• Title/Summary/Keyword: Tissue regeneration.

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Optimization of in vitro lily culture system with different treatments of taurine (타우린 처리에 의한 나리 기내 식물체 생산체계 최적화)

  • Lee, Sang-Hee;Yang, Hwan-Rae;Kim, Sun Tae;Jun, Tae Hwan;Kim, Yong Chul;Kim, Jong Bo
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.484-489
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    • 2017
  • Lilies as cut flowers are one of the most popular ornamental plants in South Korea. It is necessary to develop lily cultivars with high qualities. Therefore, highly efficient propagation systems are needed following release of elite cultivars. In this study, we used taurine treatment to improve the growth conditions including shoot and bulb formation, fresh weight gain, and reduction of rooting and browning. We experimentally evaluated the effect of taurine as a growth stimulator, at concentrations of 0, 2.5, 5, 10, 15 and 20 mg/l. The results showed that 20 mg of taurine enhanced shoot formation by 85% and increased fresh weight 5.5-fold, which was higher than the approximately four-fold increase in the control. In addition, multiple bulb formation rate was increased by 80% and rooting by 82% following exposure to 20 mg/l of taurine. The efficiency of taurine treatment was higher than that of control with 50% multiple bulb formation rate and 60% rooting rate. The browning was 10.6% at 2.5 mg/l of taurine when compared with 0.8% at 20 mg/l. Taurine showed a positive effect on the overall growth of lily plants in terms of increased fresh weight, shoot formation rate, rooting, and formation of multiple bulbs, indicating that taurine can be used as an alternative to amino acids or as an antioxidant such as citrate and vitamin C in plant tissue culture.

MANDIBULAR DISTRACTION OSTEOGENESIS WITH COMPRESSION FORCE - BONE DENSITY, HISTOLOGICAL FINDINGS AND TMJ RESPONSE (압축력을 병용한 하악골 신장술)

  • Hwang, Young-Seob;Heo, June;Kim, Uk-Kyu;Park, Seong-Jin;Hwang, Dae-Seok;Kim, Yong-Deok;Chung, In-Kyo;Kim, Kyu-Cheon
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.531-548
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    • 2006
  • The purpose of this study was to investigate the biomechanical, histologic findings of distracted regenerate and TMJ response in modified distraction osteogenesis (DO) technique combined with compression force as biomechanical stimulation method which has been suggested in 2002, and developed thereafter by authors. This study was performed with two experiments. First experiment was designed to explore the optimal ratio of compression force versus distraction force for the new DO technique. Second experiment was planned to evaluate the reaction of TMJ tissue, especially condyle, disc after application of the DO technique with compression force. Total 52 New Zealand adult male-rabbits with 3.0kg body weight were used for the study. For the first study, 30 adult male-rabbits underwent osteotomy at one side of mandibular body and a external distraction device was applied on each rabbit with same manner. In the control group of 10 rabbits, final 8 mm of distraction with 1 mm rate per day was done with conventional DO technique after 5 latency days. For the experimental group of 20 rabbits, a compression force with 1 mm rate per day was added to the distracted mandible on 3-latency day after over-distraction (over-lengthening). As the amount of the rate of compression versus distraction, experimental subgroup I (10 rabbits) was set up as 2 mm compression versus 10 mm distraction (1/5) and experimental subgroup II (10 rabbits) was set up as 3 mm compression versus 11 mm distraction (about 1/3). All 30 rabbits were set up to obtain final 8 mm distraction and sacrificed on postoperative 55 day to analysis on biomechanical, and histologic findings of the bone regenerates. For second study, 22 adult male-rabbits were used to evaluate TMJ response after the DO method application with compression force. In the control group, 10 rabbits was used to be performed with conventional DO method, on the other hand, in a experimental group of 10 rabbits, 10 mm distraction with 2 mm compression (1/5 ratio) was done. The remaining 2 rabbits served as the normal control group. Histomorphologic examinations on both condyle, histological studies on condyle, disc were done at 1, 2, 3, 4, 7 weeks after distraction force application. The results were as follows: 1. On the bone density findings, the experimental group II (force ratio - 1/3) showed higher bone density than the other experimental group (force ratio - 1/5) and control group (control group - $0,2906\;g/cm^2$, experimental group I - $0.2961\;g/cm^2$, experimental group II - $0.3328\;g/cm^2$). 2. In the histologic findings, more rapid bone maturation like as wide lamellar bone site, more trabeculae formation was observed in two experimental groups compared to the conventional DO control group. 3. In morphologic findings of condyle, there were no differences of size and architecture in the condyle in the control and experimental groups. 4. In histologic findings of condyles, there were thicker fiberous and proliferative layers in experimental group than those of control group until 2 weeks after distraction with compression force. But, no differences were seen between two groups on 3, 4, 7 weeks after compression. 5. In histologic findings of disc, more collagen contents in extracellular matrix, more regular fiber bundles, and less elastin fibers were seen in experimental group than control group until 2 weeks after distraction with compression. But, no differences were seen between two groups on 3, 4, 7 weeks after distraction with compression. From this study, we could identify that the new distraction osteogenesis technique with compression stimulation might improve the quality of bone regeneration. The no remarkable differences on TMJ response between control and experimental groups were seen and TMJ tissues were recovered similarly to normal TMJ condition after 3 weeks.

