Biomimetic Mineralization Promotes Viability and Differentiation of Human Mesenchymal Stem Cells in a Perfusion Bioreactor

In bone tissue engineering, the design of 3D systems capable of recreating composition, architecture and micromechanical environment of the native extracellular matrix (ECM) is still a challenge. While perfusion bioreactors have been proposed as potential tool to apply biomechanical stimuli, its use has been limited to a low number of biomaterials. In this work, we propose the culture of human mesenchymal stem cells (hMSC) in biomimetic mineralized recombinant collagen scaffolds with a perfusion bioreactor to simultaneously provide biochemical and biophysical cues guiding stem cell fate. The scaffolds were fabricated by mineralization of recombinant collagen in the presence of magnesium (RCP.MgAp). The organic matrix was homogeneously mineralized with apatite nanocrystals, similar in composition to those found in bone. X-Ray microtomography images revealed isotropic porous structure with optimum porosity for cell ingrowth. In fact, an optimal cell repopulation through the entire scaffolds was obtained after 1 day of dynamic seeding in the bioreactor. Remarkably, RCP.MgAp scaffolds exhibited higher cell viability and a clear trend of up-regulation of osteogenic genes than control (non-mineralized) scaffolds. Results demonstrate the potential of the combination of biomimetic mineralization of recombinant collagen in presence of magnesium and dynamic culture of hMSC as a promising strategy to closely mimic bone ECM.

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Nonmineralized and Mineralized Collagen Scaffolds Induce Differential Osteogenic Signaling Pathways in Human Mesenchymal Stem Cells

Advanced Healthcare Materials ◽

10.1002/adhm.201700641 ◽

2017 ◽

Vol 6 (23) ◽

pp. 1700641 ◽

Cited By ~ 10

Author(s):

Qi Zhou ◽

Xiaoyan Ren ◽

David Bischoff ◽

Daniel W. Weisgerber ◽

Dean T. Yamaguchi ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Signaling Pathways ◽

Human Mesenchymal Stem Cells ◽

Collagen Scaffolds ◽

Osteogenic Signaling ◽

Mineralized Collagen

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Ca2+-activated mitochondrial biogenesis and functions improve stem cell fate in Rg3-treated human mesenchymal stem cells

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01974-3 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Taeui Hong ◽

Moon Young Kim ◽

Dat Da Ly ◽

Su Jung Park ◽

Young Woo Eom ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Stem Cell ◽

Cell Fate ◽

Mitochondrial Biogenesis ◽

Biological Activities ◽

Adipogenic Differentiation ◽

Human Mesenchymal Stem Cells ◽

Stem Cell Fate ◽

Mitochondrial Functions

Abstract Although mitochondrial functions are essential for cell survival, their critical roles in stem cell fate, including proliferation, differentiation, and senescence, remain elusive. Ginsenoside Rg3 exhibits various biological activities and reportedly increases mitochondrial biogenesis and respiration. Herein, we observed that Rg3 increased proliferation and suppressed senescence of human bone marrow-derived mesenchymal stem cells. Osteogenic, but not adipogenic, differentiation was facilitated by Rg3 treatment. Rg3 suppressed reactive oxygen species production and upregulated mitochondrial biogenesis and antioxidant enzymes, including superoxide dismutase. Consistently, Rg3 strongly augmented basal and ATP synthesis-linked respiration with high spare respiratory capacity. Rg3 treatment elevated cytosolic Ca2+ concentration contributing to mitochondrial activation. Reduction of intracellular or extracellular Ca2+ levels strongly inhibited Rg3-induced activation of mitochondrial respiration and biogenesis. Taken together, Rg3 enhances capabilities of mitochondrial and antioxidant functions mainly through a Ca2+-dependent pathway, which improves the proliferation and differentiation potentials and prevents the senescence of human mesenchymal stem cells.

