Extracellular Vesicles Derived from Kefir Grain Lactobacillus Ameliorate Intestinal Inflammation via Regulation of Proinflammatory Pathway and Tight Junction Integrity.

The aim of this study was to demonstrate the anti-inflammatory effect of Lactobacillus kefirgranum PRCC-1301-derived extracellular vesicles (PRCC-1301 EVs) on intestinal inflammation and intestinal barrier function. Human intestinal epithelial cells (IECs) Caco-2 were treated with PRCC-1301 EVs and then stimulated with dextran sulfate sodium (DSS). Real-time RT-PCR revealed that PRCC-1301 EVs inhibited the expression of pro-inflammatory cytokines in Caco-2 cells. PRCC-1301 EVs enhanced intestinal barrier function by maintaining intestinal cell integrity and the tight junction. Loss of Zo-1, claudin-1, and occludin in Caco-2 cells and the colitis tissues was recovered after PRCC-1301 EVs treatment, as evidenced by immunofluorescence analysis. Acute murine colitis was induced using 4% DSS and chronic colitis was generated in piroxicam-treated IL-10-/- mice. PRCC-1301 EVs attenuated body weight loss, colon shortening, and histological damage in acute and chronic colitis models in mice. Immunohistochemistry revealed that phosphorylated NF-κB p65 and IκBα were reduced in the colon tissue sections treated with PRCC-1301 EVs. Our results suggest that PRCC-1301 EVs may have an anti-inflammatory effect on colitis by inhibiting the NF-κB pathway and improving intestinal barrier function.

The Basic Mechanism of Hair Growth Stimulation by Adipose-derived Stem Cells and Their Secretory Factors.

BACKGROUND: Adipose-derived stem cells (ADSCs) are mesenchymal stem cells (MSCs) within the stromal vascular fraction of subcutaneous adipose tissue. ADSCs secrete growth factors and other proteins, and have been used to regenerate skin with satisfactory results. OBJECTIVE: This review focuses on the effect of ADSCs and their secretory factors on the stimulation of hair growth in vitro, ex vivo and in vivo. RESULTS: The conditioned media of ADSCs (ADSC-CM) increases the proliferation rate of human follicular cells. ADSCs-derived proteins improve hair growth and protect human dermal papilla cells against cytotoxic injury caused by androgen and reactive oxygen species. Moreover, ADSC-CM induces the anagen phase and promotes hair growth in mice, and enhances the elongation of hair shafts in ex vivo human hair organ cultures. CONCLUSION: ADSC-CM promotes hair growth in vitro, ex vivo, and in vivo. Given that ADSCs are one of the most accessible sources of MSCs, ADSC-derived proteins may be feasible clinical therapeutic agents for the treatment of hair loss.

Up-to-date Clinical Trials of Hair Regeneration Using Conditioned Media of Adipose-Derived Stem Cells in Male and Female Pattern Hair Loss.

BACKGROUND: The primary roles of mesenchymal stem cells (MSCs) are to maintain the stem cell niche, facilitate recovery after injury, and ensure healthy aging and the homeostasis of organ and tissues. MSCs have recently emerged as a new therapeutic option for hair loss. OBJECTIVE: Since adipose-derived stem cells (ADSCs) are the most accessible sources of MSCs, ADSCbased hair regeneration is investigated. Besides replacing degenerated cells in affected organs, ADSCs exhibit their beneficial effects through the paracrine actions of various cytokines and growth factors. RESULTS: Several laboratory experiments and animal studies have shown that ADSC-related proteins can stimulate hair growth. In addition, we introduce our clinical pilot studies using conditioned media of ADSCs for pattern hair loss in men and women. CONCLUSION: We believe that conditioned media of ADSCs represents a promising alternative therapeutic strategy for hair loss. We also discuss practical therapeutic challenges and the direction of future research.

Clinical use of conditioned media of adipose tissue-derived stem cells in female pattern hair loss: a retrospective case series study.

