Enhanced depth-independent chondrocyte proliferation and phenotype maintenance in an ultrasound bioreactor and an assessment of ultrasound dampening in the scaffold

Thakurta, Sanjukta Guha; Kraft, Mikail; Viljoen, Hendrik J.; Subramanian, Anuradha

doi:10.1016/j.actbio.2014.07.013

Cited by 22 publications

(19 citation statements)

References 40 publications

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“…To obtain prewetted scaffolds, BM scaffolds (5 mm × 2.5 mm) were sterilized with sequential treatments of 70% ethanol for 1 h followed by sterile 1X phosphate buffered saline (PBS) rinse and then incubated for 12 h in medium (high-glucose Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS). One side of the prewetted scaffold disks was seeded with hMSCs (passage 5) at a seeding density of 2 × 10 5 cells/scaffold using procedures outlined elsewhere 29 and were transferred to a new six-well tissue culture plate (TCP) housing a cell crown ™ insert/well with 15–18 scaffolds/insert.…”

Section: Methodsmentioning

confidence: 99%

“… 18 The experimental setting was described elsewhere. 29 Automated US stimulation was provided in the near field at the following regimen: 14 kPa (5.0 MHz, 2.5 Vpp), 5 min/application and 6 applications/day at a regular interval. At indicated time points, cell-seeded scaffolds were retrieved randomly from respective groups and assayed as indicated.…”

Section: Methodsmentioning

confidence: 99%

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Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis

2015

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Ultrasound at 5.0 MHz was noted to be chondro-inductive, with improved SOX-9 gene and COL2A1 protein expression in constructs that allowed for cell-to-cell contact. To achieve tissue-engineered cartilage using macroporous scaffolds, it is hypothesized that a combination of ultrasound at 5.0 MHz and transforming growth factor-β3 induces human mesenchymal stem cell differentiation to chondrocytes. Expression of miR-145 was used as a metric to qualitatively assess the efficacy of human mesenchymal stem cell conversion. Our results suggest that in group 1 (no transforming growth factor-β3, no ultrasound), as anticipated, human mesenchymal stem cells were not efficiently differentiated into chondrocytes, judging by the lack of decrease in the level of miR-145 expression. Human mesenchymal stem cells differentiated into chondrocytes in group 2 (transforming growth factor-β3, no ultrasound) and group 3 (transforming growth factor-β3, ultrasound) with group 3 having a 2-fold lower miR-145 when compared to group 2 at day 7, indicating a higher conversion to chondrocytes. Transforming growth factor-β3–induced chondrogenesis with and without ultrasound stimulation for 14 days in the ultrasound-assisted bioreactor was compared and followed by additional culture in the absence of growth factors. The combination of growth factor and ultrasound stimulation (group 3) resulted in enhanced COL2A1, SOX-9, and ACAN protein expression when compared to growth factor alone (group 2). No COL10A1 protein expression was noted. Enhanced cell proliferation and glycosaminoglycan deposition was noted with the combination of growth factor and ultrasound stimulation. These results suggest that ultrasound at 5.0 MHz could be used to induce chondrogenic differentiation of mesenchymal stem cells for cartilage tissue engineering.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis

2015

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“…4. Основные типы биореакторов [63,64]: а -система, включающая вращающийся сосуд; б -система, основанная на комбинации гидростатического давления и перфузии; в -система, основанная на растяжении; г -система, осно-ванная на сжатии; д -незамкнутая система, основанная на перфузии; е -замкнутая система, основанная на перфузии; ж -система, основанная на воздействии электрического поля; з -система, основанная на воздействии ультразвука [63,64]: а -system comprising a rotating vessel; б -system, based on the combination of hydrostatic pressure and perfusion; в -system, based on stretching; г -system, based on compression; д -open system, based on perfusion; е -closed system, based on perfusion; ж -system, based on the infl uence of an electric fi eld; з -system, based on ultrasound Системы, основанные на применении силы сдвига Одна из основных сил, воздействующих на ткань -сила сдвига, создаваемая сжатием синови-альной жидкости гладкими поверхностями хряща при движении сустава. На силу сдвига ткань реаги-рует деформацией без изменения объема, создания градиента давления и потока жидкости.…”

