Rapid Cartilage Regeneration of Spheroids Composed of Human Nasal Septum-Derived Chondrocyte in Rat Osteochondral Defect Model

Abstract

BACKGROUND: Cell-based therapies have been studied for articular cartilage regeneration. Articular cartilage defects have little treatments because articular cartilage was limited regenerative capacity. Damaged articular cartilage is difficult to obtain a successful therapeutic effect. In additionally these articular cartilage defects often cause osteoarthritis. Chondrocyte implantation is a widely available therapy used for regeneration of articular cartilage because this tissue has poor repair capacity after injury. Human nasal septum-drived chondrocytes (hNCs) from the septum show greater proliferation ability and chondrogenic capacity than human articular chondrocytes (hACs), even across different donors with different ages. Moreover, the chondrogenic properties of hNCs can be maintained after extensive culture expansion. METHODS: In this study, 2 dimensional (2D) monolayer cultured hNCs (hNCs-2D) and 3 dimensional (3D) spheroids cultured hNCs (hNCs-3D) were examined for chondrogenic capacity in vitro by PCR and immunofluorescence staining for chondrogenic marker, cell survival during cultured and for cartilage regeneration ability in vivo in a rat osteochondral defect model. RESULTS: hNCs-3D showed higher viability and more uniform morphology than 3D spheroids cultured hACs (hACs-3D) in culture. hNCs-3D also showed greater expression levels of the chondrocyte-specific marker Type II collagen (COL2A1) and sex-determining region Y (SRY)-box 9 (SOX9) than hNCs-2D. hNCs-3D also expressed chondrogenic markers in collagen. Specially, in the osteochondral defect model, implantation of hNCs-3D led to greater chondrogenic repair of focal cartilage defects in rats than implantation of hNCs-2D. CONCLUSION: These data suggest that hNCs-3D are valuable therapeutic agents for repair and regeneration of cartilage defects.

Engineering Multi‐Cellular Spheroids for Tissue Engineering and Regenerative Medicine

“…The presence of abundant cell–cell/cell–ECM inter‐communications within 3D spheroids has been reported to regulate cell functionality (e.g., viability, stimuli responsiveness, and protein secretion), leading to a substantial difference compared to 2D cultured cells. [ 2 ] For example, an increase in albumin secretion was confirmed in HepG2 spheroids as compared to that in 2D cultured ones; [ 32 ] chondrocytes derived from human nasal septum were assembled into spheroids that demonstrated greater chondrogenic differentiation and more physiological phenotype than 2D cultured cells; [ 33 ] spheroids prepared from human gingival fibroblasts reported enhancement in collagen secretion and greater chondrogenic differentiation potential compared to 2D cultured cells; and human mesenchymal stem cells (MSCs) also showed increased gene expression of pluripotency marker and ECM protein when cultured as spheroids. [ 34,35 ] In addition, human adipose‐derived mesenchymal stem cell (ASC) spheroids were superior in their protection from cellular damage by reactive oxygen species, demonstrating a decrease in the number of apoptotic cells as compared to those cultured in 2D conditions upon treatment with 300 µ m of hydrogen peroxide.…”

The utility of biomedical scaffolds laden with spheroids in various tissue engineering applications

“…In vivo, the spheroids differentiated into both bone and cartilage ( Figure 4 A ) 73 . Human nasal septum-derived chondrocyte (hNC) spheroids repaired osteochondral defects both in vitro and in vivo , resulting in improved cell viability, proliferation, and gene expression, as well as generated more ECM components, compared to a 2D monoculture of hNCs ( Figure 4 B ) 74 .…”

“…A. Schematic illustrations of a cylindrical construct composed of a fused spheroid and implantation 73 . B. Schematic of MSC spheroid injection with collagen in a cartilage defect model 74 . C. Illustration (left) and an optical image of cartilaginous tubes fabricated using the Kenzan method (middle), and an H&E-stained image of the graft at 35 days after transplantation (right).…”

Human nasal septal chondrocytes (NSCs) preconditioned on NSC-derived matrix improve their chondrogenic potential

“…In this study, we prepare nasal septal chondrocytes (NSCs)-derived ECM (CHDM) and examine its impact on NSCs behaviors, such as cell attachment, proliferation, and differentiation. As a promising source of primary chondrocytes, NSCs are isolated from nasal septal tissues that are surgically harvested from patients undergoing septoplasty [ 19 ]. Compared to the chondrocytes from knee articular cartilage, NSCs are beginning to find applications in cartilage regeneration [ 20 ].…”