Cell attachment and proliferation on the polymer films of triblock copolymer(ester-ether)s comprising po1y (L-1actide) (PLLA) and poly (oxyethylene-co-oxypropylene)(PN) were investigated using 3T3 fibroblasts. It was found that on the tissue culture polystyrene(TCPS) and the PLLA control film the cells could spread well while on the copolymer films the cells showed a rounded morphology without spreading and proliferated weakly. Especially, little cells proliferated on the films of copolymer having a LN composition of 20 wt%. While the water absorption of the copolymer films increased with increasing PN content, the contact angle against water of copolymer films immersed in aqueous medium was almost identical, being slightly lower than that of the PLLA film. These properties were compatible with the results of cell attachment. The in vitro hydrolysis of the films of triblock and multiblock type copolymers was faster with increasing PN content. The increased hydrolyzability, the flexibility and the decreased cell attachment suggested that these copolymers may have high potential as biodegradable materials for medical use.
It is challenging to diagnose metastatic tumors whose cellular morphology is different from the primary. We characterized canine primary pulmonary adenocarcinoma (PAC) and its xenografted tumors by histological and immunohistochemical analyses for critical diagnostic and cancer stem cell (CSC) markers. To generate a tumor xenograft model, we subsequently transplanted the tissue pieces from the PAC into athymic nude mice. Immunohistochemical examination was performed for diagnostic (TTF-1, Napsin A, and SP-A) and CSC markers (CD44 and CD133). The use of CSC markers together with diagnostic markers can improve the detection and diagnosis of canine primary and metastatic adenocarcinomas.
Endometrial tissue is a known source of mesenchymal stem cells (MSCs). We isolated canine endometrial stem cells from canine endometrial tissues using an enzymatic method and confirmed the immunophenotype of mesenchymal stem cells and multilineage differentiation. Canine endometrial tissues were obtained from canine ovariohysterectomy surgery and isolated using 0.2% collagenase type I. We measured the immunophenotype of stem cells using flow cytometry. To confirm the differentiation ability, a trilineage differentiation assay was conducted. In this study, canine endometrialderived MSCs (cEM-MSCs) were isolated by enzyme treatment and showed a spindle-shaped morphology under a microscope. Moreover, cEM-MSCs showed a trilineage differentiation ability. In this study, the canine endometrium was a good source of MSCs.
Park, Se-Il;Moon, Young-Mi;Jeong, Jae-Ho;Jang, Kwang-Ho;Ahn, Myun-Hwan
Journal of Veterinary Clinics
/
v.28
no.5
/
pp.486-496
/
2011
A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.
In order to evaluate taxonomic status of Latcuca hallaisanensis H. $L{\acute{e}}v$., an endemic species of the Jeju-do Island, we investigated fruit wall structure and chromosome morphology. The fruit wall structure had 10-11 obtuse costae in the transverse section. The costa was wholly occupied by libriform fiber cells, and the underlying fibersclereid tissue was only one to three cells layers thick. Also, the intercosta lacked fiber-sclereid layers. Somatic chromosome numbers and karyotype of Latcuca hallaisanensis were recorded for the first time. This diploid species (2n=10) with the same basic number of x=5 has the total chromosome length $23.3{\mu}m$ and the length of each chromosome falls in $1.9{\mu}m-2.9{\mu}m$. It possess the karyotype complement i.e., 3sm+2st and a characteristic chromosome pair (No. 1 and 2) with a secondary constriction at the distal portion of the short arms. The overall similarity in external morphology (involucre, achene etc), chromosome morphology as well as in fruit wall anatomy between Lactuca hallaisanensis and Crepidiastrum s. lat. clearly indicated that this species should be treated as Crepidiastrum, rather than Lactuca.