The Use of MTT Assay, In Vitro and Ex Vivo, to Predict the Radiosensitivity of Colorectal Cancer (In-vitro와 Ex-vivo MTT Assay를 통한 직장암의 방사선치료 감수성 예측 가능성 검증)

  • Kim, Ji-Eun;Kim, Mi-Sook;Kang, Chang-Mo;Kim, Jong-Il;Shin, Hye-Kyung;Choi, Chul-Won;Seo, Young-Seok;Ji, Young-Hoon
    • Radiation Oncology Journal
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    • v.26 no.3
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    • pp.166-172
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    • 2008
  • Purpose: The measurement of radiosensitivity of individuals is useful in radiation therapy. Unfortunately, the measurement of radiation survival using a clonogenic assay, which is the established standard, can be difficult and time consuming. The aim of this study is to compare radiosensitivity results obtained from the MTT and clonogenic assays, and to evaluate whether the MTT assay can be used on clinical specimens. Materials and Methods: HCT-8, LoVo, CT-26, and WiDr were the colon cancer cell lines used for this study. The clonogenic assay was performed to obtain the cell survival curves and surviving fractions at a dose of 2 Gy ($SF_2$) as the standard technique for radiosensitivity. Also, the MTT assay was performed for each of the cell lines (in vitro). To simulate clinical specimens, the cell lines were inoculated into nude mice, removed when the tumors reached 1 cm in diameter, and chopped. Next, the tumors were subjected to the same process involved with the MTT assay in vitro. The inhibition rates (IR) of 10 Gy or 20 Gy of irradiation for in vitro and ex vivo were calculated based on the optical density of the MTT assay, respectively. Results: According to $SF_2$ and the cell survival curve, the HCT-8 and WiDr cell lines were more resistant to radiation than LoVo and CT-26 (p<0.05). The IR was measured by in vitro. The MTT assay IR was 17.3%, 21%, 30% and 56.5% for the WiDr, HCT-8, LoVo and CT-26 cell lines, respectively. In addition, the IR measured ex vivo by the MTT assay was 23.5%, 26%, 38% and 53% in the HCT-8, WiDr, LoVo and CT-26 tumors, respectively. Conclusion: The radiosensitivity measured by the MTT assay was correlated with the measures obtained from the clonogenic assay. This result highlights the possibility that the MTT assay could be used in clinical specimens for individual radiosensitivity assays.

