scholarly journals Induction of osteogenic differentiation of bone marrow stromal cells on 3D polyester-based scaffolds solely by subphysiological fluidic stimulation in a laminar flow bioreactor

2021 ◽  
Vol 12 ◽  
pp. 204173142110193
Author(s):  
Shuntaro Yamada ◽  
Mohammed Ahmed Yassin ◽  
Thomas Schwarz ◽  
Jan Hansmann ◽  
Kamal Mustafa
Keyword(s):  
Bone Marrow ◽  
Laminar Flow ◽  
Stromal Cells ◽  
Collagen Type ◽  
Morphological Changes ◽  
Chemical Stimuli ◽  
Collagen Type 1 ◽  
Microporous Scaffolds

The fatal determination of bone marrow mesenchymal stem/stromal cells (BMSC) is closely associated with mechano-environmental factors in addition to biochemical clues. The aim of this study was to induce osteogenesis in the absence of chemical stimuli using a custom-designed laminar flow bioreactor. BMSC were seeded onto synthetic microporous scaffolds and subjected to the subphysiological level of fluid flow for up to 21 days. During the perfusion, cell proliferation was significantly inhibited. There were also morphological changes, with F-actin polymerisation and upregulation of ROCK1. Notably, in BMSC subjected to flow, mRNA expression of osteogenic markers was significantly upregulated and RUNX2 was localised in the nuclei. Further, under perfusion, there was greater deposition of collagen type 1 and calcium onto the scaffolds. The results confirm that an appropriate level of fluid stimuli preconditions BMSC towards the osteoblastic lineage on 3D scaffolds in the absence of chemical stimulation, which highlights the utility of flow bioreactors in bone tissue engineering.

scholarly journals The Benefit of a Human Bone Marrow Stem Cells Concentrate in addition to an Inorganic Scaffold for Bone Regeneration: AnIn VitroStudy

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
J. Torres ◽  
A. Lopes ◽  
M. A. Lopes ◽  
M. Gutierres ◽  
A. T. Cabral ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Viability ◽  
Bone Regeneration ◽  
Collagen Type ◽  
Mass Ratio ◽  
Orthopedic Surgical Procedures ◽  
Hydroxyapatite Composite ◽  
Collagen Type 1 ◽  
Pellet Formulation

Background. This work compares the osteoblastic behaviour of a bone marrow (BM) aspirate and a prepared BM concentrate of nucleated cells associated with a glass reinforced hydroxyapatite composite (GRHC) in a microporous pellet formulation.Methods. BM aspirate (30 mL) was collected during 3 orthopedic surgical procedures, and a concentration system was used to achieve 3 rapid preparations of a concentrate of nucleated cells (3 mL) from the BM aspirates. The BM aspirates (53% cell viability;2.7×106nucleated cell/mL) and the BM concentrates (76% cell viability;2×107nucleated cell/mL) were cultured over glass reinforced hydroxyapatite pellets, at the same volume/mass ratio, for 30 days. Cultures performed in standard tissue culture plates were used as control.Results. The colonized BM concentrate/material constructs exhibited a representative osteoblastic proliferation/differentiation pathway, evidenced by a high alkaline phosphatase (ALP) activity, expression of collagen type 1, ALP, BMP-2, M-CSF, RANKL, and OPG, and formation of a calcium phosphate mineralized matrix. A clear improved behaviour was noticed compared to the BM aspirate/material constructs.Conclusions. The results suggest the benefit of using an autologous BM concentrate/material construct in the clinical setting, in bone regeneration applications.

scholarly journals Pro-fibrotic phenotype of bone marrow stromal cells in Modic type 1 changes

2021 ◽  
Vol 41 ◽  
pp. 648-667
Author(s):  
I Heggli ◽  
◽  
S Epprecht ◽  
A Juengel ◽  
R Schuepbach ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Bone Marrow Stromal Cells ◽  
Collagen Type ◽  
Collagen Type I ◽  
Marrow Stromal Cells ◽  
Type I ◽  
Cell Contractility ◽  
Modic Type

