Nucleolin-Targeting AS1411 Aptamer-Conjugated Nanospheres for Targeted Treatment of Glioblastoma.

Post-operative chemotherapy is still required for the treatment of glioblastoma (GBM), for which nanocarrier-based drug delivery has been identified as one of the most effective methods. However, the blood-brain barrier (BBB) and non-specific delivery to non-tumor tissues can significantly limit drug accumulation in tumor tissues and cause damage to nearby normal tissues. This study describes a targeted cancer therapy approach that uses AS1411 aptamer-conjugated nanospheres (100-300 nm in size) loaded with doxorubicin (Dox) to selectively identify tumor cells overexpressing nucleolin (NCL) proteins. The study demonstrates that the active target model, which employs aptamer-mediated drug delivery, is more effective than non-specific enhanced permeability and maintenance (EPR)-mediated delivery and passive drug delivery in improving drug penetration and maintenance in tumor cells. Additionally, the study reveals the potential for anti-cancer effects through 3D spheroidal and in vivo GBM xenograft models. The DNA-protein hybrid nanospheres utilized in this study offer numerous benefits, such as efficient synthesis, structural stability, high drug loading, dye labeling, biocompatibility, and biodegradability. When combined with nanospheres, the 1411 aptamer has been shown to be an effective drug delivery carrier allowing for the precise targeting of tumors. This combination has the potential to produce anti-tumor effects in the active targeted therapy of GBM.

Enhanced delivery to brain using sonosensitive liposome and microbubble with focused ultrasound.

Glioblastoma is considered one of the most aggressive and dangerous brain tumors. However, treatment of GBM has been still challenged due to blood-brain barrier (BBB). BBB prevents that the chemotherapeutic molecules are extravasated to brain. In this study, sonosensitive liposome encapsulating doxorubicin (DOX) was developed for enhancement of GBM penetration in combination with focused ultrasound (FUS) and microbubbles. Upon ultrasound (US) irradiation, microbubbles induce cavitation resulting in the tight junction of BBB endothelium to temporarily open. In addition, the composition of sonosensitive liposome was optimized by comparison of sonosensitivity and intracellular uptake to U87MG cells. The optimal sonosensitive liposome, IMP301-DC, resulted 123.9 ± 38.2 nm in size distribution and 98.2 % in loading efficiency. Related to sonosensitivity of IMP301-DC, US-triggered release ratio of doxorubicin was 69.2 ± 12.3 % at 92 W/cm2 of US intensity for 1 min. In the in vivo experiments, the accumulation of DiD fluorescence probe labeled IMP301-DC-shell in the brain through the BBB opening was increased more than two-fold compared to that of Doxil-shell, non-sonosensitive liposome. US exposure significantly increased GBM cytotoxicity of IMP301-DC. In conclusion, this study demonstrated that IMP301-DC could serve as an alternative solution to enhance the penetration to GBM treatment via BBB opening by non-invasive FUS combined with microbubbles.

Sandwich-structure transferable free-form OLEDs for wearable and disposable skin wound photomedicine.

Free-form optoelectronic devices can provide hyper-connectivity over space and time. However, most conformable optoelectronic devices can only be fabricated on flat polymeric materials using low-temperature processes, limiting their application and forms. This paper presents free-form optoelectronic devices that are not dependent on the shape or material. For medical applications, the transferable OLED (10 µm) is formed in a sandwich structure with an ultra-thin transferable barrier (4.8 µm). The results showed that the fabricated sandwich-structure transferable OLED (STOLED) exhibit the same high-efficiency performance on cylindrical-shaped materials and on materials such as textile and paper. Because the neutral axis is freely adjustable using the sandwich structure, the textile-based OLED achieved both folding reliability and washing reliability, as well as a long operating life (>150 h). When keratinocytes were irradiated with red STOLED light, cell proliferation and cell migration increased by 26 and 32%, respectively. In the skin equivalent model, the epidermis thickness was increased by 39%; additionally, in organ culture, not only was the skin area increased by 14%, but also, re-epithelialization was highly induced. Based on the results, the STOLED is expected to be applicable in various wearable and disposable photomedical devices.

The Co-Expression Pattern of p63 and HDAC1: A Potential Way to Disclose Stem Cells in Interfollicular Epidermis.

