• The Journal of Korean Academy of Prosthodontics
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• v.43 no.6
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• pp.751-763
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• 2005

Statement of problem. To improve a direct implant fixation to the bone, various strategies have been developed focusing on the surface of materials. The surface quality of the implant depends on the chemical, physical, mechanical and topographical properties of the surface. The different properties will interact with each other and a change in thickness of the oxide layer may also result in a change in surface energy, the surface topography and surface, chemical composition. However, there is limited the comprehensive study with regard to changed surface and biologic behavior of osteoblast by anodization. Purpose of study. The aim of this study was to analyze the characteristics of an oxide layer formed and to evaluate the cellular biologic behaviors on titanium by anodic oxidation (anodization) by cellular proliferation, differentiation, ECM formation and gene expression. And the phospholipase activity was measured on the anodized surface as preliminary study to understand how surface properties of Ti implant are transduced into downstream cellular events. Methods and Materials. The surface of a commercially pure titanium(Grade 2) was modified by anodic oxidation. The group 1 samples had a machined surface and other three experimental specimens were anodized under a constant voltage of 270 V(Group 2), 350 V(Group 3), and 450 V(Group 4). The specimen characteristics were inspected using the following five categories; the surface morphology, the surface roughness, the thickness of oxide layer, the crystallinity, and the chemical composition of the oxide layer. Cell numbers were taken as a marker for cell proliferation. While the expression of alkaline phosphatase and Runx2 (Cbfa1) was used as early differentiation marker for osteoblast. The type I collagen production was determined, which constitutes the main structural protein of the extracellular matrix. Phospholipase $A_2$ and D activity were detected. Results. (1) The anodized titanium had a porous oxide layer, and there was increase in both the size and number of pores with increasing anodizing voltage. (2) With increasing voltage, the surface roughness and thickness of the oxide film increased significantly (p<0.01), the $TiO_2$phase changed from anatase to rutile. During the anodic oxidization, Ca and P ions were more incorporated into the oxide layer. (3) The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity, type I collagen production, and Cbfa 1 gene expression increased significantly (p<0.01), while the cell proliferation decreased. (4) In preliminary study on the relation of surface property and phospholipase, PLD activity was increased but $PLA_2$ activity did not changed according to applied voltage. Conclusion. The anodized titanium shows improved surface characteristics than the machined titanium. The surface properties acquired by anodization appear to give rise more mature osteoblast characteristics and might result in increased bone growth, and contribute to the achievement of a tight fixation. The precise mechanism of surface property signaling is not known, may be related to phospholipase D.

• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.215-222
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• 1989

In order to investigate the morphological characteristics of epididymal duct of the squirrel, the histological and histochemical studies were carried out. The results obtained were summarized as follows: The epididymal duct can be divided into 9 segments by histological and histochemical features. Segments 1 to 5 were located in the head, segments 6 and 7 in the body, and segments 8 and 9 in the tail of the epididymis. The apical cells were numerous in the segment 1. Clear cells which has a compact, deeply staining nucleus and a characteristically clear cytoplasm were scattered in the epithelium throughout the duct. Interepithelial clear cells which had PAS-positive granules tended to increase in number caudally. Strong PAS-positive reaction was detected at the intralumen of the segments 3,8 and 9. Acid phosphatase activity was relatively high in the basal cytoplasm of the segment 7, and then in the supranuclear region of the segments 8 and 9. Alkaline phosphatase activity was weakly positive or negative except the segments 3 and 4. ATPase activity was strong in the free surface of the epithelium in the head and the entire cytoplasm in the body and tail, a,nd SDH activity was generally weak except for the body where it was more intense.

• Journal of Periodontal and Implant Science
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• v.25 no.3
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• pp.623-634
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• 1995

Periodontitis is characterized by gingival inflammation and results in periodontal pocket formation with loss of the supporting alveolar bone and connective tissue around the teeth. Therapeutic modalities should therefore aim not only at eliminating the gingival inflammatory process and preventing the progression of periodontal disease but also at reestablishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, progenitor cells must migrate to the denuded root surface, attach to it, proliferate and mature into an organized and functional fibrous attachment apparatus. Likewise, progenitor bone cells must also migrate, proliferate, and mature in conjunction with the regenerating periodontal ligament. Significant advances have been made during the last decade in understanding the factors controlling the migration, attachment and proliferation of cells. A group of naturally occuring molecules known as polypeptide growth factors in conjunction with certain matrix proteins are key regulators of these biological events. Of these, the fibroblast growth factor(FGF), platelet-derived growth factor(PDGF) , insulin like growth factor(CIGFs), transforming growth factor(TGFs), epidermal growth factor(EGF) and bone morphogenetic growth factor(BMPs) apper to have an important role in periodontal wound healing. The purpose of this study was to determine the effects of BMP on periodontal ligament cells. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Cultured periodontal ligament cells were treated with BMP. Cellular activities were determined by MTT(3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay and ALP(alkaline phosphatase) activity. The results were as follows ; Regardless of cultured time, cellular activities were stimulated by BMP. Also, BMP greatly increased alkaline phosphatase(ALP) in periodontal ligament cells. These results suggest that BMP not only have no cytotoxic effect on periodontal ligament cells, but also have osteogenic stimulatory effect on periodontal ligament cells.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.8
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• pp.1102-1107
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• 2003

