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The dynamic healing profile of human periodontal ligament stem cells: histological and immunohistochemical analysis using an ectopic transplantation model

 Y.-T. Kim  ;  J.-C. Park  ;  S.-H. Choi  ;  K.-S. Cho  ;  G.-I. Im  ;  B.-S. Kim  ;  C.-S. Kim 
 JOURNAL OF PERIODONTAL RESEARCH, Vol.47(4) : 514-524, 2012 
Journal Title
Issue Date
Alkaline Phosphatase/biosynthesis ; Analysis of Variance ; Animals ; Bone Regeneration/physiology ; Calcium Phosphates ; Cell Adhesion ; Cementogenesis/physiology ; Chronobiology Phenomena ; Collagen Type I/biosynthesis ; Collagen Type III/biosynthesis ; Core Binding Factor Alpha 1 Subunit/biosynthesis ; Durapatite ; Humans ; Immunoenzyme Techniques ; Male ; Mesenchymal Stem Cell Transplantation* ; Mice ; Mice, SCID ; Models, Biological ; Osteocalcin/biosynthesis ; Osteopontin/biosynthesis ; Periodontal Ligament/cytology* ; Periodontal Ligament/physiology* ; Regeneration/physiology* ; Statistics, Nonparametric ; Subcutaneous Tissue/surgery ; Tissue Engineering/methods* ; Tissue Scaffolds
periodontal ligament ; periodontium ; regeneration ; stem cell ; tissue engineering ; tri-calcium phosphate
BACKGROUND AND OBJECTIVE: Human periodontal ligament stem cells (hPDLSCs) have been reported to play the pivotal role in periodontal regeneration. However, the dynamic cellular healing process initiated by hPDLSCs still remains to be elucidated. In the present study, the sequence of regeneration by hPDLSCs was assessed using histological and immunohistochemical observation in an ectopic transplantation model, which is a well-standardized assessment tool that excludes the innate healing factors from the animals. MATERIAL AND METHODS: Human periodontal ligament stem cells that were isolated and characterized from teeth (n=12) extracted for the purpose of orthodontic treatment were transplanted with carriers into ectopic subcutaneous pouches in immunocompromised mice (n=20). Animals were killed after several different healing periods: 3 d (n=4), 1 (n=4), 2 (n=4), 4 (n=4) and 8 wk (n=4). Histological analysis for regenerated tissues formed by hPDLSCs was conducted using hematoxylin and eosin, Masson's trichrome and picrosirius red staining. In addition, immunohistochemical staining was performed to observe the sequential expression of osteogenic/cementogenic and periodontal ligament tissue-specific markers associated with periodontal regeneration. RESULTS: The whole healing process by transplanted hPDLSCs could be broadly divided into four distinctive phases. In the first phase, proliferated hPDLSCs migrated evenly all over the carrier, and collagenous tissues appeared in the form of amorphous collagen matrices. In the second phase, collagen fibers were well arranged among the carriers, and cementoid-like tissues were observed. In the third phase, the formation of mature collagen fibers, resembling Sharpey's fibers, was associated with active mineralization of cementum-like tissues, and in the fourth phase, the maturation of cementum-like tissues was observed on carrier surfaces. Various osteogenic/cementogenic markers related to the regeneration processes were expressed in a well-orchestrated time order. Interestingly, well-organized cementum-like and periodontal ligament fiber-like tissues and cells with early and late osteogenic/cementogenic markers were frequently observed in the secluded area of carrier surfaces. We termed this area the cell-rich zone. CONCLUSION: The results from this study clearly demonstrated the sequential histological changes during periodontal tissue regeneration by hPDLSCs. Understanding of this process would potentially enable us to develop better cell-based treatment techniques.
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2. College of Dentistry (치과대학) > Dept. of Periodontics (치주과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Chang Sung(김창성) ORCID logo https://orcid.org/0000-0003-3902-1071
Park, Jung Chul(박정철)
Cho, Kyoo Sung(조규성) ORCID logo https://orcid.org/0000-0002-6777-5287
Choi, Seong Ho(최성호) ORCID logo https://orcid.org/0000-0001-6704-6124
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