Injectable osteogenic microtissues containing mesenchymal stromal cells conformally fill and repair critical-size defects

AbstractRepair of complex fractures with bone loss requires a potent, space-filling intervention to promote regeneration of bone. We present a minimally-invasive strategy combining mesenchymal stromal cells (MSC) with a chitosan-collagen matrix to form modular microtissues designed for delivery through a needle to conformally fill cavital defects. Implantation of microtissues into a calvarial defect in the mouse showed that osteogenically pre-differentiated MSC resulted in complete bridging of the cavity, while undifferentiated MSC produced mineralized tissue only in apposition to native bone. Decreasing the implant volume reduced bone regeneration, while increasing the MSC concentration also attenuated bone formation, suggesting that the cell-matrix ratio is important in achieving a robust response. Conformal filling of the defect with microtissues in a carrier gel resulted in complete healing. Taken together, these results show that modular microtissues can be used to augment the differentiated function of MSC and provide an extracellular environment that potentiates bone repair.

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Mineralization of 3D Osteogenic Model Based on Gelatin-Dextran Hybrid Hydrogel Scaffold Bioengineered with Mesenchymal Stromal Cells: A Multiparametric Evaluation

Materials ◽

10.3390/ma14143852 ◽

2021 ◽

Vol 14 (14) ◽

pp. 3852

Author(s):

Federica Re ◽

Luciana Sartore ◽

Elisa Borsani ◽

Matteo Ferroni ◽

Camilla Baratto ◽

...

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Bone Repair ◽

Active Role ◽

Ionization Mass ◽

Hydrogel Scaffold ◽

Hybrid Hydrogel ◽

Hydrogel Scaffolds ◽

Human Platelet Lysate ◽

Identical Composition

Gelatin–dextran hydrogel scaffolds (G-PEG-Dx) were evaluated for their ability to activate the bone marrow human mesenchymal stromal cells (BM-hMSCs) towards mineralization. G-PEG-Dx1 and G-PEG-Dx2, with identical composition but different architecture, were seeded with BM-hMSCs in presence of fetal bovine serum or human platelet lysate (hPL) with or without osteogenic medium. G-PEG-Dx1, characterized by a lower degree of crosslinking and larger pores, was able to induce a better cell colonization than G-PEG-Dx2. At day 28, G-PEG-Dx2, with hPL and osteogenic factors, was more efficient than G-PEG-Dx1 in inducing mineralization. Scanning electron microscopy (SEM) and Raman spectroscopy showed that extracellular matrix produced by BM-hMSCs and calcium-positive mineralization were present along the backbone of the G-PEG-Dx2, even though it was colonized to a lesser degree by hMSCs than G-PEG-Dx1. These findings were confirmed by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), detecting distinct lipidomic signatures that were associated with the different degree of scaffold mineralization. Our data show that the architecture and morphology of G-PEG-Dx2 is determinant and better than that of G-PEG-Dx1 in promoting a faster mineralization, suggesting a more favorable and active role for improving bone repair.

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Bone marrow-derived mesenchymal stromal cells and platelet-rich plasma on a collagen matrix to improve fascial healing

Hernia ◽

10.1007/s10029-012-0941-2 ◽

2012 ◽

Vol 16 (6) ◽

pp. 677-687 ◽

Cited By ~ 9

Author(s):

J. J. Heffner ◽

J. W. Holmes ◽

J. P. Ferrari ◽

J. Krontiris-Litowitz ◽

H. Marie ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Platelet Rich Plasma ◽

Collagen Matrix

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Mapping the structure and biological functions within mesenchymal bodies using microfluidics

Science Advances ◽

10.1126/sciadv.aaw7853 ◽

2020 ◽

Vol 6 (10) ◽

pp. eaaw7853 ◽

Cited By ~ 6

Author(s):

Sébastien Sart ◽

Raphaël F.-X. Tomasi ◽

Antoine Barizien ◽

Gabriel Amselem ◽

Ana Cumano ◽

...

