Thanh Minh Dang

This study aimed to develop an accessible, adjustable cell culture model utilizing varying liquid column heights to evaluate the combined microenvironmental impacts of hydrostatic pressure (HP), nutrient availability, and depth‐dependent oxygen gradients on NIH/3T3 fibroblasts. The liquid column medium (LCM) system was constructed by integrating customized glass pipes with standard 48‐well cell culture plates, achieving a maximum column height of 23.3 cm (equivalent to 18 mmHg of base HP). Twenty‐four hours post‐seeding, fibroblasts were exposed to four distinct LCM heights: 0.3 cm (control), 7.7, 15.5, and 23.3 cm, and analyzed after 1 and 3 days of exposure. First‐order mathematical modeling of oxygen diffusion predicted that increasing medium depth concurrently restricts pericellular oxygen availability over extended culture durations. Exposure to LCM heights induced marked cell spreading, elevated cell density along the well margins, and significantly reduced cell proliferation in a height‐dependent manner. By day 3, the highest cell count was observed in the control group (16 × 10 ³ ± 850 cells/well), whereas total cell counts decreased progressively under increasing LCM heights (13 × 10 ³ ± 720, 10 × 10 ³ ± 610, and 6.5 × 10 ³ ± 430 cells/well at 7.7, 15.5, and 23.3 cm, respectively). Furthermore, LCM exposure expanded mean nuclear area and increased the proportion of binucleated cells. Overall, these findings demonstrate that manipulating medium column height provides a versatile, low‐cost in vitro model for exploring cell behaviors under coupled mechanical pressure and fluid depth conditions.

Phenylbutenoids belong to polyphenolic compounds that have demonstrated distinctive biological activities and are primarily characterised in the genus Zingiber (Zingiberaceae) species. From the EtOAc extract of the rhizomes of Zingiber cassumunar Roxb., three phenylbutenoid-type compounds, cassudimin B (1), cassumunol O (2), and cassumunol P (3), were isolated, along with five known compounds (4-8). Their structures were characterised through spectroscopic evidence and reference data. Biological activity investigation revealed that compounds 4, 5, and 8 exhibited promising potential for anti-α-glucosidase with IC₅₀ values of 151.5, 180.1, and 39.5 µM, respectively, surpassing the positive control acarbose (IC₅₀, 190.6 µM). Additionally, compounds 3-8 displayed cytotoxic effects on HepG2 cells ranging from 12.0 to 293.2 µM.

Fibrin gel is appropriate for maintaining quiescence characteristics in primary culture of mouse hepatic stellate cells. Embedded culture of hepatic stellate cells in fibrin gel simulates in vivo cell morphology. Stiffness and adhesion molecules of fibrin gel play a crucial role in the hepatic stellate cell’s primary culture.

Hepatic stellate cell (HSC) activation via the autophagy pathway is a critical factor in liver fibrogenesis. This study tests the hypothesis that chloroquine (CQ) treatment can prevent autophagy and HSC activation in vitro and in vivo in bile-duct-ligated (BDL) mice. Sham-operated and BDL mice were treated with either PBS or CQ in two 60 mg/kg doses the day (D) before and after surgery. On day 2 (2D), HSCs were isolated, and their biological activities were evaluated by measuring intracellular lipid content, α-sma/collagen, and expression of autophagy lc3, sqstm1/p62 markers. The treatment efficacy on liver function was evaluated with serum albumin, transaminases (AST/ALT), and hepatic histology. Primary HSCs were treated in vitro for 24 h with CQ at 0, 2.5, 5, 10, 30, and 50 µM. Autophagy and HSC activation were assessed after 2D of treatment. CQ treatment improved serum AST/ALT, albumin, and bile duct proliferation in 2D BDL mice. This is associated with a suppression of HSC activation, shown by higher HSC lipid content and collagen I staining, along with the blockage of HSC autophagy indicated by an increase in p62 level and reduction in lc3 staining. CQ 5 µM inhibited autophagy in primary HSCs in vitro by increasing p62 and lc3 accumulation, thereby suppressing their in vitro activation. The autophagy inhibitor CQ reduced HSC activation in vitro and in vivo. CQ improved liver function and reduced liver injury in BDL mice.