Section: системы основанные на ультразвукеunclassified

Tissue Engineering and Regenerative Medicine Technologies in the Treatment of Articular Cartilage Defects

Басок

Севастьянов

2017

RJTAO

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1 ФГБУ «Федеральный научный центр трансплантологии и искусственных органов имени академика В.И. Шумакова» Минздрава России, Москва, Российская Федерация 2 АНО «Институт медико-биологических исследований и технологий», Москва, Российская Федерация Важнейшими проблемами здравоохранения в индустриальном обществе являются повреждение и деге-нерация суставного хряща, что связано с ограниченными возможностями ткани к регенерации. В обзоре подробно описаны существующие и разрабатываемые технологии восстановления и замещения повреж-денных хрящевых тканей суставов. Дан анализ полученных результатов по двум основным направлени-ям: стимулирование регенерации поврежденной хрящевой ткани и выращивание элементов хрящевой ткани в биореакторах.Ключевые слова: суставной хрящ, клеточно-и тканеинженерные конструкции, регенеративная медицина, биореакторы. TISSUE ENGINEERING AND REGENERATIVE MEDICINE TECHNOLOGIES IN THE TREATMENT OF ARTICULAR CARTILAGE DEFECTS

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“…In vitro experiments have been conducted in various US bioreactor designs [4][5][6][7]. A major limitation present in the majority of commonly used bioreactor designs is the presence of an air-liquid interface in the direct path of US wave propagation.…”

Section: Introductionmentioning

confidence: 99%

Analysis of a Novel Bioreactor Designed for Ultrasound Stimulation of Cell-Seeded Scaffolds

Crapps

Rahimi²,

Case³

2018

Preprint

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Although daily low intensity pulsed ultrasound (LIPUS) treatment has been shown to induce cellular responses supporting bone repair, in vitro studies in 3D models, such as cellseeded scaffolds, are needed to further investigate the underlying cellular mechanisms. This requires well-controlled conditions in an US bioreactor. Computational studies are needed to investigate various effects on US wave propagation influenced by bioreactor configurations, such as reflections at interfaces and wave interference, and optimize the bioreactor design for experimental repeatability. In this study, an enclosed cylindrical sample holder that contained an inner well for placement of a scaffold immersed in culture medium was fabricated by stereolithography 3D printing and combined with an acoustic absorbent material to eliminate the presence of an air-liquid interface perpendicular to the wave propagation path. Finite element simulations conducted in the frequency domain demonstrated that weak standing waves were present within the culture medium, indicating the effects of reflections at solidliquid interfaces within the sample holder, as expected. Focusing on the acoustic pressure at the inner well surface, it was found that the spatially-averaged pressure varied from a maximum to a minimum value as the thickness of the water layer beneath the sample holder was changed. Average pressure values at antinode positions were 2-fold higher than at node positions. A volumeaveraged pressure was calculated within the culture medium corresponding to the region where a scaffold would be centrally located within the bioreactor. It was shown that the thickness of the volume analyzed had a minimal effect on the calculated average pressure. Time-dependent simulations for one complete pulse (i.e. 1 ms) showed that the acoustic pressure in volumes that would be occupied by scaffolds of two different thicknesses (diameter of 8.5 mm and thicknesses of 0.2 or 2.0 mm) reached a stable value after 45 µs, and then remained at that value until the active period of the pulse ceased. Once the active period ended, the acoustic pressure rapidly decreased to a low baseline pressure. Overall, this study showed that the proposed novel bioreactor design provided a controlled environment for the US treatment of a cell-seeded scaffold by removing the air-liquid interface using a custom-designed sample holder and an acoustic absorbent material.

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Enhanced depth-independent chondrocyte proliferation and phenotype maintenance in an ultrasound bioreactor and an assessment of ultrasound dampening in the scaffold

Cited by 22 publications

References 40 publications

Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis

Growth factor and ultrasound-assisted bioreactor synergism for human mesenchymal stem cell chondrogenesis

Tissue Engineering and Regenerative Medicine Technologies in the Treatment of Articular Cartilage Defects

Analysis of a Novel Bioreactor Designed for Ultrasound Stimulation of Cell-Seeded Scaffolds

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