Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen

Biomineralized collagen with intrafibrillar calcium phosphate mineral provides an excellent mimic of the composition and structure of the extracellular matrix of bone, from nano- to micro-scale. Scaffolds prepared from this material have the potential to become the next-generation of synthetic bone...

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Main Authors: Daniel de Melo Pereira, Maria Eischen-Loges, Zeinab Tahmasebi Birgani, Pamela Habibovic
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-10-01
Series:Frontiers in Bioengineering and Biotechnology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fbioe.2020.554565/full
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author Daniel de Melo Pereira
Maria Eischen-Loges
Zeinab Tahmasebi Birgani
Pamela Habibovic
author_facet Daniel de Melo Pereira
Maria Eischen-Loges
Zeinab Tahmasebi Birgani
Pamela Habibovic
author_sort Daniel de Melo Pereira
collection DOAJ
description Biomineralized collagen with intrafibrillar calcium phosphate mineral provides an excellent mimic of the composition and structure of the extracellular matrix of bone, from nano- to micro-scale. Scaffolds prepared from this material have the potential to become the next-generation of synthetic bone graft substitutes, as their unique properties make them closer to the native tissue than synthetic alternatives currently available to clinicians. To understand the interaction between biomineralized collagen and cells that are relevant in the context of bone regeneration, we studied the growth and osteogenic differentiation of bone marrow derived human mesenchymal stromal cells (hMSCs) cultured on biomineralized collagen membranes, and compared it to the cell behavior on collagen membranes without mineral. Cells proliferated normally on both biomimetic membranes, and were more triggered to differentiate toward the osteogenic lineage by the biomineralized collagen. This was shown by the elevated mRNA levels of RUNX2, SPP1, ENPP1, and OCN after 3 days of culture, and COL1A1 after 14 days of culture on mineralized collagen. The mRNA levels of the tested markers of osteogenesis were lower on collagen membranes without mineral, with the exception of OCN, which was more highly expressed on collagen than on biomineralized collagen membranes. Expression by hMSCs of OPG, a gene involved in inhibition of osteoclastogenesis, was higher on biomineralized collagen at day 3, while M-CSF, involved in osteoblast-osteoclast communication, was upregulated on both membranes at day 3 and 14 of culture. Alkaline phosphatase activity of hMSCs was high on both biomimetic membranes when compared with cells cultured on tissue culture plastic. Cell-induced mineralization was observed on collagen membranes, while the high mineral content of the biomineralized membranes prohibited a reliable analysis of cell-induced mineralization on these membranes. In conclusion, we have identified that both collagen and biomineralized collagen support proliferation, osteogenic differentiation and mineralization of hMSCs, with biomineralized membranes having a more pronounced positive effect. These findings support the existing evidence that biomineralized collagen is a promising material in the field of bone regeneration.
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spelling doaj.art-bf5c592b2ad14a749dc8533c4ac85e602022-12-21T23:17:24ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-10-01810.3389/fbioe.2020.554565554565Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized CollagenDaniel de Melo PereiraMaria Eischen-LogesZeinab Tahmasebi BirganiPamela HabibovicBiomineralized collagen with intrafibrillar calcium phosphate mineral provides an excellent mimic of the composition and structure of the extracellular matrix of bone, from nano- to micro-scale. Scaffolds prepared from this material have the potential to become the next-generation of synthetic bone graft substitutes, as their unique properties make them closer to the native tissue than synthetic alternatives currently available to clinicians. To understand the interaction between biomineralized collagen and cells that are relevant in the context of bone regeneration, we studied the growth and osteogenic differentiation of bone marrow derived human mesenchymal stromal cells (hMSCs) cultured on biomineralized collagen membranes, and compared it to the cell behavior on collagen membranes without mineral. Cells proliferated normally on both biomimetic membranes, and were more triggered to differentiate toward the osteogenic lineage by the biomineralized collagen. This was shown by the elevated mRNA levels of RUNX2, SPP1, ENPP1, and OCN after 3 days of culture, and COL1A1 after 14 days of culture on mineralized collagen. The mRNA levels of the tested markers of osteogenesis were lower on collagen membranes without mineral, with the exception of OCN, which was more highly expressed on collagen than on biomineralized collagen membranes. Expression by hMSCs of OPG, a gene involved in inhibition of osteoclastogenesis, was higher on biomineralized collagen at day 3, while M-CSF, involved in osteoblast-osteoclast communication, was upregulated on both membranes at day 3 and 14 of culture. Alkaline phosphatase activity of hMSCs was high on both biomimetic membranes when compared with cells cultured on tissue culture plastic. Cell-induced mineralization was observed on collagen membranes, while the high mineral content of the biomineralized membranes prohibited a reliable analysis of cell-induced mineralization on these membranes. In conclusion, we have identified that both collagen and biomineralized collagen support proliferation, osteogenic differentiation and mineralization of hMSCs, with biomineralized membranes having a more pronounced positive effect. These findings support the existing evidence that biomineralized collagen is a promising material in the field of bone regeneration.https://www.frontiersin.org/articles/10.3389/fbioe.2020.554565/fullbiomineralizationintrafibrillarosteogenesishMSCbone regenerationgraft substitute
spellingShingle Daniel de Melo Pereira
Maria Eischen-Loges
Zeinab Tahmasebi Birgani
Pamela Habibovic
Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen
Frontiers in Bioengineering and Biotechnology
biomineralization
intrafibrillar
osteogenesis
hMSC
bone regeneration
graft substitute
title Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen
title_full Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen
title_fullStr Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen
title_full_unstemmed Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen
title_short Proliferation and Osteogenic Differentiation of hMSCs on Biomineralized Collagen
title_sort proliferation and osteogenic differentiation of hmscs on biomineralized collagen
topic biomineralization
intrafibrillar
osteogenesis
hMSC
bone regeneration
graft substitute
url https://www.frontiersin.org/articles/10.3389/fbioe.2020.554565/full
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AT mariaeischenloges proliferationandosteogenicdifferentiationofhmscsonbiomineralizedcollagen
AT zeinabtahmasebibirgani proliferationandosteogenicdifferentiationofhmscsonbiomineralizedcollagen
AT pamelahabibovic proliferationandosteogenicdifferentiationofhmscsonbiomineralizedcollagen