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Background: Post-tooth extraction socket preservation is necessary due to alveolar bone resorptive patterns through regenerative dentistry approaches that involve the use of stem cells, scaffold and growth factor. Stem cells derived from human exfoliated deciduous teeth (SHED) are known to potentially possess the osteogenic ability. Meanwhile, carbonate apatite scaffold (CAS) can act as a biocompatible scaffold capable of supporting mesenchymal stem cells (MSCs) to proliferate and differentiate optimally. The aim of this study is to investigate the expression of bone morphogenic protein-2 and 7 (BMP2, BMP7) and Matrix Metalloproteinase-8 (MMP-8) after the transplantation of SHED-incorporated CAS during in vivo bone remodeling. Material and Methods: A total of 14 healthy, male, Wistar rats, whose mandible anterior teeth were extracted by means of sterile needle holder clamps, constituted the subjects of this study of alveolar bone defects. Two research groups were created: a control group (CAS) as group I and an experimental group (CAS + SHED) as group II. SHED with a density of 10 6 cells were incorporated into CAS before being transplanted into the experimental group. After 7 days, all the animals were sacrificed and their mandible anterior region extracted. The BMP2, BMP7 and MMP-8 expression were subsequently analyzed by means of immunostaining. An unpaired t-test was conducted to analyze the treatment and control group (p<0.01) data. Results: The expression of BMP-2 and BMP-7 was higher in group II compared to group I. Meanwhile, the level of MMP-8 was lower in group II than group I. There was greater significant increased expression of BMP-2 and BMP-7 expression in Group II compared to Group I. There was significant decreased expression of MMP-8 between group II than group I (p<0.01). Conclusion: SHED-incorporated CAS can enhance BMP-2 and BMP-7 expression while attenuating MMP-8 expression during in vivo alveolar bone remodeling.

The precise predictions of the differentiation direction and potential of mesenchymal stromal cells (MSCs) are an important key to the success of regenerative medicine. The expression levels of fate-determining genes may provide tools for predicting differentiation potential. The expression levels of 95 candidate marker genes and glycosaminoglycan (GAG) contents after chondrogenic induction in 10 undifferentiated ilium and 5 jaw MSC cultures were determined, and their correlations were analyzed. The expression levels of eight genes before the induction of chondrogenic MSC differentiation were significantly correlated with the GAG levels after induction. Based on correlation patterns, the eight genes were classified into two groups: group 1 genes (AURKB, E2F1, CDKN2D, LIF, and ACLY), related to cell cycle regulation, and group 2 genes (CD74, EFEMP1, and TGM2), involved in chondrogenesis. The expression levels of the group 2 genes were significantly correlated with the ages of the cell donors. The expression levels of CDKN2D, CD74, and TGM2 were >10-fold higher in highly potent MSCs (ilium MSCs) than in MSCs with limited potential (jaw MSCs). Three-dimensional (3D) scatter plot analyses of the expression levels of these genes showed reduced variability between donors and confirmed predictive potential. These data suggest that group 2 genes are involved in age-dependent decreases in the chondrogenic differentiation potential of MSCs, and combined 3D analyses of the expression profiles of three genes, including two group 2 genes, were predictive of MSC differentiation potential.

Abstract. Dental pulp cells (DPCs) are a promising source of transplantable cells in regenerative medicine. However, DPCs have not been fully characterized at the molecular level. The aim of the present study was to distinguish DPCs from various source-derived mesenchymal stem cells (MSCs), fibroblasts (FBs) and other cells by the expression of several DPC-characteristic genes. DPCs were isolated from human pulp tissues by the explant method or the enzyme digestion method, and maintained with media containing 10% serum or 7.5% platelet-rich plasma. RNA was isolated from the cells and from dental pulp tissue specimens. The mRNA levels were determined by DNA microarray and quantitative polymerase chain reaction analyses. The msh homeobox 1, msh homeobox 2, T-box 2 and ectonucleoside triphosphate diphosphohydrolase 1 mRNA levels in DPCs were higher than that of the levels identified in the following cell types: MSCs derived from bone marrow, synovium and adipose tissue; and in cells such as FBs, osteoblasts, adipocytes and chondrocytes.The enhanced expression in DPCs was consistently observed irrespective of donor age, tooth type and culture medium. In addition, these genes were expressed at high levels in dental pulp tissue in vivo. In conclusion, this gene set may be useful in the identification and characterization of DPCs in basic studies and pulp cell-based regeneration therapy.

