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Application of Three-Dimensional Light Microscopy for Thick Specimen Studies
Rhyu, Yeon Seung,Lee, Se Jeong,Kim, Dong Heui,Uhm, Chang-Sub Korean Society of Microscopy 2016 Applied microscopy Vol.46 No.2
The thickness of specimen is an important factor in microscopic researches. Thicker specimen contains more information, but it is difficult to obtain well focused image with precise details due to optical limit of conventional microscope. Recently, a microscope unit that combines improved illumination system, which allows real time three-dimensional (3D) image and automatic z-stack merging software. In this research, we evaluated the usefulness of this unit in observing thick samples; Golgi stained nervous tissue and ground prepared bone, tooth, and non-transparent small sample; zebra fish teeth. Well focused image in thick samples was obtained by processing z-stack images with Panfocal software. A clear feature of neuronal dendrite branching pattern could be taken. 3D features were clearly observed by oblique illumination. Furthermore, 3D array and shape of zebra fish teeth was clearly distinguished. A novel combination of two channel oblique illumination and z-stack imaging process increased depth of field and optimized contrast, which has a potential to be further applied in the field of neuroscience, hard tissue biology, and analysis of small organic structures such as ear ossicles and zebra fish teeth.
Enhanced bone formation by transforming growth factor-β1-releasing collagen/chitosan microgranules
Lee, Jue-Yeon,Kim, Kyoung-Hwa,Shin, Seung-Yoon,Rhyu, In-Chul,Lee, Yong-Moo,Park, Yoon-Jeong,Chung, Chong-Pyoung,Lee, Seung-Jin Wiley Subscription Services, Inc., A Wiley Company 2006 Journal of biomedical materials research. Part A Vol.a76 No.3
<P>Collagen/chitosan composite microgranules were fabricated as bone substitutes for the purpose of obtaining high bone-forming efficacy. The microgranules have the flexibility to fill various types of defect sites with closer packing. The interconnected pores formed spaces between the microgranules, which allowed new bone ingrowth and vascularization. In addition, the transforming growth factor-beta 1 (TGF-β1) was incorporated into the microgranules in order to improve bone-healing efficacy. The collagen/chitosan microgranules were fabricated by dropping a mixed solution into a NaOH/ethanol solution. TGF-β1 was loaded into the collagen/chitosan microgranules by soaking the microgranules in a TGF-β1 solution. Scanning electron microscopy (SEM) observations and experiments examining the release of TGF-β1 from chitosan and the collagen/chitosan microgranules were performed. SEM was used to examine the cell morphologies on the microgranules and cell proliferation was evaluated using a dimethylthiazole tetrazolium bromide assay. The differentiated cell function was assessed by measuring the alkaline phosphatase (ALPase) activity as well as detecting an osteocalcin assay. The in vivo bone-regeneration experiments were performed using a rabbit calvarial defect model. TGF-β1 was released from the collagen/chitosan microgranules at a therapeutic concentration for 4 weeks. SEM indicated that the seeded osteoblastic cells were firmly attached to the microgranules and proliferated in a multilayer manner. The proliferation of the osteoblasts on the TGF-β1-loaded microgranules was the highest among the different types of microgranules tested. The ALPase activity and osteocalcin level of all the samples increased during the culture period, and the TGF-β1-loaded microgranules had a significantly higher ALPase activity and osteocalcin content than the other microgranules. The TGF-β1-loaded microgranules demonstrated a higher bone-regenerative capacity in the rabbit calvarial defects after 4 weeks than the TGF-β1-unloaded microgranules. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2006</P>
Enhanced bone formation by transforming growth factor-β1-releasing collagen/chitosan microgranules
Lee, Jue-Yeon,Kim, Kyoung-Hwa,Shin, Seung-Yoon,Rhyu,In-Chul,Lee, Yong-Moo,Park, Yoon-Jeong,Chung, Chong-Pyoung,Lee, Seung-Jin 이화여자대학교 약학연구소 2008 藥學硏究論文集 Vol.- No.17
