• Title/Summary/Keyword: Cellular regeneration

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BIOLOGICAL EFFECTS OF pH CONCENTRATION ON CULTURED HUMAN PERIODONTAL LIGAMENT CELL ACTIVITY IN VITRO (수소이온 농도의 변화가 배양 인체 치주인대 세포의 활성에 미치는 영향)

  • Kim, Seong-Ho;Park, Kui-Woon;Yoo, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.539-556
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    • 1995
  • Periodontal therapeutic modalities should be re-establishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, periodontal ligament cells must selective migrate to the deneded root surface, attached and proliferated it. Local pH concentration is one of the most factors that periodontal regeneration. The aims of this study were to examine on biological effects of pH to the human periodontal ligament cells in vitro, especially on the cell morphology, attachment, activity, vitality and viability. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Immediately after extraction, any soft tissue adhering to the cervical parts of the roots was carefully removed with a sterile curette. To produce different pH levels in the media, Eagle's MEM was adjusted from pH 6.6 to 8.2 in 0.2 intervals with 1 M NaOH and 1 N HCl. After cultivation, Then, Periodontal ligament cells were cultured at pH ranging from 6.6-8.2. attachment assay was done at 1, 2 day incubation and activity assay was done at 1, 2, 3 day incubation. The experiments were evaluated by scaning electron microscopic techniques (HITACHIX-650 Scaning Electron Microanalyzer, Tokyo, Japan), MTT assay, and the cultured periodontal ligament cells were fixed in neutral formalin for 24 hours and immunohistochemically processed by PCNA for proliferating ability. The surviving cells in the medium showed slightly increased volume and widening intercellular distances at low concentration of pH than control group (pH 7.4), and apparently shrinkage at high concentration of pH than control group (pH 7.4). The results of the statistical analysis from the experiment on attachment, vitality and viability were as follows. Attachment of periodontal ligament cells at 1st and 2nd day, similar attachment rate of low concentration pH compared with control value (pH 7.4). But above pH 8.0, attachment rate were statistically significant decrease from control value(P<0.05). Periodontal ligament cell's activities were maximum at pH 7.6 by MTT assay. Similar with control value at low concentration of pH. But, the activities were statistically significant decrease at high concentraration of pH(P<0.05). Cellular proliferating rate (PCNA index) were statistically significant decrease from control value at low and high concentration of pH(p<0.05). This results suggested that hjgh concentration pH, in other words, alkali pH was cytotoxic effects on human periodontal ligament cells in vitro.

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Effect of Stem Cell Transplantation on Pain Behavior and Locomotor Function in Spinal Cord Contusion Model

  • Park, Hea-Woon;Kim, Su-Jeong;Cho, Yun-Woo;Hwang, Se-Jin;Lee, Won-Yub;Ahn, Sang-Ho;Jang, Sung-Ho
    • The Journal of Korean Physical Therapy
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    • v.22 no.3
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    • pp.79-85
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    • 2010
  • Purpose: Many trials for new therapeutic approaches such as stem cell-based transplantation have been conducted to improve the repair and regeneration of injured cord tissue and to restore functions following spinal cord injury (SCI) in animals and humans. Adipose tissue-derived stromal cells (ATSCs) have multi-lineage potential to differentiate into cells with neuron-like morphology. Most studies of stem cell transplantation therapy after SCI are focused on cellular regeneration and restoration of motor function, but not on unwanted effects after transplantation such as neuropathic pain. This study was focused on whether transplantation of ATSCs could facilitate or attenuate hindpaw pain responses to heat, cold and mechanical stimulation, as well as on improvement of locomotor function in a rat with SCI. Methods: A spinal cord injury rat model was produced using an NYU impactor by dropping a 10 g rod from a height of 25 mm on to the T9 segment. Human ATSCs (hATSCs; approximately $5{\times}10^5$ cells) or DMEM were injected into the perilesional area 9 days after the SCI. After transplantation, hindpaw withdrawal responses to heat, cold and mechanical allodynia were measured over 7 weeks. Motor recovery on the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale and on the inclined plane test were also evaluated. Results: The present study demonstrated that increased hindpaw withdrawal responses to cold allodynia was observed in both groups after transplantation, but the development of cold-induced allodynia in the hATSC transplantation group was significantly larger than in the control group. The difference between the two groups in locomotor functional improvement after SCI was also significant. Conclusion: Careful consideration not only of optimal functional benefits but also of unintended side effects such as neuropathic pain is necessary before stem cell transplantation therapy after SCI.