The ES cell can provide a useful system for studying differentiation and development in vitro and a powerful tool for producing transgenic animalds. To investigate the culture condition of chicken embryonic stem (CES) cells which can retain their multipotentiality or totipotency, three kinds of feeder layer cells, SNL cells, primary mice embryonic fibroblasts (PMEF) cells and primary chicken embryonic fibroblasts (PCEF) cells, were used as the feeder cells in media of DMEM supplemented with leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF) and stem cell factor (SCF) for co-culture with blastoderm cells from stage X embryos of chicken. The alkaline phosphatase (AKP) test, differentiation experiment in vitro and chimeric chicken production were carried out. The results showed that culture on feeder layer of PMEF yielded high quality CES cell colonies. The typical CES cells clone shape revealed as follows: nested aggregation (clone) with clear edge and round surface as well as close arrangement within the clone. Strong alkaline phosphatase (AKP) reactive cells were observed in the fourth passage cells. On the other hand, the fourth passage CES cells could differentiate into various cells in the absence of feeder layer cells and LIF in vitro. The third and fourth passage cells were injected into the subgerminal cavity of recipient embryos at stage X. Of 269 Hailan embryos injected with CES cells of Shouguang Chickens, 8.2% (22/269) survived to hatching, 5 feather chimeras had been produced. This suggests that an effective culture system established in this study can promote the growth of CES cells and maintain them in the state of undifferentiated and development, which lays a solid foundation for the application of CES cells and may provide an alternative tool for genetic modification of chickens.

• Korean Journal of Veterinary Research
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• v.24 no.1
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• pp.8-16
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• 1984

The work was conducted with tole purpose of investigation on the development pattern of epididymis in accordance with the growth of meat-type cockerels. 1. Histological features of various ductules in epididymis of the cockerel on the age of weeks were as follow: within 10 weeks after hatching rete testis and connecting ductules were well developed but efferent ductules were observed in immature form. During 10th to 20th week, the lining epithelium of various ductules in epididymis was in the developing stage near to the mature form. From 21th week, various ductules were abruptly matured. Lumen of rete testis was lined by simple squamous or simple columnar epithelial cells and that of efferent ductules, having many folds and being larger than any others, were lined by ciliated pseudostratified columnar epithelium with ciliated columnar cells, clear cells and basal cells were noted. Luminal epithelium of connecting ductules was composed of ciliated low pseudostratified columnar epithelial cells, ciliated columnar cells, clear cells and basal cells. The luminal surface of epididymal ducts was pseudostratified columnar epithelium and which was composed of high columnar cells and basal cells. 2. In the India ink absorption test, India ink granules were noted above the nucleus of some cells in the efferent ductules and the connecting ductules at 7 hours after administration of India ink to the mature epididymis, but not absorbed in the other ductules. The granules reactive to acid phosphatase were most abundant in some epithelial cells of efferent ductules and connecting ductules, especially above the nucleus of cells. The granules reactive to alkaline phosphatase were noted on the luminal border of efferent ductules. The granules reactive to PAS were scattered in the epithelial cells of efferent ductules and connecting ductules.

• Korean Journal of Veterinary Research
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• v.14 no.2
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• pp.159-171
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• 1974

Histological and histochemical studies were made on the lining epithelia of the various ducts in epidymis of the Rooster and absorptive function of the canal epithelial cells in the Rooster epididymis were also investigated after administration of India ink. The results obtained were summerized as follows; 1. Epihtelium lining the rate testis was mainaly composed of single later of cuboidal cells, and was partially composed of flattened squamous or low columnar cells. Efferential ductules were characterized by having many villous projections orrfolds which extened into the lumen, and were lined by stereociliated pseudostratified epithelium which consisted of manily ciliated columnar cells, a few scattered clear cells and basal cells. Connecting ductules were lined by ciliated pseudostriatified colummnar epithelium in which ciliated columnar cells, clear cells and basal cells were noted. Epididymal ducts were lined by pseudostratified epidhelium in which columnar and basal cells were noted. 2. PAS-granules, saliva resistant were noted mainly in the epithelial cells of efferential and connecting ductules. 3. Sudan black B stained heavily the granules in the epithelial cells of sufferential and connecting ductules. 4. The granules reactive to acid phosphatase most abundant in the epithelial cells of efferential ductules and were lesser amount in the epithelial cells of connecting ductules where as very few or no granules were seen in the rest of the ducts. 5. Alkaline phosphatase activity was most prominent but discontinuous in the luminal surface of the epithelium of efferential ductules and less marked in the connecting ductless. No enzyme activity was noted in the canal epithelium of epididyml duct. 6. India ink granules were most numerous in the epithlial cells of efferential ductules and were a few in connecting ductules. Very few or no granules of India ink were noted in the other types of the ducts. India ink granules in the epithelium increased gradually as the time after the administration of India ink (one up to twenty-nine hours) has proceeded. From those results it is suggested that epithelial cells of efferential and connecting ductules have active absorptive function, whereas the rest of duct system in the epididymis of the Rooster may be the mere pathway of the seminal fluid without significant modification of its constituents.

