The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.
Lee, So Jin;Kang, Su Ji;Kim, Hye Yun;Lee, Jung Hwan;Kim, Eun Young;Kwon, Soon Yong;Chung, Jin Wha;Joo, Choun-Ki;Khang, Gilson
Polymer(Korea)
/
v.38
no.1
/
pp.24-30
/
2014
Retinal pigment epithelium (RPE) plays an important role in maintaining the visual function and the degeneration of the RPE causes several retinal degeneration disease. In order to fabricate the suitable carrier for RPE transplantation, the hybrid poly(lactide-co-glycolide) (PLGA) film with hesperidin was prepared. Hesperidin has an anti-inflammatory and antioxidant characteristics. ARPE-19 was seeded on hesperidin/PLGA film and then, cell proliferation was determined by the MTT assay, and cell adhesion and cell morphology were confirmed by SEM. Also, RT-PCR was performed to confirm the expression of the specific genes, and AEC immunohistochemical staining was performed to determine the expression of RPE65. As a result, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells were more excellent on hesperidin/PLGA film than PLGA film, thereby we were able to confirm the potential applications of hesperidin/PLGA film as tissue engineering carrier for regeneration of retina.
Kim, Ji-Eun;Lee, Ho-Chan;Kang, Eun-Jeong;Choi, Jung-Wha;Kim, Jong-Han;Park, Soo-Yeon;Jung, Min-Yeong
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.32
no.3
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pp.58-76
/
2019
Objectives : This study was designed to examine the effects of Daecheonglyong-tang(DCL) on atopic dermatitis induced by DNCB in mice Methods : The Nc/Nga mice were divided into 5 groups, and four groups excluding the normal group were applied by 2,4-dinitrochlorobenzene(DNCB), to cause AD and were orally administered with distilled water(negative control), dexamethasone(positive control), and DCL 200 or 400mg/kg once a day for 4 weeks respectively. The visual changes on skin, changes in skin tissue thickness and eosinophil infiltration were observed. IgE, Histamine, Cytokines, immune cells and the amount of gene expression of filaggrin, VEGF, $TGF-{\beta}1$, EGF were measured. Results : Dermatitis score showed a gross improvement on all DCL groups, similar to or better than positive control. All DCL groups showed no significant change in the basophils, while neutrophils and eosinophils decreased. In only DCL 400 mg/kg groups, white blood cells and mononuclear cells were decreased and lymphocytes were increased. In particular, neutrophils had similar or better effects than the positive control. In all DCL groups, IgE, Histamine, $IL-1{\beta}$, IL-4, IL-5, IL-6 and $TNF-{\alpha}$ were decreased and IL-2 was increased. In only DCL 400 mg/kg groups, IL-10 decreased and $IFN-{\gamma}$ increased. In particular, $IL-1{\beta}$ and $IFN-{\gamma}$ showed a similar rate of increase and decrease comparing positive control in DCL 400 mg/kg. $TGF-{\beta}$1 was increased in all DCL groups, filaggrin and VEGF were increased in only DCL 400 mg/kg groups. EGF did not make any changes. Epidermis, dermis thickness and eosinophil infiltration were also decreased in all DCL groups. Conclusions : By increasing Th1 cytokine and decreasing Th2 cytokine, DCL extracts appear to be effective in controlling immune response imbalances, anti-inflammatory and skin regeneration and are likely to be available as a treatment for AD.
Kim, Jong Youn;Adhikari, Prakash Babu;Ahn, Chang Ho;Kim, Dong Hwi;Kim, Young Chang;Han, Jung Yeon;Kondeti, Subramanyam;Choi, Yong Eui
Journal of Ginseng Research
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v.43
no.1
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pp.38-48
/
2019
Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.
Rice (Oryza sativa L.) anthers collected from different genotypes were cultured to investigate the priority to increase callus production rate among several factors affecting on callus formation. Rice varieties, F$_1$ hybrids and F$_2$ plants differed greatly in their abilities to produce callus from anthers, and it was confirmed that the culturability of rice anther was a heritable characteristic. Cold shock treatment before plating anthers promoted callus formation rate, but combined effect with genotype having high culturability was more significant. The response to sucrose concentration in culture medium in callus induction rate was different between rice genotypes, but combined effect with genotype was not significant. Single supplement of NAA to R$_2$ medium increased callus production rate remarkably only in rice genotype having high culturability. Conclusively selection of genotypes is most important to increase callus initiation frequency from rice anthers.