Collagen/chitosan composite microgranules were fabricated as bone substitutes for the purpose of obtaining high bone-forming efficacy. The microgranules have the flexibility to fill various types of defect sites with closer packing. The interconnected pores formed spaces between the microgranules, which allowed new bone ingrowth and vascularization. In addition, the transforming growth factorbeta 1 (TGF-β1) was incorporated into the microgranules in order to improve bone-healing efficacy. The collagen/chitosan microgranules were fabricated by dropping a mixed solution into a NaOH/ethanol solution. TGF-β1 was loaded into the collagen/chitosan microgranules by soaking the microgranules in a TGF-β1 solution. Scanning electron microscopy (SEM) observations and experiments examining the release of TGF-β1 from chitosan and the collagen/chitosan microgranules were performed. SEM was used to examine the cell morphologies on the microgranules and cell proliferation was evaluated using a dimethylthiazole tetrazolium bromide assay. The differentiated cell function was assessed by measuring the alkaline phosphatase(ALPase) activity as well as detecting an osteocalcin assay. The in vivo bone-regeneration experiments were performed using a rabbit calvarial defect model. TGF-β1 was released from the collagen/chitosan microgranules at a therapeutic concentration for 4 weeks. SEM indicated that the seeded osteoblastic cells were firmly attached to the microgranules and proliferated in a multilayer manner. The proliferation of the osteoblasts on the TGF-β1-loaded microgranules was the highest among the different types of microgranules tested. The ALPase activity and osteocalcin level of all the samples increased during the culture period, and the TGF-β1-loaded microgranules had a significantly higher ALPase activity and osteocalcin content than other microgranules. The TGF-β1-loaded microgranules demonstrated a higher bone regenerative capacity in the rabbit calvarial defects after 4 weeks than the TGF-β-unloaded microgranules. ⓒ 2005 Wiley Periodicals, Inc. J Biomed Mater Res 76A: 530-539, 2006
Enhanced bone formation by transforming growth factor-β1-releasing collagen/chitosan microgranules
Lee, Jue-Yeon,Kim, Kyoung-Hwa,Shin, Seung-Yoon,Rhyu, In-Chul,Lee, Yong-Moo,Park, Yoon-Jeong,Chung, Chong-Pyoung,Lee, Seung-Jin 이화여자대학교 약학연구소 2005 藥學硏究論文集 Vol.- No.16
Collagen/chitosan composite microgranules were fabricated as bone substitutes for the purpose of obtaining high bone-forming efficacy. The microgranules have the flexibility to fill various types of defect sites with closer packing. The interconnected pores formed spaces between the microgranules, which allowed new bone ingrowth and vascularization. In addition, the transforming growth factor-beta 1 (TCF-βl) was incorporated into the microsranules in order to improve bone-healing efficacy. The collagen/chitosan microgranules were fabricated by dropping a mixedsolution into a NaOH/ethanol solucon. TCF-β1 was loaded into the collagen/chitosan microgranules by soaking the microgranules in a TCF-βl solution. Scanning electron microscopy (SEM) observations and experiments examining the release of TCF-β1 from chitosan and the collagen/chitosan microgranules were performed. SEM was used to examine the cell morphologies on the microgranules and cell proliferation was evaluated using a dimethylthiazole tetrazolium bromide assay. The differentiated cell function was assessed by measuring the alkaline phosphatase (ALPase)activity as well as detecting an osteocalcin assay. The in vivo bone-regeneration experiments were performed using a rabbit calvarial defect model. TCF-β1 was released from the collagen/chitosan microgranules at a therapeutir concentration for 4 weeks. SEM indicated that the seeded osteoblastic cells were firmly attached to the microgranules and proliferated in a multilayer manner. The proliferation of the osteoblasts on the TCF-β1-loaded microgranules was the high-est among the different types of microgranules tested. The ALPase activity and osteocalcin level of ail the samples increased during the culture period, and the TCF-β1-loaded microgranules had a significantly higher ALPase activity and osteocalcin content than the other microgranules. The TCF-β1-loaded microgranules demonstrated a higher bone-regenerative capacity in the rabbit calvarial defects after 4weeks than the TCF-01-unloaded microgranules.