BACKGROUND: Female pattern hair loss (FPHL) is a common disorder but presents severe psychosocial problems in many female patients. Adipose tissue-derived stem cells (ADSCs) and conditioned media of ADSCs (ADSC-CM) are reported to promote hair growth in vitro. However, there are no clinical reports on the treatment of alopecia using ADSC-CM. OBJECTIVES: This study evaluates our clinical experience in the use of ADSC-CM for the treatment of FPHL. METHODS: A retrospective, observational study of outcomes in 27 patients with FPHL treated with ADSC-CM was performed. To evaluate the efficacy of the treatment, patients' medical records and phototrichographic images were analyzed. RESULTS: The application of ADSC-CM showed efficacy in treating FPHL after 12 weeks of therapy. Hair density increased from 105.4 to 122.7 hairs/cm(2) (P < 0.001). Hair thickness increased from 57.5 µm to 64.0 µm (P < 0.001). None of the patients reported severe adverse reactions. CONCLUSIONS: The application of ADSC-CM is a potential treatment option for FPHL.

Hair growth stimulated by conditioned medium of adipose-derived stem cells is enhanced by hypoxia: evidence of increased growth factor secretion.

Adipose-derived stem cells (ADSCs) and their secretomes mediate diverse skin-regeneration effects, such as wound-healing and antioxidant protection, that are enhanced by hypoxia. We investigated the hair-growth-promoting effect of conditioned medium (CM) of ADSCs to determine if ADSCs and their secretomes regenerate hair and if hypoxia enhances hair regeneration. If so, we wanted to identify the factors responsible for hypoxia-enhanced hair-regeneration. We found that ADSC-CM administrated subcutaneously induced the anagen phase and increased hair regeneration in C(3)H/NeH mice. In addition, ADSC-CM increased the proliferation of human follicle dermal papilla cells (HFDPCs) and human epithelial keratinocytes (HEKs), which are derived from two major cell types present in hair follicles. We investigated the effect of hypoxia on ADSC function using the same animal model in which hypoxia increased hair regrowth. Forty-one growth factors in ADSC-CM from cells cultured under hypoxic or normoxic conditions were analyzed. The secretion of insulin-like growth factor binding protein (IGFBP)-1, IGFBP-2, macrophage colony-stimulating factor (M-CSF), M-CSF receptor, platelet-derived growth factor receptor-beta, and vascular endothelial growth factor was significantly increased by hypoxia, while the secretion of epithelial growth factor production was decreased. It is reasonable to conclude that ADSCs promote hair growth via a paracrine mechanism that is enhanced by hypoxia.

Hypoxia-enhanced wound-healing function of adipose-derived stem cells: increase in stem cell proliferation and up-regulation of VEGF and bFGF.

Adipose-derived stem cells (ADSCs) have been shown to induce wound-healing effects. Because inflammation near the wound area induces oxygen deficiency, it is interesting to elucidate the effect of hypoxia on the function of ADSCs. In this work, we asked: (1) does hypoxia alter the wound-healing function of ADSCs? and (2) what are the major factors responsible for the alteration in the wound-healing function? Effect of hypoxia on the proliferation of ADSCs was first examined that hypoxia (2% O(2)) enhanced the proliferation of ADSCs in either the presence of serum or in the absence of serum. The conditioned medium of ADSCs harvested under hypoxia (hypoCM) significantly promoted collagen synthesis and the migration of human dermal fibroblasts, compared with that in normoxia (norCM). In the animal studies, hypoCM significantly reduced the wound area compared with norCM. Furthermore, mRNA and protein measurements showed that hypoxia up-regulated growth factors such as vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Inhibition of VEGF and bFGF using neutralizing antibodies reversed the migration of the wounded human dermal fibroblasts and the healing of wounds in animal experiment. Collectively, these results suggest that hypoxia increases the proliferation of ADSCs and enhances the wound-healing function of ADSCs, at least partly, by up-regulating the secretion of VEGF and bFGF.

The wound-healing and antioxidant effects of adipose-derived stem cells.

