Usage of stem cells in bone and cartilage tissue engineering

2016 ◽  
pp. 93-114
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
And Cartilage

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.

192 BEHAVIOR OF PORCINE MESENCHYMAL STEM CELLS ON A COLLAGEN-GLYCOSAMINOGLYCAN HYDROGEL SCAFFOLD FOR BONE AND CARTILAGE TISSUE ENGINEERING

2017 ◽  
Vol 29 (1) ◽  
pp. 205 ◽  
Author(s):  
S. A. Womack ◽  
D. J. Milner ◽  
D. W. Weisgerber ◽  
B. A. C. Harley ◽  
M. B. Wheeler
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Density ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Differentiation Potential ◽  
Static Culture ◽  
A Cell ◽  
And Cartilage

The pig is an ideal species for use in tissue engineering studies of bone and cartilage defect repair. 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 models 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 if mesenchymal stem cells from fat (ASC) or bone marrow (BMSC) displayed better adherence and penetration into the CG scaffold material. The BMSC and ASC isolated from young adult Yorkshire pigs were cultured in DMEM with 10% fetal bovine serum. The ASC and BMSC were then trypsinized and used to seed ~3 mm diameter CG scaffolds with 140,000 cells/scaffold. Scaffolds were then cultured for 10 days by 3 different methods: roller culture, free-floating non-adherent dishes (floating), or attached to tissue culture-treated dishes (static). At the conclusion of the incubation period, the scaffold pieces were then fixed with 4% paraformaldehyde, embedded for cryosectioning, and sliced into 10 µm cryosections. Sections were stained for vimentin and 4’,6-diamidino-2-phenylindole (DAPI) to label cells. Stained sections were observed on a Leica DMB4200 microscope (Leica Microsystems, Wetzlar, Germany) and images acquired using ImagePro Plus software (Media Cybernetics Inc., Rockville, MD, USA). The DAPI-stained cells were counted to determine cell density and expressed as average number of nuclei per millimeter squared for each cell and culture type. Data were analysed by ANOVA utilising a post hoc Holm multiple comparison analysis. Samples from roller cultures did not display adhered cells for either BMSC or ASC. In contrast, floating and static culture allowed both ASC and BMSC to adhere to the scaffold and migrate to the centre of the scaffold equally well. However, significant differences in cell densities were noted between ASC and BMSC on CG scaffolds, with BMSC growing to higher densities than ASC in both floating and static culture. For floating cultures, BMSC-loaded scaffolds exhibited a cell density of 105.7 compared with 53.3 cells/mm2 for ASC (n = 4; P < 0.05). For static cultures, BMSC-loaded scaffolds exhibited a cell density of 128.3 compared with 36.8 cells/mm2 for ASC-loaded samples (n = 3; P < 0.01). Thus, BMSC grow to greater densities more rapidly than ASC and may be more efficient for use in forming bone and cartilage on these scaffolds. Current experiments underway will compare osteogenic and chondrogenic differentiation potential of ASC and BMSC on CG scaffolds, and will attempt to engineer osteochondral interface tissue on CG scaffolds from co-cultures of chondrocytes and stem cells.

scholarly journals Adult Stem Cells in Bone and Cartilage Tissue Engineering

2006 ◽  
Vol 1 (3) ◽  
pp. 345-364 ◽  
Author(s):  
Antonio Salgado ◽  
Joao Oliveira ◽  
Adriano Pedro ◽  
Rui Reis
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Adult Stem Cells ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
And Cartilage

Spidroin Striped Micropattern Promotes Chondrogenic Differentiation of Human Wharton’s Jelly Mesenchymal Stem Cells

2021 ◽  
Author(s):  
Anggraini Barlian ◽  
Dinda Hani’ah Arum Saputri ◽  
Adriel Hernando ◽  
Ekavianty Prajatelistia ◽  
Hutomo Tanoto
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Spider Silk ◽  
Cartilage Tissue Engineering ◽  
Wharton’S Jelly ◽  
Cartilage Tissue ◽  
Type Ii ◽  
Wharton's Jelly

Abstract Cartilage tissue engineering, particularly micropattern, can influence the biophysical properties of mesenchymal stem cells (MSCs) leading to chondrogenesis. In this research, human Wharton’s jelly MSCs (hWJ-MSCs) were grown on a striped micropattern containing spider silk protein (spidroin) from Argiope appensa. This research aims to direct hWJ-MSCs chondrogenesis using micropattern made of spidroin bioink as opposed to fibronectin that often used as the gold standard. Cells were cultured on striped micropattern of 500 µm and 1000 µm width sizes without chondrogenic differentiation medium for 21 days. The immunocytochemistry result showed that spidroin contains RGD sequences and facilitates cell adhesion via integrin β1. Chondrogenesis was observed through the expression of glycosaminoglycan, type II collagen, and SOX9. The result on glycosaminoglycan content proved that 1000 µm was the optimal width to support chondrogenesis. Spidroin micropattern induced significantly higher expression of SOX9 mRNA on day-21 and SOX9 protein was located inside the nucleus starting from day-7. COL2A1 mRNA of spidroin micropattern groups was downregulated on day-21 and collagen type II protein was detected starting from day-14. These results showed that spidroin micropattern enhances chondrogenic markers while maintains long-term upregulation of SOX9, and therefore has the potential as a new method for cartilage tissue engineering.

scholarly journals Cartilage Tissue Engineering Using Mesenchymal Stem Cells and 3D Chitosan Scaffolds – In vitro and in vivo Assays

10.5772/25085 ◽  
2011 ◽  
Author(s):  
Natalia Martins ◽  
Alessandra Arcoverde ◽  
Juliana Lott ◽  
Viviane Silva ◽  
Dawidson Gomes ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
In Vivo Assays ◽  
Chitosan Scaffolds
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