Dataset for article Targeted delivery of berberine via ROS-sensitive polymersomes enhances its hepatoprotective activity in CCl4-intoxicated mice

--------------------------------------------------------------------------------------------------------------------------- Dataset for article Targeted delivery of berberine via ROS-sensitive polymersomes enhances its hepatoprotective activity in CCl4-intoxicated mice --------------------------------------------------------------------------------------------------------------------------- ReadMe version: 1.0 (2026-06-25) Dataset version: 1.0 (2026-06-25) Dataset DOI: 10.5281/zenodo.21234283 Related article: Iva Suman, Damir Klepac, Martina Vragovic, Hrvoje Krizan, Eliezer Jager, Alessandro Jager, Ewa Pavlova, Martin Hruby and Robert Domitrovic, Targeted delivery of berberine via ROS-sensitive polymersomes enhances its hepatoprotective activity in CCl4-intoxicated mice, Nanoscale Advances (2025), DOI: 10.1039/d5na00706b -------------------------------------------------------------------- CONTACT -------------------------------------------------------------------- Dataset/repository contact (estimated from the supplied example and author list): Martin Hruby / Martin Hruby [e-mail]; [e-mail] ORCID: 0000-0002-5075-261X Institute of Macromolecular Chemistry, Czech Academy of Sciences (IMC CAS) Heyrovsky sq. 2, 162 06 Prague 6, Czech Republic Article corresponding author: Robert Domitrovic [e-mail] Department of Medical Chemistry, Biochemistry and Clinical Chemistry, Faculty of Medicine, University of Rijeka Brace Branchetta 20, 51000 Rijeka, Croatia ______Creators______ Iva Suman (ORCID 0000-0002-0815-2805) University of Rijeka (Investigation; Formal analysis; Supervision; Funding acquisition) Damir Klepac (ORCID 0000-0001-6329-4526) University of Rijeka (Funding acquisition; Methodology; Project administration; Supervision) Martina Vragovic, IMC CAS (Investigation; Methodology; Visualization) Hrvoje Krizan (ORCID 0000-0002-2922-5620) University of Rijeka (Investigation) Eliezer Jager (ORCID 0000-0001-9939-2355) IMC CAS (Resources; Methodology; Supervision) Alessandro Jager, IMC CAS (Resources; Methodology; Supervision) Ewa Pavlova, IMC CAS (Investigation; Methodology) Martin Hruby (ORCID: 0000-0002-5075-261X) IMC CAS (Supervision; Resources; Funding acquisition) Robert Domitrovic (0000-0002-1235-504X ) University of Rijeka (Conceptualization; Methodology; Formal analysis; Resources; Project administration; Funding acquisition) -------------------------------------------------------------------- DATA AVAILABILITY AND ACCESS INSTRUCTIONS -------------------------------------------------------------------- The dataset is openly accessible under the DOI listed above. The license and terms of reuse are shown in the following chapter below. -------------------------------------------------------------------- LICENSE -------------------------------------------------------------------- ______ReadMe file license______ ReadMe by Martin Hruby and co-authors is licensed under CC BY 4.0 License information: https://creativecommons.org/licenses/by/4.0/ ______Dataset license______ Dataset for article Targeted delivery of berberine via ROS-sensitive polymersomes enhances its hepatoprotective activity in CCl4-intoxicated mice by Iva Suman, Damir Klepac, Martina Vragovic, Hrvoje Krizan, Eliezer Jager, Alessandro Jager, Ewa Pavlova, Martin Hruby and Robert Domitrovic is licensed under CC BY 4.0 License information: https://creativecommons.org/licenses/by/4.0/ -------------------------------------------------------------------- DESCRIPTION AND METHODOLOGY -------------------------------------------------------------------- ______About the dataset______ Carbon tetrachloride (CCl4) metabolism generates highly reactive radicals and causes oxidative liver damage. The related study evaluated whether berberine (BER), an isoquinoline alkaloid with hepatoprotective activity but limited bioavailability, can be delivered more effectively using ROS-sensitive polymersomes (PS). The polymersomes are based on an amphiphilic block copolymer, PHPMA37-b-PbAPE42, containing a boronic ester ROS-responsive