Fabrication and Characteristic Evaluation of Three-Dimensional Blended PCL (60 wt %)/β-TCP (40 wt %) Scaffold (3 차원 Blended PCL (60 wt %)/β-TCP (40 wt %) 인공지지체의 제작 및 특성 평가)

  • Sa, Min-Woo;Kim, Jong Young
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.38 no.4
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    • pp.371-377
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    • 2014
  • In tissue engineering, a scaffold is a three-dimensional(3D) structure that serves as a template for regeneration the functions of damaged tissues or organs. Among materials for scaffolds, polycaprolactone(PCL) and ${\beta}$-tricalcium phosphate(${\beta}$-TCP) are biodegradable and biocompatible. In this study, we fabricated 3D PCL, blended PCL (60 wt %)/${\beta}$-TCP (40 wt %), and pure ${\beta}$-TCP scaffolds by a multi-head scaffold fabrication system. Scaffolds with a pore size of $600{\pm}20{\mu}m$ was observed by scanning electron microscopy. The effects of 3D PCL, blended PCL (60 wt %)/${\beta}$-TCP (40 wt %) and pure ${\beta}$-TCP scaffolds were analyzed by evaluating their mechanical characteristics. In addition, in an in-vitro study using osteoblast-like saos-2 cells, we confirmed the effects of 3D scaffolds on cellular behaviors such as cell adhesion and proliferation. In summary, the 3D blended PCL (60 wt %)/${\beta}$-TCP (40 wt %) scaffold was found to be suitable for human cancellous bone in terms of its the compressive strength, biocompatibility, and osteoconductivity. Thus, blending PCL and ${\beta}$-TCP could be a promising approach for fabricating 3D scaffolds for effective bone regeneration.

Effects of platelet-derived growth factor and epidermal growth factor on the characteristics of beagle dog's periodontal ligament and bone marrow cells (혈소판유래성장인자와 상피성장인자가 치주인대세포와 골수세포의 성상에 미치는 영향)

  • Cho, Byeong-Do;Herr, Yeek;Park, Joon-Bong;Kwon, Young-Hyuk;Lee, Man-Sup
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.491-510
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    • 1996
  • This study was performed to evaluate the effects of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) on the characteristics of beagle dog's periodontal ligament (BPD) cells and bone marrow (BBM) cells which have the important role on the early stage of periodontal tissue regeneration in vitro. In control group, the cells ($1.5{\times}10^5$cells/ml) were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu]g/ml$ ascorbic acid, and 10mM/ml ${\beta}-glycerophosphate$. In experimental groups, growth factors, PDGF or EGF(10ng/ml), were added into the above culture condition. And then each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity at 1, 5, 9, 13, 17th day after seeding of cells into the culture wells. The results were as follows: 1. Both BPD and BBM cells in PDGF-treated group proliferated more rapidly than non-treated cells. This finding also was observed in EGF-treated group but it was not as prominent as that of PDGF-treated group. The proliferation rates of both cells showed the time-dependent pattern during experimental periods in all three groups. 2. Amount of total protein synthesis was more increased in PDGF-treated group than in control group. But no significant difference between EGF-treated group and control group was observed throughout experimental periods even though the tendency of amount of protein synthesis was time-dependent pattern. 3. Alkaline phosphatase activity also more increased in PDGF-treated group than control group. But slight decrease tendency was seen in both cells of EGF-treated group. From the above results, PDGF appeared to enhance the proliferation and cellular activities including amount of total protein synthesis and alkaline phosphatase activity of BPD and BBM cell, but EGF did not show notable effects. The optimal application of these growth factors was thought to be useful as the adjunctive means in periodontal regeneration procedures.