BACKGROUND: The aim of tissue engineering is to repair and regenerate damaged organs using a combination of cells, biomaterials and growth factors. Mesenchymal stem cells within the stromal-vascular fraction of subcutaneous adipose tissue, that is adipose-derived stem cells (ADSCs) have been used in skin repair with satisfactory results. The production and secretion of growth factors has been reported to be an essential function of ADSCs, and diverse regenerative effects of ADSCs in the skin have been demonstrated. OBJECTIVE: Recent research developments concerning the wound-healing and antioxidant effects of ADSCs are briefly described. METHODS: Various experimental results regarding the wound-healing and antioxidant effect of ADSCs are introduced, and the mechanisms and identification of active proteins involved in these function are further discussed. RESULTS/CONCLUSION: Evidence of ADSC differentiation of skin has not been reported in vivo, but ADSCs accelerate wound-healing and exhibit antioxidant effects under various conditions. The wound-healing and antioxidant effects of ADSCs are mainly mediated by the activation of dermal fibroblasts and keratinocytes via the paracrine mechanism. Since ADSCs are easily obtained in large quantities and have an advantage over other stem cell sources, ADSCs and their secretory factors show promise for use in skin repair and regeneration.

Protective role of adipose-derived stem cells and their soluble factors in photoaging.

As individuals age, the skin undergoes changes, such as irregular pigmentation, thinning and loss of elasticity, that are due to both genetic and environmental factors. These changes may worsen, progressing to precancerous and cancerous diseases. Various medical treatments and topical cosmeceuticals have been used to treat some symptoms of photoaging, however, the results have been less than satisfactory. Mesenchymal stem cells within the stromal-vascular fraction of subcutaneous adipose tissue, adipose-derived stem cells (ADSCs), display multi-lineage developmental plasticity and secrete various growth factors that control and manage the damaged neighboring cells. Recently, the production and secretion of growth factors has been reported as an essential function of ADSCs, and diverse regenerative effects of ADSCs have been demonstrated in the skin. For example, conditioned medium from ADSCs (ADSC-CM) stimulated both collagen synthesis and migration of dermal fibroblasts, which improved the wrinkling and accelerated wound healing in animal models. ADSC-CM also inhibited melanogenesis in B16 melanoma cells, and protected dermal fibroblasts from oxidative stress induced by chemicals and UVB irradiation. Therefore, ADSCs and soluble factors show promise for the treatment of photoaging, and this review introduces recent research developments of the ADSCs and ADSC-derived secretory factors regarding this issue.

Prevention of thyroidectomy scar using a new 1,550-nm fractional erbium-glass laser.

BACKGROUND: Surgical scars are a common cosmetic problem that occurs in various surgical fields including dermatology. Diverse trials have been made to prevent this annoying scar formation. Recently, 585- and 595-nm pulsed dye laser irradiation presented satisfactory cosmetic outcome for the treatment of surgical scars. Other fractionated lasers or light devices were also applied for scar treatment. OBJECTIVE: To determine the effectiveness and safety of a newly developed 1,550-nm fractional erbium-glass laser in the prevention of scar formation after total thyroidectomy. MATERIALS AND METHODS: Twenty-seven ethnic South Korean patients with linear surgical suture lines after total thyroidectomy operation were treated with a 1,550-nm fractional erbium-glass laser. The same surgeon performed all of the operations using the same surgical techniques. Each patient was treated four times at 1-month intervals using the same parameters (5- x 10-mm spot size, 10 mJ, 1,500 spot/cm(2), static mode). Initiation of the first irradiation was made approximately 2 to 3 weeks after the thyroidectomy. The scar prevention effects were evaluated each month for 6 months after thyroidectomy. Two kinds of assessment methods were applied in this evaluation. First, the Vancouver Scar Scale (VSS) was used. Second, three independent physicians gave a global assessment valuation to the final cosmetic results: poor (1), fair (2), good (3), or excellent (4). These results were compared with the surgical scars of a control group (patients who denied laser treatments and had no other treatments during the 6 months after total thyroidectomy by the same surgeon). RESULTS: The average VSS score was lower in the laser treatment group. The global assessment also presented better cosmetic outcomes in the treatment group than in the controls. CONCLUSION: A new 1,550-nm fractional erbium-glass laser may efficiently repress the formation and hypertrophy of thyroidectomy scars on the neck, and it can be safely applied in relatively dark Asian skin without noticeable adverse effects.

Antiwrinkle effect of adipose-derived stem cell: activation of dermal fibroblast by secretory factors.

