Modulation of fibroblast-to-myofibroblast differentiation and fibroblast migration: in vitro assessment of the anti-fibrotic effects of human adipose derived multipotent mesenchymal stromal cells

Abstract Background Nerve injuries are debilitating, leading to long-term motor deficits. Remyelination and axonal growth are supported and enhanced by growth factor and cytokines. Combination of nerve guidance conduits (NGCs) with adipose-tissue-derived multipotent mesenchymal stromal cells (AdMSCs) has been performing promising strategy for nerve regeneration. Methods 3D-printed polycaprolactone (PCL)-NGCs were fabricated. Wistar rats subjected to critical sciatic nerve damage (12-mm gap) were divided into sham, autograft, PCL (empty NGC), and PCL + MSCs (NGC multi-functionalized with 106 canine AdMSCs embedded in heterologous fibrin biopolymer) groups. In vitro, the cells were characterized and directly stimulated with interferon-gamma to evaluate their neuroregeneration potential. In vivo, the sciatic and tibial functional indices were evaluated for 12 weeks. Gait analysis and nerve conduction velocity were analyzed after 8 and 12 weeks. Morphometric analysis was performed after 8 and 12 weeks following lesion development. Real-time PCR was performed to evaluate the neurotrophic factors BDNF, GDNF, and HGF, and the cytokine and IL-10. Immunohistochemical analysis for the p75NTR neurotrophic receptor, S100, and neurofilament was performed with the sciatic nerve. Results The inflammatory environment in vitro have increased the expression of neurotrophins BDNF, GDNF, HGF, and IL-10 in canine AdMSCs. Nerve guidance conduits multi-functionalized with canine AdMSCs embedded in HFB improved functional motor and electrophysiological recovery compared with PCL group after 12 weeks. However, the results were not significantly different than those obtained using autografts. These findings were associated with a shift in the regeneration process towards the formation of myelinated fibers. Increased immunostaining of BDNF, GDNF, and growth factor receptor p75NTR was associated with the upregulation of BDNF, GDNF, and HGF in the spinal cord of the PCL + MSCs group. A trend demonstrating higher reactivity of Schwann cells and axonal branching in the sciatic nerve was observed, and canine AdMSCs were engrafted at 30 days following repair. Conclusions 3D-printed NGCs multi-functionalized with canine AdMSCs embedded in heterologous fibrin biopolymer as cell scaffold exerted neuroregenerative effects. Our multimodal approach supports the trophic microenvironment, resulting in a pro-regenerative state after critical sciatic nerve injury in rats.

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The effects of different doses of thymulin in vivo and in vitro on some biological properties of bone marrow-derived multipotent mesenchymal stromal cells in mice of different strains

Cell and Organ Transplantology ◽

10.22494/cot.v6i1.82 ◽

2018 ◽

Vol 6 (1) ◽

pp. 66-73

Author(s):

I. Labunets ◽

◽

A. Rodnichenko ◽

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Multipotent Mesenchymal Stromal Cells ◽

Biological Properties ◽

Different Strains ◽

Different Doses

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Chelidonic Acid and Its Derivatives from Saussurea Controversa: Isolation, Structural Elucidation and Influence on the Osteogenic Differentiation of Multipotent Mesenchymal Stromal Cells In Vitro

Biomolecules ◽

10.3390/biom9050189 ◽

2019 ◽

Vol 9 (5) ◽

pp. 189 ◽

Cited By ~ 3

Author(s):

Elena Avdeeva ◽

Elvira Shults ◽

Tatyana Rybalova ◽

Yaroslav Reshetov ◽

Ekaterina Porokhova ◽

...

Keyword(s):

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Cell Mass ◽

Multipotent Mesenchymal Stromal Cells ◽

Bone Tissue Regeneration ◽

X Ray ◽

Icp Ms ◽

First Time ◽

Asteraceae Family

4-oxo-4H-pyran-2.6-dicarboxylic acid (chelidonic acid, ChA) in the native state and in the complex with calcium [Ca(ChA)(H2O)3], named saucalchelin (CaChA), was isolated from the extract of Saussurea controversa leaves for the first time for the Asteraceae family. The structure of ChA was determined by NMR, MS and confirmed by X-ray analysis of its monomethyl ester, and CaChA was described by IR, ICP-MS, CHN analysis. The yield of ChA and CaChA was 45 mg/g and 70 mg/g of extract, respectively. The osteogenic activity of ChA, n-monobutyl ester of chelidonic acid, and CaChA has been studied in vitro in a 21-day culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) in a standard nutrient medium without osteogenic supplements. CaChA significantly stimulated the growth of cell mass and differentiation of hAMMSCs into osteoblasts with subsequent mineralization of the culture and it may be a promising substance for accelerating bone tissue regeneration and engineering.

