Synthesis of scaffold-free, three dimensional, osteogenic constructs following culture of skeletal osteoprogenitor cells on glass surfaces
Synthesis of scaffold-free, three dimensional, osteogenic constructs following culture of skeletal osteoprogenitor cells on glass surfaces
Background: efficient differentiation of stem cells into three-dimensional (3D) osteogenic construct is still an unmet challenge. These constructs can be crucial for patients with bone defects due to congenital or traumatic reasons. The modulation of cell fate and function as a consequence of interaction with the physical and chemical properties of materials is well known.
Methods: the current study has examined the osteogenic differentiation potential of human skeletal populations following culture on glass surfaces, as a monolayer, or in glass tubes as a pellet culture. The 3D prosperities were assessed morphometrically and the differentiation was evaluated through molecular characterization as well as matrix formation.
Results: early temporal expression of alkaline phosphatase expression of skeletal populations was observed following culture on glass surfaces. Skeletal populations seeded on glass tubes, adhered as a monolayer to the tube base and subsequently formed 3D pellets at the air -media interface. The pellets cultured on glass displayed 4.9 ± 1.3 times the weight and 2.9 ± 0.1 the diameter of their counterpart cultured in plastic tubes and displayed enhanced production of osteogenic matrix proteins, such a collagen I and osteonectin. The size and weight of the pellets correlated with surface area in contrast to cell numbers seeded. Global DNA methylation level was decreased in pellets cultured on glass. In contrast, gene expression analysis confirmed upregulation extracellular matrix proteins and osteogenesis-related growth factors.
Conclusion: this simple approach to the culture of skeletal cells on glass tubes provides a scaffold-free, 3D construct platform for generating pellets enabling analysis and evaluation of tissue development and integration of multiple constructs with implications for tissue repair and regenerative application on scale-up.
3D culture, Differentiation, Osteogenesis, Osteoprogenitor cells, Pellets
101143
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
alghfeli, Latifa
487a4799-4abc-4c4a-9727-0dfadba51165
Parambath, D
cbdb79d8-71c9-4b1f-ae6a-fb5d271c73a8
Manzoor, S
e799ebaa-54b2-45ab-ab63-43cbe2dd9e94
Roach, Helmtrud
1c34f427-ea30-43a1-af00-48fde3a7b9fa
El-Serafi, Ahmed
41c64a7d-31d6-4166-911a-89e10ee1ced6
December 2021
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
alghfeli, Latifa
487a4799-4abc-4c4a-9727-0dfadba51165
Parambath, D
cbdb79d8-71c9-4b1f-ae6a-fb5d271c73a8
Manzoor, S
e799ebaa-54b2-45ab-ab63-43cbe2dd9e94
Roach, Helmtrud
1c34f427-ea30-43a1-af00-48fde3a7b9fa
El-Serafi, Ahmed
41c64a7d-31d6-4166-911a-89e10ee1ced6
Oreffo, Richard, alghfeli, Latifa, Parambath, D, Manzoor, S, Roach, Helmtrud and El-Serafi, Ahmed
(2021)
Synthesis of scaffold-free, three dimensional, osteogenic constructs following culture of skeletal osteoprogenitor cells on glass surfaces.
Bone Reports, 15, , [101143].
(doi:10.1016/j.bonr.2021.101143).
Abstract
Background: efficient differentiation of stem cells into three-dimensional (3D) osteogenic construct is still an unmet challenge. These constructs can be crucial for patients with bone defects due to congenital or traumatic reasons. The modulation of cell fate and function as a consequence of interaction with the physical and chemical properties of materials is well known.
Methods: the current study has examined the osteogenic differentiation potential of human skeletal populations following culture on glass surfaces, as a monolayer, or in glass tubes as a pellet culture. The 3D prosperities were assessed morphometrically and the differentiation was evaluated through molecular characterization as well as matrix formation.
Results: early temporal expression of alkaline phosphatase expression of skeletal populations was observed following culture on glass surfaces. Skeletal populations seeded on glass tubes, adhered as a monolayer to the tube base and subsequently formed 3D pellets at the air -media interface. The pellets cultured on glass displayed 4.9 ± 1.3 times the weight and 2.9 ± 0.1 the diameter of their counterpart cultured in plastic tubes and displayed enhanced production of osteogenic matrix proteins, such a collagen I and osteonectin. The size and weight of the pellets correlated with surface area in contrast to cell numbers seeded. Global DNA methylation level was decreased in pellets cultured on glass. In contrast, gene expression analysis confirmed upregulation extracellular matrix proteins and osteogenesis-related growth factors.
Conclusion: this simple approach to the culture of skeletal cells on glass tubes provides a scaffold-free, 3D construct platform for generating pellets enabling analysis and evaluation of tissue development and integration of multiple constructs with implications for tissue repair and regenerative application on scale-up.
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Accepted/In Press date: 8 October 2021
e-pub ahead of print date: 18 October 2021
Published date: December 2021
Additional Information:
© 2021 The Authors. Published by Elsevier Inc.
Keywords:
3D culture, Differentiation, Osteogenesis, Osteoprogenitor cells, Pellets
Identifiers
Local EPrints ID: 453201
URI: http://eprints.soton.ac.uk/id/eprint/453201
ISSN: 2352-1872
PURE UUID: 2a85f62f-49d5-46b8-94c0-21d5fab50983
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Date deposited: 10 Jan 2022 18:06
Last modified: 17 Mar 2024 02:50
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Contributors
Author:
Latifa alghfeli
Author:
D Parambath
Author:
S Manzoor
Author:
Helmtrud Roach
Author:
Ahmed El-Serafi
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