Osteodifferentiation of Human Preadipocytes Induced by Strontium Released from Hydrogels

In recent years, there has been an increasing interest in interactive application principles of biology and engineering for the development of valid biological systems for tissue regeneration, such as for the treatment of bone fractures or skeletal defects. The application of stem cells together with biomaterials releasing bioactive factors promotes the formation of bone tissue by inducing proliferation and/or cell differentiation. In this study, we used a clonal cell line from human adipose tissue-derived mesenchymal stem cells (hADSCs or preadipocytes), named PA2-E12, to evaluate the effects of strontium (Sr2+) released in the culture medium from an amidated carboxymethylcellulose (CMCA) hydrogel enriched with different Sr2+concentrations on osteodifferentiation. The osteoinductive effect was evaluated through both the expression of alkaline phophatase (ALP) activity and the hydroxyapatite (HA) production during 42 days of induction. Present data have shown that Sr2+released from CMCA promotes the osteodifferentiation induced by an osteogenic medium as shown by the increase of ALP activity at 7 and 14 days and of HA production at 14 days. In conclusion, the use of biomaterials able to releasein situosteoinductive agents, like Sr2+, could represent a new strategy for future applications in bone tissue engineering.

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A novel gelatin/chitooligosaccharide/demineralized bone matrix composite scaffold and periosteum-derived mesenchymal stem cells for bone tissue engineering

Biomaterials Research ◽

10.1186/s40824-021-00220-y ◽

2021 ◽

Vol 25 (1) ◽

Author(s):

Thakoon Thitiset ◽

Siriporn Damrongsakkul ◽

Supansa Yodmuang ◽

Wilairat Leeanansaksiri ◽

Jirun Apinun ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Formation ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Demineralized Bone Matrix ◽

Bone Matrix ◽

Alp Activity

Abstract Background A novel biodegradable scaffold including gelatin (G), chitooligosaccharide (COS), and demineralized bone matrix (DBM) could play a significant part in bone tissue engineering. The present study aimed to investigate the biological characteristics of composite scaffolds in combination of G, COS, and DBM for in vitro cell culture and in vivo animal bioassays. Methods Three-dimensional scaffolds from the mixture of G, COS, and DBM were fabricated into 3 groups, namely, G, GC, and GCD using a lyophilization technique. The scaffolds were cultured with mesenchymal stem cells (MSCs) for 4 weeks to determine biological responses such as cell attachment and cell proliferation, alkaline phosphatase (ALP) activity, calcium deposition, cell morphology, and cell surface elemental composition. For the in vivo bioassay, G, GC, and GCD, acellular scaffolds were implanted subcutaneously in 8-week-old male Wistar rats for 4 weeks and 8 weeks. The explants were assessed for new bone formation using hematoxylin and eosin (H&E) staining and von Kossa staining. Results The MSCs could attach and proliferate on all three groups of scaffolds. Interestingly, the ALP activity of MSCs reached the greatest value on day 7 after cultured on the scaffolds, whereas the calcium assay displayed the highest level of calcium in MSCs on day 28. Furthermore, weight percentages of calcium and phosphorus on the surface of MSCs after cultivation on the GCD scaffolds increased when compared to those on other scaffolds. The scanning electron microscopy images showed that MSCs attached and proliferated on the scaffold surface thoroughly over the cultivation time. Mineral crystal aggregation was evident in GC and greatly in GCD scaffolds. H&E staining illustrated that G, GC, and GCD scaffolds displayed osteoid after 4 weeks of implantation and von Kossa staining confirmed the mineralization at 8 weeks in G, GC, and GCD scaffolds. Conclusion The MSCs cultured in GCD scaffolds revealed greater osteogenic differentiation than those cultured in G and GC scaffolds. Additionally, the G, GC, and GCD scaffolds could promote in vivo ectopic bone formation in rat model. The GCD scaffolds exhibited maximum osteoinductive capability compared with others and may be potentially used for bone regeneration.

