Human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) are used in tissue repair and regeneration; however, the mechanisms involved are not well understood. We investigated the hair growth-promoting effects of hUCB-MSCs treatment to determine whether hUCB-MSCs enhance the promotion of hair growth. Furthermore, we attempted to identify the factors responsible for hair growth. The effects of hUCB-MSCs on hair growth were investigated in vivo, and hUCB-MSCs advanced anagen onset and hair follicle neogeneration. We found that hUCB-MSCs co-culture increased the viability and up-regulated hair induction-related proteins of human dermal papilla cells (hDPCs) in vitro. A growth factor antibody array revealed that secretory factors from hUCB-MSCs are related to hair growth. Insulin-like growth factor binding protein-1 (IGFBP-1) and vascular endothelial growth factor (VEGF) were increased in co-culture medium. Finally, we found that IGFBP-1, through the co-localization of an IGF-1 and IGFBP-1, had positive effects on cell viability; VEGF secretion; expression of alkaline phosphatase (ALP), CD133, and ${\beta}-catenin$; and formation of hDPCs 3D spheroids. Taken together, these data suggest that hUCB-MSCs promote hair growth via a paracrine mechanism.
Choi, Youn Kyung;Kang, Jung-Il;Hyun, Jin Won;Koh, Young Sang;Kang, Ji-Hoon;Hyun, Chang-Gu;Yoon, Kyung-Sup;Lee, Kwang Sik;Lee, Chun Mong;Kim, Tae Yang;Yoo, Eun-Sook;Kang, Hee-Kyoung
Biomolecules & Therapeutics
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v.29
no.2
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pp.211-219
/
2021
Alopecia is a distressing condition caused by the dysregulation of anagen, catagen, and telogen in the hair cycle. Dermal papilla cells (DPCs) regulate the hair cycle and play important roles in hair growth and regeneration. Myristoleic acid (MA) increases Wnt reporter activity in DPCs. However, the action mechanisms of MA on the stimulation of anagen signaling in DPCs is not known. In this study, we evaluated the effects of MA on anagen-activating signaling pathways in DPCs. MA significantly increased DPC proliferation and stimulated the G2/M phase, accompanied by increasing cyclin A, Cdc2, and cyclin B1. To elucidate the mechanism by which MA promotes DPC proliferation, we evaluated the effect of MA on autophagy and intracellular pathways. MA induced autophagosome formation by decreasing the levels of the phospho-mammalian target of rapamycin (phospho-mTOR) and increasing autophagy-related 7 (Atg7) and microtubule-associated protein 1A/1B-light chain 3II (LC3II). MA also increased the phosphorylation levels of Wnt/β-catenin proteins, such as GSK3β (Ser9) and β-catenin (Ser552 and Ser675). Treatment with XAV939, an inhibitor of the Wnt/β-catenin pathway, attenuated the MA-induced increase in β-catenin nuclear translocation. Moreover, XAV939 reduced MA-induced effects on cell cycle progression, autophagy, and DPC proliferation. On the other hand, MA increased the levels of phospho (Thr202/Tyr204)-extracellular signal regulated kinases (ERK). MA-induced ERK phosphorylation led to changes in the expression levels of Cdc2, Atg7 and LC3II, as well as DPC proliferation. Our results suggest that MA promotes anagen signaling via autophagy and cell cycle progression by activating the Wnt/β-catenin and ERK pathways in DPCs.
Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.
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
/
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.
Background and Objective: Coptidis Rhizoma is a medicinal herb known for its antioxidant and anti-inflammatory effect. The purpose of this study was to examine the effects of CR on the experimental burn elicitation in vitro and in vivo. Material and Methods: In order to know the antioxidant effect on skin cell of mice after burn elicitation, superoxide dismutase (SOD) activity was measured. In vitro, the RAW 264.7 macrophage cells were treated with lipopolysaccharides for experimental inflammation. iNOS mRNA expression was observed after CR-treatment. In order to know effects on the skin regeneration in the burned mice, we counted the nitric oxide (NO) in blood. We also observed the histological structure in the epidermal basal layer and the dermal section, and we studied changes of angiogenesis in the capillaries surrounding the basal layer and dermal papilla. The changes of transcription of iNOS mRNA (inducible nitric oxide synthase mRNA) and changes of NF-${\kappa}$B (nuclear factor ${\kappa}$B) p65 positive reaction were also observed to investigate the changes of the stress in the skin. Results: The results indicated that CR has significant effects on the antioxidant effect on skin cells of mice after burn elicitation by increasing SOD activity in the in vitro test. It seemed that CR decreased the amount of NF-${\kappa}$B which induced the iNOS mRNA dose-dependently and suppress activating NO and angiogenesis. Furthermore, CR facilitated the process of skin recovery after experimental burn. Conclusion: CR can be applied for burned skin via antioxidant effect and skin regeneration.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.22
no.1
/
pp.62-75
/
2009
Objective : The purpose of this study is to examine the effects of Gamisipjeontang-gamibang(GT) on an experimental bum elicitation. Method : The absorbance of the photo cell mixed with GT at the Abs 560 nm was measured after irradiation for 1 min. In order to know the antioxidant effect on skin cell of mice after burn elicitation, superoxide dismutase(SOD) activity was measured. Also, in order to know effects on the skin regeneration in the burned mice, we counted the eosinophil in blood from animals via cardiac puncture and observed the histological structure in the epidermal basal layer and the dermal section on the 3rd, 7th and 14th day after burn elicitation. We also studied changes in angiogenesis in the capillary surrounding the basal layer and dermal papilla. The changes of HSP70 distribution and changes of p53 positive reaction were observed to investigate the changes of the stress in the skin as well. Result : The results indicated that GT has a significant impact on the antioxidant effect on skin cells of mice after bum elicitation by increasing SOD activity in vitro test. GT seems to decrease MIP-2 which induces neutrophil infiltration and promotes the angiogenesis dose-dependently. Furthermore, GT decreased HSP70, the expression of which was induced by elevated temperatures, and p53 which induce a apoptosis after stress. Conclusion : GT can be applied to burned skin through its antioxidant effect and skin regeneration property.
