Comparison of Chondrocytes Produced from Adipose Tissue–Derived Stem Cells and Cartilage Tissue

Abstract Background Cartilage injury and pathological degeneration are reported in millions of patients globally. Cartilages such as articular hyaline cartilage are characterized by poor self-regeneration ability due to lack of vascular tissue. Current treatment methods adopt foreign cartilage analogue implants or microfracture surgery to accelerate tissue repair and regeneration. These methods are invasive and are associated with the formation of fibrocartilage, which warrants further exploration of new cartilage repair materials. The present study aims to develop an injectable modified gelatin hydrogel. Method The hydrogel effectively adsorbed proteoglycans secreted by chondrocytes adjacent to the cartilage tissue in situ, and rapidly formed suitable chondrocyte survival microenvironment modified by ε-poly-L-lysine (EPL). Besides, dynamic covalent bonds were introduced between glucose and phenylboronic acids (PBA). These bonds formed reversible covalent interactions between the cis−diol groups on polyols and the ionic boronate state of PBA. PBA-modified hydrogel induced significant stress relaxation, which improved chondrocyte viability and cartilage differentiation of stem cells. Further, we explored the ability of these hydrogels to promote chondrocyte viability and cartilage differentiation of stem cells through chemical and mechanical modifications. Results In vivo and in vitro results demonstrated that the hydrogels exhibited efficient biocompatibility. EPL and PBA modified GelMA hydrogel (Gel-EPL/B) showed stronger activity on chondrocytes compared to the GelMA control group. The Gel-EPL/B group induced the secretion of more extracellular matrix and improved the chondrogenic differentiation potential of stem cells. Finally, thus hydrogel promoted the tissue repair of cartilage defects. Conclusion Modified hydrogel is effective in cartilage tissue repair.

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Biomimetic composites and stem cells interaction for bone and cartilage tissue regeneration

Journal of Materials Chemistry ◽

10.1039/c1jm14401d ◽

2012 ◽

Vol 22 (12) ◽

pp. 5239 ◽

Cited By ~ 29

Author(s):

N. Naveena ◽

J. Venugopal ◽

R. Rajeswari ◽

S. Sundarrajan ◽

R. Sridhar ◽

...

Keyword(s):

Stem Cells ◽

Tissue Regeneration ◽

Cartilage Tissue ◽

And Cartilage

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Usage of stem cells in bone and cartilage tissue engineering

Biomedical Nanomaterials: From Design To Implementation ◽

10.1049/pbhe004e_ch4 ◽

2016 ◽

pp. 93-114

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Cartilage Tissue Engineering ◽

Cartilage Tissue ◽

And Cartilage

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Synovial Mesenchymal Stem Cells Derived From the Cotyloid Fossa Synovium Have Higher Self-renewal and Differentiation Potential Than Those From the Paralabral Synovium in the Hip Joint

The American Journal of Sports Medicine ◽

10.1177/0363546518794664 ◽

2018 ◽

Vol 46 (12) ◽

pp. 2942-2953 ◽

Cited By ~ 10

Author(s):

Yoichi Murata ◽

Soshi Uchida ◽

Hajime Utsunomiya ◽

Akihisa Hatakeyama ◽

Hirotaka Nakashima ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Femoroacetabular Impingement ◽

Impingement Syndrome ◽

Cartilage Tissue ◽

Gene Expressions ◽

Differentiation Potential ◽

Femoroacetabular Impingement Syndrome ◽

And Cartilage

Background: Several studies have shown the relationship between poorer clinical outcomes of arthroscopic femoroacetabular impingement syndrome surgery and focal chondral defects or global chondromalacia/osteoarthritis. Although recent studies described good outcomes after the conjunctive application of synovial mesenchymal stem cells (MSCs), none demonstrated the application of synovial MSCs for cartilaginous hip injuries. Purpose: To compare the characteristics of MSCs derived from the paralabral synovium and the cotyloid fossa synovium and determine which is the better source. Study Design: Controlled laboratory study. Methods: Synovium was harvested from 2 locations of the hip—paralabral and cotyloid fossa—from 18 donors. The number of cells, colony-forming units, viability, and differentiation capacities of adipose, bone, and cartilage were collected and compared between groups. In addition, real-time polymerase chain reaction was used to assess the differentiation capacity of adipose, bone, and cartilage tissue from both samples. Results: The number of colonies and yield obtained at passage 0 of synovium from the cotyloid fossa was significantly higher than that of the paralabral synovium ( P < .01). In adipogenesis experiments, the frequency of detecting oil red O–positive colonies was significantly higher in the cotyloid fossa than in the paralabral synovium ( P < .05). In osteogenesis experiments, the frequency of von Kossa and alkaline phosphatase positive colonies was higher in the cotyloid fossa synovium than in the paralabral synovium ( P < .05). In chondrogenic experiments, the chondrogenic pellet culture and the gene expressions of COL2a1 and SOX9 were higher in the cotyloid fossa synovium than in the paralabral synovium ( P < .05). Conclusion: MSCs from the cotyloid fossa synovium have higher proliferation and differentiation potential than do those from the paralabral synovium and are therefore a better source. Clinical Relevance: Synovial cells from the cotyloid fossa synovium of patients with femoroacetabular impingement syndrome are more robust in vitro, suggesting that MSCs from this source may be strongly considered for stem cell therapy.