Tracheal Reconstruction with Perichondrial Graft - An Experimental Study in Rabbits - (연골피막편 이식후 기관 결손부위의 재생에 관한 실험적 연구(제 1 보))

  • 이원상;서장수;이성은;홍원표;박찬일
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1982.05a
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    • pp.10.3-11
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    • 1982
  • Recently through the advancement of medical and surgical managements and the development of low pressure cuffed endotracheal tube, incidence of tracheal stenosis was decreased significantly. Though its incidence was decreased markedly, stenosis was developted unfortunately in the situations such as long term use of respirator, heavy infection, trauma of the trachea and long term intubation etc. Tracheal stenosis had been handled with various methods such as mechanical dilatation, tissue graft techniques, luminal augumentation and end to end anastomosis due to their individual advantages but their effects were not satisfactory. In 1959 Lester had been found the regenerated cartilage from the perichondrium of the rib incidentaly. Since then Skoog, Sohn and Ohlsen were reported chondrogenic potential of perichondrium through the animal experiments. Though many different materials have been tried to rebuild stenosis and gaping defect of trachea, tracheal reconstruction has been a perplexing clinical problems. We choose the perichondrium as the graft material because cartilage is the normal supporting matrix of that structure and it will be an obvious advantage to be able to position perichondrium over a defect and obtain new cartilage there. The young rabbits, which were selected as our experimental animals, were sacrified from two to eight weeks after surgery. The results of our experiment were as follows; 1) In control group, the defect site of trachea was covered with fibrosis and vessels but graft site was covered with hypertrophied perichondrium and vessels. 2) Respiratory mucosa was completely regenerated in defect sites both control and grafted groups. 3) The histologic changes of the grafted sites were as follows: 2 weeks- microvessel dilatation, inflammatory reaction, initiation of fibrosis 4 weeks- decreased microvessel engorgement, submucosal fibrosis, decreased inflammatory reaction immatured cartilage island was noted in the grafted perichondrium (one specimen) 6 weeks- mild degree vascular engorgement submucosal fibrosis. chronic inflamatory reaction cartilage island and endochondrial ossification was noted in the grafted perichondrium (Two specimens) 8 weeks- minute vascular engorgement dense submucosal fibrosis. loss of inflammatory reaction. cartilage island was noted in the grafted perichondrium (two specimens) 4) There was no significant differences in regeneration between active surface in and out groups. 5) We observed immatured cartilage islands and endochondrial ossification in the perichondrial grafted groups where as such findings were not noted in control groups except fibrosis. We concluded that perichondrium was the adequate material for the reconstruction of defected trachea but our results was not sufficient in the aspect of chondrogenic potential of perichondrium. So further research has indicated possibility of chondrogenic potential of perichondrium.

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Growth Regulators and Colchicine Treatments for Embryo Culture Efficiency in Barley (보리 배배양 효율증진을 위한 생장조절제와 콜히친처리 효과)

  • Bong Yeon, Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.6
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    • pp.757-767
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    • 1995
  • This experiment was done to determine the optimum concentration of IAA for root development in plants regenerated from the callus culture of barley embryos. Two concentrations of 2,4-D, 3ppm and 5ppm selected as an optimum among five different concentrations in the previous experiment were used for callus induction and proliferation in this experiment. For callus induction, 3ppm of 2,4-D produced 35.6% in immature embryos and 4.4% in mature embryos, while 5ppm gave 33.8% in immature and 5.6% in mature embryos. Out of 320 immature embryos cultured, 111 embryos were induced to calli and 684 plants were produced from them, while only 16 embryos were induced to calli from 320 mature embryos and 92 plants were restored. The rates of callusing and plant regeneration were 34.7%, 214% in immature embryos and 5.0%, 28.7% in mature embryos, respectively. The average root lengths and root numbers of plants restored from callus at five different IAA concentrations of 0ppm, Ippm, 5ppm, l0ppm and 30ppm were 7.9mm, 3.6; 18.4mm, 5.2; 16.1mm, 3.9; 8.5mm, 3.5 and 6.4mm, 3.4, while plants directly obtained from mature embryos were 14.8mm, 4.9; 4.9mm, 3.6; 4.3mm, 3.1; 3.6mm, 2.6 and 3.2mm, 2.1, respectively. Therefore, 1ppm gave the best result for the root. promotion in callus, whle 0ppm, a control, gave the largest root developmemt in embryos. High concentration of lAA(30ppm) in callus and any exogeneous supplement of lAA in embryos negatively affected to the root lengths and root numbers. Genotypic effect was also observed in given four varieties, Bruce, Klages, Olbori and Albori. For chromosome doubling, when 0.1% colchicine was applied on 428 plants under three different conditions such as air circulation, temperatures and growth stages, 319 plants of doubled haploids were obtained so that the rate was 74.5%

