scholarly journals Extracorporeal shockwaves (ESWs) enhance the osteogenic medium-induced differentiation of adipose-derived stem cells into osteoblast-like cells

2014 ◽  
Vol 11 (2) ◽  
pp. 390-399 ◽  
Author(s):  
Maria Graziella Catalano ◽  
Francesca Marano ◽  
Letizia Rinella ◽  
Laura de Girolamo ◽  
Ornella Bosco ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Induced Differentiation

189 The Effect of Vitamin K on the Differentiation of Adipose-Derived Stem Cells into Osteoblasts

2018 ◽  
Vol 30 (1) ◽  
pp. 234
Author(s):  
T. A. Bane ◽  
J. C. Bertels ◽  
K. M. Polkoff ◽  
M. Rubessa ◽  
M. B. Wheeler
Keyword(s):  
Stem Cells ◽  
Vitamin K ◽  
Bone Mineralization ◽  
Negative Control ◽  
Nodule Formation ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Mineral Density ◽  
Alizarin Red ◽  
Significant Difference

Tissue engineering offers a viable alternative to bone grafts in repairing large bone defects. This involves using scaffolds of various sizes and shapes that contain stem cells and other osteoinductive molecules. The aim of this project was to evaluate the effects of vitamin K in osteogenic medium and its effect on the differentiation of adipose-derived stem cells (ASC) into osteoblasts. Vitamin K has been shown to increase bone mineral density by acting as a coenzyme in the γ-carboxylation of osteocalcin, a protein involved in bone mineralization (Weber 2001 Nutrition 11–12, 1024). Our hypothesis was that the presence of vitamin K in the osteogenic medium would positively influence the number of osteoblastic nodules formed. Swine ASC were isolated as described (Monaco et al. 2009 Open Tissue Eng. Regen. Med. J. 2, 20–33). The ASC were divided into 7 different treatments: 5 concentrations of vitamin K in the osteogenic medium (10, 50 100, 500, 1000 nM) plus 2 control treatments (osteogenic medium without vitamin K and a negative control, DMEM). The media was changed twice a week for 4 weeks. The experiment was replicated 6 times. At the end of the culture period, cells were stained with Alizarin Red S and Von Kossa. In each well, we counted the nodules and then divided them in 2 categories: formed and forming nodules. Data were analysed by analysis of variance using the generalized linear model (GLM) procedure of SPSS (IBM/SPSS, Armonk, NY, USA); the least significant difference (l.s.d.) post hoc test was used to perform statistical multiple comparison, and the α-level was set at 0.05. The results showed (in Table 1) that there was no positive effect on nodule formation when vitamin K was added to the medium; however, when 1000 nM vitamin K was added, nodule formation decreased. More experiments need to be conducted to determine if vitamin K can act synergistically with other vitamins to produce a significant role in ASC differentiation into osteoblasts. This preliminary experiment is the first step towards the analysis of the behaviour of ASC on scaffolds with vitamin K incorporated into their matrix. Table 1.The average number of formed and forming osteoblast nodules compared between treatment groups (SD in parentheses)

188 The Effect of Vitamin B12 on the Differentiation of Adipose-Derived Stem Cells into Osteoblasts

2018 ◽  
Vol 30 (1) ◽  
pp. 234
Author(s):  
T. A. Bane ◽  
J. C. Bertels ◽  
K. M. Polkoff ◽  
M. Rubessa ◽  
M. B. Wheeler
Keyword(s):  
Stem Cells ◽  
Vitamin B12 ◽  
Difficult Problem ◽  
Negative Control ◽  
Nodule Formation ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Alizarin Red ◽  
Α Level

Large bone defects present a tremendous challenge to the treating surgeon. Tissue engineering using scaffolds of various sizes and shapes that contain stem cells and other osteoinductive molecules offer a potential solution to this difficult problem. The aim of this project was to evaluate if the osteogenic medium infused with vitamin B12 influences the differentiation of adipose-derived stem cells (ASC) into osteoblasts. Vitamin B12 has been shown to have a stimulatory effect on osteoclastogenesis in vitro (Vaes et al. 2009 Calcified Tissue Int. 84, 413-422). Our hypothesis was that the presence of vitamin B12 in the osteogenic medium would positively influence the number of osteoblastic nodules formed. Swine ASC were isolated as described (Monaco et al. 2009 Open Tissue Eng. Regen. Med. J. 2, 20-33). The ASC were divided in 8 different treatments: 8 concentrations of vitamin B12 in the osteogenic medium (0.1, 0.2, 1, 2, 10, and 20 μM) plus 2 control treatments (osteogenic medium without vitamin B12 and a negative control, DMEM). The medium was changed twice a week for 4 weeks. The experiment was replicated 6 times. At the end of the culture period, cells were stained with Alizarin Red and Von Kossa stains. In each well, we counted the nodules and then divided them in 2 categories: formed and forming nodules. Data was analysed using the generalized linear model (GLM) procedure in SPSS (IBM/SPSS, Armonk, NY, USA). Bonferroni’s post hoc test was used to perform statistical multiple comparison. The α-level was set at 0.01. The results showed that the concentration of 20 μM vitamin B12 was detrimental for nodule formation. Table 1 illustrates the number of formed and forming nodules in addition to their standard deviation. There was no positive effect on nodule formation when different concentrations of vitamin B12 were added to the osteogenic medium. More experiments need to be conducted to determine if vitamin B12 can act synergistically with other vitamins to produce a significant role in ASC differentiation into osteoblasts. This preliminary experiment is the first step towards the analysis of the behaviour of ASC on scaffolds with vitamin B12 incorporated into their matrix. Table 1.The average number of formed and forming osteoblast nodules compared between treatment groups (SD in parentheses)