Stem cell markers of interfollicular epidermis (IEF) have not been established thus far. The aim of this study is to suggest a new way to disclose IFE-stem cells by combining the expression of histone deacetylases (HDAC) 1 and p63. Immunohistochemical staining of HDAC1 and p63 was performed in six normal human samples. Moreover, a skin equivalent (SE) model was treated with suberoylanilohydroxamic acid (SAHA, an HDAC inhibitor) to elucidate the role of HDAC1. Finally, rapidly adhering (RA) keratinocytes to a type IV collagen, which have been identified to represent epidermal stem cells, were subjected to Western blot analysis with antibodies against HDAC1. In normal samples, there was a minor subpopulation comprised of p63-positive and HDAC1-negative cells in the basal layers. The proportion of this subpopulation was decreased with age. In the SE model, SAHA treatment increased the epidermal thickness and number of p63-positive cells in a dose dependent manner. After SAHA treatment, the expression of differentiation markers was decreased, while that of basement membrane markers was increased. In a Western blot analysis, HDAC1 was not expressed in RA cells. In conclusion, the combination of p63-positive and HDAC1-negative expressions can be a potential new way for distinguishing epidermal stem cells.

Phlorizin, an Active Ingredient of Eleutherococcus senticosus, Increases Proliferative Potential of Keratinocytes with Inhibition of MiR135b and Increased Expression of Type IV Collagen.

E. senticosus extract (ESE), known as antioxidant, has diverse pharmacologic effects. It is also used as an antiaging agent for the skin and phlorizin (PZ) is identified as a main ingredient. In this study, the effects of PZ on epidermal stem cells were investigated. Cultured normal human keratinocytes and skin equivalents are used to test whether PZ affects proliferative potential of keratinocytes and how it regulates these effects. Skin equivalents (SEs) were treated with ESE and the results showed that the epidermis became slightly thickened on addition of 0.002% ESE. The staining intensity of p63 as well as proliferating cell nuclear antigen (PCNA) is increased, and integrin α6 was upregulated. Analysis of ESE confirmed that PZ is the main ingredient. When SEs were treated with PZ, similar findings were observed. In particular, the expression of integrin α6, integrin ß1, and type IV collagen was increased. Levels of mRNA for type IV collagen were increased and levels of miR135b were downregulated. All these findings suggested that PZ can affect the proliferative potential of epidermal cells in part by microenvironment changes via miR135b downregulation and following increased expression of type IV collagen.

Novel Antioxidant Tripeptide "ACQ" Can Prevent UV-Induced Cell Death and Preserve the Number of Epidermal Stem Cells.

We found that tripeptide "ACQ: alanine-cysteine-glutamine" has significant DPPH scavenging activity compared to that of glutathione. Antioxidant effects of ACQ were tested in in vitro and in vivo models. When treated with H2O2, mock treated fibroblasts and keratinocytes showed strong staining by H2DCFA. But, ACQ showed good protective effects against hydrogen peroxide treatment. When mice were fed for 2 or 4 weeks, similar protective effects were observed. In the control group, epidermis was severely damaged by UV irradiation and apoptotic keratinocytes were observed. There were also numerous TUNEL positive cells. But in the ACQ group, epidermis became thicker and there was no sign of severe damage. Interestingly, the number of p63 cells was also higher in ACQ fed mice. To confirm the stem cell rescuing effects of ACQ, three-dimensional skin samples were constructed. Results showed that ACQ increased the expression of integrin α6 and the number of p63 positive cells. These findings showed that ACQ has good antioxidant activity and may increase stem cell activities by the regulation of integrin α6.

Histidine decarboxylase expression influences the neofolliculogenesis of newborn mouse dermal cells.

BACKGROUND: With the introduction of hair regeneration techniques using epidermal and dermal cells, hair follicle regeneration became much easier and faster. Current success has been dependent on the availability of cells from newborn or embryonic mice. We recently observed that the hair-inducing ability of newborn mouse dermal cells disappeared in the first few days of life and there was a drastic decrease of histidine decarboxylase (HDC) gene expression from postnatal day 0 (p0) to day 7 (p7). OBJECTIVE: The aim of this study was to study the role of HDC in hair follicle induction. METHODS: The mRNA levels of HDC in p0, p7 and p48 C57BL/6 mouse skin were checked with a real time reverse transcription polymerase chain reaction and immunohistochemistry. To test the effect of HDC, HDC expression in p0 mouse dermal cells was suppressed with small interfering RNA (siRNA) transfection. Mock treated and HDC siRNA treated cells were then injected with adult epidermal cells into nude mice skin. Three weeks later, the number, length and thickness of induced hairs were compared. RESULTS: Compared with p0, the mRNA level of HDC was much lower at p7 and p48. Immunohistochemical staining also revealed a marked decrease of HDC expression in p7 mice skin, compared with p0 skin. Hair patch assays showed that the HDC siRNA treated p0 dermal cells induced less hair follicle structures and shorter and thinner hair shafts than mock treated cells. CONCLUSION: HDC, whose expression is remarkably downregulated during the first few days after birth in dermal cells of mice, plays essential roles in the hair-inducing ability of newborn mouse dermal cells.