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Three-dimensional formation of bone tissue of osteoblast cells which carried out differentiation induction from the human adipose tissue stem cells

Journal of Oral and Maxillofacial Surgery ◽

10.1016/j.joms.2014.06.342 ◽

2014 ◽

Vol 72 (9) ◽

pp. e190-e191

Author(s):

A. Ohyama ◽

J. Toyomura ◽

Y. Ide ◽

T. Nakahara ◽

T. Tachibana ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Bone Tissue ◽

Three Dimensional ◽

Human Adipose Tissue ◽

Osteoblast Cells ◽

Differentiation Induction

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In VitroBehavior of Human Adipose Tissue-Derived Stem Cells on Poly(ε-caprolactone) Film for Bone Tissue Engineering Applications

BioMed Research International ◽

10.1155/2015/323571 ◽

2015 ◽

Vol 2015 ◽

pp. 1-12 ◽

Cited By ~ 6

Author(s):

Cecilia Romagnoli ◽

Roberto Zonefrati ◽

Gianna Galli ◽

Dario Puppi ◽

Alessandro Pirosa ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Human Adipose Tissue ◽

Experimental Period ◽

Suitable Model ◽

Good Biocompatibility

Bone tissue engineering is an emerging field, representing one of the most exciting challenges for scientists and clinicians. The possibility of combining mesenchymal stem cells and scaffolds to create engineered tissues has brought attention to a large variety of biomaterials in combination with osteoprogenitor cells able to promote and regenerate bone tissue. Human adipose tissue is officially recognized as an easily accessible source of mesenchymal stem cells (AMSCs), a significant factor for use in tissue regenerative medicine. In this study, we analyze the behavior of a clonal finite cell line derived from human adipose tissue seeded on poly(ε-caprolactone) (PCL) film, prepared by solvent casting. PCL polymer is chosen for its good biocompatibility, biodegradability, and mechanical properties. We observe that AMSCs are able to adhere to the biomaterial and remain viable for the entire experimental period. Moreover, we show that the proliferation process and osteogenic activity of AMSCs are maintained on the biofilm, demonstrating that the selected biomaterial ensures cell colonization and the development of an extracellular mineralized matrix. The results of this study highlight that AMSCs and PCL film can be used as a suitable model to support regeneration of new bone for future tissue engineering strategies.

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In situ chondrogenic differentiation of human adipose tissue-derived stem cells in a TGF-β1 loaded fibrin–poly(lactide-caprolactone) nanoparticulate complex

Biomaterials ◽

10.1016/j.biomaterials.2009.05.034 ◽

2009 ◽

Vol 30 (27) ◽

pp. 4657-4664 ◽

Cited By ~ 54

Author(s):

Youngmee Jung ◽

Yong-Il Chung ◽

Sang Hee Kim ◽

Giyoong Tae ◽

Young Ha Kim ◽

...

Keyword(s):

Stem Cells ◽

Adipose Tissue ◽

Chondrogenic Differentiation ◽

Human Adipose Tissue ◽

Tgf Β1

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Use of calcium phosphate cement scaffolds for bone tissue engineering: in vitro study

Acta Cirurgica Brasileira ◽

10.1590/s0102-86502011000100003 ◽

2011 ◽

Vol 26 (1) ◽

pp. 7-11 ◽

Cited By ~ 19

Author(s):

Taís Somacal Novaes Silva ◽

Bruno Tochetto Primo ◽

Aurelício Novaes Silva Júnior ◽

Denise Cantarelli Machado ◽

Christian Viezzer ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Human Donor ◽

