A Pulsatile Bioreactor for Conditioning of Tissue-Engineered Cardiovascular Constructs under Endoscopic Visualization

König, Fabian; Hollweck, Trixi; Pfeifer, Stefan; Reichart, Bruno; Wintermantel, E.; Hagl, Christian; Akra, Bassil

doi:10.3390/jfb3030480

Cited by 18 publications

(16 citation statements)

References 32 publications

(66 reference statements)

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“…Student's t-test was performed for comparison of data; the probability value p < 0.05 was considered significant. 10 B: Bioreactor for 5 day conditioning period 11 (750-1100 ml/min). B1: Mounting with polyurethane scaffold placed in the incubation chamber filled with 400 ml culture medium during the experiment.…”

Section: Evaluation Methodsmentioning

confidence: 99%

“…in the following 5 days, the valves were incubated in a novel self-made conditioning bioreactor (EU-Patent pending: EP10166094.2; Figure 2B). 11 During this period, the valves were perfused with 400 ml supplemented FGM by an increasing sinusoidal pulsatile flow for 48 hours at 750 ml/min (≈24 bpm) and 72 hours at 1100 ml/min (≈35.5 bpm). Half of the supplemented FGM was exchanged after 2 days.…”

Section: Cell Seeding and Conditioningmentioning

confidence: 99%

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In Vitro Comparison of Novel Polyurethane Aortic Valves and Homografts After Seeding and Conditioning

Thierfelder

Koenig²,

Bombien

et al. 2013

ASAIO Journal

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The aim of the study was to compare the behavior of seeded cells on synthetic and natural aortic valve scaffolds during a low-flow conditioning period. Polyurethane (group A) and aortic homograft valves (group B) were consecutively seeded with human fibroblasts (FB), and endothelial cells (EC) using a rotating seeding device. Each seeding procedure was followed by an exposure to low pulsatile flow in a dynamic bioreactor for 5 days. For further analysis, samples were taken before and after conditioning. Scanning electron microscopy showed confluent cell layers in both groups. Immunohistochemical analysis showed the presence of EC and FB before and after conditioning as well as the establishment of an extracellular matrix (ECM) during conditioning. A higher expression of ECM was observed on the scaffolds' inner surface. Realtime polymerase chain reaction showed higher inflammatory response during the conditioning of homografts. Endothelialization caused a decrease in inflammatory gene expression. The efficient colonization, the establishment of an ECM, and the comparable inflammatory cell reaction to the scaffolds in both groups proved the biocompatibility of the synthetic scaffold. The newly developed bioreactor permits conditioning and cell adaption to shear stress. Therefore, polyurethane valve scaffolds may offer a new option for aortic valve replacement. ASAIO Journal 2013;59:309-316.

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Section: Evaluation Methodsmentioning

confidence: 99%

Section: Cell Seeding and Conditioningmentioning

confidence: 99%

In Vitro Comparison of Novel Polyurethane Aortic Valves and Homografts After Seeding and Conditioning

Thierfelder

Koenig²,

Bombien

et al. 2013

ASAIO Journal

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“…For DC , PHVPs (n = 5) were conditioned in a novel pulsatile bioreactor with an endoscopic monitoring unit (Figure 2; EU-Patent pending EP10166094; manufactured in-house, [9]) for 3 d at 750 ml/min and 2 d at 1100 ml/min medium flow, after FB seeding and after EC seeding. For the conditioning of FB seeded PHVP, fibroblast growth medium supplemented with 11% FCS and 0.2% Penicillin/Streptomycin were used.…”

Section: Methodsmentioning

confidence: 99%

“…Seeded PHVPs in the group DC were conditioned in a pulsatile bioreactor with an endoscopic monitoring unit [9] for 3 d at 750 ml/min and 2 d at 1100 ml/min. Scale bar = 50 mm.…”

Section: Methodsmentioning

confidence: 99%

Use of a special bioreactor for the cultivation of a new flexible polyurethane scaffold for aortic valve tissue engineering