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Effect of Resistance Training on Skeletal Muscle Gene Expression in Rats: a Beadarray Analysis (저항성 운동이 골격근 유전자 발현에 미치는 영향: Beadarray 분석)

  • Oh, Seung-Lyul;Oh, Sang-Duk
    • Journal of Life Science
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    • v.23 no.1
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    • pp.116-124
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    • 2013
  • The aim was to examine resistance exercise-related genes after 8 weeks of resistance training. Thirty-two male Sprague-Dawley rats were divided into four groups: 4 weeks sedentary (4 wks CON, n=8), 8 weeks sedentary (8 wks CON, n=8), 4 weeks exercise training (4 wks REG, n=8), and 8 weeks exercise training (8 wks REG, n=8). The rats were trained to climb a 1-m vertical incline (85-degree), with weights secured to their tails. They climbed 10 times, 3 days per week, for 8 consecutive weeks. Skeletal muscle was taken from the flexor halucis longus after the exercise training. After separating the total RNA, large-scale gene expression was investigated by beadarray (Illumina RatRef-12 Expression BeadChip) analysis, and qPCR was used to inspect the beadarray data and to analyze the RNA quantitatively. The detection p-value for the genes was p<0.01, the M-value {M=$log_2$(condition)-$log_2$(reference)} was >1.0, and the DiffScore was >20. In total, the expression of 30 genes significantly increased 4 weeks after the exercise training, and the expression of six genes decreased. At 8 weeks, the expression of five genes significantly increased and that of 12 decreased. Several genes are potentially involved in resistance exercise and muscle hypertrophy, including 1) regulation of cell growth (IGFBP1, PLA2G2A, OKL38); 2) myogenesis (CSRP3); 3) tissue regeneration and muscle development (MUSTN1, MYBPH); 4) hypertrophy (CYR61, ATF3, NR4A3); and 5) glucose metabolism (G6PC, PCK1). These results may help to explain previously reported physiological changes of the skeletal muscle and suggest new avenues for further investigation.

Chondrogenesis of Mesenchymal Stem Cells Derived from Human Umbilical Cord Blood (사람 제대혈 유래 간엽줄기세포로부터 연골세포 분화)

  • Koh, Phil-Ok;Cho, Jae-Hyun;Nho, Kyoung-Hwan;Cha, Yun-Im;Kim, Young-Ki;Cho, Eun-Hae;Lee, Hee-Chun;Jung, Tae-Sung;Yeon, Seong-Chan;Kang, Kyung-Sun;Lee, Hyo-Jong
    • Journal of Veterinary Clinics
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    • v.26 no.6
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    • pp.528-533
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    • 2009
  • In the current study, the mesenchymal stem cells (MSCs) isolated and propagated from the human umbilical cord blood (UCB) were tested for their capabilities of differentiation into chondrocytes in vitro. The mesenchymal progenitor cells (MPCs) collected from UCB were cultured in a low glucose DMEM medium with 10% FBS, L-glutamine and antibiotics. The human MSC colonies were positively stained by PAS reaction. When the immunophenotypes of surface antigens on the MSCs were analyzed by fluorescence-activated cell sorter (FACS) analysis, these cells expressed positively MSC-related antigens of CD 29, CD44, CD 90 and CD105, whereas they did not express antigens of CD14, CD31, CD34, CD45, CD133 and HLA-DR. Following induction these MSCs into chondrocytes in the chondrogenic differentiation medium for 3 weeks or more, the cells were stained positively with safranin O. We clearly confirmed that human MSCs were successfully differentiated into chondrocytes by RT-PCR and immunofluorescent stain of type-II collagen protein. These data also indicate that the isolation, proliferation and differentiation of the hUCB-derived MSCs in vitro can be used for elucidating the mechanisms involved in chondrogenesis. Moreover this differentiation technique can be applied to developing cell-based tissue regeneration or repair damaged tissues.