Osteogenic Cells ◽

Alp Activity ◽

Proliferation And Differentiation

Purpose: To evaluate the ability of macroporous tricalcium phosphate cement (CPC) scaffolds to enable the adhesion, proliferation, and differentiation of mesenchymal stem cells derived from human bone marrow. Methods: Cells from the iliac crest of an adult human donor were processed and cultured on macroporous CPC discs. Paraffin spheres sized between 100 and 250µm were used as porogens. Cells were cultured for 5, 10, and 15 days. Next, we assessed cells' behavior and morphology on the biomaterial by scanning electron microscopy. The expression levels of the BGLA and SSP1 genes and the alkaline phosphatase (ALP) activity were quantified by the quantitative real-time polymerase chain reaction technique (QT-PCR) using the fluorophore SYBR GREEN®. Results: QT-PCR detected the expression of the BGLA and SSP1 genes and the ALP activity in the periods of 10 and 15 days of culture. Thus, we found out that there was cell proliferation and differentiation in osteogenic cells. Conclusion: Macroporous CPC, with pore sized between 100 and 250µm and developed using paraffin spheres, enables adhesion, proliferation, and differentiation of mesenchymal stem cells in osteogenic cells and can be used as a scaffold for bone tissue engineering.

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Osteoconduction of Unrestricted Somatic Stem Cells on an Electrospun Polylactic-Co-Glycolic Acid Scaffold Coated with Nanohydroxyapatite

Cells Tissues Organs ◽

10.1159/000485122 ◽

2018 ◽

Vol 205 (1) ◽

pp. 9-19 ◽

Cited By ~ 3

Author(s):

Maryam Shahi ◽

Mahmood Nadari ◽

Mehdi Sahmani ◽

Ehsan Seyedjafari ◽

Naser Ahmadbeigi ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Bone Tissue Engineering ◽

Osteogenic Differentiation ◽

Bone Tissue ◽

Glycolic Acid ◽

Somatic Stem Cells ◽

Human Cord Blood ◽

Alp Activity ◽

Unrestricted Somatic Stem Cells

The limitation of traditional bone grafts could be overcome by applying engineered bone constructs, which are mainly produced by seeding suitable stem cells on appropriate scaffolds. So far, bone marrow-derived stromal cells have been the most applied cells in bone tissue engineering, but current data show that unrestricted somatic stem cells (USSCs) from human cord blood might actually be a better stem cell source due to the accessibility and noninvasive procedure of collection. In this study, we cultured USSCs on a plasma-treated electrospun polylactic-co-glycolic acid (PLGA) scaffold coated with nanohydroxyapatite (nHA). Adhesion and proliferation of USSCs on PLGA/nHA were assessed by scanning electron microscopy and MTT assay. Osteogenic differentiation of USSCs into osteoblast lineage cells was evaluated via alkaline phosphatase (ALP) activity and real-time polymerase chain reaction. Our observation showed that USSCs attached and proliferated on PLGA/nHA. Osteogenic differentiation was confirmed by increased ALP activity and OSTEONECTIN expression in USSCs on PLGA/nHA after the 1st week of the osteogenic period. Therefore, using USSCs on electrospun PLGA/nHA is a promising approach in bone tissue engineering.

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Human Adipose Tissue-Derived Stem Cells and a Poly(ε-Caprolactone) Scaffold Produced by Computer-Aided Wet Spinning for Bone Tissue Engineering

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2017.1614 ◽

2017 ◽

Vol 7 (8) ◽

pp. 622-633 ◽

Cited By ~ 5

Author(s):

Cecilia Romagnoli ◽

Roberto Zonefrati ◽

Dario Puppi ◽

Claudio Rosati ◽

Alessandra Aldinucci ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Adipose Tissue ◽

Bone Tissue Engineering ◽

Bone Tissue ◽

Human Adipose Tissue ◽

Wet Spinning ◽

Computer Aided

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Osteogenic potency of stem cell-based genetic engineering targeting Wnt3a and Wnt9a

Open Life Sciences ◽

10.2478/s11535-011-0079-8 ◽

2011 ◽

Vol 6 (6) ◽

pp. 963-972

Author(s):

Weerachai Singhatanadgit ◽

Manatsanan Varodomrujiranon

Keyword(s):

