Chondrogenic and osteogenic potential of adipose derived stem cells from RA and OA patients

Aim. Macroporous sponge-like gelatin–hyaluronic acid (Gl–HA) scaffolds cross-linked by EDC were produced using cryogelation technology, which allows for the preparation of highly porous scaffolds without compromising their mechanical properties, and is a more cost-efficient process than freeze drying. The aim of this study is to evaluate the osteogenic potential of porcine adipose-derived stem cells (PADSCs) in GI–HA cryogel. Method. The character of the GI–HA cryogel was evaluated. The pore size and the microstructure were observed using scanning electron microscope (SEM). The swelling ratio was measured. The PADSCs were harvested and isolated from pig inguinal area. Then, the GI–HA cryogel was seeded with PADSCs. The cryogel/ASCs mixture was cultured in osteogenic medium for 0, 3, 7, 14, and 21 days. The cell proliferation was measured by MTS. The RT-PCR of specific osteogenic gene expression such as osteocalcin (OC), RUNX2 was used to assess the osteogenic ability. The SEM was used to observe the interaction between scaffold and cells. Energy dispersive spectrum (EDS) was used to analyze the mineralization around cells. Results. The pore size was variable between 200 and 369 μm. The swelling ratio was around 8.67 ± 1.669%. The cell proliferation was increasing along with the increase of induction periods. The expression of early gene of RUNX2 and late gene of OC mean that the PADSCs were differentiated well into osteoblasts within the cryogels. The SEM detailed that the PADSCs cell can proliferate well in the pore of GI–HA scaffold. The EDS also demonstrated the mineralization of PADSCs in GI–HA scaffold after induction. Conclusions. To conclude, the PADSCs can proliferate and differentiate well into osteoblasts in the three-dimensional, porous, GI–HA cryogel.

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The Osteogenic Potential of Adipose-Derived Stem Cells for the Repair of Rabbit Calvarial Defects

Annals of Plastic Surgery ◽

10.1097/01.sap.0000210629.17727.bd ◽

2006 ◽

Vol 56 (5) ◽

pp. 543-548 ◽

Cited By ~ 111

Author(s):

Jason R. Dudas ◽

Kacey G. Marra ◽

Gregory M. Cooper ◽

Virginia M. Penascino ◽

Mark P. Mooney ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Potential ◽

Adipose Derived Stem Cells ◽

Calvarial Defects

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14-3-3β paralog is inhibited by acetylation during differentiation to the osteogenic lineage.

10.21203/rs.3.rs-17059/v1 ◽

2020 ◽

Author(s):

Yesica Romina Frontini López ◽

Aldana Daniela Gojanovich ◽

Marina Uhart ◽

Diego Martin Bustos

Keyword(s):

Stem Cells ◽

Stromal Vascular Fraction ◽

Protein Family ◽

Negative Regulator ◽

Osteogenic Potential ◽

Adipose Derived Stem Cells ◽

Hairpin Rna ◽

Signaling Systems ◽

Phosphorylated Proteins ◽

Osteogenic Lineage

Abstract Background 14-3-3 protein family binds and regulate hundreds of serine/threonine phosphorylated proteins. Considered as redundant, ubiquitous and constantly expressed, this protein family was treated as an accessory for many signaling systems. Recently, specific biological functions were discovered for each of its seven paralogs in humans. Here, we studied the reversible inhibition by acetylation of its essential N- ε -lysine 49/51 residue during the osteogenic differentiation of human adipose-derived stem cells. Methods Human adipose-derived stem cells were obtained from healthy patients who underwent a dermolipectomy. The material was processed to obtain the ASCs from the stromal vascular fraction. Proteins of interest were analyzed by confocal microscopy and Western blot, and the expression level of selected proteins were modified by using lentivirus and specific shRNAs. Results We found that during the differentiation of ASC, the levels of 14-3-3 acK49/51 increased, showing that inhibition of 14-3-3 is necessary during this process. Among the 7 paralogs of this family, the inhibition by this posttranslational modification occurs mostly on the paralog β , located specifically in the nucleus, where 14-3-3 was described to bind to H3 histone and many transcription factors. Paralog specific inhibition by short hairpin RNA showed that silencing of 14-3-3 β gene, but not 14-3-3γ, increases significantly the osteogenic potential of the cells. Transdifferentiation from osteogenic to adipogenic linage of cell lines that were specifically silenced for 14-3-3 β o γ paralogs showed that 14-3-3b could be a negative regulator of the differentiation to the osteogenic lineage. Conclusion We showed that specifically 14-3-3 β is a negative regulator of osteogenesis. The levels of its inhibition by acetylation on lysine 51 is the key cellular mechanism to regulate it. The HBO1 acetyltransferase could be the enzyme responsible of this modification. Cells were this inhibition was emulated by decreasing the levels of expression of 14-3-3 β showed significant increased potential for the differentiation to the osteogenic lineage.

