Differential Chondrogenic Potential of Human and Bovine Mesenchymal Stem Cells in Agarose and Photocrosslinked Hyaluronic Acid Hydrogels

2010 ◽  
Author(s):  
Minwook Kim ◽  
Isaac E. Erickson ◽  
Jason A. Burdick ◽  
George R. Dodge ◽  
Robert L. Mauck
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Hyaluronic Acid ◽  
Strong Dependence ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Human Mscs ◽  
Biologically Relevant

Articular cartilage has a limited regenerative capacity, and there exist no methodologies to restore structure and function after damage or degeneration. This has focused intense work on cell-based therapies for cartilage repair, with considerable literature demonstrating that chondrocytes in vitro and in vivo can generate cartilage-like tissue replacements. However, use of primary cells is limited by the amount and quality of autologous donor cells and tissue. Multipotential mesenchymal stem cells (MSCs) derived from bone marrow offer an alternative cell source for cartilage tissue engineering. MSCs are easily accessible and expandable in culture, and differentiate towards a chondrocyte-like phenotype with exposure to TGF-β [1]. For example, we have shown that bovine MSCs undergo chondrogenic differentiation and mechanical maturation in agarose, self-assembling peptide, and photocrosslinkable hyaluronic acid (HA) hydrogels [2]. HA hydrogels are particularly advantageous as they are biologically relevant and easily modified to generate a range of hydrogel properties [3]. Indeed, bovine MSCs show a strong dependence of functional outcomes on the macromer density of the HA gel [4]. To further the clinical application of this material, the purpose of this study was to investigate functional chondrogenesis of human MSCs in HA compared to agarose hydrogels. To carry out this study, juvenile bovine and human MSCs were encapsulated and cultured in vitro in HA and agarose hydrogels, and cell viability, biochemical, biomechanical, and histological properties were evaluated over 4 weeks of culture.

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

Influence of Chondrocyte Zone on Co-Cultures With Mesenchymal Stem Cells in HA Hydrogels for Cartilage Tissue Engineering

2012 ◽  
Author(s):  
Minwook Kim ◽  
Jason A. Burdick ◽  
Robert L. Mauck
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Hyaluronic Acid ◽  
Articular Chondrocytes ◽  
Cartilage Tissue Engineering ◽  
3D Culture ◽  
Cartilage Tissue ◽  
Cell Type

Mesenchymal stem cells (MSCs) are an attractive cell type for cartilage tissue engineering in that they can undergo chondrogenesis in a variety of 3D contexts [1]. Focused efforts in MSC-based cartilage tissue engineering have recently culminated in the formation of biologic materials possessing biochemical and functional mechanical properties that match that of the native tissue [2]. These approaches generally involve the continuous or intermittent application of pro-chondrogenic growth factors during in vitro culture. For example, in one recent study, we showed robust construct maturation in MSC-seeded hyaluronic acid (HA) hydrogels transiently exposed to high levels of TGF-β3 [3]. Despite the promise of this approach, MSCs are a multipotent cell type and retain a predilection towards hypertrophic phenotypic conversion (i.e., bone formation) when removed from a pro-chondrogenic environment (e.g., in vivo implantation). Indeed, even in a chondrogenic environment, many MSC-based cultures express pre-hypertrophic markers, including type X collagen, MMP13, and alkaline phosphatase [4]. To address this issue, recent studies have investigated co-culture of human articular chondrocytes and MSCs in both pellet and hydrogel environments. Chondrocytes appear to enhance the initial efficiency of MSC chondrogenic conversion, as well as limit hypertrophic changes in some instances (potentially via secretion of PTHrP and/or other factors) [5–7]. While these findings are intriguing, articular cartilage has a unique depth-dependent morphology including zonal differences in chondrocyte identity. Ng et al. showed that zonal chondrocytes seeded in a bi-layered agarose hydrogel construct can recreate depth-dependent cellular and mechanical heterogeneity, suggesting that these identities are retained with transfer to 3D culture systems [8]. Further, Cheng et al. showed that differences in matrix accumulation and hypertrophy in zonal chondrocytes was controlled by bone morphogenic protein [9]. To determine whether differences in zonal chondrocyte identity influences MSC fate decisions, we evaluated functional properties and phenotypic stability in photocrosslinked hyaluronic acid (HA) hydrogels using distinct, zonal chondrocyte cell fractions co-cultured with bone marrow derived MSCs.

