Osteogenic differentiation of dura mater stem cells cultured in vitro on three-dimensional porous scaffolds of poly(ε-caprolactone) fabricated via co-extrusion and gas foaming

Osteogenic differentiation of osteoprogenitor cells in three-dimensional (3D) in vitro culture remains poorly understood. Using quantitative real-time RT-PCR techniques, we examined mRNA expression of alkaline phosphatase, osteocalcin, and vascular endothelial growth factor (VEGF) in murine preosteoblastic MC3T3-E1 cells cultured for 48 h and 14 days on conventional two-dimensional (2D) poly(l-lactide-co-glycolide) (PLGA) films and 3D PLGA scaffolds. Differences in VEGF secretion and function between 2D and 3D culture systems were examined using Western blots and an in vitro Matrigel-based angiogenesis assay. Expression of both alkaline phosphatase and osteocalcin in cells cultured on 3D scaffolds was significantly downregulated relative to 2D controls in 48 h and 14 day cultures. In contrast, elevated levels of VEGF expression in 3D culture were noted at every time point in short- and long-term culture. VEGF protein secretion in 3D cultures was triple the amount of secretion observed in 2D controls. Conditioned medium from 3D cultures induced an enhanced level of angiogenic activity, as evidenced by increases in branch points observed in in vitro angiogenesis assays. These results collectively indicate that MC3T3-E1 cells commit to osteogenic differentiation at a slower rate when cultured on 3D PLGA scaffolds and that VEGF is preferentially expressed by these cells when they are cultured in three dimensions.

Download Full-text

Mesenchymal stem cells cultured on a collagen scaffold: In vitro osteogenic differentiation

Archives of Oral Biology ◽

10.1016/j.archoralbio.2006.07.007 ◽

2007 ◽

Vol 52 (1) ◽

pp. 64-73 ◽

Cited By ~ 125

Author(s):

E. Donzelli ◽

A. Salvadè ◽

P. Mimo ◽

M. Viganò ◽

M. Morrone ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Osteogenic Differentiation ◽

Collagen Scaffold ◽

Cells Cultured

Download Full-text

Stem cells in tissue engineering – dynamic cultivation requirement

Stomatoloski glasnik Srbije ◽

10.2478/sdj-2018-0005 ◽

2018 ◽

Vol 65 (1) ◽

pp. 37-44

Author(s):

Dijana Trišić ◽

Vukoman Jokanović ◽

Đorđe Antonijević ◽

Dejan Marković

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Three Dimensional ◽

Future Research ◽

Porous Scaffolds ◽

High Quality ◽

Culture Techniques ◽

Dynamic Cultivation

Summary Stem cells have shown great potential for in vitro tissue engineering, regenerative medicine, cell therapy and pharmaceutical applications. All these applications, especially in clinical trials, will require guided production of high-quality cells. Traditional culture techniques and applications have been performed for the majority of primary and established cell lines and standardized for various analyses. Still, these culture conditions are unable to mimic dynamic and specialized three-dimensional microenvironment of the stem cells’ niche from in vivo conditions. In an attempt to provide biomimetic microenvironments for stem cells in vitro growth, three-dimensional culture techniques have been developed. In our study advantages of newly developed porous scaffolds as the most promising in vitro imitation of niche that provides physical support, enables cell growth, regeneration and neovascularization, while they are replaced in time with newly created tissue was explained. Furthermore, dynamic cultivation techniques have been described, as new way of cell culturing that will be the main subject of our future research. In that manner, by developing an optimal dynamic culturing method, high-quality new cells and tissues would be possible to obtain, for any future clinical application.

Download Full-text

In Vitro Characterization of Periodontal Ligament Stem Cells Derived from Supernumerary Teeth in Three-Dimensional Culture Method

Applied Sciences ◽

10.3390/app11136040 ◽

2021 ◽

Vol 11 (13) ◽

pp. 6040

Author(s):

Yun Yeong Jeong ◽

Mi Sun Kim ◽

Ko Eun Lee ◽

Ok Hyung Nam ◽

Ji-Hyun Jang ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Osteogenic Differentiation ◽

Periodontal Ligament ◽

Three Dimensional ◽

3D Culture ◽

Culture Method ◽

Supernumerary Teeth ◽

Periodontal Ligament Stem Cells

Objective: The aim of this study was to compare the characteristics of periodontal ligament stem cells derived from supernumerary teeth (sPDLSCs), cultured using a three-dimensional (3D) method and a conventional two-dimensional (2D) method. Methods: The morphology, viability, and osteogenic differentiation of the cells were analyzed. In addition, gene expression was analyzed by RNA sequencing, to characterize the functional differences. Results: The diameter of the 3D-cultured sPDLSCs decreased over time, but the spheroid shape was maintained for 7 days. The osteogenic differentiation was similar in the 2D and 3D. The gene expression related to the extracellular matrix (7.3%), angiogenesis (5.6%), cell proliferation (4.6%), inflammatory response (3.7%), and cell migration (3.5%) differed (p < 0.05). Conclusions: Within the limitations of this study, sPDLSCs varied in formation and function, depending on the culture method. In future, it is necessary to study tissue engineering using the advantages of 3D culture and the fewer ethical problems of supernumerary teeth.