BACKGROUND: Adipose-derived stem cells (ADSC) have wound-healing and antioxidant effects on human skin via secretion of growth factors and activation of dermal fibroblasts. OBJECTIVE: Paracrine mechanism reducing ultraviolet-B (UVB)-induced wrinkles by ADSC is investigated in this study. METHODS AND RESULTS: Wrinkles were induced by an eight-week UVB irradiation, and were significantly improved by the subcutaneous injection of ADSC in hairless mice. In a replica analysis, parameters involving wrinkles were improved with mid-level and high doses of ADSC (1x10(4) and 1x10(5) cells). Dermal thickness and collagen contents in the dermis also were increased in the ADSC-injected groups. To characterize the paracrine mechanism involving the antiwrinkle effect of ADSC, a conditioned medium of ADSC (ADSC-CM) was directly incubated in human dermal fibroblasts (HDF). UVB irradiation reduced the proliferation of HDF, but this was reversed by the pretreatment of ADSC-CM in a dose-dependent manner. In a cell cycle analysis, ADSC-CM decreased the UVB-induced apoptotic cell death, which was demonstrated by the reduced sub-G1 phase of HDF. In addition, the ADSC-CM increased the protein expression of collagen type I and decreased the protein level of matrix metalloproteinase 1 in HDF, which may account for the increased collagen contents in the dermis. CONCLUSIONS: Collectively, these results indicate that the ADSC and its secretory factors are effective for UVB-induced wrinkles, and the antiwrinkle effect is mainly mediated by reducing UVB-induced apoptosis and stimulating collagen synthesis of HDF.

Novel porous matrix of hyaluronic acid for the three-dimensional culture of chondrocytes.

A novel three-dimensional (3D) scaffold of chemically unmodified hyaluronic acid (HA) with minimum cross-linkage was developed for the culture of chondrocytes, thereby to promote cartilage repair. The porous structure of the scaffold was observed by scanning electron microscopy (SEM), and the pore size was controlled by fabrication conditions including swelling time and composition of the HA matrix. Rabbit primary chondrocytes and human chondrocytic cell lines (C-20/A4) were cultured in the HA matrix to investigate whether they can be applied to construct the cartilage tissue in vitro. The chondrocytes retained chondrocytic spherical morphology in this HA matrix. Moreover, results from the MTT assay showed good cellular viability within the HA matrix; optical density increased for up to 28 days, demonstrating that the cells continued to proliferate inside the HA matrix. Phenotypic analysis (RT-PCR, Alcian blue staining and quantification of s-GAG) showed that chondrocytes, when three-dimensionally cultured within the HA matrix, expressed transcripts encoding collagen type II and aggrecan, and produced sulfated glycosaminoglycans (s-GAG), indicating chondrogenic differentiation. The new HA matrix therefore appears as a potentially promising scaffold for the three-dimensional culture of chondrocytes for cartilage tissue engineering.

Adipose-derived stem cells and their secretory factors as a promising therapy for skin aging.

BACKGROUND: Adipose-derived stem cells (ADSCs) and their secretory factors can stimulate collagen synthesis and migration of fibroblasts during the wound healing process. Conventional treatments for skin aging, such as lasers and topical regimens, induce new collagen synthesis via activation of dermal fibroblasts or growth factors. Considering the results of our previous study, ADSCs can also be used for the treatment of skin aging. OBJECTIVE: The aim was to verify the effectiveness of ADSCs for the treatment of skin aging. METHODS: We analyzed secretory factors of ADSCs and intradermally injected ADSCs (1 x 10(6) cells in 1 mL of Hanks' buffered salt solution) and conditioned media of ADSCs on the back of a micropig. In addition, as a pilot study, intradermal injections of purified autologous processed lipoaspirate (PLA) cells were tried with the photoaged skin of one patient. RESULTS: We demonstrated that ADSCs produce many useful growth factors, increase collagen production in animal study, and reverse skin aging in human trial. CONCLUSION: ADSCs and their secretory factors show promise for application in cosmetic dermatology, especially in the treatment of skin aging.

Whitening effect of adipose-derived stem cells: a critical role of TGF-beta 1.