Stem Cells ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Osteoblast Differentiation ◽

Genetically Modified ◽

Differentiation Markers ◽

Alp Activity ◽

Promising Therapeutic Approach ◽

Skeletal Defects

AbstractBone engineering is a promising therapeutic approach to correct skeletal defects, and genetically-modified stem cells have been implicated in engineering new bone. However, the use of genetically-modified human mesenchymal stem cells targeting an osteogenic growth factor Wnt is not yet investigated. In the present study, a proliferation assay and the alkaline phosphatase (ALP) activity and expression of runt-related transcription factor 2 (Runx2) and osteocalcin (OC) transcripts were investigated to examine the effect of Wnt2 overexpression, Wnt3a overexpression, and Wnt9a knockdown on cell proliferation and osteoblast differentiation of bone marrowderived mesenchymal stem cells (BMSCs). The results showed that the expression of Wnt2 and Wnt3a was up-regulated throughout the osteoblast differentiation period of BMSCs, whereas that of Wnt9a was down-regulated. Overexpression of Wnt3a stimulated cell proliferation while knockdown of Wnt9a increased the ALP activity and the expression of Runx2 and OC. Double transfection producing Wnt3a overexpression and Wnt9a knockdown simultaneously resulted in up-regulation of osteoblast differentiation markers, i.e., the ALP activity and the Runx2 expression. In conclusion, simultaneous genetic modification of Wnt3a overexpression and Wnt9a knockdown enhances osteoblast differentiation of BMSCs, suggesting its osteogenic potency to regenerate new bone in vivo.

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Differential effects of hypoxia on osteochondrogenic potential of human adipose-derived stem cells

AJP Cell Physiology ◽

10.1152/ajpcell.00398.2009 ◽

2010 ◽

Vol 298 (2) ◽

pp. C355-C364 ◽

Cited By ~ 89

Author(s):

Christophe Merceron ◽

Claire Vinatier ◽

Sophie Portron ◽

Martial Masson ◽

Jérôme Amiaud ◽

...

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Oxygen Tension ◽

Chondrogenic Differentiation ◽

Human Adipose Tissue ◽

Type Ii Collagen ◽

Osteogenic Potential ◽

Osteogenic Medium ◽

Alizarin Red ◽

Alp Activity

Human adipose tissue-derived stem cells (hATSC) have been contemplated as reparative cells for cartilage engineering. Chondrogenic differentiation of hATSC can be induced by an enriched culture medium and a three-dimensional environment. Given that bone is vascularized and cartilage is not, oxygen tension has been suggested as a regulatory factor for osteochondrogenic differentiation. Our work aimed at determining whether hypoxia affects the osteochondrogenic potential of hATSC. hATSC were cultured in chondrogenic or osteogenic medium for 28 days, in pellets or monolayers, and under 5% or 20% oxygen tension. Cell differentiation was monitored by real-time PCR (COL2A1, aggrecan, Runx2, and osteocalcin). The chondrogenic differentiation was further evaluated by Alcian blue and immunohistological staining for glycosaminoglycans (GAGs) and type II collagen, respectively. Osteogenic differentiation was also assessed by the staining of mineralized matrix (Alizarin Red) and measurement of alkaline phosphatase (ALP) activity. The expression of chondrogenic markers was upregulated when hATSC were exposed to hypoxia in chondrogenic medium. Conversely, osteocalcin expression, mineralization, and ALP activity were severely reduced under hypoxic conditions even in the presence of osteogenic medium. Our data strongly suggest that hypoxia favors the chondrogenic differentiation of hATSC as evidenced by the expression of the chondrogenic markers, whereas it could alter their osteogenic potential. Our results highlight the differential regulatory role of hypoxia on the chondrogenic and osteogenic differentiation processes of hATSC. These data could help us exploit the potential of tissue engineering and stem cells to replace or restore the function of osteoarticular tissues.

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