Lee, Jue-Yeon,Seol, Yang-Jo,Kim, Kyoung-Hwa,Lee, Yong-Moo,Park, Yoon-Jeong,Rhyu, In-Chul,Chung, Chong-Pyoung,Lee, Seung-Jin 이화여자대학교 약학연구소 2004 藥學硏究論文集 Vol.- No.14
Purpose. Tricalcium phosphate (TCP)/chitosan composite microgranules were developed as bone substitutes and tissue engineering scaffolds with the aim of obtaining a high bone forming efficacy. The microgranules have the ability to fill various types of defect sites with closer packing. In addition. the transforming growth factor-beta I(TGF-β1) was added to the microgranules in order to improve bonehealing efficacy. Methods. TCP/chitosan microgranules were fabricated by dropping a TCP suspended chitosan solution into a NaOH/ethanol solution. TGF-β1 was incorporated into the TCP/chitosan microgranules by soaking the microgranules into the TGF-β1 solution. Scanning electron microscopy(SEM) observations as well as experiments examining the release of TGF-β1 from chitosan and TCP/chitosan microgranules were performed. SEM was used to examine the cell morphologies on the microgranules, and the extent of cell proliferation was evaluated using a dimethyl-thiazol tetrazolium bromide (MTT) assay. The differentiated cell function was assessed by measuring the alkaline phosphatase activity as well as performing an osteocalcin assay. Results. The size of the prepared microgranules was 350-500㎛ and TCP powders were observed on the surface of the microgranules. TGF-β1 was released from the TCP/chitosan microgranules at a therapeutic concentration for 4 weeks. The proliferation of osteoblasts on the TGF-β1 loaded microgranules was the highest among the microgranules. SEM indicated that the seeded osteoblastice cells were firmly attached to the microgranules and proliferated in a multilayer fashion. The ALPase activity and osteocalcin content of all the samples increased during the culture period Conclusions. These results suggest that the TCP/chitosan microgranules are potential bone substitutes with a drug releasing capacity and a osteoblastic cells culture scaffold.
Study on the Peptide Engineered Synthetic Oligopeptide Domain for Bone Regeneration
( Jun Beom Park ),( Jue Yeon Lee ),( Yoon Jeong Park ),( Sang Hoon Rhee ),( Sang Cheol Lee ),( Tae Il Kim ),( Yang Jo Seol ),( Yong Moo Lee ),( Young Ku ),( In Chul Rhyu ),( Soo Boo Han ),( Chong Pyou 대한치주과학회 2007 대한치주과학회 학술대회자료집 Vol.2007 No.2
Downregulation of Intraepidermal nerve fiber densities in psoriasis skin
( Dai Hyun Kim ),( Se Jeong Lee ),( Im Joo Rhyu ),( Yeon Seok Lee ),( Tae Young Han ),( June Hyunkyung Lee ),( Jae Eun Choi ) 대한피부과학회 2021 대한피부과학회 학술발표대회집 Vol.73 No.-
Background: How the cutaneous nervous system is changed in psoriasis skin has been still remained unclear because of previously reported conflicting results. Objective: The aim was to verify the changes of intraepidermal nerve fiber densities in psoriasis skin. Methods: Fourteen patients (thirteen men and one woman, mean age of 51 years, ranged from 26 to 77 years) with psoriasis were enrolled for the study. The documented paraffin blocks of skin biopsies obtained from lesional and adjacent non-lesional sites were sectioned with 8-㎛ thickness. The paraffin sections were proceeded with antigen retrieval processes and immunostained with protein gene product (PGP) 9.5 to quantify intraepidermal nerve fiber densities (IENFDs). The immunolabelled results were imaged with a confocal microscope and serial optical sections taken at intervals of 1-㎛ were reconstructed. The maximum projection images were used for quantification of cutaneous nerve fibers. Results: The average values of IENFDs in psoriasis and non-lesional skin were 0.93 ± 0.44 and 1.92 ± 0.90, respectively. The IENFD in psoriasis skin was meaningfully downregulated compared with the results obtained from non-lesional skin with statistical significance (p < 0.05). However, there was no significant correlation between IENFD and clinical parameters such as psoriasis area and severity index (PASI) and pruritus visual analogue scale (VAS). Conclusion: In conclusion, our results showed statistically significant reduction in epidermal innervation in psoriatic lesions compared with non-lesional skin. In addition, the results also presented the possible usage of documented paraffin blocks in the quantification of IENFDs instead of cryo-blocks.