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The Effect of Pore Sizes on Poly(L-lactide-co-glycolide) Scaffolds for Annulus Fibrosus Tissue Regeneration (조직공학적 섬유륜재생을 위한 PLGA 지지체 제조시 다공크기의 영향에 관한 연구)

  • So, Jeong-Won;Jang, Ji-Wook;Kim, Soon-Hee;Choi, Jin-Hee;Rhee, John-M.;Min, Byung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.32 no.6
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    • pp.516-522
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    • 2008
  • Biodegradable polymers have been used extensively as scaffolding materials to regenerate new tissues and the ingrowth of tissue have been reported to be dependent directly of the porosity, pore diameter, pore shape, and porous structure of the scaffold. In this study, porous poly (L-lactide-co-glycolide) (PLGA) scaffolds with five different pore sizes were fabricated to investigate the effect of pore sizes for AF tissue regeneration. Cellular viability and proliferation were assayed by MTT test. Hydroxyproline/DNA content of AF cells on each scaffold was measured. sGAG analyses were performed at each time point of 2 and 6 weeks. Scaffold seeded AF cells were implanted into the back of athymic nude mouse to observe the difference of formation of disc-like tissue depending on pore size in vivo. We confirmed that scaffold with $180{\sim}250{\mu}m$ pores displayed high cell viability in vitro and produced higher ECM than scaffold with other pore sizes in vivo.

Fine Needle Aspiration Cytology of Hepatocellular Carcinoma - A Study on 247 Cases - (간세포암종의 세침흡인 세포학적 소견 - 247예에 대한 연구 -)

  • Lee, Kwang-Gil;Lee, Jong-Tae;Choi, Soo-Im;Park, Chan-Il
    • The Korean Journal of Cytopathology
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    • v.1 no.1
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    • pp.1-17
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    • 1990
  • Hepatocellular carcinoma (HCC) is malignant tumor frequently occurring in Koreans. There have been few reports regarding the cytologic findings of fine needle aspiration (FNA) of HCC. Most have suggested a diagnostic problem in the cytology distinguishing HCC from some benign hepatic lesions-for example, a regeneration nodule in cirrhosis and liver cell adenoma. In spite of its high frequency in Korea, no cytologic study has been reported, concerning the FNA of HCC. In an attempt to achieve cytologic criteria for the diagnosis of HCC, the authors studied retrospectively cytopathologic findings of 247 cases of HCC. These cases were confirmed either by histoiogic examination including lobectomy, biopsy, or ceil block material, or, when tissue diagnosis was unavailable, by a high serum alpha-fetoprotein level (over 400 I. U.). All aspiration smears were stained by the Papanicolaou method. In each case, the smears were analyzed for cell patterns and various cytomorphology of the tumor cells. The smear background was assessed for the presence of tumor cell necrosis and inflammatory components and compared to that of metastatic carcinomas. The cell patterns were classified as trabecular, acinar, dispersed, and irregular. The cytologic parameters analyzed included the degree of nuclear atypia and the presence of mitoses, intranuclear cytoplasmic inclusions, nucleolar prominency, endothelial lining, multinucleated giant cells, eosinophiic globules, bile, and Mallory body. Most of the FNA of HCC showed markedly cellular smears. The tumor cells were most frequently arranged in a trabecular pattern (80.3%). The irregular (12.6%), the acinar (5.5%), and the dispersed patterns (1.7%) followed in decreasing frequency. Individual hepatoma cells were larger than normal liver cells. However, they had morphologic features characteristic of the hepatic cells the cells were round or polygonal, their cytoplasm was abundant and granular with eosinophilic or amphophilic stainability, and their nuclei were round to oval, located centrally, and tended to have prominent nucleoli. Anaplasia and pleomorphism of tumor cells were generally mild to moderate. These findings existed even in very well differentiated cases. Mitotic figures were present in about 85% of the cases. Prominent nucleoli were observed only in about half the cases. The frequency of other cytologic features was as follows intranuclear cytoplasmic inclusion in 86.8% : endothelial lining in 56.1% : bile in 19.8% : and giant cells in 60.1%. Clear cells were often present in 11.7%, Most aspiration smears of HCC displayed clean background without necrosis or inflammatory material in contrast to the dirty, necrotic background of metastatic cancers and cholangiocarcinomas. Based on the above mentioned features, it is suqqested that the cytologic critieria most important for the diagnosis of HCC include a markedly cellular smear, trabecular pattern, hepatocytoid appearance of tumor cells, endothelial lining, the presence of bile, giant cells, intranuclear cytoplasmic inclusions, and prominent nucleoli, Among these, trabecular pattern, endothelial lining, giant cells and clean smear background are points to be considered in differentiating HCC from metastatic and cholangiocellular carcinoma.