Download Full-text

Evaluation of the Impact of Different Pain Medication and Proton Pump Inhibitors on the Osteogenic Differentiation Potential of hMSCs Using 99mTc-HDP Labelling

Life ◽

10.3390/life11040339 ◽

2021 ◽

Vol 11 (4) ◽

pp. 339

Author(s):

Tobias Grossner ◽

Uwe Haberkorn ◽

Tobias Gotterbarm

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Osteogenic Differentiation ◽

Bone Metabolism ◽

Negative Impact ◽

Pain Medication ◽

Group 3 ◽

Group 5 ◽

The Impact

First-line analgetic medication used in the field of musculoskeletal degenerative diseases, like Nonsteroidal anti-inflammatory drugs (NSAIDs), reduces pain and prostaglandin synthesis, whereby peptic ulcers are a severe adverse effect. Therefore, proton pump inhibitors (PPI) are frequently used as a concomitant medication to reduce this risk. However, the impact of NSAIDs or metamizole, in combination with PPIs, on bone metabolism is still unclear. Therefore, human mesenchymal stem cells (hMSCs) were cultured in monolayer cultures in 10 different groups for 21 days. New bone formation was induced as follows: Group 1 negative control group, group 2 osteogenic differentiation media (OSM), group 3 OSM with pantoprazole (PAN), group 4 OSM with ibuprofen (IBU), group 5 OSM with diclofenac (DIC), group 6 OSM with metamizole (MET), group 7 OSM with ibuprofen and pantoprazole (IBU + PAN), group 8 OSM with diclofenac and pantoprazole (DIC + PAN), group 9 OSM with metamizole and pantoprazole (MET + PAN) and group 10 OSM with diclofenac, metamizole and pantoprazole (DIC + MET + PAN). Hydroxyapatite content was evaluated using high-sensitive radioactive 99mTc-HDP labeling. Within this study, no evidence was found that the common analgetic medication, using NSAIDs alone or in combination with pantoprazole and/or metamizole, has any negative impact on the osteogenic differentiation of mesenchymal stem cells in vitro. To the contrary, the statistical results indicate that pantoprazole alone (group 3 (PAN) (p = 0.016)) or diclofenac alone (group 5 (DIC) (p = 0.008)) enhances the deposition of minerals by hMSCS in vitro. There is an ongoing discussion between clinicians in the field of orthopaedics and traumatology as to whether post-surgical (pain) medication has a negative impact on bone healing. This is the first hMSC in vitro study that investigates the effects of pain medication in combination with PPIs on bone metabolism. Our in vitro data indicates that the assumed negative impact on bone metabolism is subsidiary. These findings substantiate the thesis that, in clinical medicine, the patient can receive every pain medication needed, whether or not in combination with PPIs, without any negative effects for the osteo-regenerative potential.

Download Full-text

Expression Profile of New Marker Genes Involved in Differentiation of Canine Adipose-Derived Stem Cells into Osteoblasts

International Journal of Molecular Sciences ◽

10.3390/ijms22136663 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6663

Author(s):

Maurycy Jankowski ◽

Mariusz Kaczmarek ◽

Grzegorz Wąsiatycz ◽

Claudia Dompe ◽

Paul Mozdziak ◽

...

Keyword(s):

Stem Cells ◽

In Vitro Culture ◽

Osteogenic Differentiation ◽

Molecular Mechanisms ◽

Marker Genes ◽

P Value ◽

Illumina Platform

Next-generation sequencing (RNAseq) analysis of gene expression changes during the long-term in vitro culture and osteogenic differentiation of ASCs remains to be important, as the analysis provides important clues toward employing stem cells as a therapeutic intervention. In this study, the cells were isolated from adipose tissue obtained during routine surgical procedures and subjected to 14-day in vitro culture and differentiation. The mRNA transcript levels were evaluated using the Illumina platform, resulting in the detection of 19,856 gene transcripts. The most differentially expressed genes (fold change >|2|, adjusted p value < 0.05), between day 1, day 14 and differentiated cell cultures were extracted and subjected to bioinformatical analysis based on the R programming language. The results of this study provide molecular insight into the processes that occur during long-term in vitro culture and osteogenic differentiation of ASCs, allowing the re-evaluation of the roles of some genes in MSC progression towards a range of lineages. The results improve the knowledge of the molecular mechanisms associated with long-term in vitro culture and differentiation of ASCs, as well as providing a point of reference for potential in vivo and clinical studies regarding these cells’ application in regenerative medicine.

Download Full-text