• Carbon letters
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• v.22
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• pp.42-47
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• 2017

In this paper, the in vitro biocompatibility of graphene film (GF) with osteoblasts was evaluated through cell adhesion, viability, alkaline phosphatase activity, F-actin and vinculin expressions, versus graphite paper as a reference material. The results showed that MG-63 cells exhibited stronger cell adhesion, better proliferation and viability on GF, and osteoblasts cultured on GF exhibited vinculin expression throughout the cell body. The rougher and wrinkled surface morphology, higher elastic modulus and easy out-of-plane deformation associated with GF were considered to promote cell adhesion. Also, the biomineralization of GF was assessed by soaking in simulated body fluid, and the GF exhibited enhanced mineralization ability in terms of mineral deposition, which almost pervaded the entire GF surface. Our results suggest that graphene promotes cell adhesion, activity and the formation of bone-like apatite. This research is expected to facilitate a better understanding of graphene-cell interactions and potential applications of graphene as a promising toughening nanofiller in bioceramics used in load-bearing implants.

• Korean Journal of Materials Research
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• v.17 no.2
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• pp.107-114
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• 2007

Porous Ti compacts were fabricated by spark plasma sintering (SPS) method and their in vitro and in vivo biocompatibilities were investigated. Alkaline phosphatase (ALP) activity representing the activity of osteoblast was increased when osteoblast-like MG-63 cells were cultured on the Ti powder surface. Some genes related to cell growth were over-expressed through microarray analysis. The porous Ti compact with 32.2% of porosity was implanted in the subcutaneous tissue of rats to confirm in vivo cytotoxicity. 12 weeks post-operation, outer surface and inside the porous body was fully filled with fibrous tissue and the formation of new blood vessels were observed. No inflammatory response was confirmed. To investigate the osteoinduction, porous Ti compact was implanted in the femur of NZW rabbits for 4 months. Active in-growth of new bone from the surrounded compact bone was observed around the porous body. From the results, The porous Ti compacts fabricated by spark plasma sintering might be available for the application of the stem part of artificial hip joint.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.419-428
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• 2019

Phytases are enzymes that can hydrolyze phytate and its salts into inositol and phosphoric acid, and have been utilized to increase the availability of nutrients in animal feed and mitigate environmental pollution. However, the enzymes' low thermostability has limited their application during the feed palletization process. In this study, a combination of B-value calculation and protein surface engineering was applied to rationally evolve the heat stability of Escherichia coli phytase. After systematic alignment and mining for homologs of the original phytase from the histidine acid phosphatase family, the two models 1DKL and 1DKQ were chosen and used to identify the B-values and spatial distribution of key amino acid residues. Consequently, thirteen potential amino acid mutation sites were obtained and categorized into six domains to construct mutant libraries. After five rounds of iterative mutation screening, the thermophilic phytase mutant P56214 was finally yielded. Compared with the wild-type, the residual enzyme activity of the mutant increased from 20% to 75% after incubation at $90^{\circ}C$ for 5 min. Compared with traditional methods, the rational engineering approach used in this study reduces the screening workload and provides a reference for future applications of phytases as green catalysts.

• Korean Journal of Food Science and Technology
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• v.31 no.5
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• pp.1392-1396
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• 1999

This work was to apply the microwave energy to HTST pasteurization of milk in order to prevent undesirable quality changes due to the fouling and overheating on the surface of heat exchanger. A continuous tubulartype microwave pasteurization system was designed using a domestic microwave oven(800w and 2,450MHz). Raw milk was HTST pasteurized$(at\;72^{circ}C\;for\;15\;sec)$ by three methods; by heating in a stainless steel tube immersed in a hot water bath(MP0), by heating in a microwave cavity to a desired temperature and then holding in a hot water bath(MP1) and by both heating and holding in a microwave cavity(MP2). The microbial quality based on the total plate count and Psychotrophic bacterial count was in the order MP0, MP2 and MP1 ; however, the quality difference was not significant(p<0.05) when the initial microbial numbers were involved in the statistical analysis. In addition, the three samples pasteurized by different methods showed the similar microbial quality based on the coliform count and phosphatase activity. The similar microbial quality of the three samples supports the potential use of microwave energy for the pasteurization of milk and other fluid food products.