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Osteogenic activity of an adenovirus expressing BMP-2 on Human Periodontal Ligament cells (Adenovirus에 의해서 발현된 BMP-2가 치주인대세포의 분화에 미치는 영향)

  • Kim, Kyoung-Hwa;Park, Yoon-Jeong;Lee, Sang-Cheul;Kim, Tae-Il;Seol, Yang-Jo;Lee, Yong-Moo;Ku, Young;Han, Soo-Boo;Chung, Chong-Pyoung;Rhyu, In-Chul
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.511-524
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    • 2005
  • The regeneration of lost periodontal tissue is a major goal of therapy. Periodontal ligament cell(PDL) is a specialized connective tissue that connects cementum and alveolar bone to maintain and support teeth in situ and preserve tissue homoeostasis. Bone morphogenetic proteins(BMPs) have shown much potential in the reconstruction of the periodontum by stimulate new bone and new cementum formation. Limitiations of BMP administration to periodontal lesions is high dose delivery, BMP transient biological activity, and low bioavailability of factors at the wound site. Gene delivery method can be alternative treatment strategy to deliver BMPs to periodontal tissue. The purpose of this study is to investigate efficiency of BMP-2 gene delivery with cell-based therapy using PDL cells. PDL cell were transduced with adenoviruses encoding either BMP-2 or Lac-Z gene. To evaluate osteogenic activity of expressed BMP-2 on PDL cells, we investigated secreted BMP-2, cellular activity, ALPase, produced mineralized nodules. To evaluate collagen scaffold as carrier for transduced cell delivery, we examined morphology and secreted BMP-2 of transducd PDL cells on it. BMP-2 transducd PDL cells produced higher levels of BMP-2, ALPase, mineralized nodules than non transduced cells. Cellular activity of transduced cells was showed similar activity to non transduced cells. Transduce cells attached on collagen scaffold secreted BMP-2 at 7day and was showed similar morphology to non transduced cells. These results demonstrated that transduced PDL cells produced biologically active BMP-2 and collagen scaffold could be carrier of transducd cells.

Long-Term Priming by Three Small Molecules Is a Promising Strategy for Enhancing Late Endothelial Progenitor Cell Bioactivities

  • Kim, Yeon-Ju;Ji, Seung Taek;Kim, Da Yeon;Jung, Seok Yun;Kang, Songhwa;Park, Ji Hye;Jang, Woong Bi;Yun, Jisoo;Ha, Jongseong;Lee, Dong Hyung;Kwon, Sang-Mo
    • Molecules and Cells
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    • v.41 no.6
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    • pp.582-590
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    • 2018
  • Endothelial progenitor cells (EPCs) and outgrowth endothelial cells (OECs) play a pivotal role in vascular regeneration in ischemic tissues; however, their therapeutic application in clinical settings is limited due to the low quality and quantity of patient-derived circulating EPCs. To solve this problem, we evaluated whether three priming small molecules (tauroursodeoxycholic acid, fucoidan, and oleuropein) could enhance the angiogenic potential of EPCs. Such enhancement would promote the cellular bioactivities and help to develop functionally improved EPC therapeutics for ischemic diseases by accelerating the priming effect of the defined physiological molecules. We found that preconditioning of each of the three small molecules significantly induced the differentiation potential of $CD34^+$ stem cells into EPC lineage cells. Notably, long-term priming of OECs with the three chemical cocktail (OEC-3C) increased the proliferation potential of EPCs via ERK activation. The migration, invasion, and tube-forming capacities were also significantly enhanced in OEC-3Cs compared with unprimed OECs. Further, the cell survival ratio was dramatically increased in OEC-3Cs against $H_2O_2$-induced oxidative stress via the augmented expression of Bcl-2, a pro-survival protein. In conclusion, we identified three small molecules for enhancing the bioactivities of ex vivo-expanded OECs for vascular repair. Long-term 3C priming might be a promising methodology for EPC-based therapy against ischemic diseases.

