Effect of Cell Seeding Conditions on the Efficiency of In Vivo Bone Formation

Hori, Akiko; Agata, Hideki; Takaoka, Megumi; Tojo, Arinobu; Kagami, Hideaki

doi:10.11607/jomi.4729

Cited by 4 publications

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“…A number of different prevascularization strategies have been developed over the past decades, which can be differentiated according to in vitro vs. in vivo and extrinsic vs. intrinsic approaches.Scaffolds can be fabricated with tubular-shaped structures and seeded with vascular cells such as endothelial cells or progenitor cells in vitro On the other hand, for in vivo prevascularization, scaffolds are implanted in a highly vascularized area such as subcutaneous or muscle tissue21 .…”

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confidence: 99%

The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

Weigand

Beier

Arkudas

et al. 2016

JoVE

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A functional blood vessel network is a prerequisite for the survival and growth of almost all tissues and organs in the human body. Moreover, in pathological situations such as cancer, vascularization plays a leading role in disease progression. Consequently, there is a strong need for a standardized and well-characterized in vivo model in order to elucidate the mechanisms of neovascularization and develop different vascularization approaches for tissue engineering and regenerative medicine. We describe a microsurgical approach for a small animal model for induction of a vascular axis consisting of a vein and artery that are anastomosed to an arteriovenous (AV) loop. The AV loop is transferred to an enclosed implantation chamber to create an isolated microenvironment in vivo, which is connected to the living organism only by means of the vascular axis. Using 3D imaging (MRI, micro-CT) and immunohistology, the growing vasculature can be visualized over time. By implanting different cells, growth factors and matrices, their function in blood vessel network formation can be analyzed without any disturbing influences from the surroundings in a well controllable environment. In addition to angiogenesis and antiangiogenesis studies, the AV loop model is also perfectly suited for engineering vascularized tissues. After a certain prevascularization time, the generated tissues can be transplanted into the defect site and microsurgically connected to the local vessels, thereby ensuring immediate blood supply and integration of the engineered tissue. By varying the matrices, cells, growth factors and chamber architecture, it is possible to generate various tissues, which can then be tailored to the individual patient's needs.

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confidence: 99%

The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

Weigand

Beier

Arkudas

et al. 2016

JoVE

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“…The vast majority of these optimisation studies usually investigate one or two variables, commonly cell seeding density, static versus dynamic conditions, various dynamic conditions, incubation time and cell culture plate surface chemistry. 25 – 28 , 51 , 52 In our study, we used a full factorial design where we looked at four different variables that were simultaneously varied to observe not only their effect on cell seeding efficiency but also their interactions, if any, with each other. If it was not for the factorial DOE approach, we would not have been able to study the interaction of the four variables.…”

Section: Discussionmentioning

confidence: 99%

“…Traditionally, optimisation of cell seeding efficiency has been done by varying one-factor-at-a-time (OFAT) where it is assumed that the different factors are independent of each other. 25 – 28 Therefore, interaction of factors is not studied. Moreover, OFAT experiments are time-consuming.…”

Section: Introductionmentioning

confidence: 99%

The importance of factorial design in tissue engineering and biomaterials science: Optimisation of cell seeding efficiency on dermal scaffolds as a case study

Levin¹,

Sharma²,

Hook³

et al. 2018

J Tissue Eng

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This article presents a case study to show the usefulness and importance of using factorial design in tissue engineering and biomaterials science. We used a full factorial experimental design (2 × 2 × 2 × 3) to solve a routine query in every biomaterial research project: the optimisation of cell seeding efficiency for pre-clinical in vitro cell studies, the importance of which is often overlooked. In addition, tissue-engineered scaffolds can be cellularised with relevant cell type(s) to form implantable tissue constructs, where the cell seeding method must be reliable and robust. Our results show the complex relationship between cells and scaffolds and suggest that the optimum seeding conditions for each material may be different due to different material properties, and therefore, should be investigated for individual scaffolds. Our factorial experimental design can be easily translated to other cell types and three-dimensional biomaterials, where multiple interacting variables can be thoroughly investigated for better understanding of cell–biomaterial interactions.

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“…To seed the cells, BM-MNCs (1 × 10 6 cells) in 20 µl of the culture medium were put directly onto the scaffold as reported previously (Hori, Agata, Takaoka, Tojo, & Kagami, 2016). To confirm cell attachment after seeding, some of the scaffolds with added cells were immersed in the same culture media for 24 hr, then fixed with 4% buffered paraformaldehyde solution (Wako Pure Chemical Corporation, Osaka, Japan), washed in PBS, and incubated with 0.2% crystal violet (Wako, Osaka, Japan) at room temperature for 5 min.…”

Section: Cell Seeding and Visualization Of The Attached Cells On The Scaffoldmentioning

confidence: 99%

Tooth transplantation with a β‐tricalcium phosphate scaffold accelerates bone formation and periodontal tissue regeneration

Uchikawa

Yoshizawa

et al. 2020

Oral Diseases

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Effect of Cell Seeding Conditions on the Efficiency of In Vivo Bone Formation

Cited by 4 publications

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The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

The Arteriovenous (AV) Loop in a Small Animal Model to Study Angiogenesis and Vascularized Tissue Engineering

The importance of factorial design in tissue engineering and biomaterials science: Optimisation of cell seeding efficiency on dermal scaffolds as a case study

Tooth transplantation with a β‐tricalcium phosphate scaffold accelerates bone formation and periodontal tissue regeneration

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