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Hidemitsu Harada - One of the best experts on this subject based on the ideXlab platform.

  • Regenerated teeth: the future of Tooth replacement. An update
    Regenerative medicine, 2015
    Co-Authors: Thimios A. Mitsiadis, Hidemitsu Harada
    Abstract:

    A plethora of dental materials are successfully used for partial dental tissue repair, while innovative dental implants are used for Tooth replacement. Cell-based Tooth Regeneration is an attractive approach that complements traditional restorative or surgical techniques for replacement of damaged dental tissues. Recent efforts focus mostly on partial Tooth Regeneration for the treatment of diseases that commonly affect dental tissues. However, several attempts have been also made for the Regeneration of entire teeth.

  • Stem cell sources for Tooth Regeneration: current status and future prospects
    Frontiers in physiology, 2014
    Co-Authors: Keishi Otsu, Mika Kumakami-sakano, Naoki Fujiwara, Kazuko Kikuchi, Laetitia Keller, Hervé Lesot, Hidemitsu Harada
    Abstract:

    Stem cells are capable of renewing themselves through cell division and have the remarkable ability to differentiate into many different types of cells. They therefore have the potential to become a central tool in regenerative medicine. During the last decade, advances in tissue engineering and stem cell-based Tooth Regeneration have provided realistic and attractive means of replacing lost or damaged teeth. Investigation of embryonic and adult (tissue) stem cells as potential cell sources for Tooth Regeneration has led to many promising results. However, technical and ethical issues have hindered the availability of these cells for clinical application. The recent discovery of induced pluripotent stem (iPS) cells has provided the possibility to revolutionize the field of regenerative medicine (dentistry) by offering the option of autologous transplantation. In this article, we review the current progress in the field of stem cell-based Tooth Regeneration and discuss the possibility of using iPS cells for this purpose.

  • Differentiation of induced pluripotent stem cells into dental mesenchymal cells.
    Stem cells and development, 2011
    Co-Authors: Keishi Otsu, Naoki Fujiwara, Ryota Kishigami, Ai Oikawa-sasaki, Satoshi Fukumoto, Aya Yamada, Kiyoto Ishizeki, Hidemitsu Harada
    Abstract:

    Similar to embryonic stem cells, induced pluripotent stem (iPS) cells can differentiate into various cell types upon appropriate induction, and thus, may be valuable cell sources for regenerative medicine. However, iPS cells have not been reported to differentiate into odontogenic cells for Tooth Regeneration. Here we demonstrated that neural crest-like cells (NCLC) derived from mouse iPS cells have the potential to differentiate into odontogenic mesenchymal cells. We developed an efficient culture protocol to induce the differentiation of mouse iPS cells into NCLC. We confirmed that the cells exhibited neural crest (NC) cell markers as evidenced by immunocytochemistry, flow cytometry, and real-time reverse transcription-polymerase chain reaction. Further, in recombination cultures of NCLC and mouse dental epithelium, NCLC exhibited a gene expression pattern involving dental mesenchymal cells. Some NCLC also expressed dentin sialoprotein. Conditioned medium of mouse dental epithelium cultures further enhanced the differentiation of NCLC into odontoblasts. These results suggest that iPS cells are useful cell sources for Tooth Regeneration and Tooth development studies.

Tzong-fu Kuo - One of the best experts on this subject based on the ideXlab platform.

  • Tooth germ like construct transplantation for whole Tooth Regeneration an in vivo study in the miniature pig
    Artificial Organs, 2016
    Co-Authors: Kai Chiang Yang, Hao-hueng Chang, Yutaka Kitamura, Thai-yen Ling, Tzong-fu Kuo
    Abstract:

