Enhanced Cartilage Formation via Three-Dimensional Cell Engineering of Human Adipose-Derived Stem Cells

Abstract

Autologous chondrocyte implantation is an effective treatment for damaged articular cartilage. However, this method involves surgical procedures that may cause further cartilage degeneration, and in vitro expansion of chondrocytes can result in dedifferentiation. Adipose-derived stem cells (ADSCs) may be an alternative autologous cell source for cartilage regeneration. In this study, we developed an effective method for large-scale in vitro chondrogenic differentiation, which is the procedure that would be required for clinical applications, and the subsequent in vivo cartilage formation of human ADSCs (hADSCs). The spheroid formation and chondrogenic differentiation of hADSCs were induced on a large scale by culturing hADSCs in three-dimensional suspension bioreactors (spinner flasks). In vitro chondrogenic differentiation of hADSCs was enhanced by a spheroid culture compared with a monolayer culture. The enhanced chondrogenesis was probably attributable to hypoxia-related cascades and enhanced cell-cell interactions in hADSC spheroids. On hADSCs loading in fibrin gel and transplantation into subcutaneous space of athymic mice for 4 weeks, the in vivo cartilage formation was enhanced by the transplantation of spheroid-cultured hADSCs compared with that of monolayer-cultured hADSCs. This study shows that the spheroid culture may be an effective method for large-scale in vitro chondrogenic differentiation of hADSCs and subsequent in vivo cartilage formation.

Adipose-Derived Stem Cells in Tissue Regeneration: A Review

“…Neocartilage formation with integration with the surrounding host cartilage and bone has also been found in a larger model system using pig ASCs and full thickness articular defects [213], a model system that could provide important information on the way to the use of ASCs in the clinic. As a precursor to clinical application, successful cartilage formation in vivo has recently been reported using large-scale in vitro differentiation methods for the production of multiple spheroids suitable for implantation from human ASCS [214]. Finally, chondrogenic differentiation by ASCs in vivo cannot only be induced through their preinduction with TGF 1, but induced in vivo by using the scaffold itself.…”

Monitoring Cartilage Tissue Engineering Using Magnetic Resonance Spectroscopy, Imaging, and Elastography

“…Thus, much research is currently underway to improve cartilage tissue-engineering techniques. 10 Chondrocytes derived from natural sources or from MSCs differentiating toward chondrocytes are commonly used as a cell source in cartilage tissue engineering; however, there are unmet challenges in expanding and maintaining chondrocytes (e.g., the composition of the construct and the regulation of culture conditions need careful monitoring). 13,14,50,56,57 In addition, chondrogenic growth factors play a key role in chondrocyte differentiation and cartilage generation.…”

“…These techniques combine biocompatible scaffolds (natural or synthetic), appropriate cell types, and cell signaling molecules to create a tissue that mimics the functional native tissue. 10 Scaffolds are selected based on their ability to promote adhesion and proliferation of the desired cell types, although scaffold-free methods have also been reported in the literature. 11,12 As a cell source, either chondrocytes harvested from cartilage or mesenchymal stem cells (MSCs) differentiating toward chondrocytes are used.…”

Monitoring Tissue Engineering and Regeneration by Magnetic Resonance Imaging and Spectroscopy

“…For example, engineered cartilage aims to provide a long-term relief to patients with osteoarthritis or to individuals with acute sports injuries [3], whereas engineered bone can provide a cure for large bone defects [4]. Cartilage tissue engineering uses chondrocytes or mesenchymal stem cells as the cell source, hydrogel or fibrin based scaffolds and biomimetic or growth factor delivery based growth conditions to produce a native like functional cartilage engineered tissue [5][6][7][8]. The growth conditions such as the cell density, the mechanical properties of scaffolds, the oxygen concentration and the growth factor strength are varied to test the effectiveness of engineered tissue for its target biochemical and mechanical properties [5,9,10].…”

“…Cartilage tissue engineering uses chondrocytes or mesenchymal stem cells as the cell source, hydrogel or fibrin based scaffolds and biomimetic or growth factor delivery based growth conditions to produce a native like functional cartilage engineered tissue [5][6][7][8]. The growth conditions such as the cell density, the mechanical properties of scaffolds, the oxygen concentration and the growth factor strength are varied to test the effectiveness of engineered tissue for its target biochemical and mechanical properties [5,9,10]. Bone tissue engineering uses similar but bone-specific strategies, such as bone marrow derived mesenchymal stem cells (MSCs), suitable scaffolds and growth conditions to stimulate osteogenic differentiation of cells, to generate engineered bone with target biochemical and mechanical properties like a native bone [11][12][13][14][15][16].…”