Abstract
ObjectiveTo explore the role of a three-dimensional (3D)-printed porous titanium alloy scaffold (3D scaffold) in the osteogenic differentiation of human adipose-derived mesenchymal stem cells (hADMSCs) and the underlying mechanism.MethodshADMSCs were divided into control and 3D scaffold groups. The osteogenic differentiation of hADMSCs and expression of osteogenic makers were estimated. Based on the information from published articles, five candidate circular RNAs were selected, and among them, hsa_circ_0019142 showed the most promising results. Finally, control group cells were overexpressed or silenced with the hsa_circ_0019142. Then, Alizarin red S (ARS) staining, calcium content analysis and estimation of alkaline phosphatase (ALP), osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), and collagen-1 (COL1) were performed to evaluate the role of hsa_circ_0019142 on osteogenic differentiation.ResultsOsteogenic differentiation of the hADMSCs was significantly higher in the 3D scaffold group than in the control group, as evidenced by ARS staining, increased calcium concentration, and elevated expression of above four osteogenic factors. qPCR revealed that the expression of hsa_circ_0019142 was significantly higher in the 3D scaffold group. Overexpression of hsa_circ_0019142 promoted the osteogenic differentiation of hADMSCs, while knockdown of hsa_circ_0019142 caused the opposite results.ConclusionThe 3D-printed scaffold promoted osteogenic differentiation of hADMSCs by upregulating hsa_circ_0019142.
Highlights
Replacement and regeneration of bone defects that occur following trauma, osteoarthritis, tumors, metabolic disorders, and osteoporosis poses significant challenges [1]
To assess the osteogenic activity of the two cell groups, we measured the mRNAs of the osteogenesis markers, including runt-related transcription factor 2 (RUNX2), collagen protein 1 (COL1), osteocalcin (OCN), and alkaline phosphatase (ALP), using Quantitative polymerase chain reaction (qPCR)
Expression of the mRNAs was significantly increased in the 3D scaffold group compared to the control group (P < 0.001 for ALP and RUNX2 and P < 0.01 for OCN and COL1) (Figure 2c)
Summary
Replacement and regeneration of bone defects that occur following trauma, osteoarthritis, tumors, metabolic disorders, and osteoporosis poses significant challenges [1]. Bone grafting (autologous and allografting) plays an important role in the field of orthopedics; the chance of disease transmission (in allografting) and donor-related issues, such as impaired mobility and unavailability of a suitable donor, limit the widespread application of grafting [2,3]. The focus is on the maintenance and improvement in specific organ functions through the replacement and/or repair of specific organs [4,5]. The conventional two-dimensional (2D) cell culture does not allow interaction between the cells and extracellular matrix [6]. Three-dimensional (3D) scaffolds composed of stem cells and other biomolecules are gaining popularity [7]
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