    The purpose of this study was to demonstrate the feasibility of whole-Tooth Regeneration using a Tooth germ-like construct. Dental pulp from upper incisors, canines, premolars, and molars were extracted from sexually mature miniature pigs. Pulp tissues were cultured and expanded in vitro to obtain dental pulp stem cells (DPSCs), and cells were differentiated into odontoblasts and osteoblasts. Epithelial cells were isolated from gingival epithelium. The epithelial cells, odontoblasts, and osteoblasts were seeded onto the surface, upper, and lower layers, respectively, of a bioactive scaffold. The lower first and second molar Tooth germs were removed bilaterally and the layered cell/scaffold constructs were transplanted to the mandibular alveolar socket of a pig. At 13.5 months postimplantation, seven of eight pigs developed two teeth with crown, root, and pulp structures. Enamel-like tissues, dentin, cementum, odontoblasts, and periodontal tissues were found upon histological inspection. The regenerated Tooth expressed dentin matrix protein-1 and osteopontin. All pigs had regenerated molar teeth regardless of the original Tooth used to procure the DPSCs. Pigs that had Tooth germs removed or who received empty scaffolds did not develop teeth. Although periodontal ligaments were generated, ankylosis was found in some animals. This study revealed that implantation of a Tooth germ-like structure generated a complete Tooth with a high success rate. The implant location may influence the morphology of the regenerated Tooth.

  • Tooth Germ‐Like Construct Transplantation for Whole‐Tooth Regeneration: An In Vivo Study in the Miniature Pig
    Artificial organs, 2015
    Co-Authors: Kai Chiang Yang, Hao-hueng Chang, Yutaka Kitamura, Thai-yen Ling, Tzong-fu Kuo
    Abstract:

    The purpose of this study was to demonstrate the feasibility of whole-Tooth Regeneration using a Tooth germ-like construct. Dental pulp from upper incisors, canines, premolars, and molars were extracted from sexually mature miniature pigs. Pulp tissues were cultured and expanded in vitro to obtain dental pulp stem cells (DPSCs), and cells were differentiated into odontoblasts and osteoblasts. Epithelial cells were isolated from gingival epithelium. The epithelial cells, odontoblasts, and osteoblasts were seeded onto the surface, upper, and lower layers, respectively, of a bioactive scaffold. The lower first and second molar Tooth germs were removed bilaterally and the layered cell/scaffold constructs were transplanted to the mandibular alveolar socket of a pig. At 13.5 months postimplantation, seven of eight pigs developed two teeth with crown, root, and pulp structures. Enamel-like tissues, dentin, cementum, odontoblasts, and periodontal tissues were found upon histological inspection. The regenerated Tooth expressed dentin matrix protein-1 and osteopontin. All pigs had regenerated molar teeth regardless of the original Tooth used to procure the DPSCs. Pigs that had Tooth germs removed or who received empty scaffolds did not develop teeth. Although periodontal ligaments were generated, ankylosis was found in some animals. This study revealed that implantation of a Tooth germ-like structure generated a complete Tooth with a high success rate. The implant location may influence the morphology of the regenerated Tooth.

  • Tooth Regeneration WITH DENTAL STEM CELL RESEARCH IN MINIATURE PIG MODEL
    Taiwan Veterinary Journal, 2015
    Co-Authors: Tzong-fu Kuo, Shi-yuan Sheu, Ching-chuan Jiang, Hao-hueng Chang, Shuo-tsung Chen, Rung-shu Chen, Chin-hsiung Hsieh, Min-huey Chen
    Abstract:

    To develop a cell-based approach to Tooth Regeneration, we isolated dental stem cells from Tooth buds of mini pigs and culture-expanded them for about three weeks. Flow cytometry revealed that the majority of the proliferating cells were positive for stem cell surface markers, including CD105, CD29, CD44, CD71. These dental stem cells were then seeded in biodegradable agarose gel for autograft in various areas of the jaw bone. Mini pigs with Tooth buds removed without transplantation of dental stem cells served as control. After one year, well differentiated Tooth crown and root sheath were formed in mini pigs in the experimental group. The Tooth formed by dental stem cells transplanted in alveolar bone was similar in size to original Tooth crown (approximately 2 cm × 2 cm × 2 cm). The teeth formed by dental stem cells transplanted under the roots of the anterior teeth and in the cortical bone area were smaller. In the control group, the space where the Tooth bud was extracted without cell transplantation was seen to be closed by forward movement of the posterior teeth. Histological analysis showed the structures of enamel, dentin and pulp in the regenerated Tooth crown. Immunohistochemical analysis of amelogenin, type I collagen and bone sialoprotein confirmed enamel and dentin formation. Enamel and dentin structures on the regenerated Tooth were also identified using scanning electron microscopy. These findings demonstrate a successful model of Tooth Regeneration from dental stem cells in mini pigs. In addition, local environment and available space also significantly affect the extent of tissue Regeneration. Our study demonstrates the feasibility of using dental stem cells for Tooth Regeneration in clinical applications.

