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The Effect of Platelet Derived Growth Factor - BB Loaded Chitosan/Calcium Metaphosphate on Bone Regeneration (혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구)

  • Lee, Seung-Yeol;Seol, Yang-Jo;Lee, Yong-Moo;Lee, Ju-Yeon;Lee, Seung-Jin;Kim, Suk-Young;Ku, Young;Rhyu, In-Chul;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.31 no.1
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    • pp.1-23
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    • 2001
  • Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

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Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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The Effects of Gunyuljejo-tang on the CCl4-induced Liver Damage in Rat (건율제조탕이 CCl4로 유발(誘發)된 간손상(肝損傷) 백서(白鼠)에 미치는 영향(影響))

  • Kim, Jung-Yul;Kim, Hyuk;Yang, Sang-Mook;Kim, Dal-Rae;Jeon, Jong-Weon
    • Journal of Sasang Constitutional Medicine
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    • v.16 no.3
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    • pp.96-107
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    • 2004
  • 1. Objectives This study was carried out to investigate the effects of Gunyuljejo-tang on the $CCl_4$-induced Liver Damage in Rats. 2. Methods Sprague-Dawley rats were devided into 5 experimental groups : Normal, $NS+CCl_4$(Solid extract of $CCl_4$ injection group after Normal Saline feed), $GYJJT+CCl_4$(Solid extract of $CCl_4$ injection group after Gunyuljejo-tang feed), $CCl_4+NS$(Normal Saline feed group after $CCl_4$ injection), $CCl_4+GYJJT$(Solid extract of Gunyuljejo-tang feed group after $CCl_4$ injection). Biochemical assays for serum enzyme activities such as AST, ALT, ALP, BUN, Creatinine, Uric Acid, Total Protein, Albumin, Total Cholesterol, Triglyceride, Glucose, and mRNA Revelation of Cytochrome p450 and activities such as LPO, GSH, GST, Glutathione Reductase, Glutathione Peroxidase, SOD, Catalase, Hydroxyproline, and ${\beta}$-Glucuronidase were performed. 3. Results (1) $GYJJT+CCl_4$ showed lower revelation of Cytochrome p450. (2) $GYJJT+CCl_4$ showed higher GSH activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher GSH activity than $CCl_4+NS$ injection significantly. (3) $GYJJT+CCl_4$ showed higher GST activity than $NS+CCl_4$. $CCl_4+GYJJT$ showed higher GST activity than $CCl_4+NS$ significantly. (4) $GYJJT+CCl_4$ showed higher Glutathione Peroxidase activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher Glutathione Peroxidase activity than $CCl_4+NS$ significantly. (5) $CCl_4+GYJJT$ showed higher SOD activity than $CCl_4+NS$ significantly. (6) $CCl_4+GYJJT$ showed higher Catalase activity than $CCl_4+NS$ significantly. (7) $GYJJT+CCl_4$ showed lower Hydroxyproline than $NS+CCl_4$ significantly, $CCl_4+GYJJT$ showed higher Hydroxyproline than $CCl_4+NS$ significantly. (8) $GYJJT+CCl_4$ showed higher ${\beta}$-Glucuronidase activity than $NS+CCl_4$, $CCl_4+GYJJT$ showed higher ${\beta}$-Glucuronidase activity than $CCl_4+NS$ significantly. 4. Conclusions Gunyuljejo-tang has the recovering effects on the $CCl_4$-induced Liver Damage significantly.

