A systematic study for optimal cell seeding and culture conditions in a perfusion mode bone-tissue bioreactor

Beşkardeş, Işıl Gerçek; Aydın, Gizem; Bektaş, Şeyma; Cengiz, Alper; Gümüşderelı́oğlu, Menemşe

doi:10.1016/j.bej.2018.01.006

Cited by 28 publications

(20 citation statements)

References 27 publications

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“…In another study, MC3TC3 (pre-osteoblast) cells were cultured on chitosan-hydroxyapatite (HA) scaffolds (6 mm diameter × 4 mm thickness circular discs) using static method. The perfusion culture showed greater cell viability and osteogenic differentiation of MC3TC3 (pre-osteoblast cells) compared to static culture [32] . Though, we investigated the osteogenic potential of MSCs under perfusion culture, proved that FF could enhance the osteogenesis.…”

Section: Resultsmentioning

confidence: 95%

Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

2018

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Background3D cell culture is an appropriate method to develop engineered bone tissue, where different bioreactors have been designed to mitigate the challenges in 3D culture. Currently, we tailored a perfusion reactor to witness human mesenchymal stem cells (MSCs) proliferation and differentiation over polylactic acid-polyethylene glycol (PLA/PEG) composite scaffolds.MethodsThe composite scaffolds with different weight ratios of PLA and PEG were prepared using solvent casting-particulate leaching technique. Human umbilcal card blood MSCs were cultured under dynamic and static conditions to elucidate the role of dynamic fluid flow in osteogenesis of MSCs.ResultsThe human MSCs distribution over the scaffolds was confirmed with fluorescent microscopy. Alkaline phosphatase (ALP), calcium mineralization, and collagen formation were found to be higher in PLA90 scaffolds than PLA100 and PLA75. PLA90 scaffolds with better cell adhesion/proliferartion were considered for bioreactor studies and they exhibited enhanced ALP, Ca+2 mineralization and collagen formation under dynamic perfusion than static culture. We further confirmed our observation by looking at expression levels of osteogenic marker (Runx2 and osteonectin) in differentiated MSCs subjected to perfusion culture compared to static culture.ConclusionThe results of the current investigation once again proves that dynamic perfusion cultures improve the osteogenic differentiation of MSCs over hybrid polymer scaffolds (PLA90) for effective bone regeneration.

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Section: Resultsmentioning

confidence: 95%

Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

2018

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“…Comparison between static and dynamic cell seeding in different culture systems sometimes led to controversial results. For example, Beskardes et al compared dynamic and static cell seeding systems in bone tissue cultures, and they concluded that exclusively the static seeding method combined with perfusion culture could enhance cell viability and osteogenic differentiation [31,32].…”

Section: Discussionmentioning

confidence: 99%

Tissue Engineered Esophageal Patch by Mesenchymal Stromal Cells: Optimization of Electrospun Patch Engineering

Pisani

Croce

Chiesa

et al. 2020

IJMS

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Aim of work was to locate a simple, reproducible protocol for uniform seeding and optimal cellularization of biodegradable patch minimizing the risk of structural damages of patch and its contamination in long-term culture. Two seeding procedures are exploited, namely static seeding procedures on biodegradable and biocompatible patches incubated as free floating (floating conditions) or supported by CellCrownTM insert (fixed conditions) and engineered by porcine bone marrow MSCs (p-MSCs). Scaffold prototypes having specific structural features with regard to pore size, pore orientation, porosity, and pore distribution were produced using two different techniques, such as temperature-induced precipitation method and electrospinning technology. The investigation on different prototypes allowed achieving several implementations in terms of cell distribution uniformity, seeding efficiency, and cellularization timing. The cell seeding protocol in stating conditions demonstrated to be the most suitable method, as these conditions successfully improved the cellularization of polymeric patches. Furthermore, the investigation provided interesting information on patches’ stability in physiological simulating experimental conditions. Considering the in vitro results, it can be stated that the in vitro protocol proposed for patches cellularization is suitable to achieve homogeneous and complete cellularizations of patch. Moreover, the protocol turned out to be simple, repeatable, and reproducible.

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“…The flow into scaffold pores is very important for osteogenic differentiation [ 115 ]. In bioreactors for bone formation, perfusion and shear stress should be ensured.…”

Section: Manufacturing Technology Of Composite Scaffoldsmentioning

confidence: 99%

Fabrication of Scaffolds for Bone-Tissue Regeneration

2019

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The present article describes the state of the art in the rapidly developing field of bone tissue engineering, where many disciplines, such as material science, mechanical engineering, clinical medicine and genetics, are interconnected. The main objective is to restore and improve the function of bone tissue by scaffolds, providing a suitable environment for tissue regeneration and repair. Strategies and materials used in oral regenerative therapies correspond to techniques generally used in bone tissue engineering. Researchers are focusing on developing and improving new materials to imitate the native biological neighborhood as authentically as possible. The most promising is a combination of cells and matrices (scaffolds) that can be fabricated from different kinds of materials. This review summarizes currently available materials and manufacturing technologies of scaffolds for bone-tissue regeneration.

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A systematic study for optimal cell seeding and culture conditions in a perfusion mode bone-tissue bioreactor

Cited by 28 publications

References 27 publications

Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

Improved osteogenic differentiation of umbilical cord blood MSCs using custom made perfusion bioreactor

Tissue Engineered Esophageal Patch by Mesenchymal Stromal Cells: Optimization of Electrospun Patch Engineering

Fabrication of Scaffolds for Bone-Tissue Regeneration

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