Modic type 1 changes (MC1) are painful vertebral bone marrow lesions frequently found in patients suffering from chronic low-back pain. Marrow fibrosis is a hallmark of MC1. Bone marrow stromal cells (BMSCs) are key players in other fibrotic bone marrow pathologies, yet their role in MC1 is unknown. The present study aimed to characterise MC1 BMSCs and hypothesised a pro-fibrotic role of BMSCs in MC1. BMSCs were isolated from patients undergoing lumbar spinal fusion from MC1 and adjacent control vertebrae. Frequency of colony-forming unit fibroblast (CFU-F), expression of stem cell surface markers, differentiation capacity, transcriptome, matrix adhesion, cell contractility as well as expression of pro-collagen type I alpha 1, α-smooth muscle actin, integrins and focal adhesion kinase (FAK) were compared. More CFU-F and increased expression of C-X-C-motif-chemokine 12 were found in MC1 BMSCs, possibly indicating overrepresentation of a perisinusoidal BMSC population. RNA sequencing analysis showed enrichment in extracellular matrix proteins and fibrosis-related signalling genes. Increases in pro-collagen type I alpha 1 expression, cell adhesion, cell contractility and phosphorylation of FAK provided further evidence for their pro-fibrotic phenotype. Moreover, a leptin receptor high expressing (LEPRhigh) BMSC population was identified that differentiated under transforming growth factor beta 1 stimulation into myofibroblasts in MC1 but not in control BMSCs. In conclusion, pro-fibrotic changes in MC1 BMSCs and a LEPRhigh MC1 BMSC subpopulation susceptible to myofibroblast differentiation were found. Fibrosis is a hallmark of MC1 and a potential therapeutic target. A causal link between the pro-fibrotic phenotype and clinical characteristics needs to be demonstrated.

scholarly journals Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Stephan Payr ◽  
Elizabeth Rosado-Balmayor ◽  
Thomas Tiefenboeck ◽  
Tim Schuseil ◽  
Marina Unger ◽  
...  
Keyword(s):  
Gene Expression ◽  
Collagen Type ◽  
3D Culture ◽  
Osteogenic Potential ◽  
Donor Age ◽  
Collagen Type 1 ◽  
2D And 3D ◽  
Human Primary Osteoblasts ◽  
Gene Expression Levels

Abstract Background The aim of this study was the investigation of the osteogenic potential of human osteoblasts of advanced donor age in 2D and 3D culture. Methods Osteoblasts were induced to osteogenic differentiation and cultivated, using the same polystyrene material in 2D and 3D culture for 2 weeks. Samples were taken to evaluate alkaline phosphatase (ALP) activity, mineralization and gene expression. Results Osteoprotegerin (OPG) levels were significantly increased (8.2-fold) on day 7 in 3D compared to day 0 (p < 0.0001) and 11.6-fold higher in 3D than in 2D (p < 0.0001). Both culture systems showed reduced osteocalcin (OC) levels (2D 85% and 3D 50% of basic value). Collagen type 1 (Col1) expression was elevated in 3D on day 7 (1.4-fold; p = 0.009). Osteopontin (OP) expression showed 6.5-fold higher levels on day 7 (p = 0.002) in 3D than in 2D. Mineralization was significantly higher in 3D on day 14 (p = 0.0002). Conclusion Advanced donor age human primary osteoblasts reveal significantly higher gene expression levels of OPG, Col1 and OP in 3D than in monolayer. Therefore, it seems that a relatively high potential of bone formation in a natural 3D arrangement is presumably still present in osteoblasts of elderly people. Trial registration 5217/11 on the 22nd of Dec. 2011.

scholarly journals Determination of the membrane hydraulic permeability of MSCs

2016 ◽  
Vol 2 (1) ◽  
pp. 323-327
Author(s):  
Jennifer Contreras Lopez ◽  
Lothar Lauterböck ◽  
Birgit Glasmacher
Keyword(s):  
Activation Energy ◽  
Bone Marrow ◽  
Cooling Rate ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Hydraulic Permeability ◽  
Osmotic Behaviour ◽  
Additional Influence ◽  
Dimethyl Sulfoxid

AbstractA successful cryopreservation is based on knowledge of the optimal cooling rate. So far, this is often determined by way of complex parameter studies. Alternatively, the identification of cell specific characteristics, such as osmotic behaviour, membrane hydraulic permeability and activation energy could be used to calculate the optimal cooling rate. These parameters should be determined for supra-zero and sub-zero temperatures. In this study cryomicroscopy was used. Mesenchymal stromal cells (MSCs) from bone marrow were analysed. The determined membrane hydraulic permeability for sub-zero temperatures is significantly lower than that for supra-zero temperatures. On the contrary the activation energy is significantly higher in the presence of ice. The addition of a cryoprotective agent (CPA) such as dimethyl sulfoxid (DMSO) shows an additional influence on the characteristics of the membrane of the cell. The optimal cooling rate was determined with these parameters. For cryopreservation without DMSO the optimal cooling rate was found to be 12.82 K/min. If the MSCs were frozen with 5% (v/v) DMSO the optimal cooling rate is 16.25 K/min.

scholarly journals Evaluation of Collagen Type-1 Production and Anti-Inflammatory Activities of Human Placental Extracts in Human Gingival Fibroblasts

2016 ◽  
Vol 25 (3) ◽  
pp. 277-281 ◽  
Author(s):  
Homare Akagi ◽  
Yasuhiro Imamura ◽  
Yoshimasa Makita ◽  
Hiroe Nakamura ◽  
Naomi Hasegawa ◽  
...  
Keyword(s):  
Collagen Type ◽  
Human Gingival Fibroblasts ◽  
Gingival Fibroblasts ◽  
Anti Inflammatory ◽  
Collagen Type 1
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