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Molecular Image Analysis: Quantitative Description and Classification of the Nuclear Lamina in Human Mesenchymal Stem Cells

International Journal of Molecular Imaging ◽

10.1155/2011/723283 ◽

2011 ◽

Vol 2011 ◽

pp. 1-11 ◽

Cited By ~ 4

Author(s):

Christiaan H. Righolt ◽

Vered Raz ◽

Bart J. Vermolen ◽

Roeland W. Dirks ◽

Hans J. Tanke ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Fate ◽

3D Structure ◽

Quantitative Description ◽

Human Mesenchymal Stem Cells ◽

Nuclear Lamina ◽

Cellular Functions ◽

Molecular Image ◽

Structural Framework

The nuclear lamina is an intermediate filament network that provides a structural framework for the cell nucleus. Changes in lamina structure are found during changes in cell fate such as cell division or cell death and are associated with human diseases. An unbiased method that quantifies changes in lamina shape can provide information on cells undergoing changes in cellular functions. We have developed an image processing methodology that finds and quantifies the 3D structure of the nuclear lamina. We show that measurements on such images can be used for cell classification and provide information concerning protein spatial localization in this structure. To demonstrate the efficacy of this method, we compared the lamina of unmanipulated human mesenchymal stem cells (hMSCs) at passage 4 to cells activated for apoptosis. A statistically significant classification was found between the two populations.

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Covalent conjugation of mechanically stiff graphene oxide flakes to three-dimensional collagen scaffolds for osteogenic differentiation of human mesenchymal stem cells

Carbon ◽

10.1016/j.carbon.2014.11.029 ◽

2015 ◽

Vol 83 ◽

pp. 162-172 ◽

Cited By ~ 74

Author(s):

Seokyung Kang ◽

Jong Bo Park ◽

Tae-Jin Lee ◽

Seungmi Ryu ◽

Suk Ho Bhang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Graphene Oxide ◽

Osteogenic Differentiation ◽

Three Dimensional ◽

Human Mesenchymal Stem Cells ◽

Collagen Scaffolds

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Graphene Nano-Fiber Composites for Enhanced Neuronal Differentiation of Human Mesenchymal Stem Cells†

10.21203/rs.3.rs-115775/v1 ◽

2020 ◽

Author(s):

Sujata Mohanty ◽

Krishan Gopal Jain ◽

Sonali Rawat ◽

Deepika Gupta ◽

Pawan Raghav ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Fate ◽

Human Mesenchymal Stem Cells ◽

Cell Fate Decisions ◽

Physiological Relevance ◽

Nano Fiber ◽

Intracellular Ca ◽

Degree Of Differentiation ◽

Poly Caprolactone

Abstract Graphene-based nanocomposites have been extensively employed to design biomimetic platforms epitomizing the structural and functional complexity of the tissue with increased robustness and physiological relevance. The adhesive and mechanical cues provided by such nanocomposite microenvironment kindles the cell fate decisions. Owing to their differentiation and regenerative potential, Human Mesenchymal Stem Cells (hMSCs) have proven to be a promising candidate for treating several neurodegenerative disorders. However, their degree of differentiation and its reproducibility is often jeopardized by multiple levels of heterogeneity, thereby compromising their translational utilization. Baffled at this crossroad, we designed a one-step approach to electrospin Poly-caprolactone (PCL) nanocomposite, with varying graphene concentrations, to capture, for the first time, the realms of their biocompatible and anisotropic characteristics, providing biomimetic platforms for improved differentiation of human bone marrow-derived MSCs (hMSCs) into neurons. Interestingly, PCL having 0.05% graphene (PCL-G0.05) showcased an ideal nano-topography with an unprecedented combination of guidance stimuli and substrates cues, aiding in enhanced differentiation of hMSCs into dopaminergic neurons (DA). These newly differentiated DA neurons were characterized at gene, protein, and functional levels and were seen to exhibit unique neuronal arborization, enhanced intracellular Ca2+ influx, and dopamine secretion, thereby opening new horizons for pre-clinical and clinical applications.

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