Park, Ju-Hyun;Kim, Do-Kyung;Park, Jong-Tae;Kim, Su-Young;Yu, Sun-Kyoung;Cho, Kwang-Hee;Kim, Heung-Joong
International Journal of Oral Biology
/
v.35
no.3
/
pp.91-97
/
2010
The effects of chitosan upon the experimentally induced differentiation of MDPC-23 cells, derived from mouse dental papilla cells, were investigated by RT-PCR, observations of cell morphology and Alizaline red-S staining. Chitosan was found to significantly increase and accelerate the expression of ALP mRNA but decrease the ColI transcript levels, as compared with the control, in a time-dependent manner during the differentiation of MDPC-23 cells. Chitosan also significantly downregulated ON mRNA expression and accelerated mineralization in differentiating MDPC-23 cells. These results suggest that chitosan facilitates odontoblast differentiation and mineralization and may have potential clinical applications as a dentin regeneration material.
This study was carried out to investigate the effect of Origanum vulgare extracts on cell proliferation of human hair dermal papilla cell (HHDPC) using sulforhodamine B (SRB) assay, antioxidant activity by 1,1-diphenyl-2-picryl hydrazyl (DPPH) method, expression of insulin-like growth factor-1 (IGF-1) by analyzing reverse transcriptase polymerase chain reaction (RT-PCR) and hair growth in a shaving animal model of C57BL/6 mice topically applying with an amount of 0.1 mL once a day for 3 weeks. The mice were divided into 4 groups including normal group (saline, N), negative control group (dimethyl sulfoxide, NC), positive control group (5 mg/mL minoxidil, PC), and experimental group (Origanum vulgare extracts, OV). Treatment of OV didn't show cytotoxicity in HHDPC up to 10 ${\mu}g/mL$ and exhibited antioxidant activity with $IC_{50}$ of 31.0 ${\mu}g/mL$. IGF-1 expression in the skin was significantly (p<0.05) increased in the PC and OV compared to the N or NC. PC and OV also showed a prominently promoted hair regrowth compared to the N or NC in hair growth observation. The hair regrowth of OV was significantly higher than that of PC (p<0.05). Therefore, these results indicate that O. vulgare extracts effectively stimulated hair growth in an animal model.
Yerim Oh;Jae-Kwan Lee;Heung-Sik Um;Beom-Seok Chang;Jong-bin Lee
Journal of Dental Rehabilitation and Applied Science
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v.39
no.4
/
pp.276-284
/
2023
After tooth extraction, alveolar bone is resorbed over time. Loss of alveolar bone and reduction of upper soft tissue poses difficulties in future implant placement and long-term survival of the implant. This case report focuses on increasing the soft and hard tissues at the implant placement site by using alveolar ridge augmentation and a xenogeneic collagen matrix as a soft tissue substitute in an extraction socket affected by periodontal disease. In each case, the width of the alveolar bone increased to 6 mm, 8 mm, and 4 mm, and regeneration of the interdental papilla around the implant was shown, as well as buccal keratinized gingiva of 4 mm, 6 mm, and 4 mm, respectively. Enlarged alveolar bone facilitates implant surgery, and interdental papillae and keratinized gingiva enable aesthetic prosthesis. This study performed alveolar ridge augmentation on patients with extraction sockets affected by periodontal disease and additionally used soft tissue substitutes to provide a better environment for implant placement and have positive effects for aesthetic and predictive implant surgery.
Journal of Dental Rehabilitation and Applied Science
/
v.33
no.3
/
pp.230-237
/
2017
Regenerative therapy in an interproximal intrabony defect is a challenge due to unaesthetic appearance after surgery. In this article, we introduce a case series of additional use of autogenous periosteal barrier membrane combined with bovine bone mineral and enamel matrix derivative (EMD) in interproximal periodontal intrabony defects to overcome an aforementioned shortcoming. During the periodontal regenerative surgery, autogenous periosteal membrane was additionally adopted besides xenograft material and EMD. Clinical and radiographic examinations were performed before surgery and 6 months after surgical treatment. All clinical parameters were improved and the intrabony defects were resolved on the radiography 6 months after surgery. Moreover, soft tissue esthetics such as the contour of interdental papilla was better than that of conventional regenerative therapy. Periodontal regenerative therapy using several graft materials and bioactive materials was effective in the treatment of periodontal intrabony defect. Moreover, using of autogenous periosteal barrier membrane combined with xenograft and EMD has additional effect for the treatment of an interproximal intrabony defect in terms of augmentation of interdental soft tissue volume.
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