• International Journal of Oral Biology
• /
• v.36 no.2
• /
• pp.51-57
• /
• 2011

Runx2 and Osterix, the transcription factors for osteoblast differentiation, are known as fundamental factors to regulate the development of calcified tissues. However, the biological functions of these factors in the development of the periodontal tissues remain unclear. In this study, we investigated the distribution of Runx2 and Osterix during periodontal tissue development of the mice. Mandibles from 14-day-old mice were prepared for paraffin section. Serial sections of the mandible containing $1^{st}$ molar tooth germs were obtained as a thickness of $7\;{\mu}m$. Some sections were stained with hematoxylin and eosin. Others were used for immunohistochemistry for PCNA, Runx2, and Osterix. Epithelial cells in growing end of Hertwig's epithelial root sheath (HERS) and mesenchymal cells adjacent to the growing end of HERS expressed PCNA. Undifferentiated mesenchymal cells and hard tissue forming cells like cementoblasts and osteoblasts in early stage of differentiation expressed Runx2. Fully differentiated cementoblasts and osteoblasts secreting matrix proteins expressed Osterix. However, the cells terminated the matrix formation did not express Osterix. Periodontal ligament cells expressed Runx2 and Osterix. Pulp cells expressed Runx2 only. These results suggest that Runx2 and Osterix might regulate the differentiation of cementoblasts in the same manner as osteoblasts. Runx2 might participate in the process of cementoblast differentiation in early stage, whether Osterix might regulate the maturation and matrix synthesis of the cells.

• The Journal of the Korean dental association
• /
• v.48 no.1
• /
• pp.56-62
• /
• 2010

In recent years, a number of special treatment procedures have been introduced to reestablish new tooth supporting tissues with varying degrees of success including guided tissue regeneration(GTR), bone grafting(BG) and the use of enamel matrix derivative(EMD). EMD is an extract of enamel matrix and contains amelogenins of various molecular weights. Emdogain(EMD) might have some advantages over other methods of regenerating the tissue supporting teeth lost by gum disease, such as less postoperative complications. Emdogain contains proteins(derived from developing pig teeth) believed to regenerate tooth attachment. The decrease in probing depth after EMD treatment is achieved primarily by clinical attachment gain and bone regeneration and only to a minor extent by gingival recession. In conclsion, EMD seems to be safe, was able to regenerate lost periodontal tissues in previously diseased sites based on clinical parameters.

• Journal of Periodontal and Implant Science
• /
• v.32 no.1
• /
• pp.1-12
• /
• 2002

The periodontal ligament(PDL) is a unique tissue that is crucial for tooth function. However, little is known of the molecular mechanisms controlling PDL function. PDL-specific protein;PDLs22 had been previously identified as a novel protein isolated from cultured human PDL fibroblasts using subtraction hybridization between human gingival fibroblasts and PDL fibroblasts. The aim of this study was to examine the expression pattern and tissue localization of PDLs22 protein in embryonic and various postnatal stages of developing mouse using immunohistochemical staining. Embryos (E18) and postnatal (P1, P4, P5, P15, P18) were decapitated and the heads were fixed overnight in a freshly prepared solution of 4% paraformaldehyde. Some specimens were decalcified for $2{\sim}4$ weeks in a solution containing 10% of the disodium salt of ethylenediamine-tetraacetic acid (EDTA). Next, tissues were dehydrated, embedded in paraffin and sectioned serially at $6{\mu}m$ in thickness. Polyclonal antiserum raised against PDLs22 peptides, ISNKYLVKRQSRD, were made. The localization of PDLs22 in tissues was detected by polyclonal antibody against PDLs22 by means of immunohistochemical staining. The results were as follows; 1. Expression of PDLs22 protein was not detected in the tooth germ of bud and cap stage. 2. At the late bell stage and root formation stage, strong expression of PDLs22 protein was observed in developing tooth follicle, osteoblast-like cells, and subodontoblastic cells in the tooth pulp, but not in gingival fibroblasts, ameloblasts and odontoblasts of tooth germ 3. In erupted tooth, PDLs22 protein was intensely expressed in PDL and osteoblast-like cells of alveolar bone, but not in gingival fibroblasts, mature osteocytes and adjacent salivary glands. 4. In the developing alveolar bone and mid-palatal suture, expression of PDLs22 protein was seen in undifferentiated mesenchymal cells and osteoblast-like cells of developing mid-palatal suture, but not in mature osteocytes and chondrocytes. These results suggest that PDLs22 protein may play an important role in the differentiation of undifferentiated mesenchymal cells in the bone marrow and PDL cells, which can differentiate into multiple cell types including osteoblasts, cementoblasts, and PDL fibroblasts. However, more researches should be performed to gain a better understanding of the exact function of PDLs22 protein which related to the PDL cell differentiation.