Dongseob Lee;Jungwon Lee;Ki-Tae Koo;Yang-Jo Seol;Yong-Moo Lee
Journal of Periodontal and Implant Science
/
v.53
no.2
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pp.157-169
/
2023
Purpose: The aim of this study was to evaluate the impact of polydeoxyribonucleotide (PDRN) on histologic outcomes when implant placement and lateral sinus floor elevation are performed simultaneously. Methods: Three bimaxillary premolars (P2, P3, and P4) were extracted from 4 beagle dogs 2 months before lateral sinus floor elevation. After lateral elevation of the sinus membrane, each sinus was allocated to either the test or control group. Sinuses underwent either 1) collagenated synthetic bone graft with PDRN following lateral sinus floor elevation (test group) or 2) collagenated synthetic bone graft without PDRN after lateral sinus floor elevation (control group). Eight weeks after the surgical procedure, all animals were euthanised for a histologic and histomorphometric assessment. Augmented height (AH), protruding height (PH), and bone-to-implant contact in pristine (BICp) and augmented (BICa) bone were measured. The composition of the augmented area, which was divided into 3 areas of interest located in coronal, middle and apical areas (AOI_C, AOI_M, and AOI_A), was calculated with 3 parameters: the area percentage of new bone (pNB), residual bone graft particle (pRBP), and fibrovascular connective tissue (pFVT). Results: AH, PH, BICp, BICa total, BICa coronal, and BICa middle values were not significantly different between sinuses in the control and test groups (all P>0.05). The BICa apical of sinuses in the test group (76.7%±9.3%) showed statistically higher values than those of sinuses in the control group (55.6%±22.1%) (P=0.038). pNB, pRBP, and pFVT showed statistically significant differences between the 2 groups in AOI_A (P=0.038, P=0.028, and P=0.007, respectively). pNB, pRBP, and pFVT in AOI_C and AOI_M were not significantly different between samples in the control and test groups (all P>0.05). Conclusions: The histologic findings revealed that lateral sinus floor elevation with PDRN might improve early new bone formation and enable higher bone-to-implant contact.
Tae Gyeom Kim;Jongsoo Kim;Joonhaeng Lee;Jisun Shin;Mi Ran Han;Jongbin Kim;Yujin Kim;Jae Hee Park
Journal of the korean academy of Pediatric Dentistry
/
v.51
no.2
/
pp.149-164
/
2024
This study compared the solubility, water absorption, dimensional stability, release of various ions (hydroxyl, calcium, sulfur, strontium, and silicon), and cytotoxicity of light-cured resin-modified pulp-capping materials. Resin-modified calcium hydroxide (Ultra-blendTM plus, UBP), light-cured resin-modified calcium silicate (TheraCal LCTM, TLC), and dual-cure resin-modified calcium silicate (TheraCal PTTM, TPT) were used. Each material was polymerized; solubility, 24-hour water absorption, and 30- day dimensional stability experiments were conducted to test its physical properties. Solubility was assessed according to the ISO 6876 standard, and 24 hours of water absorption, 30 days of dimensional stability were assessed by referring to the previous protocol respectively. Eluates at 3 and 24 hours and on 7, 14, and 28 days were analyzed according to the ISO 10993-12 standard. And the pH, Ion-releasing ability, cell proliferation rate, and cell viability were assessed using the eluates to evaluate biochemical characteristics. pH was measured with a pH meter and Ion-releasing ability was assessed using inductively coupled plasma atomic emission spectrometry (ICP-AES). Cell proliferation rate and cell viability were assessed using human dental pulp cells (hDPCs). The former was assessed by an absorbance assay using the CCK-8 solution, and the latter was assessed by Live and Dead staining. TPT exhibited lower solubility and water absorption than TLC. UBP and TPT demonstrated higher stability than TLC. The release of sulfur, strontium, calcium, and hydroxyl ions was higher for TLC and TPT than for UBP. The 28-day release of hydroxyl and silicon ions was similar for TLC and TPT. TLC alone exhibited a lower cell proliferation rate compared to the control group at a dilution ratio of 1 : 2 in cell proliferation and dead cells from Live and Dead assay evaluation. Thus, when using light-cure resin-modified pulp-capping materials, calcium silicate-based materials can be considered alternatives to calcium hydroxide-based materials. Moreover, when comparing physical and biochemical properties, TPT could be prioritized over TLC as the first choice.