STUDY OF RAT EPIGASTRIC VESSELS ACCORDING TO THE FREEZING TIME : HISTOLOGIC, HISTOMORPHOMETRIC, IMMUNOHISTOCHEMICAL & SCANNING ELECTRON MICROSCOPIC STUDY (백서 상복부 혈관의 동결시간에 따른 변화에 대한 연구)

  • Kim, Woo-Chan;Lee, Chong-Heon;Kim, Kyung-Wook;Kim, Chang-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.2
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    • pp.89-109
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    • 1999
  • Vascular spasm which has been reported to occur in 25% of clinical cases continues to be a problem in microvascular surgery; When prolonged and not corrected, it can lead to low flow, thrombosis, and replant or free flap failure. Ischemia, intimal damage, acidosis and hypovolemia have been implicated as contributors to the vascular spasm. Although much work has been done on the etiology and prevention of vasospasm, a spasmolytic agent capable of firmly protecting against or reversing vasospasm has not been found. Therefore vascular freezing was introduced as a new safe method that immediately and permanently relieves the vasospasm and can be applied to microsurgical transfers. Cryosurgery can be defined as the deliberate destruction of diseased tissue or relief the vascular spasm in microvascular surgery by freezing in a controlled manner. 96 Sprague Dawley rats each weighing within 250g were used and divided into 2 group, experimental 1 and 2 group. In the experimental 1 group, right epigastric vessels (artery and vein) were freezed with a cryoprobe using $N_2O$ gas for 1 min. In the experimental 2 group, after freezing for 1 min, thawing for 30 secs and repeat freezing for 30 secs. Left side was chosen as control group in both group. We sacrified the experimental animals by 1 day, 3 days, 1 week, 2 weeks, 4 weeks & 5 months and observed the sequential change that occur during regeneration of epigastric vessels using a histologic, histomorphometric, immunohistochemical and SEM study after the vascular freezing. The results were as follows1. In epigastric arteries, internal diameters had statistically significant enlargement in 1 day, 3 days of Exp-1 group and 1 day, 3 days, 1 week & 2 weeks of Exp-2 group. Wall thickness had statistically significant thinning in 2 weeks of Exp-2 group. 2. In epigastric veins, internal diameters had enlargement of statistical significance in 1 day of Exp-1 and Exp-2 group. 3. The positive PCNA reactions in smooth muscle appeared in 1 week and increased until 2 weeks, decreased in 4 weeks. There was no statistical significance between Exp-1 and Exp-2 group. 4. The positive ${\alpha}$-SMA reaction in smooth muscles showed weak responses until 1 week and slowly increased in 2 weeks and showed almost control level in 4 weeks. 5. The positive S-100 reactions in the perivascular nerve bundles showed markedly decrease in 1 day, 3 days and increased after 1 week and showed almost control level in 4 weeks. Exp-1 group had stronger response than Exp-2 group. 6. In SEM, we observed defoliation of endothelial cell and flattening of vessel wall. Exp-2 group is more destroyed and healing was slower than Exp-1 group. To sum up, relief of vasospasm (vasodilatation) by freezing with cryoprobe was originated from the damage of smooth muscle layer and perivascular nerve bundle and the enlargement of internal diameter in vessels was similar to expeimental groups, but Exp-2 group had slower healing course and therefore vessel freezing in microsurgery can be clinically used, but repeat freezing time needs to be studied further.

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CHANGE OF TASTE PREFERENCE AND TASTE BUD AFTER UNILATERAL LINGUAL NERVE TRANSECTION IN RAT (백서 편측 설신경 손상 후 미각 및 설유두의 변화에 대한 연구)

