• Title/Summary/Keyword: oral culture

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Culture of Rat Salivary Acinar Cells (백서 타액선 선포 세포의 배양)

  • Lee, Sung-Woo;Han, Song;Kho, Hong-Seop
    • Journal of Oral Medicine and Pain
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    • v.24 no.2
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    • pp.163-169
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    • 1999
  • We investigated the culture condition and effects of various growth factors on the culture of salivary gland acinar cells. Male, Sprague-Dawley rats (about 6 weeks old) were sacrificed and their submandibular, sublingual, and parotid glands were used as specimens. High oxygen level more than 90% and coating of Matrigel on culture dish were important factors to help increase the survival time of acinar cells, Proper concentration of enzymes such as collagenase and hyaluronidase during isolation steps was also important. Addition of various growth factors such as dexamethasone, insulin, transferrin, selenous acid, reduced glutathione, epidermal growth factor, isoproterenol, and putrescine in culture medium helped to increase lifetime of cultured salivary acinar cells.

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TISSUE-ENGINEERED RECONSTITUTION OF ORAL MUCOSA USING POLYDIOXANONE MESH (Polydioxanone mesh를 이용한 구강점막의 조직 공학적 재건)

  • Moon, Seon-Jae;Joo, So-Yeon;Kim, Jin;Kim, Hak-Yong;Park, Jung-Keug;Cha, In-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.4
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    • pp.249-256
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    • 2003
  • The lack of sufficient oral mucosa available for intra-oral reconstruction has been dealt with by the use of skin or oral mucosa grafts harvested from donor sites but grafts requires more than one surgical procedures and could cause donor site morbidity. Many investigators have attempted to increase available soft tissue by tissue engineered skin or oral mucosa replacements for clinical applications. But, reconstructed mucosa by several methods have low physical properties such as rolling and contraction. The aims of this study were to develope an in vitro experimental model that maintains an epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally cultured oral mucosa embedded with Polydioxanone mesh by histological and immunohistochemical analysis. The results were as follows; 1. Oral mucosa reconstructed by three-dimensional organotypic culture revealed similar morphologic characteristics to equvalent normal oral mucosa in the point that they show stratification and differentiation. 2. The expression of cytokeratin 10/13 and involucrin in the cultured tissue showed the same pattern with normal oral mucosa suggesting that organotypic co-culture condition is able to induce cellular differentiation. 3. After insertion of polydioxanone mesh, increased tensile strength were observed. These results suggest that three-dimensional organotypic co-culture of the oral mucosa cell lines with the dermal equvalent consisting type I collagen and fibroblasts reproduce the morphologic and immunohistochemical characteristics similar to those in vivo condition. And increased physical properties by use of polydioxanone mesh will helpful for clinical applications.

THE MOST APPROPRIATE ANTIMITOTIC TREATMENT OF ARA-C IN SCHWANN CELL-ENRICHED CULTURE FROM DORSAL ROOT GANGLIA OF NEW BORN RAT (신생 백서 척수후근절의 슈반세포 배양을 위한 Ara-C 분열억제제의 최적 효과에 대한 연구)

  • Kim, Soung-Min;Lee, Jong-Ho;Ahn, Kang-Min;Kim, Nam-Yeol;Sung, Mi-Ae;Hwang, Soon-Jeong;Kim, Ji-Hyuck;Jahng, Jeong-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.2
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    • pp.100-107
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    • 2004
  • Schwann cell, one of important components of peripheral nervous system, interact with neurons to mutually support the growth and replication of embryonal nerves and to maintain the different functions of adult nerves. The Ara-C, known as an antimitotic agent, have been used to have high effectiveness in eliminating fibroblasts during Schwann cell culture period. This enrichment effect is also known to be cummulative with each successive pulse of Ara-C applied and is due to a progressive loss of fibroblasts. But the cytotoxicity by Ara-C is also cummulative and noticeable over the period. To determine the most effective application time and interval of Ara-C in the Schwann cell culture, we observed the Schwann cell purity and density with the Ara-C treatment in plain and three-dimensional culture from dorsal root ganglion of new born rat. By culturing dispersed dorsal root ganglia, we can repeatedly generate homogenous Schwann cells, and cellular morphology and cell count with mean percentages were evaluated in the plain culture dishes and in the immunostainings of S-100 and GFAP in the three-dimensional culture. The Ara-C treated cultures showed a higher Schwann cell percentage ($31.0%{\pm}8.09%$ in P4 group to $65.5%{\pm}24.08%$ in P2 group), compared with that obtained in the abscence of Ara-C ($17.6%{\pm}6.03%$) in the plain culture after 2 weeks. And in the three-dimensional culture, S-100 positive cells increased to $56.22%{\pm}0.67%$ and GFAP positive cells to $66.46%{\pm}1.83%$ in G2 group (p<0.05), higher yield than other groups with Ara-C application. Therefore, we concluded that the Ara-C treatment is effective for the proliferation of Schwann cells contrast to the fibroblasts in vitro culture, and the first application after 24 hours from cell harvesting and subsequent 2 pulse treatment (P2 group in plain culture and G2 group in three-dimensional culture) was more effective than other application protocols.