scholarly journals Effects of melatonin on the proliferation and differentiation of rat adipose-derived stem cells

2008 ◽  
Vol 41 (01) ◽  
pp. 08-14 ◽  
Author(s):  
Arash Zaminy ◽  
Iraj Ragerdi Kashani ◽  
Mohammad Barbarestani ◽  
Azim Hedayatpour ◽  
Reza Mahmoudi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Culture Media ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Adult Rats ◽  
Alizarin Red ◽  
Alp Activity ◽  
Proliferation And Differentiation

ABSTRACT Background: Osteogenesis driven by adipose-derived stem cells (ADSCs) is regulated by physiological and pathological factors. Accumulating evidence from in vitro and in vivo experiments suggests that melatonin may have an influence on bone formation. However, little is known about the effects of melatonin on osteogenesis, which thus remains to be elucidated. This study was performed to determine whether melatonin at physiological concentrations (0.01-10 nM) could affect the in vitro proliferation and osteogenic differentiation of rat ADSCs.Materials and Methods: ADSCs were isolated from the fat of adult rats. After cell expansion in culture media and through three passages, osteogenesis was induced in a monolayer culture using osteogenic medium with or without melatonin at physiological concentrations (0.01-10 nM). After four weeks, the cultures were examined for mineralization by Alizarin Red S and von Kossa staining and for alkaline phosphatase (ALP) activity using an ALP kit. Cell viability and apoptosis were also assayed by 3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTT) assay and flow cytometry, respectively.Results: The results indicated that at physiological concentrations, melatonin suppressed proliferation and differentiation of ADSCs. These data indicate that ADSCs exposed to melatonin, had a lower ALP activity in contrast to the cells exposed to osteogenic medium alone. Similarly, mineral deposition (calcium level) also decreased in the presence of melatonin. Flow cytometry confirmed that cell growth had decreased and that the numbers of apoptotic cells had increased.Conclusion: These results suggest that the physiological concentration of melatonin has a negative effect on ADSC osteogenesis.

208 The effect of copper on the differentiation of adipose-derived stem cells into osteoblasts

2019 ◽  
Vol 31 (1) ◽  
pp. 229
Author(s):  
T. Bane ◽  
L. Siegel ◽  
J. Bertels ◽  
K. Ratz ◽  
M. Rubessa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Lysyl Oxidase ◽  
Collagen Matrix ◽  
Negative Control ◽  
Nodule Formation ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Alizarin Red ◽  
Significant Difference ◽  
Positive Effect

Large bone defects present a tremendous challenge to the treating surgeon. Tissue engineering using scaffolds of various sizes and shapes that contain stem cells and other osteoinductive molecules offer a potential solution to this difficult problem. The aim of this project was to evaluate whether osteogenic medium infused with copper influences the differentiation of adipose-derived stem cells (ASC) into osteoblasts. Copper is a key cofactor for lysyl oxidase, an enzyme involved in producing a collagen matrix through which bone can grow. Lysyl oxidase expression is up-regulated in bone marrow stromal cells (Khosravi et al. 2014 PLoS One 9, e100669). Our hypothesis was that the presence of copper in the osteogenic medium would positively influence the number of osteoblastic nodules formed. Swine ASC were isolated as described (Monaco et al. 2009 Open Tissue Eng. Regen. Med. J. 2, 20-33). The ASC were divided in 7 different treatments: 5 concentrations of copper in the osteogenic medium (0.1, 1, 10, 50, and 100 µM) plus 2 control treatments (osteogenic medium without copper and a negative control, DMEM). The medium was changed twice a week for 4 weeks. The experiment was replicated 6 times. At the end of the culture period, cells were stained with Alizarin Red S and Von Kossa stains. In each well, we counted the total number of nodules that were either formed or forming. Data were analysed using the generalized linear model (GLM) procedure (SPSS Inc./IBM Corp., Armonk, NY). The least significant difference (l.s.d.) post hoc test was used to perform statistical multiple comparison. The α-level was set at 0.05. The results showed that more nodules were formed in the 0.1 and 1 µM copper groups compared with the osteogenic control, but there was no statistical difference between those 2 treatments. Table 1 illustrates the total number of formed and forming nodules in addition to their standard deviation. There was a positive effect on nodule formation when copper concentrations of 0.1 and 1 µM were added to the osteogenic medium. In contrast, copper concentrations of 50 and 100 µM had a cytotoxic effect. These results confirm that low concentrations of copper have a positive effect on osteogenesis. This preliminary experiment is the first step towards the analysis of the behaviour of ASC on scaffolds with copper incorporated into their matrix. Table 1.The average number (standard deviations in parentheses) of total formed and forming osteoblast nodules compared between treatment groups

