Abstract
BackgroundMesenchymal stem cell (MSC)-based cartilage tissue regeneration is a treatment with great potential. How to enhance the MSC chondrogenic differentiation is a key issue involved in cartilage formation. In the present study, we seek to expound the phenotypes and mechanisms of DLX5 in chondrogenic differentiation function in MSCs.MethodsStem cells from apical papilla (SCAPs) were used. The Alcian Blue staining, pellet culture system, and cell transplantation in rabbit knee cartilage defect were used to evaluate the chondrogenic differentiation function of MSCs. Western blot, real-time RT-PCR, and ChIP assays were used to evaluate the molecular mechanisms.ResultsDLX5 and HOXC8 expressions were upregulated during chondrogenic differentiation. In vitro results showed that DLX5 and HOXC8 enhanced the expression of chondrogenic markers including collagen II (COL2), collagen V (COL5), and sex-determining region Y box protein 9 (SOX9) and promoted the chondrogenic differentiation and the formation of cartilage clumps in the pellet culture system. Mechanically, DLX5 and HOXC8 formed protein complexes and negatively regulated the LncRNA, LINC01013, via directly binding its promoter. In vivo transplantation experiment showed that DLX5 and HOXC8 could restore the cartilage defect in the rabbit knee model. In addition, knock-down of LINC01013 enhanced the chondrogenic differentiation of SCAPs.ConclusionsIn conclusion, DLX5 and HOXC8 enhance the chondrogenic differentiation abilities of SCAPs by negatively regulating LINC01013 in SCAPs, and provided the potential target for promoting cartilage tissue regeneration.
Highlights
As a hypocellular and hypovascular tissue, the articular cartilage is formed by a specific extracellular matrix surrounding rare chondrocytes, and defects caused by natural degeneration or trauma may cause irreversible damage to its structure and function, which are leading sources of disability worldwide [1]
Distal-less homeobox 5 (DLX5) enhanced the chondrogenic differentiation potential of Stem cells from apical papilla (SCAPs) The expression of DLX5 was examined at the early stage of chondrogenic differentiation in SCAPs
In order to purify the infected cells, the transduced SCAPs were treated with 2 μg/ml puromycin for 3 days
Summary
As a hypocellular and hypovascular tissue, the articular cartilage is formed by a specific extracellular matrix surrounding rare chondrocytes, and defects caused by natural degeneration or trauma may cause irreversible damage to its structure and function, which are leading sources of disability worldwide [1] Conventional treatment strategies such as abrasion arthroplasty, bone marrow-stimulating repair, arthrocentesis, microfracture, and arthroscopic debridement have been used to solve this problem [2]. Research has confirmed that a new type of MSCs was isolated from dental tissue (nonbone marrow tissue) These dental MSCs include dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDs), periodontal ligament stem cells (PDLSCs), and stem cells from apical papilla (SCAPs) [6]. We seek to expound the phenotypes and mechanisms of DLX5 in chondrogenic differentiation function in MSCs
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