Purpose: Deproteinized bovine bone or synthetic hydroxyapatite are 2 prevalent bone grafting materials used in the clinical treatment of peri-implant bone defects. However, the differences in bone formation among these materials remain unclear. This study evaluated osteogenesis kinetics in peri-implant defects using 2 types of deproteinized bovine bone (Bio-Oss® and Bio-Oss/Collagen®) and 2 types of synthetic hydroxyapatite (Apaceram-AX® and Refit®). We considered factors including newly generated bone volume; bone, osteoid, and material occupancy; and bone-to-implant contact. Methods: A beagle model with a mandibular defect was created by extracting the bilateral mandibular third and fourth premolars. Simultaneously, an implant was inserted into the defect, and the space between the implant and the surrounding bone walls was filled with Bio-Oss, Bio-Oss/Collagen, Apaceram-AX, Refit, or autologous bone. Micro-computed tomography and histological analyses were conducted at 3 and 6 months postoperatively (Refit and autologous bone were not included at the 6-month time point due to their rapid absorption). Results: All materials demonstrated excellent biocompatibility and osteoconductivity. At 3 months, Bio-Oss and Apaceram-AX exhibited significantly greater volumes of formation than the other materials, with Bio-Oss having a marginally higher amount. However, this outcome was reversed at 6 months, with no significant difference between the 2 materials at either time point. Apaceram-AX displayed notably slower bioresorption and the largest quantity of residual material at both time points. In contrast, Refit had significantly greater bioresorption, with complete resorption and rapid maturation involving cortical bone formation at the crest at 3 months, Refit demonstrated the highest mineralized tissue and osteoid occupancy after 3 months, albeit without statistical significance. Conclusions: Overall, the materials demonstrated varying post-implantation behaviors in vivo. Thus, in a clinical setting, both the properties of these materials and the specific conditions of the defects needing reinforcement should be considered to identify the most suitable material.
Because demineralized bone particle (DBP) contains various bioactive molecules such as cytokines, it is widely used biomaterials in the field of tissue engineering. In this study, we investigated the effect of 2-dimensional DBP/PLGA hybrid films on adhesion, proliferation and phenotype maintenance of intervertebral disc cells. PLGA films incorporated with different amount (0, 10, 20, 40 and 80 wt%) of DBP were prepared by the solvent evaporation method and characterized by scanning election microscopy (SEM). PLGA film has a flat and smooth surface. According to the increase of content of DBP, the surface of DBP/PLGA film exhibited few agglomerates and increased the roughness of the surface. Annulus fibrosus (AF) and nucleus pulposus (NP) cells were cultured on PLGA and DBP/PLGA film surface, and then examined the cell adhesion and proliferation by the cell count and SEM observation. The result of cell count and SEM observation revealed that 10 and 20% DBP in DBP/PLGA films were superior to adhesion and proliferation of both AF and NP cells. We confirmed that specific gene expression of disc cells on DBP/PLGA film based on the cell count result. Disc cells seeded on 20% DBP/PLGA film expressed the gene of type I and II collagen continuously. Therefore, pertinent content of biomaterials could provide more appropriate condition on adhesion and proliferation of cell. And this results may be used as a basic data for the intervertebral disc regeneration using tissue engineering.
Kim, A Ram;Kim, Hye Min;Lee, Jung Keun;Lee, Ji Hye;Song, Jeong Eun;Yoon, Kun Ho;Lee, Dongwon;Khang, Gilson
Polymer(Korea)
/
v.36
no.6
/
pp.768-775
/
2012
Alginate, obtained from the seaweeds, is a widely used biomaterial for cell transplantation, since its positive effect on viability of capsulized cells and its easier encapsulation capability of living cells. Demineralized bone powder (DBP), derived from the natural bone tissue, is widely applied for clinical trials for its low rate of reaction and antigenicity. A chondrocyte was seeded into an alginate with DBP of different contents, and a microcapsule was produced. The adhesion and proliferation of cells was observed through the MTT analysis, and the PCR was applied to estimate the content of the glycosaminoglycan (sGAG) and collagen, and confirm the specific genetic pattern of the chondrocytes. Also, the alginate microcapsule where the chondrocyte is seeded was extracted after transplantation under the skin of a nude mouse, and was immunochemically stained. The experimental result confirmed that the alginate microcapsule containing 1% of DBP not only showed the highest proliferation of cell but had a positive effect of chondrocytes by the interaction between the alginates and the growth factor in DBP. It can be expected that the microcapsule with application of the alginates and DBP might be an appropriate scaffold for tissue engineering.
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