Enhanced Bone-Regenerative Performance of Porous Hybrid Scaffolds by Surface Immobilization of Nano-Hydroxyapatite

  • Lee, Sang-Cheon
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2009.11a
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    • pp.12.1-12.1
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    • 2009
  • Nano-hydroxyapatite (N-HAp)has shown the pivotal role in producing bone-regenerative materials since it has similarity to natural bone minerals in terms of size, morphology, and the composition. Currently, the combination of biopolymers and N-HAp is recognizedas an attractive approach in generating hybrid scaffolds for bone tissueengineering. Surface engineering is an important issue since it determines whether cells can effectively adhere and proliferate on porous scaffolds. We aim to develop a synthetic approach to porous 3D scaffolds by immobilizing N-HAp on pore surfaces. The discrete nano-level anchoring of N-HAp on the scaffold pore surface is achieved using surface-repellent stable colloidal N-HAp with surface phosphate functionality. This rational surface engineering enables surface-anchored N-HAp to express its overall intrinsic bioactivity,since N-HAp is not phase-mixed with the polymers. The porous polymer scaffolds with surface-immobilized N-HAp provide more favorable environments thanconventional bulk phase-mixed polymer/N-HAp scaffolds in terms of cellular interaction and growth. In vitro biological evaluation using alkalinephosphatase activity assay supports that immobilized N-HAp on pore surfaces of polymer scaffolds contributed to the more enhanced in vitro osteogenicpotential. Besides, the scaffolds with surface-exposed N-HAp provide favorable environments for enhanced in vivo bone tissue growth, estimated by characteristic biomarkers of bone formation such as collagen. The results suggest that newly developed hybrid scaffolds with surface-immobilized N-HApmay serve as a useful 3D substrate with pore surfaces featuring excellent bonetissue-regenerative properties. Acknowledgement. This research was supported by a grant (code #: 2009K000430) from 'Center for Nanostructured Materials Technology' under '21st Century Frontier R&D Programs' of the Ministry of Education, Science and Technology, Korea.

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The Effect of Fibrillar Collagen on Bony Healing of Calvarial Defect in Rats (골 조직 치유과정에서 Collagen 막의 효과)

  • Kim, Jae-Bung;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.29 no.2
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    • pp.355-373
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    • 1999
  • Many researches have been reported that collagen as cellular stroma, matrix of grafting materials, mediator of agents for the purpose of promoting healing process invivo, but the responses in vivo were seen various. The goal of this experiment is to assess the effect of collagen on bony healing, through histological evaluation of implanted collagen on the calvarial defect in rats. 2-month-old Sprague-Dawley, 24 rats were used and 12 rats assigned to each group of control and test. Defect of 5mm in diameter was made on the calvarial bone with trephine bur. Following thorough saline rinse, defect of control group was left in empty and that of experimental group was filled with fibrillar collagen($COLLATAPE^{(R)}$, COLLA-TEC. INC. U.S.A.) soaked in saline. 3 rats in each group were sacrificed at 3, 7, 14, 21 days after operation respectively, and the tissue blocks were prepared for light microscope with H-E for evaluation of overall healing, with TRAP(tartrate resistant acid phosphatase) for evaluation of osteoclastic activity and with immunohistochemical staining for macrophages. The results were as follows : 1. In the control group, inflammatory responses were disappeared at day 14, but, in the experimental group inflammatory infiltrates were reduced at day 21. Thus, the experimental group showed more severe soft tissue inflammation than control group. 2. Both control and experimental group showed slight appositional growth at day 7 and gradual bony growth to 21th day. But, complete bony healing of the defect was not shown. There was no significant difference in bony healing between control and experimental group 3. Specific response of macrophages for implanted collagen was observed at day 14 in the experimental group. In conclusion, although fibrillar collagen caused inflammation of soft tissue during initial healing period, inflammatory responses by fibrillar collagen didn't inhibit bony regeneration and implanted collagen was biodegradaded by macrophages. Thus, we expect that fibrillar collagen can be used for useful mediator of graft materials or growth factors.

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