scholarly journals 14-3-3ε protein-loaded 3D hydrogels favor osteogenesis

2020 ◽  
Vol 31 (11) ◽  
Author(s):  
Ana A. Aldana ◽  
Marina Uhart ◽  
Gustavo A. Abraham ◽  
Diego M. Bustos ◽  
Aldo R. Boccaccini
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
3D Printing ◽  
Osteogenic Differentiation ◽  
Alginate Hydrogel ◽  
Hydrogel Scaffolds ◽  
Future Developments ◽  
Positive Effect ◽  
Cell Adhesion And Proliferation

Abstract3D printing has emerged as vanguard technique of biofabrication to assemble cells, biomaterials and biomolecules in a spatially controlled manner to reproduce native tissues. In this work, gelatin methacrylate (GelMA)/alginate hydrogel scaffolds were obtained by 3D printing and 14-3-3ε protein was encapsulated in the hydrogel to induce osteogenic differentiation of human adipose-derived mesenchymal stem cells (hASC). GelMA/alginate-based grid-like structures were printed and remained stable upon photo-crosslinking. The viscosity of alginate allowed to control the pore size and strand width. A higher viscosity of hydrogel ink enhanced the printing accuracy. Protein-loaded GelMA/alginate-based hydrogel showed a clear induction of the osteogenic differentiation of hASC cells. The results are relevant for future developments of GelMA/alginate for bone tissue engineering given the positive effect of 14-3-3ε protein on both cell adhesion and proliferation.

scholarly journals Fluid Flow Mechanical Stimulation-Assisted Cartridge Device for the Osteogenic Differentiation of Human Mesenchymal Stem Cells

Micromachines ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 927
Author(s):  
Ki-Taek Lim ◽  
Dinesh-K. Patel ◽  
Sayan-Deb Dutta ◽  
Keya Ganguly
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Fluid Flow ◽  
Osteogenic Differentiation ◽  
Flow Condition ◽  
Cultured Cells ◽  
Human Mesenchymal Stem Cells ◽  
Proliferation And Differentiation ◽  
Static Conditions

Human mesenchymal stem cells (hMSCs) have the potential to differentiate into different types of mesodermal tissues. In vitro proliferation and differentiation of hMSCs are necessary for bone regeneration in tissue engineering. The present study aimed to design and develop a fluid flow mechanically-assisted cartridge device to enhance the osteogenic differentiation of hMSCs. We used the fluorescence-activated cell-sorting method to analyze the multipotent properties of hMSCs and found that the cultured cells retained their stemness potential. We also evaluated the cell viabilities of the cultured cells via water-soluble tetrazolium salt 1 (WST-1) assay under different rates of flow (0.035, 0.21, and 0.35 mL/min) and static conditions and found that the cell growth rate was approximately 12% higher in the 0.035 mL/min flow condition than the other conditions. Moreover, the cultured cells were healthy and adhered properly to the culture substrate. Enhanced mineralization and alkaline phosphatase activity were also observed under different perfusion conditions compared to the static conditions, indicating that the applied conditions play important roles in the proliferation and differentiation of hMSCs. Furthermore, we determined the expression levels of osteogenesis-related genes, including the runt-related protein 2 (Runx2), collagen type I (Col1), osteopontin (OPN), and osteocalcin (OCN), under various perfusion vis-à-vis static conditions and found that they were significantly affected by the applied conditions. Furthermore, the fluorescence intensities of OCN and OPN osteogenic gene markers were found to be enhanced in the 0.035 mL/min flow condition compared to the control, indicating that it was a suitable condition for osteogenic differentiation. Taken together, the findings of this study reveal that the developed cartridge device promotes the proliferation and differentiation of hMSCs and can potentially be used in the field of tissue engineering.

Dextran-coated fluorapatite nanorods doped with lanthanides in labelling and directing osteogenic differentiation of bone marrow mesenchymal stem cells

2014 ◽  
Vol 2 (23) ◽  
pp. 3609-3617 ◽  
Author(s):  
Haifeng Zeng ◽  
Xiyu Li ◽  
Fang Xie ◽  
Li Teng ◽  
Haifeng Chen
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Tissue Engineering ◽  
Osteogenic Differentiation ◽  
Bone Tissue ◽  
Novel Approach ◽  
Marrow Mesenchymal Stem Cells

A novel approach for labelling and tracking BMSCs in bone tissue engineering by using dextran-coated fluorapatite nanorods doped with lanthanides.

scholarly journals 3D Printed SiOC(N) Ceramic Scaffolds for Bone Tissue Regeneration: Improved Osteogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells

2021 ◽  
Vol 22 (24) ◽  
pp. 13676
Author(s):  
Yuejiao Yang ◽  
Apoorv Kulkarni ◽  
Gian Domenico Soraru ◽  
Joshua M. Pearce ◽  
Antonella Motta
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
3D Printing ◽  
Pore Size ◽  
Osteogenic Differentiation ◽  
Bone Tissue ◽  
Low Cost ◽  
Human Mesenchymal Stem Cells ◽  
Bone Tissue Regeneration ◽  
Gene Expressions

Bone tissue engineering has developed significantly in recent years as there has been increasing demand for bone substitutes due to trauma, cancer, arthritis, and infections. The scaffolds for bone regeneration need to be mechanically stable and have a 3D architecture with interconnected pores. With the advances in additive manufacturing technology, these requirements can be fulfilled by 3D printing scaffolds with controlled geometry and porosity using a low-cost multistep process. The scaffolds, however, must also be bioactive to promote the environment for the cells to regenerate into bone tissue. To determine if a low-cost 3D printing method for bespoke SiOC(N) porous structures can regenerate bone, these structures were tested for osteointegration potential by using human mesenchymal stem cells (hMSCs). This includes checking the general biocompatibilities under the osteogenic differentiation environment (cell proliferation and metabolism). Moreover, cell morphology was observed by confocal microscopy, and gene expressions on typical osteogenic markers at different stages for bone formation were determined by real-time PCR. The results of the study showed the pore size of the scaffolds had a significant impact on differentiation. A certain range of pore size could stimulate osteogenic differentiation, thus promoting bone regrowth and regeneration.

178 Differential Behavior of Porcine Mesenchymal Stem Cells from Bone Marrow and Adipose During Osteogenic Differentiation on a Glycosaminoglycan Hydrogel Scaffold for Bone and Cartilage Tissue Engineering

2018 ◽  
Vol 30 (1) ◽  
pp. 229 ◽  
Author(s):  
S. A. Womack ◽  
D. J. Milner ◽  
D. W. Weisgerber ◽  
B. A. Harley ◽  
M. B. Wheeler
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Scaffold Material ◽  
Alizarin Red ◽  
And Cartilage

The pig is an ideal species for use in tissue engineering studies targeted towards repair of bone and cartilage defects. Novel collagen-glycosaminoglycan hydrogel (CG) scaffolds have shown promise for supporting bone and cartilage growth from mesenchymal stem cells. In order to determine the suitability of these scaffolds for use in porcine model systems for bone and cartilage tissue engineering, we have begun to investigate the behaviour of porcine mesenchymal stem cells on this material. The purpose of this study was to determine whether mesenchymal stem cells from adipose (ASC) and bone marrow (BMSC) form bone on a CG scaffold material. Primary BMSC and ASC from 6-month-old Yorkshire pigs were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% fetal bovine serum. The ASC and BMSC were then trypsinized at passage 4 or 5 and used to seed ~4-mm-diameter CG scaffolds with 2 million cells/scaffold. Scaffolds were seeded by suspending the cells in medium that had been equilibrated for 30 min, and then placing the CG scaffold into the medium. This method of seeding was determined to be most effective in previous experiments. Scaffolds were then cultured for 7 days in DMEM followed by 21 days in osteogenic media. At the conclusion of the incubation period, the diameter of the scaffolds was measured, and they were fixed with 4% paraformaldehyde and cryosectioned. Then, 10-µm sections were stained with Alizarin Red to assay for mineralization, a hallmark of osteogenic differentiation. Both ASC- and BMSC-loaded scaffolds showed Alizarin Red staining throughout the section after incubation, demonstrating that both undergo osteogenesis on the scaffold material (n = 4). During osteogenic differentiation, scaffolds seeded with both ASC and BMSC showed a decrease in diameter. Unseeded scaffolds showed no decrease in size when in media. The BMSC scaffolds demonstrated a more extensive decrease in size than ASC. The average diameter of ASC loaded scaffolds after differentiation was 2.49 ± 0.39 mm, and that of BMSC-loaded scaffolds was 1.47 ± 0 0.18 mm (n = 3, P < 0.05, Student’s t-test). This suggests a differential ability of ASC and BMSC to break down and metabolize the scaffold matrix, and may indicate that one cell type may be preferable to the other for repairing osteogenic defects using these scaffolds. Current experiments underway will analyse expression of matrix-degrading enzymes to determine the source of the difference between cell types in scaffold shrinkage during differentiation. We will also quantify mineralization in ASC- v. BMSC-loaded scaffolds and assay gene expression of osteogenic markers to determine if there is a difference in osteogenic potential between sources of mesenchymal stem cells on these scaffolds.