  • fibrin glue mixed with platelet rich fibrin as a scaffold seeded with dental bud cells for Tooth Regeneration
    Journal of Tissue Engineering and Regenerative Medicine, 2012
    Co-Authors: Kai Chiang Yang, Hao-hueng Chang, Chun Hao Wang, Wing P Chan, Chauhwa Chi, Tzong-fu Kuo
    Abstract:

    Odontogenesis is a complex process with a series of epithelial-mesenchymal interactions and odontogenic molecular cascades. In tissue engineering of teeth from stem cells, platelet-rich fibrin (PRF), which is rich in growth factors and cytokines, may improve Regeneration. Accordingly, PRF was added into fibrin glue to enrich the microenvironment with growth factors. Unerupted second molar Tooth buds were harvested from miniature swine and cultured in vitro for 3 weeks to obtain dental bud cells (DBCs). Whole blood was collected for the preparation of PRF and fibrin glue before surgery. DBCs were suspended in fibrin glue and then enclosed with PRF, and the DBC-fibrin glue-PRF composite was autografted back into the original alveolar sockets. Radiographic and histological examinations were used to identify the regenerated Tooth structure 36 weeks after implantation. Immunohistochemical staining was used to detect proteins specific to Tooth Regeneration. One pig developed a complete Tooth with crown, root, pulp, enamel, dentin, odontoblast, cementum, blood vessels, and periodontal ligaments in indiscriminate shape. Another animal had an unerupted Tooth that expressed cytokeratin 14, dentin matrix protein-1, vascular endothelial growth factor, and osteopontin. This study demonstrated, using autogenic cell transplantation in a porcine model, that DBCs seeded into fibrin glue-PRF could regenerate a complete Tooth. Copyright © 2011 John Wiley & Sons, Ltd.

  • Bone marrow combined with dental bud cells promotes Tooth Regeneration in miniature pig model.
    Artificial organs, 2010
    Co-Authors: Tzong-fu Kuo, Min-huey Chen, H.c. Lin, Kai Chiang Yang, Feng-huei Lin, Hao-hueng Chang
    Abstract:

    Growth factors and morphogens secreted by bone marrow mesenchymal stem cells (BMSCs) of bone marrow fluid may promote Tooth Regeneration. Accordingly, a tissue engineering approach was utilized to develop an economical strategy for obtaining the growth factors and morphogens from BMSCs. Unerupted second molar Tooth buds harvested from miniature pigs were cultured in vitro to obtain dental bud cells (DBCs). Bone marrow fluid, which contains BMSCs, was collected from the porcine mandible before operation. DBCs suspended in bone marrow fluid were seeded into a gelatin/chondoitin-6-sulfate/hyaluronan tri-copolymer scaffold (GCHT scaffold). The DBCs/bone marrow fluid/GCHT scaffold was autografted into the original alveolar sockets of the pigs. Radiographic and histological examinations were applied to identify the structure of regenerated Tooth at 40 weeks postimplantation. The present results showed that one pig developed a complete Tooth with crown, root, pulp, enamel, dentin, odontoblast, cementum, blood vessel, and periodontal ligament in indiscriminate shape. Three animals had an unerupted Tooth that expressed dentin matrix protein-1, vascular endothelial growth factor, and osteopontin; and two other pigs also had dental-like structure with dentin tubules. This study reveals that DBCs adding bone marrow fluid and a suitable scaffold can promote the Tooth Regeneration in autogenic cell transplantation.