TRENDS AND CHALLENGES OF CARTILAGE TISSUE ENGINEERING

2009 ◽  
Vol 21 (03) ◽  
pp. 149-155 ◽  
Author(s):  
Hsu-Wei Fang
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Growth Factors ◽  
Bone Cells ◽  
Cartilage Tissue Engineering ◽  
Bioactive Molecules ◽  
In Vivo Studies ◽  
Cartilage Tissue

Cartilage injuries may be caused by trauma, biomechanical imbalance, or degenerative changes of joint. Unfortunately, cartilage has limited capability to spontaneous repair once damaged and may lead to progressive damage and degeneration. Cartilage tissue-engineering techniques have emerged as the potential clinical strategies. An ideal tissue-engineering approach to cartilage repair should offer good integration into both the host cartilage and the subchondral bone. Cells, scaffolds, and growth factors make up the tissue engineering triad. One of the major challenges for cartilage tissue engineering is cell source and cell numbers. Due to the limitations of proliferation for mature chondrocytes, current studies have alternated to use stem cells as a potential source. In the recent years, a lot of novel biomaterials has been continuously developed and investigated in various in vitro and in vivo studies for cartilage tissue engineering. Moreover, stimulatory factors such as bioactive molecules have been explored to induce or enhance cartilage formation. Growth factors and other additives could be added into culture media in vitro, transferred into cells, or incorporated into scaffolds for in vivo delivery to promote cellular differentiation and tissue regeneration.Based on the current development of cartilage tissue engineering, there exist challenges to overcome. How to manipulate the interactions between cells, scaffold, and signals to achieve the moderation of implanted composite differentiate into moderate stem cells to differentiate into hyaline cartilage to perform the optimum physiological and biomechanical functions without negative side effects remains the target to pursue.

scholarly journals Comparison of Chondral Defects Repair with In Vitro and In Vivo Differentiated Mesenchymal Stem Cells

2007 ◽  
Vol 16 (8) ◽  
pp. 823-832 ◽  
Author(s):  
Hongbin Fan ◽  
Haifeng Liu ◽  
Rui Zhu ◽  
Xusheng Li ◽  
Yuming Cui ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cartilage Tissue ◽  
Burst Release ◽  
In Vitro Differentiation ◽  
Chondral Defects ◽  
In Vivo Differentiation ◽  
Repair Group

The purpose of this study was to compare chondral defects repair with in vitro and in vivo differentiated mesenchymal stem cells (MSCs). A novel PLGA-gelatin/chondroitin/hyaluronate (PLGA-GCH) hybrid scaffold with transforming growth factor-β1 (TGF-β1)-impregnated microspheres (MS-TGF) was fabricated to mimic the extracellular matrix. MS-TGF showed an initial burst release (22.5%) and a subsequent moderate one that achieved 85.1% on day 21. MSCs seeded on PLGA-GCH/MS-TGF or PLGA-GCH were incubated in vitro and showed that PLGA-GCH/MS-TGF significantly augmented proliferation of MSCs and glycosaminoglycan synthesis compared with PLGA-GCH. Then MSCs seeded on PLGA-GCH/MS-TGF were implanted and differentiated in vivo to repair chondral defect on the right knee of rabbit (in vivo differentiation repair group), while the contralateral defect was repaired with in vitro differentiated MSCs seeded on PLGA-GCH (in vitro differentiation repair group). The histology observation demonstrated that in vivo differentiation repair showed better chondrocyte morphology, integration, and subchondral bone formation compared with in vitro differentiation repair 12 and 24 weeks postoperatively, although there was no significant difference after 6 weeks. The histology grading score comparison also demonstrated the same results. The present study implies that in vivo differentiation induced by PLGA-GCH/MS-TGF and the host microenviroment could keep chondral phenotype and enhance repair. It might serve as another way to induce and expand seed cells in cartilage tissue engineering.