Stimulated Osteogenic Differentiation of Human Mesenchymal Stem Cells by Reduced Graphene Oxide

2015 ◽  
Vol 15 (10) ◽  
pp. 7966-7970 ◽  
Author(s):  
Linhua Jin ◽  
Jong Ho Lee ◽  
Oh Seong Jin ◽  
Yong Cheol Shin ◽  
Min Jeong Kim ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Graphene Oxide ◽  
Osteogenic Differentiation ◽  
Reduced Graphene Oxide ◽  
Human Mesenchymal Stem Cells ◽  
Water Soluble ◽  
Alizarin Red ◽  
Reduced Graphene

Osteoprogenitor cells play a significant role in the growth or repair of bones, and have great potential as cell sources for regenerative medicine and bone tissue engineering, but control of their specific differentiation into bone cells remains a challenge. Graphene-based nanomaterials are attractive candidates for biomedical applications as substrates for stem cell (SC) differentiation, scaffolds in tissue engineering, and components of implant devices owing to their biocompatible, transferable and implantable properties. This study examined the enhanced osteogenic differentiation of human mesenchymal stem cells (hMSCs) by reduced graphene oxide (rGO) nanoparticles (NPs), and rGO NPs was prepared by reducing graphene oxide (GO) with a hydrazine treatment followed by annealing in argon and hydrogen. The cytotoxicity profile of each particle was examined using a water-soluble tetrazolium-8 (WST-8) assay. At different time-points, a WST-8 assay, alkaline phosphatase (ALP) activity assay and alizarin red S (ARS) staining were used to determine the effects of rGO NPs on proliferation, differentiation and mineralization, respectively. The results suggest that graphene-based materials have potential as a platform for stem cells culture and biomedicalapplications.

scholarly journals Nanotopography in directing osteogenic differentiation of mesenchymal stem cells: potency and future perspective

2021 ◽  
Author(s):  
Anggraini Barlian ◽  
Katherine Vanya
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
Osteogenic Differentiation ◽  
Future Perspective ◽  
Osteogenic Markers

Severe bone injuries can result in disabilities and thus affect a person's quality of life. Mesenchymal stem cells (MSCs) can be an alternative for bone healing by growing them on nanopatterned substrates that provide mechanical signals for differentiation. This review aims to highlight the role of nanopatterns in directing or inducing MSC osteogenic differentiation, especially in bone tissue engineering. Nanopatterns can upregulate the expression of osteogenic markers, which indicates a faster differentiation process. Combined with growth factors, nanopatterns can further upregulate osteogenic markers, but with fewer growth factors needed, thereby reducing the risks and costs involved. Nanopatterns can be applied in scaffolds for tissue engineering for their lasting effects, even in vivo, thus having great potential for future bone treatment.

scholarly journals 3D Printed SiOC(N) Ceramic Scaffolds for Bone Tissue Regeneration: Improved Osteogenic Differentiation of Human Bone Marrow‐Derived Mesenchymal Stem Cells

2021 ◽  
Author(s):  
Yuejiao Yang ◽  
Apoorv Kulkarni ◽  
Gian Domenico Soraru ◽  
Joshua M Pearce ◽  
Antonella Motta
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
3D Printing ◽  
Pore Size ◽  
Osteogenic Differentiation ◽  
Bone Tissue ◽  
Low Cost ◽  
Human Mesenchymal Stem Cells ◽  
Bone Tissue Regeneration ◽  
Gene Expressions

Bone tissue engineering has developed significantly in recent years as the increasing demand for bone substitutes due to trauma, cancer, arthritis, and infections. The scaffolds for bone regeneration need to be mechanically stable and have a 3D architecture with interconnected pores. With the advances in additive manufacturing technology, these requirements can be fulfilled by 3D printing scaffolds with controlled geometry and porosity using a low-cost multistep process. The scaffolds, however, must also be bioactive to promote the environment for the cells to regenerate into bone tissue. To determine if a low-cost 3D printing method for bespoke SiOC(N) porous structures can regenerate bone these structures were tested for osteointegration potential by using human mesenchymal stem cells (hMSCs). This includes checking the general biocompatibilities under the osteogenic differentiation environment (cell proliferation and metabolism). Moreover, cell morphology was observed by confocal microscopy and gene expressions on typical osteogenic markers at different stages for bone formation were determined by real-time PCR. The results of the study showed the pore size of the scaffolds had a significant impact on differentiation. A certain range of pore size could stimulate osteogenic differentiation, thus promoting bone regrowth and regeneration.