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Study of growth parameters and potentialities of differentiation of multipotent mesenchymal stromal cells from rat bone marrow in vitro

Bulletin of Experimental Biology and Medicine ◽

10.1007/s10517-006-0214-0 ◽

2006 ◽

Vol 141 (4) ◽

pp. 530-535 ◽

Cited By ~ 3

Author(s):

N. S. Sergeeva ◽

I. K. Sviridova ◽

V. A. Kirsanova ◽

S. A. Akhmedova ◽

N. V. Marshutina ◽

...

Keyword(s):

Bone Marrow ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Growth Parameters ◽

Multipotent Mesenchymal Stromal Cells ◽

Rat Bone

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In Vitro Characterization of Multipotent Mesenchymal Stromal Cells Isolated from Palatal Subepithelial Tissue Grafts

Microscopy and Microanalysis ◽

10.1017/s143192761201433x ◽

2013 ◽

Vol 19 (2) ◽

pp. 370-380 ◽

Cited By ~ 13

Author(s):

Alexandra Roman ◽

Andrada Şoancă ◽

Adrian Florea ◽

Emőke Páll

Keyword(s):

Stem Cells ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Embryoid Body ◽

Basic Research ◽

Multipotent Mesenchymal Stromal Cells ◽

Positive Staining ◽

Basic Characteristics ◽

Tissue Grafts

AbstractThe aim of this study was to analyze whether the mesenchymal stromal cells (MSCs) isolated from palatal tissue grafts harvested in order to cover gingival recessions have the basic characteristics of stem cells. The palatal tissue cells were processed using a special culture medium that stimulated the development of only undifferentiated cellular lines. Cells at passage 4 were evaluated by flow cytometry to examine the expression of specific surface markers and were tested for multilineage differentiation capacity. These cells collected at passage 4 were also investigated for the capacity to cluster into embryoid body aggregates. Palatal MSCs displayed positive staining for the mesenchymal markers CD29, CD73, CD105, CD 49e, and CD44, but did not express hematopoietic markers CD34/45. The palatal MSCs successfully differentiated into osteogenic, adipogenic, and chondrogenic lineages. When seeded in special conditions, palatal MSCs propagated into unattached spheres resembling embryoid body aggregates consisting both of differentiated and undifferentiated cells as revealed at the ultrastructural evaluation. It is concluded that the isolated palatal MSCs fulfilled the basic criteria defining the stem cells. This new source of stem cells characterized here for the first time opens new perspectives on possible applications in basic research and in regenerative medicine.

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Allogeneic Adipose-Derived Mesenchymal Stromal Cells Ameliorate Experimental Autoimmune Encephalomyelitis by Regulating Self-Reactive T Cell Responses and Dendritic Cell Function

Stem Cells International ◽

10.1155/2017/2389753 ◽

2017 ◽

Vol 2017 ◽

pp. 1-15 ◽

Cited By ~ 18

Author(s):

Per Anderson ◽

Elena Gonzalez-Rey ◽

Francisco O’Valle ◽

Francisco Martin ◽

F. Javier Oliver ◽

...

Keyword(s):

T Cell ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Cell Function ◽

Multipotent Mesenchymal Stromal Cells ◽

Brain Inflammation ◽

Inos Inhibitor ◽

Cox 1

Multipotent mesenchymal stromal cells (MSCs) have emerged as a promising therapy for autoimmune diseases, including multiple sclerosis (MS). Administration of MSCs to MS patients has proven safe with signs of immunomodulation but their therapeutic efficacy remains low. The aim of the current study has been to further characterize the immunomodulatory mechanisms of adipose tissue-derived MSCs (ASCs) in vitro and in vivo using the EAE model of chronic brain inflammation in mice. We found that murine ASCs (mASCs) suppress T cell proliferation in vitro via inducible nitric oxide synthase (iNOS) and cyclooxygenase- (COX-) 1/2 activities. mASCs also prevented the lipopolysaccharide- (LPS-) induced maturation of dendritic cells (DCs) in vitro. The addition of the COX-1/2 inhibitor indomethacin, but not the iNOS inhibitor L-NAME, reversed the block in DC maturation implicating prostaglandin (PG) E2 in this process. In vivo, early administration of murine and human ASCs (hASCs) ameliorated myelin oligodendrocyte protein- (MOG35-55-) induced EAE in C57Bl/6 mice. Mechanistic studies showed that mASCs suppressed the function of autoantigen-specific T cells and also decreased the frequency of activated (CD11c+CD40high and CD11c+TNF-α+) DCs in draining lymph nodes (DLNs). In summary, these data suggest that mASCs reduce EAE severity, in part, through the impairment of DC and T cell function.

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