Abstract
Three-dimensional clumps of mesenchymal stem cells (MSCs)/extracellular matrix (ECM) complexes (C-MSCs) can be transplanted into tissue defect site with no artificial scaffold. Importantly, most bone formation in the developing process or fracture healing proceeds via endochondral ossification. Accordingly, this present study investigated whether C-MSCs generated with chondro-inductive medium (CIM) can induce successful bone regeneration and assessed its healing process. Human bone marrow-derived MSCs were cultured with xeno-free/serum-free (XF) growth medium. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and then torn off. The sheet was rolled to make a round clump of cells. The cell clumps, i.e., C-MSCs, were maintained in XF-CIM. C-MSCs generated with XF-CIM showed enlarged round cells, cartilage matrix, and hypertrophic chondrocytes genes elevation in vitro. Transplantation of C-MSCs generated with XF-CIM induced successful bone regeneration in the SCID mouse calvaria defect model. Immunofluorescence staining for human-specific vimentin demonstrated that donor human and host mouse cells cooperatively contributed the bone formation. Besides, the replacement of the cartilage matrix into bone was observed in the early period. These findings suggested that cartilaginous C-MSCs generated with XF-CIM can induce bone regeneration via endochondral ossification.
Highlights
Bone plays a significant role in supporting the body structure, shielding the vital organs, and providing minerals and blood cells to maintain homeostasis [1,2]
For the scaffold-free bone regenerative cell therapy, we have recently developed three-dimensional clumps of mesenchymal stem cells (MSCs)/extracellular matrix (ECM) complexes (C-MSCs), which consisted of cells and self-produced ECM [15]
One cells (MSCs)/extracellular matrix (ECM) complexes (C-MSCs) cultured with GM or chondroinductive medium (CIM) for 5, 10, or 15 days was transplanted into the defect with no artificial scaffold, respectively
Summary
Bone plays a significant role in supporting the body structure, shielding the vital organs, and providing minerals and blood cells to maintain homeostasis [1,2]. The process of combining biomaterials and MSCs degrades the cell-cell or cell-extracellular matrix (ECM) contact, which may result in the disruption of the appropriate exercise of cellular function To overcome these problems, great scientific efforts have been made to develop novel biomaterials that can mimic cellular microenvironment and bring out both grafted and host cells’ function [12,13,14]. Great scientific efforts have been made to develop novel biomaterials that can mimic cellular microenvironment and bring out both grafted and host cells’ function [12,13,14] These promising biomaterials will be applied to the clinical settings in the near future
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