  • Kim, Yoon-Tae;Jeon, Seung-Ho;Yeom, Hak-Ryol;Kang, Jin-Han;Ahn, Kang-Min;Kim, Sung-Min;Jahng, Jeong-Won;Park, Kyung-Pyo;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.6
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    • pp.515-525
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    • 2005
  • Purpose of study: Lingual nerve damage can be caused by surgery or trauma such as physical irriatation, radiation, chemotherapy, infection and viral infection. Once nerve damage occurred, patients sometimes complain taste change and loss of taste along with serious disturbance of tongue. The purpose of this study was to evaluate the effects of unilateral lingual nerve transection on taste as well as on the maintenance of taste buds. Materials & Methods: Male Sprague-Dawley rats weighing 220-250g received unilateral transection of lingual nerve, subjected to the preference test for various taste solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) with two bottle test paradigm at 2, 4, 6, or 8 weeks after the operation. Tongue was fixed with 8% paraformaldehyde. After fixation, they were observed with scanning electron microscope(JSM-$840A^{(R)}$, JEOL, JAPAN) and counted the number of the dorsal surface of the fungiform papilla for changes of fungiform papilla. And, Fungiform papilla were obtained from coronal sections of the anterior tongue(cryosection). After cryosection, immunostaining with $G{\alpha}gust$(I-20)(Santa Cruz Biotechnology, USA), $PLC{\beta}2$(Q-15)(Santa Cruz Biotechnology, USA), and $T_1R_1$(Alpha Diagnostic International, USA) were done. Immunofluorescence of labeled taste bud cells was examined by confocal microscopy(F92-$300^{(R)}$, Olympus, JAPAN). Results: The preference score for salty and sweet tended to be higher in the operated rats with statistical significance, compared to the sham rats. Fungiform papilla counting were decreased after lingual nerve transaction. In 2 weeks, maximum differences occurred. Gustducin and $T_1R_1$ expressions of taste receptor in 2 and 4 weeks were decreased. $PLC{\beta}2$ were not expressed in both experimental and control group. Conclusion: This study demonstrated that the taste recognition for sweet and salty taste changed by week 2 and 4 after unilateral lingual nerve transection. However, regeneration related taste was occurred in the presence of preserving mesoneurial tissue and the time was 6 weeks. Our results demonstrated that unilateral lingual nerve damage caused morphological and numerical change of fungiform papilla. It should be noted in our study that lingual nerve transection resulted in not only morphological and numerical change but also functional change of fungiform papillae.

THE ROLE OF TYPE 2 DIABETES AS A PREDISPOSING RISK FACTOR ON THE PULPO-PERIAPICAL PATHOGENESIS: REVIEW ARTICLE (치수 치근단 병소의 전구 위험요인으로서의 제 2 형 당뇨의 역할에 관한 소고)

  • Kim, Jin-Hee;Bae, Kwang-Shik;Seo, Deog-Gyu;Hong, Sung-Tae;Lee, Yoon;Hong, Sam-Pyo;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.34 no.3
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    • pp.169-176
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    • 2009
  • Diabetes Mellitus (DM) is a syndrome accompanied with the abnormal secretion or function of insulin, a hormone that plays a vital role in controlling the blood glucose level (BGL). Type land 2 DM are most common form and the prevalence of the latter is recently increasing, The aim of this article was to assess whet her Type 2 DM could act as a predisposing risk factor on the pulpo-periapical pathogenesis. Previous literature on the pathologic changes of blood vessels in DM was thoroughly reviewed. Furthermore, a histopathologic analysis of artificially-induced periapical specimens obtained from Type 2 diabetic and DM-resistant rats was compared. Histopathologic results demonstrate that the size of periapical bone destruction w as larger and the degree of pulpal inflammation was more severe in diabetic rats, indicating that Type 2 D M itself can be a predisposing risk factor that makes the host more susceptible to pulpal infection. The possible reasons may be that in diabetic state the lumen of pulpal blood vessels are thickened by atheromatous deposits, and microcirculation is hindered, The function of polymorphonuclear leukocyte is also impair ed and the migration of immune cells is blocked, leading to increased chance of pulpal infection. Also, lack of collateral circulation of pulpal blood vessels makes the pulp more susceptible to infection. These decrease the regeneration capacity of pulpal cells or tissues, delaying the healing process, Therefore, when restorative treatment is needed in Type 2 DM patients, dentists should minimize irritation to the pulpal tissue un der control of BGL.