197 THE EFFECT OF ZINC ON THE DIFFERENTIATION OF ADIPOSE-DERIVED STEM CELLS INTO OSTEOBLASTS

2017 ◽  
Vol 29 (1) ◽  
pp. 207 ◽  
Author(s):  
J. C. Bertels ◽  
M. Rubessa ◽  
S.R. Schreiber ◽  
M. B. Wheeler
Keyword(s):  
Stem Cells ◽  
Bone Formation ◽  
Negative Control ◽  
Nodule Formation ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Alizarin Red ◽  
The Media ◽  
Positive Effect

The aim of this project was to evaluate the effects of zinc in osteogenic media and its effect on the differentiation of adipose-derived stem cells (ASC) into osteoblasts. Zinc has a stimulatory effect on bone formation and mineralization in vivo and vitro (Seo et al. 2010 Nutr. Res. Pract. 4, 356–361). Our hypothesis was that the presence of zinc in the osteogenic media would positively influence both the speed of formation and the number of osteoblastic nodules formed. Swine ASC were isolated as described (Monaco et al. 2009 Open Tissue Eng. Regen. Med. J. 2, 20–33). The ASC were divided in 8 different treatments: 6 different concentrations of zinc in the osteogenic medium (8, 4, 0.8, 0.4, 0.08, and 0.04 mM) plus 2 control treatments (osteogenic medium without zinc and a negative control, DMEM). The media was changed twice a week for 4 weeks. The experiment was replicated 4 times. At the end of the culture period, cells were stained with Alizarin Red S. In each well, we counted the nodules and divided them in 2 categories: formed and forming nodules. The second evaluation that we did was to evaluate the diameter of the largest nodules (2/well) in each group. Data were analysed by ANOVA using the Generalized Linear Model procedure (SPSS, IBM Corp., Armonk, NY, USA). Bonferroni’s post-hoc test was used to perform statistical multiple comparison. The α-level was set at 0.05. The results showed that the doses of zinc of both 4 and 8 mM were toxic to the whole cell populations in this treatment, which was indicated by cell death, whereas the concentrations of 0.8 and 0.4 mM were not cytotoxic but no nodules formed. Here we report the results that are greater than zero in Table 1. There is a positive effect on nodule formation when the zinc is added to the media. It is clear that the total number of nodules is different between the 0.08 mM zinc group and the control (P < 0.003). When we evaluated nodule diameter we found a direct correlation between the zinc concentration and the diameter of the nodules: 292.7 (±136.6) v. 366.8 (±218.7) v. 423.7 (±267.7) µm for the control, 0.04 mM zinc, and 0.08 mM zinc, respectively. The largest nodule was found in the 0.08 mM zinc treatment at 886.6 µm. These results confirmed the positive effect of this mineral on bone formation. This preliminary experiment is the first step towards the analysis of the behaviour of ASC on scaffolds with zinc incorporated into their matrix. Table 1. The average number (SD in parentheses) of formed and forming osteoblast nodules compared between treatment groups

scholarly journals Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method

2021 ◽  
Vol 22 (22) ◽  
pp. 12392
Author(s):  
Kiyofumi Takabatake ◽  
Masakazu Matsubara ◽  
Eiki Yamachika ◽  
Yuki Fujita ◽  
Yuki Arimura ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteogenic Potential ◽  
Fat Cells ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Alizarin Red ◽  
Dedifferentiated Fat Cells ◽  
Dfat Cells

Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin. Results: The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference (p < 0.05) at 6 and 9 days. The ALP levels of DFATs were higher than those of ASCs after 12 days. The expression levels of osteoblast marker genes (osterix and osteocalcin) of DFAT cells and ASCs were higher after osteogenic differentiation culture. Conclusion: DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies.