• Journal of Periodontal and Implant Science
• /
• v.51 no.5
• /
• pp.329-341
• /
• 2021

Purpose: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

• Proceedings of the KACD Conference
• /
• 2001.05a
• /
• pp.247-251
• /
• 2001

;A dental developmental anomaly is defined as an isolated aberration in tooth form, caused by a disturbance or abnormality which occurred during tooth development. There are numerous types of dental anomalies, and a considerable variation in the extent of the defects occurs with each type. Teeth with these anomalies pose unique challenges. Since the defects are not always apparent clinically, they can confuse diagnosticians investigating the etiology of pulpal pathosis. When endodontic treatment is required, the defects often hinder access cavity preparation and canal instrumentation. Treatment planning also becomes more challenging, since the defects can create complicated periodontal problems, and the malformed teeth can be difficult to restore, particularly those weakened by endodontic therapy. Fusion is defined as the joining of two developing tooth germs resulting in a single large tooth structure. The incidence of fusion is < 1% in the Caucasian population, and it is believed that physical force or pressure produces contact of the developing teeth. Clinically and radiographically, a fused tooth usually appears as one large crown with at least partially separated roots and root canals. There may be a vertical groove in the tooth crown delineating the originally separate crowns. Dens invaginatus is a deep surface invagination of the crown or root that is lined by enamel. Teeth in both maxillary and mandibular arches may be affected, but the permanent maxillary lateral incisor is the tooth most commonly involved. Studies have revealed an incidence ranging from 0.25% to as high as 10%. The invagination ranges from a slight pitting to an anomaly occupying most of the crown and root. The invagination frequently communicates with the oral cavity, allowing the entry of irritants and microorganism either directly into pulpal tissues or into an area that is deparated from pulpal tissues by only a thin layer of enamel and dentin. This continuous ingress of irritants and the subsequent inflammation usually lead to necrosis of the adjacent pulp tissue and then to periapical or periodontal abscesses. If the invagination extends from the crown to the periradicular tissue and has no communication with the root canal system, the pulp may remain vital. Recommended treatment of fused tooth and dens invaginatus has been reported in the endodontic literature. This case report describes the endodontic treatment of a maxillary laterl incisors having fused crown and dens invaginatus.natus.

• Journal of Society of Preventive Korean Medicine
• /
• v.18 no.1
• /
• pp.125-138
• /
• 2014

Objective : Polycan, exopolymers purified from Aureobasidium pullulans SM-2001 and calcium gluconate have been showed favorable inhibitory effects on the periodontitis and related alveolar bone losses through antioxidant and anti-inflammatory activities, respectively. In the present study, we intended to observe the possible synergic effects of mixed formula consisted of Polycan and calcium gluconate on ligation-induced experimental periodontitis and related alveolar bone losses in rats, and to select the fittest compositions for further developing as effective agents to ameliorate periodontal diseases. Method : Experiments were conducted as two separated two tests - first is synergic effects of Polycan and calcium gluconate 1:1, 1:9 and 9:1 mixtures, and second is 1:99, 2:98, 4:96, 8:92 and 1:9 mixtures. Experimental periodontal diseases were induced by ligature placed around the cervix of upper left incisior teeth of rats. One day after ligation placements, 200mg/kg of each single or mixed formulas of Polycan or/and calcium gluconate were orally administered for 10 days. The changes on the alveolar bone loss index and maxillary bone mineral density (BMD) were observed for detecting alveolar bone losses, and for anti-inflammatory effects, myeloperoxidase (MPO) activities and proinflammatory cytokine (tumor necrosis factor; TNF-${\alpha}$) contents were also evaluated in gingival tissues around ligature placed incisior teeth. The results of mixtures were compared with those of singe Polycan and calcium gluconate treated rat. Results : Each single or mixed formulas of Polycan or/and calcium gluconate favorably and significantly inhibited the inflammatory changes. The inhibitory effects of mixed formula consisted of Polycan and calcium gluconate 1:9 showed against periodontitis and related alveolar bone losses as compared with those of each Polycan and calcium gluconate single formula (p<0.05). In second experiment, Polycan and calcium gluconate 2:98, 4:96, 8:92 and 1:9 mixed formulas also showed significant increased anti-inflammatory and inhibitory effects against alveolar bone losses as compared with those of each single formula. Among them, Polycan and calcium gluconate 2:98 showed the highest efficacy against to ligation-induced experimental periodontitis and related alveolar bone losses. Conclusion : The results obtained in this study suggest that appropriated mixtures of Polycan and calcium gluconate showed synergic inhibitory effects against ligation-induced experimental periodontitis and related alveolar bone losses in rats. Moreover, Polycan and calcium gluconate 2:98 showed the highest efficacies in this experiment, suggesting the fittest composition for further developing as effective agents to ameliorate periodontal diseases.