The effects of organizational culture and self-leadership on organizational effectiveness in oral health professional (치과의료기관 종사자의 조직문화와 셀프리더십 유형이 조직유효성에 미치는 영향)

  • Jang, Jong-Hwa;Lee, Young-Soo;Moon, Ae-Eun
    • Journal of Korean society of Dental Hygiene
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    • v.11 no.3
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    • pp.395-404
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    • 2011
  • Objectives : This study was to investigate the level of organizational culture and self-leadership on organizational effectiveness and the significant factors influencing organizational effectiveness in oral health professional. Methods : Data were obtained from a cross-sectional survey of 297 oral health professional(mean age=$29.95{\pm}8.10$). Participants in the study were recruited when they attended the dental health form in Gwangju. Organizational culture was classified into four factors: relation-oriented, innovation-oriented, hierarchy-oriented, task-oriented culture. Self-readership consisted of five factors: rehearsal/self-observation, self-goal establishment, self-esteem, self-criticism, self-reward. Organizational effectiveness included organizational commitment and job satisfaction. The data was analysed using t-test, one-way ANOVA, Pearson correlation and multiple regression. Results : The dominant organizational culture was rank-oriented culture. The mean score of self-readership score was 3.51 out of a maximum 5 points. Organizational commitment was positively correlated with age, relation-oriented culture, self-goal establishment, self-reward explained 41.9% and job satisfaction was positively correlated with relation-oriented culture and self-esteem explained 48.6%. Conclusions : Based on the findings, the organizational culture and self-readership was correlated with organizational effectiveness. These results suggest that further development should be continued to develop the effective self-readership and organizational culture to improve the organizational effectiveness of oral healthcare professional.

Differentiation of CD31-Positive Vascular Endothelial Cells from Organoid Culture of Dental Pulp Stem Cells

  • Seo, Eun Jin;Park, Jae Kyung;Jeong, Hoim;Kang, Jung Sook;Kim, Hyung-Ryong;Jang, Il Ho
    • International Journal of Oral Biology
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    • v.43 no.2
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    • pp.77-82
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    • 2018
  • The mesenchymal stem cells (MSCs) that reside in dental tissues hold a great potential for future applications in regenerative dentistry. In this study, we used human dental pulp cells, isolated from the molars (DPCs), in order to establish the organoid culture. DPCs were established after growing pulp cells in an MSC expansion media (MSC-EM). DPCs were subjected to organoid growth media (OGM) in comparison with human dental pulp stem cells (DPSCs). Inside the extracellular matrix in the OGM, the DPCs and DPSCs readily formed vessel-like structures, which were not observed in the MSC-EM. Immunocytochemistry analysis and flow cytometry analysis showed the elevated expression of CD31 in the DPCs and DPSCs cultured in the OGM. These results suggest endothelial cell-prone differentiation of the DPCs and DPSCs in organoid culture condition.

ROLE OF FIBROBLASTS IN ORGANOTYPIC CULTURES OF IMMORTALIZED HUMAN ORAL KERATINOCYTES (섬유모세포의 종류에 따른 불멸화된 구강 각화세포의 삼차원적 배양에 관한 연구)

  • Cheong, Jeong-Kwon;Yoon, Kyu-Ho;Kim, Eun-Cheol
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.3
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    • pp.211-220
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    • 2007
  • Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.

BACTERIOLOGIC FEATURES AND ANTIBIOTIC SUSCEPTIBILITY IN ISOLATES FROM ORAL AND MAXILLOFACIAL INFECTIONS (구강악안면 감염 질환에서 배양된 세균의 양상과 항생제 감수성)

  • Kim, Sun-Kook;Kook, Min-Suk;Han, Chang-Hun;Ryu, Sun-Youl
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.4
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    • pp.322-328
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    • 2005
  • Oral and maxillofacial infections are most commonly odontogenic in origin. The present study was implemented for patients with oral and maxillofacial infections in order to determine what differences were present in cultured bacteria, depending upon the different types of infection. For the present study, the epidemiological characteristics, the state of infection, and the results of the pus culture and antibiotic susceptibility tests were analyzed for the 159 cases where pus culture tests were performed. The patients were treated at the Oral and Maxillofacial Surgical Department of Chonnam National University Hospital during an 18-months period from March 2003 to August 2004. Among the total 159 pus culture specimens, bacteria were cultured in 111 cases (69.8%). In the 111 pus culture specimens, Streptococcus species, Neisseria species, and Staphylococcus species were cultured from 69 cases (51.1%), 21 cases (15.6%), and 15 cases (11.1%), respectively and were determined to be bacterial strains the predominant bacteria responsible for oral and maxillofacial infectious diseases. Twenty four cases (15.1%) among the 159 specimens showed mixed infections. The mostly isolated bacteria from each of the space abscess, dentoalveolar abscess, inflammatory cyst, and pericoronitis cases were the Viridans streptococci. There was little relevance between the type of infection and the type of cultured bacteria. Antibiotic susceptibility tests showed a high level of susceptibility to teicoplanin(100%), vancomycin(100%), chloramphenicol(96.4%), ofloxacin(88.3%), imipenem(83.3%), erythromycin(82.5%) and a low susceptibility to cefazolin(40.0%), oxacillin(44.7%), ampicillin(49.4%), penicillin(51.1%). These results indicate that there was no significant difference among the cultured bacteria depending on the type of infections and their susceptibility to cephalosporin and penicillin G was low.