184 Effect of selenium on the differentiation of porcine adipose-derived stem cells into osteoblasts

2019 ◽  
Vol 31 (1) ◽  
pp. 216
Author(s):  
L. Siegel ◽  
T. Bane ◽  
J. Bertels ◽  
K. Ratz ◽  
M. Rubessa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Statistical Difference ◽  
Bone Growth ◽  
Negative Control ◽  
Nodule Formation ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Alizarin Red ◽  
Differentiation Potential ◽  
Cell Therapies

Bone damage is a common occurrence and although traditional bone healing methods work well, they are not always able to fix all issues pertaining to the injury. However, recent research on stem cell therapies using adipose-derived stem cells (ASC) has shown that ASC that differentiate into osteoblasts can be used as a potentially better solution for bone injuries. Selenium is a nutritional trace element that has been found to be essential in the production of selenoproteins and bone growth. Selenium plays an integral role in osteoblast cell differentiation and proliferation. The aim of this experiment was to test whether adding selenium to osteogenic influences the differentiation potential of ASC into osteoblasts. Porcine ASC were isolated as described (Monaco et al. 2009, Open Tissue Eng. Regen. Med. J. 2, 20-33). Seven different treatments were given to the cells: a negative control of DMEM, a positive control of osteogenic medium, and 5 concentrations of selenium in the osteogenic medium (10, 5, 1, 0.5, and 0.1 µM). Medium was changed twice a week for 4 weeks, at the end of which the cells were stained with Alizarin Red S stain. The number of osteoblastic nodules in each well were counted and divided into the categories of “forming” and “formed.” A generalized linear model (GLM) procedure (SPSS Inc./IBM Corp., Chicago, IL, USA) was used to analyse the data. The least statistical difference (l.s.d.) post hoc test was used to perform statistical multiple comparison with an α-level of 0.05. The results showed (Table 1) that medium containing selenium concentrations of 0.1, 0.5, and 1.0 µM were able to develop more nodules than the osteogenic medium. There was no statistical difference in the total amount of forming and formed nodules in those 4 groups; however, there was a statistical tendency for a difference (P=0.06) between the medium with 1 µM selenium and the osteogenic medium. Medium containing 10 or 5 µM selenium had lower nodule counts (both forming and formed) than osteogenic medium. There was no statistical difference between 10 µM and DMEM. These results suggest that low concentrations of selenium have a positive effect on nodule formation, whereas higher concentrations are detrimental. Table 1.The average number (standard deviations in parentheses) of formed, forming, and total osteoblastic nodules in each of the different treatments

scholarly journals Cytokine, Chemokine, and Growth Factor Profile Characterization of Undifferentiated and Osteoinduced Human Adipose-Derived Stem Cells

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
F. Mussano ◽  
T. Genova ◽  
M. Corsalini ◽  
G. Schierano ◽  
F. Pettini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Growth Factor ◽  
Expression Profile ◽  
Conditioned Media ◽  
High Expression ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Cell Models

Bone is the second most manipulated tissue after blood. Adipose-derived stem cells (ASCs) may become a convenient source of MSC for bone regenerative protocols. Surprisingly, little is known about the most significant biomolecules these cells produce and release after being osteoinduced. Therefore, the present study aimed at dosing 13 candidates chosen among the most representative cytokines, chemokines, and growth factors within the conditioned media of osteodifferentiated and undifferentiated ASCs. Two acknowledged osteoblastic cell models, that is, MG-63 and SaOs-2 cells, were compared. Notably, IL-6, IL-8, MCP-1, and VEGF were highly produced and detectable in ASCs. In addition, while IL-6 and IL-8 seemed to be significantly induced by the osteogenic medium, no such effect was seen for MCP-1 and VEGF. Overall SaOS-2 had a poor expression profile, which may be consistent with the more differentiated phenotype of SaOs-2 compared to ASCs and MG-63. Instead, in maintaining medium, MG-63 displayed a very rich production of IL-12, MCP-1, IP-10, and VEGF, which were significantly reduced in osteogenic conditions, with the only exception of MCP-1. The high expression of MCP-1 and VEGF, even after the osteogenic commitment, may support the usage of ASCs in bone regenerative protocols by recruiting both osteoblasts and osteoclasts of the host.

scholarly journals Induced differentiation of macaque adipose-derived stem cells in vitro

2021 ◽  
Vol Publish Ahead of Print ◽  
Author(s):  
Jun-Liang Jiang ◽  
Tao Li ◽  
Xin Bi ◽  
Zhao-Xiang Wu ◽  
Kai-Yu Hou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Derived Stem Cells ◽  
Induced Differentiation
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