osteoporosisrelated fractures and high medical costs (2, 3). While millions of people are affected by these conditions, patients subjected to prolonged immobility or bed-rest (e.g., due to spinal cord injury, and nonunion), as well as (6,12). Moreover, more complex 3D systems have been used by tissue engineers to explore the role of mechanical forces on MSC adhesion and differentiation, as well as promoting the growth of both bone and cartilage (13)(14)(15)(16)(17)(18)(19). In vivo studies using mechanical stimulation, such as whole body vibration, have also demonstrated their effects on MSC proliferation and differentiation toward osteogenesis (20)(21)(22).Mechanical signals direct MSCs toward osteoblastogenesis and are anabolic to bone, while a reduction in mechanical forces on bone leads to adverse outcomes. In vitro simulated microgravity conditions, mimicking the reduced mecha...

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Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds

Cited by 110 publications

References 64 publications

Evaluation of different culture techniques of osteoblasts on 3D scaffolds

Evaluation of different culture techniques of osteoblasts on 3D scaffolds

Bioreactor Strategy in Bone Tissue Engineering: Pre-Culture and Osteogenic Differentiation Under Two Flow Configurations

Dynamic Fluid Flow Mechanical Stimulation Modulates Bone Marrow Mesenchymal Stem Cells

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