Supplemental_material_v0.1 – Supplemental material for Bioprinting of three-dimensional dentin–pulp complex with local differentiation of human dental pulp stem cells

Supplemental material, Supplemental_material_v0.1 for Bioprinting of three-dimensional dentin–pulp complex with local differentiation of human dental pulp stem cells by Jonghyeuk Han, Da Sol Kim, Ho Jang, Hyung-Ryong Kim and Hyun-Wook Kang in Journal of Tissue Engineering</p

Additional file 1: of Porcine placenta hydrolysates regulate calcium disturbance in MC3T3-E1 osteoblastic cells

Figure S1. Protective effects of PPHs on Tg-induced cell death in MC3T3-E1 osteoblastic cells (PDF 119 KB

Isolation of amylase regulators from the leaves of <i>Ixeridium dentatum</i>

Two new compounds, one sesquiterpene lactone (1) and one phenylethanoid tautomer (2), together with eleven known compounds (3–13) were isolated from the leaves of Ixeridium dentatum. Their structures were determined by extensive spectroscopic methods, including 1D-, 2D-NMR, and mass spectrometry. All compounds were evaluated for their amylase secretion activity in human salivary gland cells after treatment in 40 mM of high glucose. All compounds showed increased amylase secretion activity. Moreover, previously undescribed compounds (1–2), luteolin 7-O-β-D-glucopyranoside (10), quercimeritrin (11), and quercetin 3-O-β-D-xylopyranoside (13) exhibited significant amylase activity, which is comparable to the positive control.</p

Visualization 1.avi

Real-time movie of a circulating RBC in the blood vessel of mouse buccal mucos

EUE active components geniposide and aucubin regulate palmitate-induced cell death.

<p>Cells were treated with 500 µM palmitate in the presence or absence of 2.5, 5, or 10 µg/mL aucubin or geniposide for 24 hours. Cell viability (A) and caspase-3 activity (B) were analyzed. Immunoblotting was performed with antibody against active caspase-3, caspase-9, or β-actin (C). Cells were incubated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 0, 6, 12, 24, or 48 hours. Cell viability (D) and caspase-3 (E) activity were analyzed. Cells were incubated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 24 or 48 hours. Immunoblotting was performed with antibody against active caspase-3, caspase-9, or β-actin (F). Cells were incubated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 24 hours and stained with Hoechst (G). Cells were incubated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 24 or 48 hours. Immunoblotting was carried out with antibody against BAX, cathepsin B, LAMP-1, or tubulin (H). Cathepsin B activity in the medium was measured (I). Cells were incubated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 24 hours. Immunostaining was performed with anti-LAMP-1 antibody and subsequently with anti-cathepsin B antibody. The degree of overlap in staining was quantified (J). ^*<i>p</i><0.05, significantly different from palmitate-treated condition Pal.; palmitate.</p

Bending Sensor Based on Controlled Microcracking Regions for Application toward Wearable Electronics and Robotics

A soft bending sensor based on the inverse pyramid structure is demonstrated, revealing that it can effectively suppress microcrack formation in designated regions, thus allowing the cracks to open gradually with bending in a controlled manner. Such a feature enabled the bending sensor to simultaneously have a wide dynamic range of bending strain (0.025–5.4%), high gauge factor (∼74), and high linearity (R2 ∼ 0.99). Furthermore, the bending sensor can capture repeated instantaneous changes in strain and various types of vibrations, owing to its fast response time. Moreover, the bending direction can be differentiated with a single layer of the sensor, and using an array of sensors integrated on a glove, object recognition was demonstrated via machine learning. Finally, a self-monitoring proprioceptive ionic electroactive polymer (IEAP) actuator capable of operating in liquid was demonstrated. Such features of our bending sensor will enable a simple and effective way of detecting sophisticated motion, thus potentially advancing wearable healthcare monitoring electronics and enabling proprioceptive soft robotics

EUE reduces hepatic lipotoxicity in rats fed a high-fat diet.

<p>Rats were given a normal diet or a high-fat diet with or without 0.25, 0.5, or 1 g/kg EUE for 10 weeks, and serum and livers were harvested. Liver tissues were loaded with 5 µM dihydroethidium and fluorescence image acquisition was performed (A). Liver tissue was subjected to lipid peroxidation assay (B), caspase-3 activity assay (C), and immunoblotting with antibody against caspase-3, -9, or β-actin (D). Serum levels of AST and ALT were <b>measured (E)</b>. Following subcellular fractionation, immunoblotting with antibody against BAX, t-Bid, PDI, COX II, or LAMP-1 was performed (F). ^*<i>p</i><0.05, significantly different from high-fat diet. HFD; high fat diet, EUE<i>; Eucommia ulmoides Oliver extract.</i></p

EUE regulates palmitate-reduced lysosomal activity.

<p>Cells were treated with 500 µM palmitate in the presence or absence of 100 µg/mL EUE for 24 hours followed by exposure to 5 µM LysoTracker and image acquisition (A). Lysosomal fluorescence was quantified (A; lower). Lysosomal V-ATPase activity was measured as described in Materials and Methods (B). Acridine orange solution and valinomycin were added to cell monolayers and intravesicular H⁺ uptake was initiated by the addition of Mg-ATP (C); fluorescence was quantified at 24 hours (C; right). Cells were treated with 500 µM palmitate in the presence or absence of 100 µg/mL EUE for 0, 6, 12, 24, or 48 hours, and levels of α-galactosidase, α-mannosidase, and acid phosphatase were measured (D). ^*<i>p</i><0.05, significantly different from palmitate-treated condition. DIC; differential interference contrast microscopy, Pal.; palmitate, EUE<i>; Eucommia ulmoides</i> Oliver extract.</p

Lysosomal V-ATPase inhibitor bafilomycin blocks the effect of EUE on lysosomal BAX location and cell death.

<p>Cells were treated with 500 µM palmitate in the presence or absence of 100 µg/mL EUE after pretreatment with 1 µM bafilomycin for 24 hours. Lysosomal V-ATPase activity was measured (A). Acridine orange solution and valinomycin were added to cell monolayers and intravesicular H⁺ uptake was initiated by the addition of Mg-ATP (B); the fluorescence was quantified at 24 hours (B; right). Cell viability assay (C) and caspase-3 activity analysis (D) were performed. Immunostaining was performed with anti-BAX or LAMP-1 antibody and the co-localized BAX was quantified as the percent of lysosomal-translocated BAX (E). Immunoblot analysis of lysosome fractions with antibody against BAX, t-Bid, PDI, COX II, or LAMP-1 (F). ^*<i>p</i><0.05, significantly different from EUE-treated condition in the presence of palmitate. Con; control, Pal.; palmitate, EUE<i>; Eucommia ulmoides</i> Oliver extract, Bafi<i>; Bafilomycin.</i></p

EUE active components geniposide and aucubin enhance lysosomal enzyme activation.

<p>Cells were treated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 24 hours. Lysosomal V-ATPase activity was measured as described in Materials and Methods (A). Cells were treated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 24 hours followed by exposure to 5 µM LysoTracker and image acquisition. The fluorescence was quantified (B). Acridine orange solution and valinomycin were added to cell monolayers and intravesicular H⁺ uptake was initiated by the addition of Mg-ATP (C); the fluorescence was quantified at 24 hours (C; right). Cells were treated with 500 µM palmitate in the presence or absence of 10 µg/mL aucubin or geniposide for 0, 12, 24, or 48 hours and the level of α-galactosidase, α-mannosidase, or acid phosphatase was measured (D). ^*<i>p</i><0.05, significantly different from palmitate-treated condition. Con; control, Pal.; palmitate.</p