Chondrogenesis of human umbilical cord-derived mesenchymal stem cells in silk-fibroin/hyaluronic acid scaffolds for cartilage tissue engineering

2014 ◽  
Vol 185 ◽  
pp. S125 ◽  
Author(s):  
Jirayut Jaipaew ◽  
Piyanun Wangkulangkul ◽  
Chitkasem Suwanrath ◽  
Jirut Meesane ◽  
Supaporn Krivimol ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Hyaluronic Acid ◽  
Umbilical Cord ◽  
Silk Fibroin ◽  
Cartilage Tissue Engineering ◽  
Cartilage Tissue ◽  
Human Umbilical Cord

scholarly journals Mesenchymal stem cells are injured by complement after their contact with serum

Blood ◽  
2012 ◽  
Vol 120 (17) ◽  
pp. 3436-3443 ◽  
Author(s):  
Yan Li ◽  
Feng Lin
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Surface ◽  
Complement Activation ◽  
Human Mscs ◽  
Cellular Injury ◽  
Allogeneic Mscs ◽  
Complement Regulators

Abstract Despite the potent immunosuppressive activity that mesenchymal stem cells (MSCs) display in vitro, recent clinical trial results are disappointing, suggesting that MSC viability and/or function are greatly reduced after infusion. In this report, we demonstrated that human MSCs activated complement of the innate immunity after their contact with serum. Although all 3 known intrinsic cell-surface complement regulators were present on MSCs, activated complement overwhelmed the protection of these regulators and resulted in MSCs cytotoxicity and dysfunction. In addition, autologous MSCs suffered less cellular injury than allogeneic MSCs after contacting serum. All 3 complement activation pathways were involved in generating the membrane attack complex to directly injure MSCs. Supplementing an exogenous complement inhibitor, or up-regulating MSC expression levels of CD55, one of the cell-surface complement regulators, helped to reduce the serum-induced MSC cytotoxicity. Finally, adoptively transferred MSCs in complement deficient mice or complement-depleted mice showed reduced cellular injury in vivo compared with those in wild type mice. These results indicate that complement is integrally involved in recognizing and injuring MSCs after their infusion, suggesting that autologous MSCs may have ad-vantages over allogeneic MSCs, and that inhibiting complement activation could be a novel strategy to improve existing MSC-based therapies.

Distribution of Seeded Mesenchymal Stem Cells on Hydroxyapatite Porous Ceramics

2007 ◽  
Vol 330-332 ◽  
pp. 1141-1144 ◽  
Author(s):  
Mika Tadokoro ◽  
Noriko Kotobuki ◽  
Akira Oshima ◽  
Hajime Ohgushi
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
In Vitro Culture ◽  
Porous Ceramic ◽  
Porous Ceramics ◽  
Human Mscs ◽  
Ceramic Scaffold ◽  
Before And After

This study focused on in vivo osteogenic capability of bone marrow mesenchymal stem cells (MSCs) seeded on ceramic scaffold. Human MSCs from a single donor were seeded on hydroxyapatite porous ceramic (HAP) and were induced to the osteogenic lineage during in vitro culture condition, then the MSCs/HAP composites were implanted subcutaneously into immunodeficient rats. The cellular activities of the composites were assayed in order to evaluate the distribution and differentiation capability of seeded MSCs before and after implantation. These results showed that the new bone, after implantation, was derived from the donor MSCs, which adhered to the surface of the ceramics pore areas during in vitro culture. Therefore, the engrafted donor cells proliferated and showed continuous osteogenic differentiation within the recipients. Consequently, our study demonstrates the usefulness of MSCs/HAP composites for clinical applications.

