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1.
J Periodontal Res ; 57(1): 162-172, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34826339

ABSTRACT

BACKGROUND AND OBJECTIVE: Exosomes are small vesicles secreted from many cell types. Their biological effects largely depend on their cellular origin and the physiological state of the originating cells. Exosomes secreted by mesenchymal stem cells exert therapeutic effects against multiple diseases and may serve as potential alternatives to stem cell therapies. We previously established and characterized human leukocyte antigen (HLA) haplotype homo (HHH) dental pulp cell (DPC) lines from human wisdom teeth. In this study, we aimed to investigate the effect of local administration of HHH-DPC exosomes in a mouse model of periodontitis. METHODS: Exosomes purified from HHH-DPCs were subjected to particle size analysis, and expression of exosome markers was confirmed by western blotting. We also confirmed the effect of exosomes on the migration of both HHH-DPCs and mouse osteoblastic MC3T3-E1 cells. A mouse experimental periodontitis model was used to evaluate the effect of exosomes in vivo. The morphology of alveolar bone was assessed by micro-computed tomography (µCT) and histological analysis. The effect of exosomes on osteoclastogenesis was evaluated using a co-culture system. RESULTS: The exosomes purified from HHH-DPCs were homogeneous and had a spherical membrane structure. HHH-DPC exosomes promoted the migration of both human DPCs and mouse osteoblastic cells. The MTT assay showed a positive effect on the proliferation of human DPCs, but not on mouse osteoblastic cells. Treatment with HHH-DPC exosomes did not alter the differentiation of osteoblastic cells. Imaging with µCT revealed that the exosomes suppressed alveolar bone resorption in the mouse model of periodontitis. Although no change was apparent in the dominance of TRAP-positive osteoclast-like cells in decalcified tissue sections upon exosome treatment, HHH-DPC exosomes significantly suppressed osteoclast formation in vitro. CONCLUSIONS: HHH-DPC exosomes stimulated the migration of human DPCs and mouse osteoblastic cells and effectively attenuated bone loss due to periodontitis.


Subject(s)
Alveolar Bone Loss , Exosomes , Periodontitis , Alveolar Bone Loss/diagnostic imaging , Alveolar Bone Loss/therapy , Animals , Cell Differentiation , Dental Pulp , Mice , Periodontitis/therapy , X-Ray Microtomography
2.
Sci Rep ; 7(1): 13500, 2017 10 18.
Article in English | MEDLINE | ID: mdl-29044129

ABSTRACT

Human dental pulp cells (DPCs), adherent cells derived from dental pulp tissues, are potential tools for cell transplantation therapy. However, little work has been done to optimize such transplantation. In this study, DPCs were treated with fibroblast growth factor-2 (FGF2) for 5-6 consecutive serial passages and were transplanted into the injury site immediately after complete transection of the rat spinal cord. FGF2 priming facilitated the DPCs to promote axonal regeneration and to improve locomotor function in the rat with spinal cord injury (SCI). Additional analyses revealed that FGF2 priming protected cultured DPCs from hydrogen-peroxide-induced cell death and increased the number of DPCs in the SCI rat spinal cord even 7 weeks after transplantation. The production of major neurotrophic factors was equivalent in FGF2-treated and untreated DPCs. These observations suggest that FGF2 priming might protect DPCs from the post-trauma microenvironment in which DPCs infiltrate and resident immune cells generate cytotoxic reactive oxygen species. Surviving DPCs could increase the availability of neurotrophic factors in the lesion site, thereby promoting axonal regeneration and locomotor function recovery.


Subject(s)
Dental Pulp/cytology , Fibroblast Growth Factor 2/pharmacology , Mesenchymal Stem Cell Transplantation/methods , Nerve Regeneration , Spinal Cord Injuries/therapy , Animals , Axon Guidance , Cells, Cultured , Female , Humans , Locomotion , Mesenchymal Stem Cells/drug effects , Rats , Rats, Wistar
3.
PLoS One ; 9(12): e115392, 2014.
Article in English | MEDLINE | ID: mdl-25521610

ABSTRACT

Human dental pulp cells (hDPCs) are a promising resource for regenerative medicine and tissue engineering and can be used for derivation of induced pluripotent stem cells (iPSCs). However, current protocols use reagents of animal origin (mainly fetal bovine serum, FBS) that carry the potential risk of infectious diseases and unwanted immunogenicity. Here, we report a chemically defined protocol to isolate and maintain the growth and differentiation potential of hDPCs. hDPCs cultured under these conditions showed significantly less primary colony formation than those with FBS. Cell culture under stringently defined conditions revealed a donor-dependent growth capacity; however, once established, the differentiation capabilities of the hDPCs were comparable to those observed with FBS. DNA array analyses indicated that the culture conditions robustly altered hDPC gene expression patterns but, more importantly, had little effect on neither pluripotent gene expression nor the efficiency of iPSC induction. The chemically defined culture conditions described herein are not perfect serum replacements, but can be used for the safe establishment of iPSCs and will find utility in applications for cell-based regenerative medicine.