Aleksieva¹,

Hollweck²,

Thierfelder³

et al. 2012

BioMed Eng OnLine

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BackgroundTissue engineering represents a promising new method for treating heart valve diseases. The aim of this study was evaluate the importance of conditioning procedures of tissue engineered polyurethane heart valve prostheses by the comparison of static and dynamic cultivation methods.MethodsHuman vascular endothelial cells (ECs) and fibroblasts (FBs) were obtained from saphenous vein segments. Polyurethane scaffolds (n = 10) were primarily seeded with FBs and subsequently with ECs, followed by different cultivation methods of cell layers (A: static, B: dynamic). Group A was statically cultivated for 6 days. Group B was exposed to low flow conditions (t1= 3 days at 750 ml/min, t2= 2 days at 1100 ml/min) in a newly developed conditioning bioreactor. Samples were taken after static and dynamic cultivation and were analyzed by scanning electron microscopy (SEM), immunohistochemistry (IHC), and real time polymerase chain reaction (RT-PCR).ResultsSEM results showed a high density of adherent cells on the surface valves from both groups. However, better cell distribution and cell behavior was detected in Group B. IHC staining against CD31 and TE-7 revealed a positive reaction in both groups. Higher expression of extracellular matrix (ICAM, Collagen IV) was observed in Group B. RT- PCR demonstrated a higher expression of inflammatory Cytokines in Group B.ConclusionWhile conventional cultivation method can be used for the development of tissue engineered heart valves. Better results can be obtained by performing a conditioning step that may improve the tolerance of cells to shear stress. The novel pulsatile bioreactor offers an adequate tool for in vitro improvement of mechanical properties of tissue engineered cardiovascular prostheses.

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“…A digital image correlation technique (DIC) can acquire strain fields with high precision by applying marker fields in a random pattern and comparing deformed marker fields to the original based on changes in pixel intensity or gray scale value . Commonly used imaging techniques for monitoring engineered tissues include endoscopic surveillance, magnetic resonance imaging (MRI), or real‐time laser scanning (RTLS) . In addition, a multimodal nonlinear optical microscopy‐optical coherence microscopy (NLOM‐OCM) system can be used to noninvasively delineate relative spatial distributions of collagen, fibrin, and cells …”

Section: Design Of Stretch Bioreactorsmentioning

confidence: 99%

Design considerations and challenges for mechanical stretch bioreactors in tissue engineering

Lei

Ferdous

2016

Biotechnology Progress

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With the increase in average life expectancy and growing aging population, lack of functional grafts for replacement surgeries has become a severe problem. Engineered tissues are a promising alternative to this problem because they can mimic the physiological function of the native tissues and be cultured on demand. Cyclic stretch is important for developing many engineered tissues such as hearts, heart valves, muscles, and bones. Thus a variety of stretch bioreactors and corresponding scaffolds have been designed and tested to study the underlying mechanism of tissue formation and to optimize the mechanical conditions applied to the engineered tissues. In this review, we look at various designs of stretch bioreactors and common scaffolds and offer insights for future improvements in tissue engineering applications. First, we summarize the requirements and common configuration of stretch bioreactors. Next, we present the features of different actuating and motion transforming systems and their applications. Since most bioreactors must measure detailed distributions of loads and deformations on engineered tissues, techniques with high accuracy, precision, and frequency have been developed. We also cover the key points in designing culture chambers, nutrition exchanging systems, and regimens used for specific tissues. Since scaffolds are essential for providing biophysical microenvironments for residing cells, we discuss materials and technologies used in fabricating scaffolds to mimic anisotropic native tissues, including decellularized tissues, hydrogels, biocompatible polymers, electrospinning, and 3D bioprinting techniques. Finally, we present the potential future directions for improving stretch bioreactors and scaffolds. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:543-553, 2016.

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A Pulsatile Bioreactor for Conditioning of Tissue-Engineered Cardiovascular Constructs under Endoscopic Visualization

Cited by 18 publications

References 32 publications

In Vitro Comparison of Novel Polyurethane Aortic Valves and Homografts After Seeding and Conditioning

In Vitro Comparison of Novel Polyurethane Aortic Valves and Homografts After Seeding and Conditioning

Use of a special bioreactor for the cultivation of a new flexible polyurethane scaffold for aortic valve tissue engineering

Design considerations and challenges for mechanical stretch bioreactors in tissue engineering

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