scholarly journals Rheb1-Deficient Neutrophils Promote Hematopoietic Stem/Progenitor Cell Proliferation via Mesenchymal Stem Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Juan Gao ◽  
Shuaibing Hou ◽  
Shengnan Yuan ◽  
Yuxia Wang ◽  
Yanan Gao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Myeloid Cells ◽  
Human Mscs ◽  
Hematopoietic Stem ◽  
Progenitor Cell Proliferation ◽  
High Level ◽  
Lower Expression

Myeloid cells have been identified as hematopoietic stem cell (HSC)-regulating cells. However, the mechanisms by which myeloid cells regulate the function of HSCs are not fully defined. Our previous study indicated that the HSCs are over-expanded in Vav1-Cre;Rheb1fl/fl mice. Here, using in vivo and in vitro models, we found that Rheb1-deficient neutrophils remodeled the bone marrow environment and induced expansion of HSCs in vivo. Further studies showed that loss of Rheb1 impaired neutrophils’ ability to secrete IL-6, led mesenchymal stem cells (MSCs) to produce more SCF, and promote HSC proliferation. We further found that IL-6 suppressed SCF mRNA expression in human MSCs. Interesting, the high level of IL-6 was also related with poor survival of chronic myeloid leukemia (CML) patients, and higher expression of IL-6 in CML cells is associated with the lower expression of SCF in MSCs in patients. Our studies suggested that blocking IL-6 signaling pathway might stimulate MSCs to secrete more SCF, and to support hematopoietic stem/progenitor cells proliferation.

Transient Exposure to TGF-β3 Improves the Functional Properties of MSC-Seeded Photocrosslinked Hyaluronic Acid Hydrogels

2011 ◽  
Author(s):  
Minwook Kim ◽  
Isaac E. Erickson ◽  
Jason A. Burdick ◽  
Robert L. Mauck
Keyword(s):  
Hyaluronic Acid ◽  
Functional Properties ◽  
Cartilage Tissue Engineering ◽  
Compressive Load ◽  
Cartilage Tissue ◽  
Attractive Alternative ◽  
Biologically Relevant ◽  
Encapsulated Cells ◽  
Chondrogenic Phenotype

Articular cartilage is the primary compressive load bearing soft tissue in diarthrodial joints. While the tissue can function remarkably well in a demanding environment over a lifetime of use, focal defects and other trauma can initiate progressive degeneration. Cartilage tissue engineering approaches have been developed with the goal of forming biologic replacement materials with functional mechanical properties [1]. While chondrocytes are a popular cell source for such approaches, and can produce constructs with near-native functional properties [2], mesenchymal stem cells (MSCs) derived from bone marrow have emerged as an attractive alternative cell type. MSCs are multi-potent and easy to expand, and so are available in a nearly unlimited supply, and in an autologous fashion. While MSCs can undergo functional chondrogenesis in a variety of 3D contexts [3], we are particularly interested in the translational capacity of hyaluronic acid (HA). Hydrogels formed from this natural constituent of the cartilage extracellular matrix provide a biologically relevant interface for encapsulated cells and gel properties are readily tunable [4, 5]. Indeed, using a methacrylated (and so photo-crosslinkable) HA macromer, we have optimized gel formation and functional matrix production by MSCs with variations in both macromer (1%, [5]) and MSC (∼60 million cells/mL, [6]) concentration, consistently producing cartilage-like constructs with near native compressive properties. Additionally, we have reported that transient exposure of TGF-β3 (for three weeks) to MSCs in agarose constructs at a high-density induced a stable chondrogenic phenotype, with functional properties at six weeks greater than continual exposure to this pro-chondrogenic factor [7]. Transient exposure presents an interesting paradigm with clinical relevance, in vivo defect filling will require robust maturation of the engineered tissue driven by TGF-β3 delivered from the material itself in a controlled and sustained fashion. The purpose of this study was to determine the minimal TGF-β3 dosage and duration of exposure required to promote the most robust chondrogenesis and functional maturation of MSCs in this HA hydrogel system.

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