Background: Stem cells from human exfoliated deciduous teeth (SHED) seeded in carbonate apatite scaffold (CAS) may have multiple functions that could be used to regenerate the alveolar bone defects. The purpose of this study is to examine the ability of SHED and CAS in alveolar bone defects using an immunohistochemical analysis. Methods: ten three-month-old healthy male Wistar rats (R. novergicus) that weighed between 150–250 grams (g) were used as animal models. A simple blind random sampling method was used to select the sample that was assigned to the study group for CAS and SHED seeded in CAS (n=5). The animal study model of the alveolar bone was established by extracting the anterior mandible teeth. Rodent anesthesia was applied to relieve the pain during the procedure for all test animals. Immunohistochemistry was performed after seven days to facilitate the examination of the receptor activator of NF-κβ ligand (RANKL), osteoprotegrin (OPG), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin, and osteopontin expression. The data was analyzed using the unpaired t-test (p<0.01) and Pearson’s correlation test (p<0.05). Results: The OPG, RUNX2, TGF-β, VEGF, ALP, osteocalcin, and ostepontin expressions were higher in SHED seeded in CAS than CAS only with a significant difference between the groups (p<0.01). Furthermore, the RANKL expression was lower in SHED seeded in CAS compared to CAS only. There was a strong reverse significant correlation between OPG and RANKL expression (p<0.05). Conclusions: The number of osteogenic marker expressing cells, such as OPG, RUNX2, TGF-β, VEGF, ALP, osteocalcin, and ostepontin, increased. However, RANKL expression in the alveolar bone defects that were implanted with SHED seeded in CAS did not increase after seven days.

Background. Autism is a neurological and developmental disorder. Children with autism have problems related to physical, psychological, and mental barriers that can hinder their ability to achieve optimal dental health status. Maintaining the dental health of children with autism is influenced by parents' teaching skills and habits. From previous study, there were about 17.4% children with autism in Saudi Arabia suffering from bleeding of the gingiva. Periodontal disease is often found in children with autism. Objective. This study analyzes the relationship between mother’s behavior with periodontal status and periodontal treatment needs of children with autism. Methods. Analytical observational study with cross-sectional approach at AGCA Centre Surabaya with a total sample of 34 pairs of children with autism and their mothers. This study used the HU-DBI questionnaire which consisted of knowledge, attitude, and mother’s action and oral examination of children with autism with the CPITN index. Results. Of the children with autism, 55.8% had healthy periodontal status. The knowledge, attitudes, and actions of mothers were high. Statistical results with Spearmen correlation test obtained a value of p>0.05 on aspects of knowledge, attitudes, and actions towards the CPITN index and periodontal treatment needs. Conclusion. There was no significant correlation between the mother’s behavior and the periodontal status and periodontal treatment needs of children with autism in managing their oral health.

Background: Stem cells from human exfoliated deciduous teeth (SHED) seeded in carbonate apatite scaffold (CAS) may have multiple functions that could be used to regenerate the alveolar bone defects. The purpose of this study is to examine the ability of SHED and CAS in alveolar bone defects using an immunohistochemical analysis. Methods: ten three-month-old healthy male Wistar rats (R. novergicus) that weighed between 150–250 grams (g) were used as animal models. A simple blind random sampling method was used to select the sample that was assigned to the study group for CAS and SHED seeded in CAS (n=5). The animal study model of the alveolar bone was established by extracting the anterior mandible teeth. Rodent anesthesia was applied to relieve the pain during the procedure for all test animals. Immunohistochemistry was performed after seven days to facilitate the examination of the receptor activator of NF-κβ ligand (RANKL), osteoprotegrin (OPG), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin, and osteopontin expression. The data was analyzed using the unpaired t-test (p<0.01) and Pearson’s correlation test (p<0.05). Results: The OPG, RUNX2, TGF-β, VEGF, ALP, osteocalcin, and ostepontin expressions were higher in SHED seeded in CAS than CAS only with a significant difference between the groups (p<0.01). Furthermore, the RANKL expression was lower in SHED seeded in CAS compared to CAS only. There was a strong reverse significant correlation between OPG and RANKL expression (p<0.05). Conclusions: The number of osteogenic marker expressing cells, such as OPG, RUNX2, TGF-β, VEGF, ALP, osteocalcin, and ostepontin, increased. However, RANKL expression in the alveolar bone defects that were implanted with SHED seeded in CAS did not increase after seven days.

Background Periodontitis progression is characterized by alveolar bone loss, and its prevention is a major clinical problem in periodontal disease management. Matrix metalloproteinase-8 (MMP-8) has been shown to adequately monitor the treatment of chronic periodontitis patients as gingival crevicular fluid MMP-8s were positively associated with the severity of periodontal disease. Moreover, modulating the vascular endothelial growth factor (VEGF) levels in bones could be a good way to improve bone regeneration and cure periodontitis as VEGF promotes endothelial cell proliferation, proteolytic enzyme release, chemotaxis, and migration; all of which are required for angiogenesis. Purpose The aim of this study was to determine the effect of hydroxyapatite incorporated with stem cells from exfoliated deciduous teeth (SHED) in Wistar rats’ initial alveolar bone remodeling based on the findings of MMP-8 and VEGF expressions. Methods A hydroxyapatite scaffold (HAS) in conjunction with SHED was transplanted into animal models with alveolar mandibular defects. A total of 10 Wistar rats ( Rattus norvegicus ) were divided into two groups: HAS and HAS + SHED. Immunohistochemistry staining was performed after 7 days to facilitate the examination of MMP-8 and VEGF expressions. Results The independent t -test found significant downregulation of MMP-8 and upregulation VEGF expressions in groups transplanted with HAS in conjunction with SHED compared with the HAS group ( p < 0.05). Conclusion The combination of SHED with HAS on alveolar bone defects may contribute to initial alveolar bone remodeling as evident through the assessments of MMP-8 and VEGF expressions.