Dong-woo Cho - One of the best experts on this subject based on the ideXlab platform.

  • analysis of the soluble human Tooth proteome and its ability to induce dentin Tooth Regeneration
    Tissue Engineering Part A, 2011
    Co-Authors: So Young Chun, Hyo Jung Lee, Young Ae Choi, Kyung-min Kim, Sang Heum Baek, Hyo-sang Park, Jae-young Kim, Jungmo Ahn, Je-yeol Cho, Dong-woo Cho
    Abstract:

    While the soluble proteins of human teeth consist of various extracellular matrix and bioactive proteins, they have not yet been characterized fully. Moreover, the role they play in Tooth Regeneration is not clear. Analysis of the soluble proteins in human teeth by liquid chromatography-mass spectrometry revealed 147 different ethylenediaminetetraacetic acid-soluble Tooth proteins (ESTPs). Of these, 29 had not been shown previously to be present in human teeth. To determine their effect on the in vitro responses of dental pulp stem cells (DPSCs), DPSCs were cultured in ESTP-coated culture plates and three-dimensional scaffolds. The ESTPs significantly enhanced DPSC odontoblast differentiation and mineralization in vitro, but had only partial effect on bone marrow stem cells or adipose tissue stem cells. To test the effect of ESTPs on in vivo dentin and Tooth formation, mouse embryonic Tooth-forming primordia and xenogenic murine apical bud epithelium/human DPSC composites were treated with ESTPs before implantation under the renal capsule of ICR mice. ESTP treatment promoted the formation of morphologically normal teeth by the Tooth-forming primordium regions and enhanced the development of a regular and large dentin structure by the composites. These observations suggest that human ESTPs contain dentinogenic proteins and can promote dentin and Tooth formation.

  • Analysis of the soluble human Tooth proteome and its ability to induce dentin/Tooth Regeneration.
    Tissue engineering. Part A, 2010
    Co-Authors: So Young Chun, Hyo Jung Lee, Young Ae Choi, Kyung-min Kim, Sang Heum Baek, Hyo-sang Park, Jae-young Kim, Jungmo Ahn, Je-yeol Cho, Dong-woo Cho
    Abstract:

    While the soluble proteins of human teeth consist of various extracellular matrix and bioactive proteins, they have not yet been characterized fully. Moreover, the role they play in Tooth Regeneration is not clear. Analysis of the soluble proteins in human teeth by liquid chromatography-mass spectrometry revealed 147 different ethylenediaminetetraacetic acid-soluble Tooth proteins (ESTPs). Of these, 29 had not been shown previously to be present in human teeth. To determine their effect on the in vitro responses of dental pulp stem cells (DPSCs), DPSCs were cultured in ESTP-coated culture plates and three-dimensional scaffolds. The ESTPs significantly enhanced DPSC odontoblast differentiation and mineralization in vitro, but had only partial effect on bone marrow stem cells or adipose tissue stem cells. To test the effect of ESTPs on in vivo dentin and Tooth formation, mouse embryonic Tooth-forming primordia and xenogenic murine apical bud epithelium/human DPSC composites were treated with ESTPs before im...

Lei Liu - One of the best experts on this subject based on the ideXlab platform.

  • Bone marrow-derived stromal cells are more beneficial cell sources for Tooth Regeneration compared with adipose-derived stromal cells.
    Cell biology international, 2015
    Co-Authors: Lin Chen, Junhui Cui, Fan Feng, Lei Liu
    Abstract:

    Tooth loss is presently a global epidemic and Tooth Regeneration is thought to be a feasible and ideal treatment approach. Choice of cell source is a primary concern in Tooth Regeneration. In this study, the odontogenic differentiation potential of two non-dental-derived stem cells, adipose-derived stromal cells (ADSCs) and bone marrow-derived stromal cells (BMSCs), were evaluated both in vitro and in vivo. ADSCs and BMSCs were induced in vitro in the presence of Tooth germ cell-conditioned medium (TGC-CM) prior to implantation into the omentum majus of rats, in combination with inactivated dentin matrix (IDM). Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of odontogenic-related genes. Immunofluorescence and immunohistochemical assays were used to detect the protein levels of odontogenic-specific genes, such as DSP and DMP-1 both in vitro and in vivo. The results suggest that both ADSCs and BMSCs have odontogenic differentiation potential. However, the odontogenic potential of BMSCs was greater compared with ADSCs, showing that BMSCs are a more appropriate cell source for Tooth Regeneration.