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Stem Cells from New Sources for Cartilage Tissue Engineering

Otolaryngology ◽

10.1016/j.otohns.2008.05.485 ◽

2008 ◽

Vol 139 (2_suppl) ◽

pp. P87-P87

Author(s):

Ulrich R. Goessler ◽

Jens Stern-Straeter ◽

Gregor Bran ◽

Haneen Sadick ◽

Karl Hoermann ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Adipose Tissue ◽

Chondrogenic Differentiation ◽

Cartilage Tissue ◽

Molecular Characteristics ◽

Fibronectin Receptor ◽

Bone Marrow Biopsies ◽

Adult Mesenchymal Stem Cells ◽

Signaling Components

Problem The use of adult mesenchymal stem cells (MSC) – especially from new sources including adipose tissue - offers new perspectives in the generation of transplants for reconstructive surgery. The extracellular matrix (ECM) plays a key role in modulating function and phenotype of the embedded cells and contains the integrins as adhesion receptors mediating cell-cell- and cell-matrix-interactions. In our study, characteristic changes in integrin expression during the course of chondrogenic differentiation of MSC from bone marrow and adipose tissue were compared. Methods MSC were isolated from bone marrow biopsies and adipose tissue. During cell culture, chondrogenic differentiation was performed. The expression of Integrins and their signaling components were analyzed with microarray and immunohistochemistry in freshly isolated MSC and after chondrogenic differentiation. Results The Fibronectin-Receptor (Integrin a5b1) was expressed by undifferentiated MSC, expression rose during chondrogenic differentiation in both types of MSC. The components of the Vitronectin/Osteopontin-Receptors (avb5) were not expressed by freshly isolated MSC. Expression rose with ongoing differentiation. Receptors for Collagens (a1b1, a2b1, a3b1) were weakly expressed by undifferentiated MSC and were activated during differentiation. As intracellular signaling components integrin linked kinase (ILK) and CD47 showed increasing expression with ongoing differentiation. For all integrins, no significant differences could be found in the 2 types of MSC. Conclusion Integrin-mediated signaling seems to play an important role in the generation and maintenance of the chondrocytic phenotype during chondrogenic differentiation. The receptors for Fibronectin, Vitronectin, Osteopontin and Collagens in particular might be involved in the generation of the ECM. Intracellularly, their signals might be transduced by ILK and CD47. Significance To fully harness the potential of these cells, future studies should be directed to ascertain their cellular and molecular characteristics for optimal identification, isolation, and expansion.

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Chondrogenesis of Adult Stem Cells from Adipose Tissue and Bone Marrow: Induction by Growth Factors and Cartilage-Derived Matrix

Tissue Engineering Part A ◽

10.1089/ten.tea.2009.0398 ◽

2010 ◽

Vol 16 (2) ◽

pp. 523-533 ◽

Cited By ~ 165

Author(s):

Brian O. Diekman ◽

Christopher R. Rowland ◽

Donald P. Lennon ◽

Arnold I. Caplan ◽

Farshid Guilak

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Adipose Tissue ◽

Growth Factors ◽

Adult Stem Cells ◽

And Cartilage

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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

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab178 ◽

2018 ◽

Vol 30 (1) ◽

pp. 229 ◽

Cited By ~ 1

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.

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

Reproduction Fertility and Development ◽

10.1071/rdv29n1ab192 ◽

2017 ◽

Vol 29 (1) ◽

pp. 205 ◽

Cited By ~ 1

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.

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