It has been demonstrated that adipose-derived stem cells (ADSCs) secrete cytokines and exhibit diverse pharmacological actions. The present study examined the unknown pharmacological action of ADSCs regarding whitening effects. A conditioned medium of ADSCs (ADSC-CM) was harvested and the whitening effect of ADSC-CM was studied in melanoma B16 cells. ADSC-CM treatment inhibited the synthesis of melanin and the activity of tyrosinase in a dose dependent manner. To clarify the underlying mechanisms of the whitening action of ADSCs, protein levels of melanogenic proteins were measured by Western blot. Although expressions of microphthalmia-associated transcription factor and tyrosinase-related protein 2 (TRP2) remained unchanged, those of tyrosinase and TRP1 were down-regulated. Transforming growth factor-beta1 (TGF-beta 1), a potent regulator of melanogenic proteins, was neutralized by the addition of a blocking antibody to ADSC-CM, and down-regulated expression of tyrosinase and TRP1 was almost reversed. Collectively, these results indicate that secretary factors of ADSC inhibit melanin synthesis by down-regulating the expression of tyrosinase and TRP1, which are mainly mediated by TGF-beta1.

Effect of quilting sutures on hematoma formation after liposuction with dermal curettage for treatment of axillary hyperhidrosis: a randomized clinical trial.

BACKGROUND: Liposuction provides further reduction of axillary sweating and malodor when combined with dermal curettage with sharp rasping cannulas. This aggressive approach is associated with relatively higher rate of hematoma formation when compared to the conventional simple liposuction. OBJECTIVE: The aim of this prospective, randomized, controlled trial was to evaluate the effect of quilting sutures on the incidence of hematoma formation after liposuction-curettage for treatment of axillary hyperhidrosis (AH). MATERIALS AND METHODS: The trial randomized 59 male patients (118 axillae) undergoing liposuction-curettage for AH to quilting procedures (61 axillae) or control group (57 axillae) for intention-to-treat analysis. Outcome measures included the incidence of hematoma formation, operative time, degree of postoperative pain, and amount of analgesics consumption. RESULTS: Quilting sutures significantly reduced the incidence of axillary hematoma from 28.1% to 4.9%. Quilting was associated with the lengthening of operative time but did not affect the postoperative pain. CONCLUSION: Considering its efficacy in reducing postoperative hematoma, quilting is recommended in combination with aggressive liposuction-curettage procedure for treating AH and osmidrosis.

Evidence supporting antioxidant action of adipose-derived stem cells: protection of human dermal fibroblasts from oxidative stress.

BACKGROUND: Mesenchymal stem cells within the stromal-vascular fraction of subcutaneous adipose tissue, adipose-derived stem cells (ADSCs), produced soluble factors and they exhibit diverse pharmacological effects in skin biology. OBJECTIVE: The present study examines the protective effect of ADSCs for human dermal fibroblasts (HDFs) through anti-oxidation in a tert-butyl hydroperoxide (tbOOH) induced oxidative injury model. METHODS AND RESULTS: The conditioned medium of ADSCs (ADSC-CM) was harvested and tested for antioxidant action. ADSC-CM had an antioxidant effect as potent as 100 microM ascorbic acid and various antioxidant proteins were detected in ADSC-CM by proteomic analysis. Morphological change and cell survival assay revealed that incubation with ADSC-CM aided HDFs to resist free radicals induced by tbOOH. In addition, activities of superoxide dismutase and glutathione peroxidase were enhanced in the ADSC-CM treated HDFs which confirmed the study hypothesis that ADSCs protect HDFs through antioxidant action. In a cell cycle analysis, ADSC-CM treatment reversed the apoptotic cell death induced by tbOOH and caused a decrease of sub-G1 cells with respect to untreated cells. The anti-apoptotic effect of ADSC-CM was also reproduced by caspase-3 activity assay. CONCLUSION: These results suggest that ADSCs have potent antioxidant activity and protect HDFs from oxidative injury by decreasing apoptotic cells. Therefore, ADSCs and ADSC-CM are good candidates for control and prevention of skin damage from free radicals in various skin conditions.

Wound healing effect of adipose-derived stem cells: a critical role of secretory factors on human dermal fibroblasts.