  • Human Umbilical Cord Mesenchymal Stem Cells: A New Therapeutic Option for Tooth Regeneration
    Stem cells international, 2015
    Co-Authors: Yuanwei Chen, Lin Chen, Junhui Cui, Quan Sun, Lei Liu
    Abstract:

    Tooth Regeneration is considered to be an optimistic approach to replace current treatments for Tooth loss. It is important to determine the most suitable seed cells for Tooth Regeneration. Recently, human umbilical cord mesenchymal stem cells (hUCMSCs) have been regarded as a promising candidate for tissue Regeneration. However, it has not been reported whether hUCMSCs can be employed in Tooth Regeneration. Here, we report that hUCMSCs can be induced into odontoblast-like cells in vitro and in vivo. Induced hUCMSCs expressed dentin-related proteins including dentin sialoprotein (DSP) and dentin matrix protein-1 (DMP-1), and their gene expression levels were similar to those in native pulp tissue cells. Moreover, DSP- and DMP-1-positive calcifications were observed after implantation of hUCMSCs in vivo. These findings reveal that hUCMSCs have an odontogenic differentiation potency to differentiate to odontoblast-like cells with characteristic deposition of dentin-like matrix in vivo. This study clearly demonstrates hUCMSCs as an alternative therapeutic cell source for Tooth Regeneration.

  • Odontogenic differentiation of adipose-derived stem cells for Tooth Regeneration: necessity, possibility, and strategy
    Medical hypotheses, 2007
    Co-Authors: Wei Jing, Lei Liu, Yunfeng Lin, Wei Tang, Weidong Tian
    Abstract:

    Tooth Regeneration using tissue engineering concepts is a promising biological approach to solving problems of Tooth loss in elderly patients. The seeding cells, however, for Tooth Regeneration such as odontoblasts from dental germ, stem cells from dental pulp and deciduous teeth, and ectomesenchymal cells from the first branchial arch are difficult, even impossible to harvest in clinic. Bone marrow mesenchymal stem cells have odontogenic capacity, but their differentiation abilities significantly decrease with the increasing age of the donors. Therefore, the cells mentioned above are not practical in the clinical application of Tooth Regeneration in the old. Adipose derived stem cells have many clinical advantages over bone marrow mesenchymal stem cells, and their differentiation potential can be maintained with aging. Here we propose the hypothesis that adipose derived stem cells could be induced into odontogenic lineage and might be used as suitable seeding cells for Tooth Regeneration to replace the lost Tooth of elderly patients.

  • Odontogenic potential of bone marrow mesenchymal stem cells.
    Journal of oral and maxillofacial surgery : official journal of the American Association of Oral and Maxillofacial Surgeons, 2007
    Co-Authors: Ling Chen, Lei Liu, Yunfeng Lin, Weidong Tian
    Abstract:

    Purpose This study aimed to investigate the odontogenic potential of bone marrow mesenchymal stem cells (BM-MSCs) for seeding in Tooth Regeneration. Materials and Methods In this study, BM-MSCs were co-cultured with oral epithelial cells derived from rat embryos. Expression of the odontogenic genes Pax9 , DMP1 , and DSPP was detected by the reverse-transcription polymerase chain reaction (RT-PCR) technique. To further characterize the odontogenic potential of BM-MSCs, the gold standard in vivo transplantation system was used. Results The results revealed that Pax9 , DMP1 , and DSPP expression was detected by RT-PCR only after co-culture of BM-MSCs and oral epithelial cells derived from embryos age E11.5. Histological analyses of the BM-MSCs/epithelial cell mass demonstrated the presence of Tooth-like structures. Conclusions The series of experiments both in vitro and in vivo demonstrated that BM-MSCs can differentiate into functional odontoblast-like cells. This implies that BM-MSCs may become a novel source of cells for seeding in Tooth Regeneration research.