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A Study on the Actual Conditions of Smoking in Middle and High School Students in One Region (일 지역 중·고등학생의 흡연실태)

  • Kim, Hyeon-Ok
    • Journal of the Korean Society of School Health
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    • v.12 no.1
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    • pp.149-167
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    • 1999
  • To investigate the actual conditions of smoking in middle and high school students in Chinan County, I used a sturctured questionnaire for 1,579 students attending twelve middle-high schools from December 1, 1998 to December 20, 1998. I collected and data correlated the using an $SPSS-PC^+$ 1. The smoking rate of middle-high schoo1 students in Chinan County was 17.9%, relatively high. This smoking rate was different according to the gender, grade, religion, and economic situation. In mals, high school students, non-religious, students low income family students, the smoking rate was higher. The smoking rate of high school students was almost the same as the smoking rate of adults, generally higher than that of foreign teenagers. Because the smoking rat of studinets in the third grade of middle school and in the first grade of high School was six times higher, increased education should be conducted during this time in an attempt to curb the sudden increase of the smoking rate. The smoking rate of girl students was 5.0%; this has increased mor than three times from ten years ago. Consequently, counter measures should be taken against the smoking of female students as well as juvenile smoking in general. In addition, the smoking rate of middle-high school students showed interesting differences when correnated with enviornmental factors. Students with low grades, who are not satisfied with school life, who don't have both parents, who have uncaring parents who nare too strict or too arbitary, who have smoking parents, or who have experienced smoking commonly smoked. Therefore, to lower the smoking rate we should improve the school environment, improve a student's interest in school life. And parents or siblings should lead by example and quit smoking at home. Schools should educate students more effectively concerning the harmful effects of smoking and create an accurate understanding of its dancers. From the beginning, we should teach students never ever to touch cigaretts. 2. The surve discovered that most students started out of curiosity, or solicitantion from friends or elders at middle school, and had been smoking one to five cigarettes for more than a year. They obtained cigarettes at stores and most of them have friends who smoke. As a result anti-smoking education should be conducted at elementary schools prior to middle school. More than 95% of the teenagers who smoke had friends who smote and smoked out of curiosity or the recommendation of elders. Thus, we must focus on teenagers who smoke in group, rather than individually. Fuyrthermore, the strict application of the regulation of tobacco sales as well as tobacco cooperation from retailers are needed. While students did not show any mood or academic achievement difference after beginning smoking, 58.1% of the students a health situdation that was worse. Juvenile smoking is more harmful to the juvenile than adult smoking is to the adult. This should be focused on in an anti-smoking campaign. 3. Students who smoke hada more positive attitude toward smoking than students who don't smoke. Students who smoke had a tendency to have a nuetral position and are not concerned about smoking compared to non-smoking students. The survey showed that the great number of students had a nuetral position. Because this nuetrality may increase Juvenile smoking, education that provides an exact understanding of smoking should be performed to build the correct attidude toward smoking. 4. Middle school students smoke when angry, gloomy, anxious, a lone and when they have some problems to solve, on when they feel inconveniened in other wores, they smoke to reliver stress. They also smok due to addiction. Because smoking is not a praetical method to relieve stress, a program which helps to acquire positive relief stress should be provided to help reduce smoking. 5. About 65% of students who smoke want to quit smoking because of health problem, 78% of them have tried mor than once to quit but failed due to weak will power and peer pressure from friends who smoke. Juvenile smoking is group, oriented. Thus, the program that advances less smoking will be the one that focuseds on groups. 6. As for advice to students who want to quit smoking, "persuasion" was used most commonly, followed by a "presentation on how to quit smoking". Another method were severe punishment. About 70% of the students wanted the anti-smoking guide at school. 7. Most students (73.5%) had a position that more anti-smoking education at school is needed. Obriously, then, anti-smoking education at middle-high schools should be reinfoced. Although the education which explains the harmful influence of tobacco is known as an efficient way prevent smoking; it does not influence students who already smoke. Therefore, for students who smoke, multi-dimensional approaches must be attempted that include physical training, phychokogical approache, consultation and discussion, medical chek-ups, audio-visual education technigues, and professonal instructors, in addition, because smoking students have more negative on lukewarm attitude to anti-smoking education anti-smoking education should be conducted through a communicative style by dedicated teachers who care about students. In order to increase the effectiveness of this program.

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