Biliary atresia is a rare congenital bile duct disease that is the leading cause of liver fibrosis in neonates. Granulocyte colony-stimulating factor (GCSF) is a potential therapy for hepatocellular diseases, but data on GCSF for cholestatic conditions remain limited. The current study examines the role of GCSF in improving bile duct obstruction in mice. Two doses were administered: 10.0 μg/kg/day and 61.5 μg/kg/day, which is the animal equivalent dose of 5.0 μg/kg in humans. Seven days (D7) after bile duct ligation (BDL), Swiss mice were treated with phosphate buffered saline or GCSF for 5 days. The intrahepatic adaptive response of BDL mice was evaluated on postsurgical days D12, D19, and D26. Treatment with 61.5 μg/kg of GCSF resulted in a significant increase in circulating leukocytes and neutrophils on D12. Amelioration of liver injury, as shown by reduced aspartate aminotransferase levels, increased albumin levels and survival rate, as well as reduced intrahepatic inflammation and hepatic myeloperoxidase expression, downregulated ductular proliferation, periportal fibroblast activation, and fibrosis, enhanced expressions of hepatocyte growth factor, peroxisome proliferator-activated receptor-alpha, and ki67, and suppressed expression of cleaved caspase-3 protein, was noted after treatment with 61.5 μg/kg of GCSF. Additionally, GCSF treatment was associated with an increased number of intrahepatic cd3^-Sca1⁺c-Kit⁺ bone marrow cells. Treatment with 61.5 μg/kg of GCSF resulted in liver regeneration and survival in BDL mice was seen, suggesting its potential use for human liver diseases.

Background and Aim: Hepatic stellate cells (HSCs) activation, a critical event in liver fibrosis, has been recently shown to be related to autophagy. Determine whether chloroquine (CQ) could affect (i) the activation of HSC in vivo and (ii) the hepatic damage in a mice acute liver injury model.Methods: The acute liver injury was induced in BALB/c mice by carbon tetrachloride (CCl₄ group); 24 h before and after CCl₄ administration animals were treated by CQ (CCl₄ + CQ group). As control, mice treated by olive oil were considered. After 48 h from CCl₄/olive oil administration, blood samples, liver tissues, and HSCs were harvested for analysis.Results: In vivo, CQ attenuates CCl₄‐induced acute liver damage as evidenced by (i) the reduction of liver enlargement, (ii) the reduction of liver swelling and necrosis also supported by a certain decrease of circulating transaminases level, and (iii) the reduction of liver fibrosis evaluated by collagen deposition and α‐sma protein expression. In HSCs isolated from CQ treated group, we observed the inhibition of autophagy proved by the increase in p62 protein and the decrease of lc3 protein. In addition, CQ reduced the expression of the HSCs activation markers α‐sma/collagen‐I and down‐regulated the expression of the proliferative marker ki67.Conclusion: The autophagy attenuation exerted by CQ together with the reduction of the expression of the proliferation marker in HSCs can lessen the acute liver damage potentially opening the way to novel therapeutic approaches for hepatic fibrosis.

Background: Because of their ease of isolation, high proliferation capacity in vitro, as well as their ability to differentiate into liver cells, Adipose-derived stem cell (ADSCs), are considered to be a promised candidate for liver disease treatment, including liver cirrhosis treatment. Recent studies show that hepatocyte growth factor (HGF) can stimulate ADSC to migrate to the injured areas and platelet rich plasma (PRP) can increase the stemness of ADSCs. Method: In this study, we cultured ADSCs in the medium supplemented 20 ng/ml and 10% PRP; then transplanted them into the cirrhosis mouse model. After 11 weeks of CCl4 induction (1ml/kg, three times per week), mice were divided into 3 groups: (1) PBS group which were injected 0.2 ml PBS; (2) ADSC group which were transplanted 5x105 ADSCs non-pretreated with HGF and PRP; (3) ADSC/HGF+PRP group which were transplanted 5x105 ADSCs pretreated with HGF and PRP via the tail vein. We evaluated the effectiveness of the therapeutic at the day 7th and 14th after transplantation. Results: The results show that the transplantation of ADSC pretreated with HGF and PRP after seven days improves the body weight (increase 4.673%); decreases the ALT level (p < 0.05), the total leukocyte number (p < 0.05) and the expression of Pro-collagen (decrease 4.1 times) as well as α-SMA (decrease 5.1 times), in comparison to the ADSC group. ADSCs pretreated with HGF and PRP also help to improve the liver tissue structure. Conclusion: The therapy using ADSCs pretreated with HGF and PRP is considered to be a promising therapy.