The purpose of this study was to determine the effects of Taxol pre-treatment to in vitro matured bovine oocytes, and sucrose and trehalose added to vitrification solution on spindle morphology and embryonic development following cryopreservation. Bovine oocytes were collected from ovaries and matured in tissue culture medium 199 (TCM 199) supplemented with 10% Fetal Bovine Serum (FBS), 0.05ng/ml epidermal growth factor, 0.01 IU/ml luteinizing hormone and $1{\mu}g/ml$ estradiol for 22h in $39^{\circ}C$, 5% $CO_2$, TCM 199-HEPES containing 20% FBS was used as basic medium (BM) to prepare vitrification solution. Oocytes were pre-treated with $1\;{\mu}M$ Taxol in maturation medium for 15 min prior to vitrification. Oocytes were exposed to 1.6 M ethylene glycol (EG) and 1.3M dimethyl sulfoxide (DMSO) in BM and then were exposed to 3.2 M EG, 2.6 M DMSO and 0.5 M sucrose in BM or 3.2 M EG, 2.6 M DMSO and 0.5 M trehalose in BM. Oocytes with cumulus cells and oocytes without cumulus cells were considered as control 1 and control 2, respectively and held in TCM 199-HEPES at $39^{\circ}C$. Oocytes were frozen using modified solid surface vitrification and were stored in cryotubes in liquid nitrogen for more than 1 week. Frozen oocytes were thawed in TCM 199-HEPES containing 0.5 M, 0.25 M and 0.1 M sucrose in BM for 2 min, respectively or 0.5 M, 0.25 M and 0.1 M trehalose in BM for 2 min, respectively. Immunoflurorescence staining of oocytes was performed to assess spindle morphology and chromosome configuration of oocytes. The rates of cleavage and blastocyst were examined following in vitro fertilization. Normal spindle morphology rate of oocytes pre-treated with Taxol prior to vitrification was not higher than that of other vitrified groups. Taxol pre-treatment did not increase cleavage and blastocyst formation rates, although control groups showed significantly higher rates (p<0.05). Percentages of normal spindle and embryonic development were not significantly different among vitrified groups regardless of type of sugar. In conclusion, Taxol pre-treatment of oocytes before cryopreservation did not reduce the damage induced by vitrification and subsequently did not improve embryonic development following vitrification. Trehalose may be used as an alternative non-permeating cryoprotectant in vitrification solution.
Kim, Hong-Jun;Kim, Ja-Young;Choi, Go-Ya;Jeong, Seung-Il;Ju, Young-Sung
Korean Journal of Oriental Medicine
/
v.12
no.3
s.18
/
pp.101-115
/
2006
The purpose of this study was to introduce the differential standard of the types of Angelicae Pubescentis Radix. We established the classificatory standard according to the external and internal morphology and the physicochemistrical pattern for the four types of Aucklandiae Radix. The results follow: 1. At the external shape, Angelica pubescens f. biserrata has rising lenticel and dark brown surface, Its section is light gray and its xylem is yellow gray. Aralia continentalis has big stem mark in upper portion, its section has no dense tissue with yellow gray. Heracleum hemsleyanum has dark brown surface and little lenticel, its cortical layer is white yellow and xylem is isabella and powdered. 2. At the internal shape, Angelica pubescens f. biserrata has phloem with half of the root and rare vessel. Aralia continentalis has phloem with two-thirds of the root and it has broad cutting and putting vessel that formed 1-2 row. Heracleum hemsleyanum also has phloem with half of the root and wide scattered latex tube and many large vessel. 3. At the TLC pattern, Heracleum hemsleyanum has remarkable dark spot at $R_{f}$ 0.23 on the sulphuric acid color pattern test, but others have faint. 4. At the HPLC pattern, all samples have generally patterns. But Angelica pubescens f. biserrata shows diminutive continentalic acid content and the peak at Rt 20.278min comes out on Heracleum hemsleyanum, but do not come out on Aralia continentalis and Angelica pubescens f. biserrata. Heracleum hemsleyanum has remarkable peak at Rt 20.278min, but shows no peak at Rt 29.023min unlike Aralia continentalis or Angelica pubescens f. biserrata. Also Aralia continentalis and Angelica pubescens f. biserrata show one remarkable peak at Rt 29.023, but Heracleum hemsleyanum do not show. Consequently, Aralia continentalis and Angelica pubescens f. biserrata are comparable whit continentalic acid content and Heracleum hemsleyanum is comparable with the peak at Rt 20.273 and Rt 29.023. So it is thought that content of continentalic acid and the peaks at Rt 20.278 and Rt 29.023 can apply to differentiate a species from other. It is considered the results of this study will be furnished the basis to succeeding studies and it is needed to extensive comparative study for the same genus-degree of relatedness.
The morphology and cellular characteristics of adventitious roots, viz aerial roots, in the epiphytic American Ivy were examined to reveal structural changes of the aerial root upon surface attachment. Immature aerial roots were composed of parenchyma cells with dense cytoplasm containing plastids, however, the upper and lower epidermis were not distinguished. At early development, electron-dense substances (EDS) were constituents of much of the aerial root tissue, but the distribution of EDS varied within the tissue. The deposits appeared most concentrated in the superficial cell layers, with lesser amounts in cell layers closer to the cortex. Electron micrographs revealed that EDS deposits were always found in the vacuole, and were mainly associated with the tonoplast. While most of them occurred in the vacuole as small spherical deposits adjacent to the tonoplast, some deposits were oddly shaped or larger in size. Many of the vacuoles eventually filled with EDS, but the EDS content in those vacuoles decreased substantially after initial attachment to the surface. When the vacuoles became almost empty, cells near the epidermis already exhibited irregularity in outline. Subsequent breakdown of cellular components took place in the cells while they were still attached to the surface. This study suggests the potential role of EDS as substances involved in the surface attachment of the plant, however, further studies must be conducted to reveal the nature of EDS and the effects of EDS storage within these vacuoles.