scholarly journals Osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells encapsulated in Thai silk fibroin/collagen hydrogel: a pilot study in vitro

2019 ◽  
Vol 12 (6) ◽  
pp. 273-279 ◽  
Author(s):  
Jirun Apinun ◽  
Sittisak Honsawek ◽  
Somsak Kuptniratsaikul ◽  
Jutarat Jamkratoke ◽  
Sorada Kanokpanont
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Pilot Study ◽  
Osteogenic Differentiation ◽  
Collagen Hydrogel ◽  
Collagen Hydrogels ◽  
Rat Bone

AbstractBackgroundSilk fibroin (SF) can be processed into a hydrogel. SF/collagen hydrogel may be a suitable biomaterial for bone tissue engineering.ObjectivesTo investigate in vitro biocompatibility and osteogenic potential of encapsulated rat bone marrow-derived mesenchymal stem cells (rat MSCs) in an injectable Thai SF/collagen hydrogel induced by oleic acid–poloxamer 188 surfactant mixture in an in vitro pilot study.MethodsRat MSCs were encapsulated in 3 groups of hydrogel scaffolds (SF, SF with 0.05% collagen [SF/0.05C], and SF with 0.1% collagen [SF/0.1C]) and cultured in a growth medium and an osteogenic induction medium. DNA, alkaline phosphatase (ALP) activity, and calcium were assayed at periodically for up to 5 weeks. After 6 weeks of culture the cells were analyzed by scanning electron microscopy and energy dispersive spectroscopy.ResultsAlthough SF hydrogel with collagen seems to have less efficiency to encapsulate rat MSCs, their plateau phase growth in all hydrogels was comparable. Inability to maintain cell viability as cell populations declined over 1–5 days was observed. Cell numbers then plateaued and were maintained until day 14 of culture. ALP activity and calcium content of rat MSCs in SF/collagen hydrogels were highest at day 21. An enhancing effect of collagen combined with the hydrogel was observed for proliferation and matrix formation; however, benefits of the combination on osteogenic differentiation and biomineralization are as yet unclear.ConclusionRat MSCs in SF and SF/collagen hydrogels showed osteogenic differentiation. Accordingly, these hydrogels may serve as promising scaffolds for bone tissue engineering.

scholarly journals Culture of rat mesenchymal stem cells on PHBV-PCL scaffolds: analysis of conditioned culture medium by FT-Raman spectroscopy

2023 ◽  
Vol 83 ◽  
Author(s):  
V. A. Nascimento ◽  
S. M. Malmonge ◽  
A. R. Santos Jr.
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Raman Spectroscopy ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Functional Groups ◽  
Culture Media ◽  
Cell Types ◽  
Ft Raman Spectroscopy ◽  
Ft Raman

Abstract Mesenchymal stem cells (MSCs) have great potential for application in cell therapy and tissue engineering procedures because of their plasticity and capacity to differentiate into different cell types. Given the widespread use of MSCs, it is necessary to better understand some properties related to osteogenic differentiation, particularly those linked to biomaterials used in tissue engineering. The aim of this study was to develop an analysis method using FT-Raman spectroscopy for the identification and quantification of biochemical components present in conditioned culture media derived from MSCs with or without induction of osteogenic differentiation. All experiments were performed between passages 3 and 5. For this analysis, MSCs were cultured on scaffolds composed of bioresorbable poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) polymers. MSCs (GIBCO®) were inoculated onto the pure polymers and 75:25 PHBV/PCL blend (dense and porous samples). The plate itself was used as control. The cells were maintained in DMEM (with low glucose) containing GlutaMAX® and 10% FBS at 37oC with 5% CO2 for 21 days. The conditioned culture media were collected and analyzed to probe for functional groups, as well as possible molecular variations associated with cell differentiation and metabolism. The method permitted to identify functional groups of specific molecules in the conditioned medium such as cholesterol, phosphatidylinositol, triglycerides, beta-subunit polypeptides, amide regions and hydrogen bonds of proteins, in addition to DNA expression. In the present study, FT-Raman spectroscopy exhibited limited resolution since different molecules can express similar or even the same stretching vibrations, a fact that makes analysis difficult. There were no variations in the readings between the samples studied. In conclusion, FT-Raman spectroscopy did not meet expectations under the conditions studied.

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