Keyword(s):

Cell Fate ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Actin Polymerization ◽

Bone Repair ◽

Spatial Organization ◽

Molecular Signaling ◽

Correlative Analysis

Organoids that recapitulate the functional hallmarks of anatomic structures comprise cell populations able to self-organize cohesively in 3D. However, the rules underlying organoid formation in vitro remain poorly understood because a correlative analysis of individual cell fate and spatial organization has been challenging. Here, we use a novel microfluidics platform to investigate the mechanisms determining the formation of organoids by human mesenchymal stromal cells that recapitulate the early steps of condensation initiating bone repair in vivo. We find that heterogeneous mesenchymal stromal cells self-organize in 3D in a developmentally hierarchical manner. We demonstrate a link between structural organization and local regulation of specific molecular signaling pathways such as NF-κB and actin polymerization, which modulate osteo-endocrine functions. This study emphasizes the importance of resolving spatial heterogeneities within cellular aggregates to link organization and functional properties, enabling a better understanding of the mechanisms controlling organoid formation, relevant to organogenesis and tissue repair.

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Mesenchymal stromal cells genetically engineered to overexpress IGF-I enhance cell-based gene therapy of renal failure-induced anemia

AJP Renal Physiology ◽

10.1152/ajprenal.00044.2008 ◽

2008 ◽

Vol 295 (2) ◽

pp. F488-F496 ◽

Cited By ~ 21

Author(s):

Terrence Kucic ◽

Ian B. Copland ◽

Jessica Cuerquis ◽

Daniel L. Coutu ◽

Lorraine E. Chalifour ◽

...

Keyword(s):

Gene Therapy ◽

Renal Failure ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Heart Function ◽

Collagen Matrix ◽

Genetically Engineered ◽

Functional Protein ◽

Igf I

We previously demonstrated that erythropoietin (EPO)-secreting mesenchymal stromal cells (MSC) can be used for the long-term correction of renal failure-induced anemia. The present study provides evidence that coimplantation of insulin-like growth factor I (IGF-I)-overexpressing MSC (MSC-IGF) improves MSC-based gene therapy of anemia by providing paracrine support to EPO-secreting MSC (MSC-EPO) within a subcutaneous implant. IGF-I receptor RNA expression in murine MSC was demonstrated by RT-PCR. Functional protein expression was confirmed by immunoblots and MSC responsiveness to IGF-I stimulation in vitro. IGF-I was also shown to improve MSC survival following staurosporin-induced apoptosis in vitro. A cohort of C57Bl/6 mice was rendered anemic by right kidney electrocoagulation and left nephrectomy. MSC-EPO were subsequently admixed in a bovine collagen matrix and implanted, in combination with MSC-IGF or MSC null, by subcutaneous injection in renal failure mice. In mice receiving MSC-EPO coimplanted with MSC-IGF, hematocrit elevation was greater and enhanced compared with control mice; heart function was also improved. MSC-IGF coimplantation, therefore, represents a promising new strategy for enhancing MSC survival within implanted matrices and for improving cell-based gene therapy of renal anemia.

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Mapping Structure and Biological Functions within Mesenchymal Bodies using Microfluidics

10.1101/514158 ◽

2019 ◽

Cited By ~ 1

Author(s):

Sébastien Sart ◽

Raphaël F.-X. Tomasi ◽

Antoine Barizien ◽

Gabriel Amselem ◽

Ana Cumano ◽

...

Keyword(s):

Cell Fate ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Actin Polymerization ◽

Bone Repair ◽

Spatial Organization ◽

Molecular Signaling ◽

Correlative Analysis

AbstractOrganoids that recapitulate the functional hallmarks of anatomic structures comprise cell populations able to self-organize cohesively in 3D. However, the rules underlying organoid formation in vitro remain poorly understood because a correlative analysis of individual cell fate and spatial organization has been challenging. Here, we use a novel microfluidics platform to investigate the mechanisms determining the formation of organoids by human mesenchymal stromal cells that recapitulate the early steps of condensation initiating bone repair in vivo. We find that heterogeneous mesenchymal stromal cells self-organize in 3D in a developmentally hierarchical manner. We demonstrate a link between structural organization and local regulation of specific molecular signaling pathways such as NF-κB and actin polymerization, which modulate osteo-endocrine functions. This study emphasizes the importance of resolving spatial heterogeneities within cellular aggregates to link organization and functional properties, enabling a better understanding of the mechanisms controlling organoid formation, relevant to organogenesis and tissue repair.

Download Full-text