Expression of ssrA in non-pathogen-induced adaptation in the oral cavity through signal exchange with oral pathogens

  • Kim, Sung-Ryoul;Kwak, Jae-Woo;Lee, Sung-Ka;Jung, Seung-Gon;Han, Man-Seung;Kim, Bang-Sin;Kook, Min-Suk;Oh, Hee-Kyun;Park, Hong-Ju
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.38 no.1
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    • pp.14-19
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    • 2012
  • Introduction: This study was conducted to evaluate ssrA expression resulting from adaptation of Escherichia coli (E. coli) to oral pathogens through signal exchange. Materials and Methods: Human cell lines Hep2 and HT29, wild-type E. coli (WT K-12), ssrA knock-out E. coli (${\Delta}K$-12), and Scleropages aureus (S. aureus) were used. A single culture consisting of Hep2, HT29, WT K-12, and ${\Delta}K$-12, and mixed cultures consisting of Hep2 and WT K-12, Hep2 and ${\Delta}K$-12, WT K-12 and S. aureus, ${\Delta}K$-12 and S. aureus, and Hep2, WT K-12, and S. aureus were prepared. For HT29, a mixed culture was prepared with WT K-12 and with WT K-12 and S. aureus. Total RNA was extracted from each culture with the resulting expression of ssrA, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$), and p53 was evaluated by Reverse transcription polymerase chain reaction (RT-PCR). Results: The expression of ssrA in a single culture of WT K-12 was lower than that observed in the mixed culture of WT K-12 with S. aureus. Greater ssrA expression was observed in the mixed culture of WT K-12 with Hep2 than in the single culture of WT K-12. The expression of NF-${\kappa}B$ was higher in the mixed culture of Hep2 with ${\Delta}K$-12 than that in the mixed culture of Hep2 with WT K-12, and was lowest in the single culture of Hep2. The expression of ssrA was higher in the mixed culture of WT K-12 with Hep2 and S. aureus than in the mixed culture of WT K-12 with Hep2. Conclusion: These results suggest that ssrA plays an important role in the mechanism of E. coli adaptation to a new environment.

FORMATION OF BASEMENT MEMBRANE AND STRATIFICATION OF RABBIT ORAL KERATINOCYTES CULTURED ON HUMAN ACELLULAR DERMAL MATRIX (인간 무세포성 진피기질 위에 배양한 가토 구강각화상피세포의 중충화와 기저막 형성에 관한 연구)

  • Kim, Yong-Deok;Ahn, Kang-Min;Yum, Hak-Yeol;Chung, Hun-Jong;Kim, Soung-Min;Jang, Jeong-Won;Sung, Mi-Ae;Park, Hee-Jung;Hwang, Soon-Jung;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.6
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    • pp.510-522
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    • 2005
  • To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

Treatment and diagnosis of oral pain without identifiable oral mucosa lesion (정상 구강 점막 소견의 구강 통증 환자의 진단 및 치료)

  • Kim, Tae-Su;Kim, Sang-Yoon;Nam, Soon-Yuhl;Roh, Jong-Lyel;Choi, Seung-Ho
    • Korean Journal of Bronchoesophagology
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    • v.14 no.1
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    • pp.29-33
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    • 2008
  • Objectives : Oral pain without identifiable oral mucosa lesion is probably multifactorial origin, which include burning mouth syndrome (BMS), oral candidiasis and so on. The aim of this study was to analyze the characteristics of oral pain without identifiable oral mucosa lesion and to evaluate treatment outcome of those patients. Materials and Methods : We reviewed 50 patients without identifiable oral mucosa lesion who were complaint of oral pain. The patients were analyzed according tothe sites, associated symptoms, laboratory tests and fungus culture. The questionnaire included questions on their current diseases, smoking and alcoholic history, psychological factors, and symptoms. Results : The average age of patients was 60 years old. The most frequently involved site was tongue (92%), followed by palate, lower lip, oropharynx, and gingiva. 60% of the patients has psychological disorder as self reported. Culture for Candida was positive in 36% of patients and serum zinc deficiency was present in 60% of patients. Serum iron, vitamin B12, hemoglobin, folic acid deficiency were present in 6-2% of patients. Seventeen patients (65%) with BMS and twelve patients (66%) with oral candidiasis were improved after treatment. Conclusion : We recommend oral candida culture to oral pain patients without oral mucosa lesion. Zinc supplementation of zinc depletion patients may be helpful whereas other laboratory tests have no diagnostic values.

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