• Journal of Periodontal and Implant Science
• /
• v.31 no.4
• /
• pp.787-801
• /
• 2001

The molecular mechanisms control the function of PDL(periodonta1 ligament) cells and/or fibroblasts remain unclear. PDLsl7, PDL-specific gene, had previousely　identified the cDNA for a novel protein from cultured　PDL fibroblasts using subtraction hybridization between gingival fibroblasts and　PDL fibroblasts. The purpose　of this study was to determine the regulation by growth factors and cytokines on　PDLsl7 gene expression in　cultured human periodontal ligament cells and observe the immunohistochemical　localization of PDLsl7 protein in various tissues of mouse. Primary PDL fibroblasts isolated by scraping the root of the extracted human mandibular third molars. The cells were incubated with various concentration of human recombinant $IL-1{\beta}$, PDGF-BB and TGF\;${\beta}$ for 48h nd 2 weeks. At each time point total RNA was extracted and the levels of transcription ere assessed by reverse transcription-polymerase chain reaction (RT-PCR assay). polyclonal antiserum raised against PDLsl7 peptides, CLSVSYNRSYQINE and SEAVHETDLHDGC, were made, and stained the tooth, periodontium, developing bone, bone marrow and mid-palatal suture of the mouse. The results were as follows. 1. PDLsl7 mRNA levels were increased in response to PDGF (10ng/ml) and $TGF\;{\beta}$(20ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF{\beta}$for 48 h. 2. PDLsl7 was up-regulated only by $TGF{\beta}$(20 ng/ml) after treatment of the $IL-1{\beta}$, PDGF-BB and $TGF\;{\beta}$ for 2 weeks and unchanged by the other stimulants. 3. PDLsl7 was a novel protein coding the 142 amino acid peptides in the ORF and the nucleotide sequences of the obtained cDNA from RT-PCR was exactly same as the nucleotides of the database. 4. Immunohistochemical analysis showed that PDLsl7 is preferentially expressed in the PDL, differentiating osteoblast-like cells and stromal cells of the bone marrow in the adult mouse. 5. The expression of PDLsl7 protein was barely detectable in gingival fibroblasts, hematopoetic cells of the bone marrow and mature osteocytes of the alveolar bone. These results suggest that PDLsl7 might upregulated by PDGF-BB or $TGF{\beta}$ and acts at the initial stage of differentiation when the undifferentiated mesenchymal cells in the bone marrow and PDL differentiate into multiple cell types. However, more research needs to be performed to gain a better understanding of the exact function of PDLsl7 during the differentiation of bone marrow mesenchymal and PDL cells.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.27 no.4
• /
• pp.449-454
• /
• 2011

Recently, jawbone osteonecrosis has been largely reported as a potential adverse effect of bisphosphonate (BP)administration. Currently available published incidence data for BRONJ are based on retrospective studies and estimates of cumulative incidence range from 0.8 to 12%. The mandible is more commonly affected than the maxilla (2:1 ratio), and 60-70% of cases are preceded by a dental surgical procedure. The signs and symptoms that may occur before the appearance of clinical evident osteonecrosis include changes in the health of periodontal tissues, non-healing mucosal ulcers, loose teeth and unexplained soft-tissue infection. Tooth extraction as a precipitating event is a common observation. The significant benefits that bisphosphonates offer to patients clearly surpass the risk of potential side effects; however, any patient for whom prolonged bisphosphonate therapy is indicated, should be provided with preventive dental care in order to minimize the risk of developing this severe condition.