Effects of murine dermal cells on the regulation of hair growth is dependent on the cell number and post-natal age of newborn mice.

Dermal cells from neonatal mice can initiate the formation of hair follicles (HFs) when combined with adult mouse epidermal cells and transplanted subcutaneously into athymic mice. In the present study, the effects of dermal cells on HF formation were tested in terms of total cell number and the time course of cell harvest. Results demonstrated that the number of dermal cells is critical to the formation of HF. Furthermore, hair forming ability is rapidly decreasing as the neonatal mice age. To examine potential differences in gene expression, cDNA array was performed. Results demonstrate that numerous molecules which are directly involved in receptor and signaling correlated with decreased hair inductivity in early time points after delivery. It is reported that bone morphogenic protein (BMP)-6 and Wnt3a treatment increased hair inductivity of dermal papilla cells. But in our study, no changes were observed in the expression levels of BMP-6 and Wnt3a. However, several Wnt related genes demonstrate increased or decreased expression levels. Thus, our results suggest that co-ordinated regulation of these molecules will be important in hair neogenesis within our model system.

Erythrocyte malondialdehyde and glutathione levels in vitiligo patients.

BACKGROUND: Vitiligo is an acquired and progressive hypomelanotic disease that manifests as circumscribed depigmented patches on the skin. Although the precise mechanism remains to be elucidated, an imbalance of the oxidant/antioxidant system has been proposed as an important etiologic mechanism. OBJECTIVE: The objective of this study was to evaluate the oxidant/antioxidant status of vitiligo patients at the erythrocyte level. METHODS: Fifty-three vitiligo patients and 65 phototype-, age-, and sex-matched healthy controls were included in this study. Blood samples were collected from all subjects, and all patients were instructed to answer a questionnaire. RESULTS: Erythrocyte levels of malondialdehyde (MDA) and glutathione (GSH) were measured. All patients were told to answer a questionnaire regarding their habitual behavior, including frequency of smoking and type of diet. We observed significantly lower levels of GSH in vitiligo patients, but the levels of MDA did not differ between patients and controls. Vitiligo patients who smoked showed significantly lower GSH levels compared to non-smoking patients, but the levels of MDA were unchanged between the 2 groups. CONCLUSION: From our results, we conclude that reduced erythrocytic or systemic GSH levels constitute a distinctive feature in vitiligo patients regardless of disease activity.

Relationships between visceral adipose tissue measurement site and the metabolic syndrome in the Korean population.

OBJECTIVE: Visceral adipose tissue (VAT) is presumed to play an important role in the development of metabolic syndrome (MS). The purpose of this study was to evaluate the influence of measurement location of VAT on the cardiometabolic risk factors and the MS in the Korean population. METHODS: To assess abdominal fat distribution, 5 single-slice computed tomography (CT) images were obtained in 470 healthy subjects. The five CT images were obtained at the intervertebral space from L1 to S1 using known anatomical landmarks. Multiple logistic regression analysis was performed to assess the relationship between regional adipose tissue areas and MS. RESULTS: All risk factors were more closely correlated with VAT than subcutaneous adipose tissue (SAT), except waist circumference and blood pressure. Images located at L2-L3 or L3-L4 provided high correlations between VAT area and all cardiometabolic risk factors. The highest adjusted odds (per SD) between VAT and MS were the L2-L3 image in men (OR 4.53) and the L1-L2 in women (OR 4.87), which was higher than measures at L4-L5 (OR 3.22 in men, OR 4.71 in women). However, differences in OR between L1-L2 VAT (OR 4.87) and L4-L5 (OR 4.71) were not great in women. CONCLUSIONS: The results of this study suggest that VAT has a stronger association with MS than ASAT in Korean population regardless of measurement site, and an image located in the upper abdomen (L2-L3 or L3-L4) would be a better predictor of the relationship between VAT and MS in Korean men.