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Clumps of mesenchymal stem cells/extracellular matrix complexes directly reconstruct the functional periodontal tissue in a rat periodontal defect model

Periodontitis is an inflammatory disease characterized by tooth‐supporting periodontal tissue destruction, including the cementum, periodontal ligament, and alveolar bone. To regenerate the damaged periodontal tissue, mesenchymal stem cells (MSCs) have attracted much scientific and medical attention. Recently, we generated clumps of MSCs/extracellular matrix (ECM) complexes (C‐MSCs), which consist of cells and self‐produced ECM. C‐MSCs can be transplanted into lesion areas without artificial scaffold to induce tissue regeneration. To develop reliable scaffold‐free periodontal tissue regenerative cell therapy by C‐MSCs, this study investigated the periodontal tissue regenerative capacity of C‐MSCs and the behavior of the transplanted cells. Rat bone marrow‐derived MSCs were isolated from rat femur. Confluent cells were scratched using a micropipette tip and then torn off. The sheet was rolled to make a three‐dimensional round clump of cells, C‐MSCs. Then, ten C‐MSCs were grafted into a rat periodontal fenestration defect model. To trace the grafted cells in the defect, PKH26‐labeled cells were also employed. Micro‐CT and histological analyses demonstrated that transplantation of C‐MSCs induced successful periodontal tissue regeneration in the rat periodontal defect model. Interestingly, the majority of the cells in the reconstructed tissue, including cementum, periodontal ligaments, and alveolar bone, were PKH26 positive donor cells, suggesting the direct tissue formation by MSCs. This study demonstrates a promising scaffold‐free MSCs transplantation strategy for periodontal disease using C‐MSCs and offers the significance of multipotency of MSCs to induce successful periodontal tissue regeneration.