Assessment of the proliferation of human mesenchymal stromal cells in the presence of human demineralised bone matrix

Biologia ◽  
2009 ◽  
Vol 64 (6) ◽  
Author(s):  
Sylvia Gromošová ◽  
Ján Rosocha ◽  
Peter Horňák ◽  
Richard Magin ◽  
Denisa Harvanová ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Stromal Cells ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Demineralised Bone Matrix ◽  
Human Mesenchymal Stromal Cells ◽  
Demineralised Bone

AbstractTissue engineering integrates discoveries from biochemistry, cell and molecular biology, material science and biomedical engineering to produce innovative three-dimensional composites that can be used to replace or correct damaged tissues and organs. Precise classification of osteoinductive properties of human demineralised bone is often the problem, because it varies from batch to batch. An in vitro assay using bone marrow derived human mesenchymal stromal cells (hMSCs) was developed to improve the classification of the osteoinductive quality of demineralised bone matrix. In this study, three-dimensional, partially demineralised bone scaffolds were investigated for their ability to induce osteogenic differentiation of hMSCs in vitro. Proliferation of the hMSCs was measured by the CelTiter 96® AQueous One Solution Cell assay. Chemical structure was evaluated using quantitative and qualitative X-ray analysis. Scanning electron microscopy revealed primary proliferation of the cells cultivated 14 days and showed elevations in the content of Ca2+. These results demonstrate that partially demineralised human bone material supports osteogenic differentiation of hMSCs in vitro. This study documents that in vitro test using hMSCs can be used for classification of the osteoinductive quality of human demineralised bone matrix.

scholarly journals Gradients in pore size enhance the osteogenic differentiation of human mesenchymal stromal cells in three-dimensional scaffolds

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Andrea Di Luca ◽  
Barbara Ostrowska ◽  
Ivan Lorenzo-Moldero ◽  
Antonio Lepedda ◽  
Wojcech Swieszkowski ◽  
...  
Keyword(s):  
Pore Size ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Three Dimensional ◽  
Human Mesenchymal Stromal Cells

scholarly journals Impact of Tranexamic Acid on Chondrocytes and Osteogenically Differentiated Human Mesenchymal Stromal Cells (hMSCs) In Vitro

2020 ◽  
Vol 9 (12) ◽  
pp. 3880
Author(s):  
Mike Wagenbrenner ◽  
Tizian Heinz ◽  
Konstantin Horas ◽  
Axel Jakuscheit ◽  
Joerg Arnholdt ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Tranexamic Acid ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Topical Application ◽  
Stromal Cells ◽  
Specific Marker ◽  
Human Mesenchymal Stromal Cells

The topical application of tranexamic acid (TXA) helps to prevent post-operative blood loss in total joint replacements. Despite these findings, the effects on articular and periarticular tissues remain unclear. Therefore, this in vitro study examined the effects of varying exposure times and concentrations of TXA on proliferation rates, gene expression and differentiation capacity of chondrocytes and human mesenchymal stromal cells (hMSCs), which underwent osteogenic differentiation. Chondrocytes and hMSCs were isolated and multiplied in monolayer cell cultures. Osteogenic differentiation of hMSCs was induced for 21 days using a differentiation medium containing specific growth factors. Cell proliferation was analyzed using ATP assays. Effects of TXA on cell morphology were examined via light microscopy and histological staining, while expression levels of tissue-specific genes were measured using semiquantitative RT-PCR. After treatment with 50 mg/mL of TXA, a decrease in cell proliferation rates was observed. Furthermore, treatment with concentrations of 20 mg/mL of TXA for at least 48 h led to a visible detachment of chondrocytes. TXA treatment with 50 mg/mL for at least 24 h led to a decrease in the expression of specific marker genes in chondrocytes and osteogenically differentiated hMSCs. No significant effects were observed for concentrations beyond 20 mg/mL of TXA combined with exposure times of less than 24 h. This might therefore represent a safe limit for topical application in vivo. Further research regarding in vivo conditions and effects on hMSC functionality are necessary to fully determine the effects of TXA on articular and periarticular tissues.

scholarly journals Chitosan-Hydrogel Polymeric Scaffold Acts as an Independent Primary Inducer of Osteogenic Differentiation in Human Mesenchymal Stromal Cells

Materials ◽  
2020 ◽  
Vol 13 (16) ◽  
pp. 3546
Author(s):  
Simona Bernardi ◽  
Federica Re ◽  
Katia Bosio ◽  
Kamol Dey ◽  
Camillo Almici ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Three Dimensional ◽  
Digital Pcr ◽  
Hydrogel Scaffold ◽  
Hybrid Hydrogel ◽  
Human Mesenchymal Stromal Cells ◽  
Polymeric Scaffold ◽  
Significant Difference

Regenerative medicine aims to restore damaged tissues and mainly takes advantage of human mesenchymal stromal cells (hMSCs), either alone or combined with three-dimensional scaffolds. The scaffold is generally considered a support, and its contribution to hMSC proliferation and differentiation is unknown or poorly investigated. The aim of this study was to evaluate the capability of an innovative three-dimensional gelatin–chitosan hybrid hydrogel scaffold (HC) to activate the osteogenic differentiation process in hMSCs. We seeded hMSCs from adipose tissue (AT-hMSCs) and bone marrow (BM-hMSCs) in highly performing HC of varying chitosan content in the presence of growing medium (GM) or osteogenic medium (OM) combined with Fetal Bovine Serum (FBS) or human platelet lysate (hPL). We primarily evaluated the viability and the proliferation of AT-hMSCs and BM-hMSCs under different conditions. Then, in order to analyse the activation of osteogenic differentiation, the osteopontin (OPN) transcript was absolutely quantified at day 21 by digital PCR. OPN was expressed under all conditions, in both BM-hMSCs and AT-hMSCs. Cells seeded in HC cultured with OM+hPL presented the highest OPN transcript levels, as expected. Interestingly, both BM-hMSCs and AT-hMSCs cultured with GM+FBS expressed OPN. In particular, BM-hMSCs cultured with GM+FBS expressed more OPN than those cultured with GM+hPL and OM+FBS; AT-hMSCs cultured with GM+FBS presented a lower expression of OPN when compared with those cultured with GM+hPL, but no significant difference was detected when compared with AT-hMSCs cultured with OM+FBS. No OPN expression was detected in negative controls. These results show the capability of HC to primarily and independently activate osteogenic differentiation pathways in hMCSs. Therefore, these scaffolds may be considered no more as a simple support, rather than active players in the differentiative and regenerative process.

Osteogenesis induced by a three-dimensional bioimplant composed of demineralised bone matrix, collagen, hydroxyapatite, and bone marrow-derived cells in massive bone defects: An experimental study

2018 ◽  
Vol 50 ◽  
pp. 69-78 ◽  
Author(s):  
Carlos E. Cuervo-Lozano ◽  
Adolfo Soto-Domínguez ◽  
Odila Saucedo-Cárdenas ◽  
Roberto Montes-de-Oca-Luna ◽  
Sergio Alonso-Romero ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Experimental Study ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Bone Defects ◽  
Demineralised Bone Matrix ◽  
Demineralised Bone ◽  
Bone Marrow Derived Cells ◽  
Massive Bone
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