Subject(s)
Dental Pulp/cytology , Induced Pluripotent Stem Cells/cytology , Adolescent , Cell Differentiation , Cells, Cultured , Humans , Primary Cell Culture/methods
4.
Arch Oral Biol ; 55(9): 648-54, 2010 Sep.
Article in English | MEDLINE | ID: mdl-20630496

ABSTRACT

The hypoxia condition was expected to be suitable for the establishment and maintenance of human dental pulp cells (hDPCs), because they reside in a low-oxygen environment in vivo. Therefore, we presently examined the effects of hypoxia on the proliferation and differentiation of hDPCs in vitro. hDPCs grown under 3% O(2) showed a significantly higher proliferation rate than those under 21% O(2). Then, we prepared hypoxic cultures of hDPCs from older patients' teeth having inflammation and succeeded in recovering and expanding a small number of hDPCs. These cells were confirmed to have capability for osteo/odontogenic differentiation. Hypoxia suppressed the osteo/odontogenic differentiation of hDPCs in vitro and increased the number of cells expressing STRO-1, an early mesenchymal stem cell marker. This simple method will increase the possibility to obtain living hDPCs from damaged and/or aged tissues, from which it is ordinarily difficult to isolate living stem cells with differentiation capability.


Subject(s)
Adult Stem Cells/cytology , Dental Pulp/cytology , Epithelial Cells/cytology , Hypoxia/pathology , Mesenchymal Stem Cells/cytology , Adolescent , Adult Stem Cells/metabolism , Aged , Antigens, Surface/biosynthesis , Cell Culture Techniques , Cell Differentiation , Cell Proliferation , Cells, Cultured , Dental Pulp/metabolism , Epithelial Cells/metabolism , Extracellular Matrix Proteins/biosynthesis , Flow Cytometry , Humans , Hypoxia/metabolism , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/metabolism , Middle Aged , Osteocalcin/biosynthesis , Phosphoproteins/biosynthesis , Polymerase Chain Reaction , Sialoglycoproteins/biosynthesis
5.
J Oral Pathol Med ; 39(5): 407-19, 2010 May.
Article in English | MEDLINE | ID: mdl-20202092

ABSTRACT

BACKGROUND: Recently, oral sensory complaints (OSC) were proposed as a disease entity to represent idiopathic sensory disturbances of dry mouth, burning mouth, and taste disturbance, even though neither the status of OSC in the general population nor its underlying mechanism has yet been elucidated. Moreover, these three OSC-related complaints have not been assessed in combination by means of a visual analog scale (VAS) in a large-scale, community-dwelling population of a broad age range. METHODS: In a 1188-member community-dwelling adult population, comprised of 373 males and 815 females, aged 20-90 years, the three OSC-related complaints and stimulated salivary flow rate (SSFR) were assessed by means of a VAS and modified Saxon test, respectively. Association of each complaint with age, gender, SSFR, and other complaints was analyzed. RESULTS: Increases in both prevalence and intensity of subjective dry mouth and burning mouth were associated closely with decreasing SSFR. Even for taste disturbance, which may be affected less significantly by salivation status than the other two complaints, a significant association was suggested between decreasing SSFR and especially severe taste disturbance. However, these oral complaints were found in considerable prevalence even in the individuals with high SSFR. Often overlapping presentation of these complaints and a close association in intensity between the complaints to each other were also found. CONCLUSIONS: Hyposalivation may be a significant and common etiology for the three oral complaints, although the considerable prevalence of complaints without hyposalivation suggests other etiologies, including those related to the OSC.


Subject(s)
Burning Mouth Syndrome/etiology , Saliva/metabolism , Taste Disorders/etiology , Xerostomia/etiology , Adult , Age Factors , Aged , Aged, 80 and over , Analysis of Variance , Chi-Square Distribution , Female , Humans , Japan , Male , Middle Aged , Pain Measurement , Physical Stimulation , Regression Analysis , Secretory Rate , Sex Factors , Surveys and Questionnaires , Xerostomia/complications , Young Adult
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