BACKGROUND: Adipose-derived stem cells (ADSCs) are a population of pluripotent cells, which have characteristics similar to bone marrow-derived mesenchymal stem cells. Whereas ADSCs have potential applications for the repair and regeneration of various damaged tissues, few studies have dealt with the effect of ADSCs on fibroblasts, which play a key role in skin biology. OBJECTIVE: In this study, we investigated the possible roles of ADSCs in skin wound healing process, especially in the aspect of fibroblast activation-proliferation, collagen synthesis and migratory properties. METHODS AND RESULTS: ADSCs promoted human dermal fibroblast (HDF) proliferation, not only by cell-to-cell direct contact, which was confirmed by co-culture experiment, but also by paracrine activation through secretory factors, resolved by transwell co-culture and culturing with conditioned medium of ADSCs (ADSC-CM). ADSC-CM enhanced the secretion of type I collagen in HDFs by regulating the mRNA levels of extracellular matrix (ECM) proteins: up-regulation of collagen type I, III and fibronectin and down-regulation of MMP-1. Moreover, ADSC-CM showed stimulatory effect on migration of HDFs in in vitro wound healing models. Additional to those in vitro evidences, wound healing effect of ADSCs was also verified with in vivo animal study, resulted that ADSCs significantly reduced the wound size and accelerated the re-epithelialization from the edge. CONCLUSION: Collectively, these data suggest that ADSC is constitutionally well suited for dermal wound healing and secretory factors derived from ADSCs promote wound healing via HDFs and ADSCs can be used for the treatment of photoaging and wound healing.

Mechanism for increased bioavailability of tacrine in fasted rats.

The mechanism associated with the increased bioavailability of tacrine as a result of a 24-h period of fasting was examined in rats. The AUC value for tacrine after the fasting was 52% higher compared with normal feeding when 4 mg kg(-1) of tacrine was orally administered, but the value for velnacrine, a hydroxylated metabolite of tacrine, was reduced by 10%. The relative metabolic ratio of tacrine in urinary excretion (Au(tacrine) divided by Au(velnacrine)) was lower in fasted rats compared with normally fed rats. This clearly shows that metabolism of tacrine is reduced with 24-h fasting after oral administration. Altered intestinal permeation in the fasting state was hypothesized, and the transport of tacrine across the rat intestine was studied. When a fasted intestine was mounted in an Ussing chamber, the mucosal-to-serosal permeability of tacrine was increased to double that for a fed rat intestine. To examine the effect of absorption rate on the hepatic metabolism of tacrine, a direct pyloric vein infusion study was carried out. Compared with an infusion of tacrine for 5 min, a slow infusion of tacrine over a period of 30 or 60 min increased the hepatic metabolism of tacrine and decreased its systemic clearance in rats. Collectively, these results suggest that rapid transport across the intestine aids tacrine in avoiding hepatic first-pass metabolism and enhances its bioavailability in fasted rats. From these findings, we conclude that both oral administration before a meal and a reduction in the dose might be recommended in tacrine therapy considering the serious hepatotoxicity of tacrine in clinical use.

Vitamin D receptor genotypes are not associated with clinical response to calcipotriol in Korean psoriasis patients.

Conflicting results have been reported on the association between BsmI restriction fragment length polymorphism (RFLP) at the vitamin D receptor gene (VDR) locus and the clinical response of psoriasis patients to calcitriol or calcipotriol therapy. We evaluated RFLPs of the VDR gene by analyzing the restriction pattern of polymerase chain reaction products in 55 Korean psoriasis patients receiving topical calcipotriol therapy, and evaluated the clinical response. Of the 55 patients, 43 completed the 8-week treatment protocol, and the response was evaluated as excellent in 9 patients, good in 20, and poor in 14. Thus, in our 43 patients BsmI and ApaI polymorphism in the VDR gene did not correlate with response to calcipotriol. The marked predominance of the b allele in the Korean population precludes the possibility that BsmI polymorphism is associated with clinical response to calcipotriol. The pattern of prevalence of the VDR genotypes in the Korean population is very different from that in Western populations. There were no differences in VDR genotype between controls and psoriasis patients at the BsmI site, but there were significant difference in terms of ApaI RFLP as previously reported. In conclusion, polymorphism analysis of the VDR gene with BsmI and ApaI restriction enzymes in psoriasis patients was not helpful in predicting clinical response to calcipotriol.