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In Vitro Studies on Nanoporous, Nanotubular and Nanosponge-Like Titania Coatings, with the Use of Adipose-Derived Stem Cells

Materials ◽

10.3390/ma13071574 ◽

2020 ◽

Vol 13 (7) ◽

pp. 1574 ◽

Cited By ~ 2

Author(s):

Michalina Ehlert ◽

Aleksandra Radtke ◽

Tomasz Jędrzejewski ◽

Katarzyna Roszek ◽

Michał Bartmański ◽

...

Keyword(s):

Stem Cells ◽

Cell Line ◽

Osteogenic Potential ◽

Biological Research ◽

Adipose Derived Stem Cells ◽

Differentiation Potential ◽

Nanoporous Surface ◽

Titania Coatings ◽

The Impact

In vitro biological research on a group of amorphous titania coatings of different nanoarchitectures (nanoporous, nanotubular, and nanosponge-like) produced on the surface of Ti6Al4V alloy samples have been carried out, aimed at assessing their ability to interact with adipose-derived mesenchymal stem cells (ADSCs) and affect their activity. The attention has been drawn to the influence of surface coating architecture and its physicochemical properties on the ADSCs proliferation. Moreover, in vitro co-cultures: (1) fibroblasts cell line L929/ADSCs and (2) osteoblasts cell line MG-63/ADSCs on nanoporous, nanotubular and nanosponge-like TiO2 coatings have been studied. This allowed for evaluating the impact of the surface properties, especially roughness and wettability, on the creation of the beneficial microenvironment for co-cultures and/or enhancing differentiation potential of stem cells. Obtained results showed that the nanoporous surface is favorable for ADSCs, has great biointegrative properties, and supports the growth of co-cultures with MG-63 osteoblasts and L929 fibroblasts. Additionally, the number of osteoblasts seeded and cultured with ADSCs on TNT5 surface raised after 72-h culture almost twice when compared with the unmodified scaffold and by 30% when compared with MG-63 cells growing alone. The alkaline phosphatase activity of MG-63 osteoblasts co-cultured with ADSCs increased, that indirectly confirmed our assumptions that TNT-modified scaffolds create the osteogenic niche and enhance osteogenic potential of ADSCs.

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A Novel Method to Apply Osteogenic Potential of Adipose Derived Stem Cells in Orthopaedic Surgery

PLoS ONE ◽

10.1371/journal.pone.0088874 ◽

2014 ◽

Vol 9 (2) ◽

pp. e88874 ◽

Cited By ~ 11

Author(s):

Xiang Fang ◽

Hideki Murakami ◽

Satoru Demura ◽

Katsuhiro Hayashi ◽

Hidenori Matsubara ◽

...

Keyword(s):

Stem Cells ◽

Orthopaedic Surgery ◽

Osteogenic Potential ◽

Adipose Derived Stem Cells ◽

Novel Method

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Exosomes from miRNA-378-modified adipose-derived stem cells prevent glucocorticoid-induced osteonecrosis of the femoral head by enhancing angiogenesis and osteogenesis via targeting miR-378 negatively regulated suppressor of fused (Sufu)

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02390-x ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Kai Nan ◽

Yuankai Zhang ◽

Xin Zhang ◽

Dong Li ◽

Yan Zhao ◽

...

Keyword(s):

Stem Cells ◽

Femoral Head ◽

Signaling Pathway ◽

Umbilical Vein ◽

Osteogenic Potential ◽

Adipose Derived Stem Cells ◽

Shh Signaling ◽

Suppressor Of Fused ◽

Alkaline Phosphatase Staining

Abstract Background Local ischemia and defective osteogenesis are implicated in the progression of glucocorticoid (GC)-induced osteonecrosis of the femoral head (ONFH). Recent studies have revealed that exosomes released from adipose-derived stem cells (ASCs) play important roles in ONFH therapy. The present study aimed to investigate whether exosomes derived from miR-378-overexpressing ASCs (miR-378-ASCs-Exos) could promote angiogenesis and osteogenesis in GC-induced ONFH. Methods In vitro, we investigated the osteogenic potential of miR-378-ASCs-Exos on bone marrow stromal cells (BMSCs) by alkaline phosphatase staining and western blotting. The angiogenic effects of miR-378-ASCs-Exos on human umbilical vein endothelial cells (HUVECs) were examined by evaluating their proliferation, migration, and tube-forming analyses. We identified the underlying mechanisms of miR-378 in osteogenic and angiogenic regulation. In addition, an ONFH rat model was established to explore the effects of miR-378-ASCs-Exos through histological and immunohistochemical staining and micro-CT in vivo. Results Administration of miR-378-ASCs-Exos improved the osteogenic and angiogenic potentials of BMSCs and HUVECs. miR-378 negatively regulated the suppressor of fused (Sufu) and activated Sonic Hedgehog (Shh) signaling pathway, and recombinant Sufu protein reduced the effects triggered by miR-378-ASCs-Exos. In vivo experiments indicated that miR-378-ASCs-Exos markedly accelerated bone regeneration and angiogenesis, which inhibited the progression of ONFH. Conclusion Our study indicated that miR-378-ASCs-Exos enhances osteogenesis and angiogenesis by targeting Sufu to upregulate the Shh signaling pathway, thereby attenuating GC-induced ONFH development.

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