These biologic test procedures are designed to test the suitability of P.V.C. made in Korea intended for parenteral preparation, which were based on the U.S. Pharmacopeia XIX "Biologic Test-Plastic Container", Official from July 1, 1975. Healthy adult human blood and rabbits weighing 2\ulcorner.2Kg were used for test materials. Sample P.V.C. were sampled from the medical equipments made in Korea randomly and Control P.V.C. were sampled from the standardized Cobe and Polystan P.V.C. tubes. P.V.C. extract was prepared from a homogeneous P.V.C. samples by incubating 60 square centimeters of the sample per 20 millimeters of sterile pyrogen-free saline at 70\ulcorner for 72 hours or autoclaving at 120\ulcorner for 1 hour. The Implantation Test was designed to evaluate the reaction of living tissue to the plastic by the method of the implantation of the Sample itself into animal tissue. The Systemic Injection Test, the Intracutaneous Test, and the remainders were designed to determine the biological response of animals to plastics by the single-dose injection of specific extracts prepared from a Sample. The results are as follows; 1.Implantation Test - No significant difference for reactions was noted between the Sample treated animal and the Control after 72 hours of implantation. 2.Systemic Toxicity Injection Test - No sign of toxicity and/or death immediately after injection and at 4, 24, 48 hours respectfully after injection. 3.Intracutaneous Test - None of the animals treated with the Sample showed a significantly greater reaction than the observed in the animals treated with Blank. 4.Pyrogen Assay-Only one animal treated with the Sample showed the maximal rise of rectal temperature about 0.2\ulcorner after 3 hours of injection, but remainders showed no change. 5.Hemolytic Index - The positive Control tube of distilled water exhibited complete hemolysis while the negative Control tube and P.V.C. extract were negative demonstrating no hemolysis. 6.Cell Morphology of Erythrocytes and Leukocytes on Stored, Heparinized Human Blood -- There was no significant difference in the morphology of either the Control or Sample extract. 7.Clotting Mechanism of Human Blood in vitro - After allowing to the P.V.C. extract at room temperature for 5 Hours and at 10\ulcorner for 24 hours, there was no appreciable difference in Prothrombin Time under these conditions. 8.Clotting Mechanism of Rabbit in vivo - At the termination of 5 days after intraperitoneal injection of the P.V.C. extract, no significant changes in Clotting Time were observed. According to the above results, it could be concluded that the P.V.C. made in Korea was acceptable for parenteral preparation, especially treated with physiologic saline and/or human blood.man blood.
The most important object of periodontal treatment is the perfect regeneration of destructed periodontal tissue. The healing of periodontal lesion is affected by several cells & factors, which result in formation of long juntional epithelium, root resorption, bony ankylosis or connective tissue attachment. And ideal healing is enhanced by epithilial exclusion or periodontal ligament cell activation. In this investigation, I studied the effect of Zizyphus Fructus extract which enhances biologic activity& collagen synthesis, on the chemotaxis & cell nature. The cells were obtained from interdental area & middle third area of the freshly extracted teeth for the orthodontic purpose. And they were fully incubated in${\alpha}-MEM$ solution containing $100{\mu]g/ml$ penicillin & $100{\mu]g/ml$ streptomycin followed by 6 generation incubation. The test cells were collected by trypsin-EDTA & centrifuge in the fully incubated cells, counted by Hernacyotmeter, incbated $5{\times}10^5/ml$ cells for 24 hours, re-incubated 24 hours in media containing natural extract and photographed. The cells were incubated for 4 hours in 48 well microchemotaxis chamber bisecting upper & lower chamber by 8ug/m pore polycarbonate membrane coating 5mg/ml gelatin solution. The migrated cells in microscope were counted, which meaned cell chemotaxis activity. The study had shown that the morphology of cell was spindle-shaped as the control group, and the subextract test groups were not significantly different. In gingival fibroblasts, the chemotaxis effect of PDGF was statistically significant compared to control group. The Zizyphus Fructus extract was more or less enhanced chemotaxis effect and in $1{\mu}g/ml$ concentration the chemotaxis effect was slightly elevated compared with $10{\mu}g/ml$ concentration. But, among the subextracts, it was not significantly defferent. In PDL cells, the chemotaxis effect of PDGF in statistically significant, and the zizyphus Fructus extract had shown the enhanced effect. The effect was slightly higher in $1{\mu}g/ml$ concentration than 10g/ml concentration,and no significance among the subextracts.
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