Yan Jin - One of the best experts on this subject based on the ideXlab platform.

  • Dental Stem Cells and Tooth Regeneration.
    Advances in experimental medicine and biology, 2018
    Co-Authors: Yi Shuai, Wenjia Liu, Tao Guo, Liqiang Zhang, Rui Yang, Yan Jin
    Abstract:

    Dental stem cells are a minor population of mesenchymal stem cells existing in specialized dental tissues, such as dental pulp, periodontium, apical papilla, dental follicle and so forth. Standard methods have been established to isolate and identify these stem cells. Due to their differentiation potential, these mesenchymal stem cells are promising for Tooth repair. Dental stem cells have been emerging to regenerated teeth and periodontal tissues, ascribe to their self-renewal, multipotency and tissue specific differentiation potential. Therefore, dental stem cells based Regeneration medicine highlights a promising access to repair damaged dental tissues or generate new teeth. In this review, we provide an overview of human dental stem cells including isolation and identification, involved pathways and outcomes of regenerative researches. A number of basic researches, preclinical studies and clinical trials have investigated that dental stem cells efficiently improve formation of dental specialized structure and healing of periodontal diseases, suggesting a great feasibility and prospect of these approaches in translational medicine of dental Regeneration.

  • Dental stem cell and dental tissue Regeneration
    Frontiers of medicine, 2018
    Co-Authors: Qiming Zhai, Zhiwei Dong, Wei Wang, Yan Jin
    Abstract:

    The teeth are highly differentiated chewing organs formed by the development of Tooth germ tissue located in the jaw and consist of the enamel, dentin, cementum, pulp, and periodontal tissue. Moreover, the teeth have a complicated regulatory mechanism, special histologic origin, diverse structure, and important function in mastication, articulation, and aesthetics. These characteristics, to a certain extent, greatly complicate the research in Tooth Regeneration. Recently, new ideas for Tooth and tissue Regeneration have begun to appear with rapid developments in the theories and technologies in tissue engineering. Numerous types of stem cells have been isolated from dental tissue, such as dental pulp stem cells (DPSCs), stem cells isolated from human pulp of exfoliated deciduous teeth (SHED), periodontal ligament stem cells (PDLSCs), stem cells from apical papilla (SCAPs), and dental follicle cells (DFCs). All these cells can regenerate the tissue of Tooth. This review outlines the cell types and strategies of stem cell therapy applied in Tooth Regeneration, in order to provide theoretical basis for clinical treatments.

  • Skin epithelial cells as possible substitutes for ameloblasts during Tooth Regeneration
    Journal of tissue engineering and regenerative medicine, 2012
    Co-Authors: Yihan Liu, Ming Jiang, Wei Hao, Wenjia Liu, Liang Tang, Hongchen Liu, Yan Jin
    Abstract:

    The disappearance of ameloblasts in erupted teeth hampers the implementation of tissue engineering-based Tooth Regeneration. We aimed at utilizing skin epithelial cells as the appropriate substitute for ameloblasts. The conversion potential of 1 day postnatal rat skin epithelial cells to ameloblasts was investigated under the induction of dental papillae mesenchymal cells (DPMCs). Induction strategies had been designed both in vitro and in vivo. Markers for ameloblasts had been detected in skin epithelial cells, which showed a columnar appearance with the nuclei located at one side, under indirect co-culture with DPMCs in vitro. An enamel–dentine-like and Tooth germ-like structure was formed by recombining skin epithelial pieces or cells with DPMCs after 14 days of implantation in rat renal capsule. Immunohistochemistry and cell labelling analysis further demonstrated that the enamel-forming cells were skin epithelium-derived. These results indicated that the skin epithelium-derived cells from postnatal rats have the potential to convert to functional ameloblasts under effective induction. Copyright © 2012 John Wiley & Sons, Ltd.