hydrophobic block. BER-loaded ROS-sensitive polymersomes (BER-PS) were compared with free BER in a mouse model of CCl4-induced liver injury. The attached raw/primary data support characterization and biological evaluation of the BER-loaded polymer systems. They include DLS size-distribution reports and loading/encapsulation calculations for non-responsive PS and ROS-sensitive PS, release curves for BER from non-responsive PS, preliminary cytotoxicity and histology images, and full uncropped western-blot images for oxidative-stress, apoptosis, autophagy, MAPK, and Akt markers. Article-level results reported in the supplied PDF include BER-PS size around 117.8 nm, zeta potential around -12.5 mV, good physical stability, enhanced BER release in ROS-rich medium, reduced ALT/AST and histopathological liver damage after CCl4 intoxication, and stronger amelioration of oxidative stress, apoptosis, autophagy, and inflammation markers by BER-PS compared with free BER. ______Ethics______ The dataset includes animal-study images and western blots derived from liver samples of Balb/C mice, as well as cell-line data. No personal, donor-level, or directly identifiable human data are present in the PowerPoint file. The animal procedures were approved by the Ethics Committee of the Faculty of Medicine, University of Rijeka, and followed European Council Directive 2010/63/EU. ______Materials and sample preparation______ Berberine was purchased from Polyphenols Laboratories AS. The ROS-responsive block copolymer poly[N-(2-hydroxypropyl)methacrylamide]-b-poly[4-(4,4,5,5-tetra-methyl-1,3,2-dioxaborolan-2-yl)benzyl methacrylate] (PHPMA37-b-PbAPE42) was synthesized as previously reported by the authors. A non-responsive polymersome control mentioned in PrimaryData.pdf was based on a non-responsive poly[N-(4-isopropylphenylacetamide)ethyl methacrylate] (PPPhA) block, with reference DOI 10.1021/acs.biomac.4c00282. BER-loaded polymersomes were manufactured using a Dolomite microfluidic setup with a glass micromixer chip. The organic phase was ROS-responsive block copolymer in THF/MeOH (80/20 v/v) at 5.0 mg/mL. Water for injection at pH 7.4 containing 1 mg BER was used as the aqueous phase. Both phases were pumped at 100 microL/min. Non-encapsulated BER and solvents were removed using Amicon Ultra-4 centrifugal filter units, and the product was concentrated to 1 mL. ______Methods of data collection______ *Dynamic light scattering (DLS) and electrophoretic light scattering (ELS) Instrument: Zetasizer Nano ZS, Model ZEN3600 (Malvern Instruments, UK), equipped with a 633 nm He-Ne laser and operating at 173 degrees. Software: Dispersion Technology Software version 6.01 according to the article; the PowerPoint slides contain Zetasizer-style size-distribution reports and result-quality fields. Conditions from the article: 1 mL of PS dispersion (0.2 mg) was measured in disposable polystyrene half-micro cuvettes at controlled temperatures of 25 and 37 degrees C. Each sample was measured as one 45 s run with 10 repetitions. Z-average diameter and PDI were obtained using general-purpose mode. Zeta potential was calculated from electrophoretic mobility using Henry's equation with the Smoluchowski approximation. DLS values visible in MVshortResults.pdf: non-responsive PSs Berberine Z-average 96.57 nm, PDI 0.163, peak 116.9 nm, result quality Good; ROS-PSs Berberine Z-average 85.73 nm, PDI 0.070, peak 93.35 nm, result quality Good. These are values transcribed from the slide and may not be identical to final article Table 1 values because they appear to be working-data slides. *HPLC/UV-vis determination of BER loading and release BER content loaded into PS was determined by HPLC at UV = 270 nm according to the article. The article defines loading content, LC(%), as BER loaded in PS divided by mass of PSs times 100, and encapsulation efficiency, EE(%), as BER loaded in PS divided by BER feeding times 100. Values visible in MVshortResults.pdf: non-responsive PSs Berberine concentration 902.45 microg/mL, sample volume 4.8 mL, LC 18.8%, LE/EE 30.08%; ROS-PSs Berberine concentration 562.753 microg/mL, sample volume 4.8 mL, LC 11.73%, LE/EE 34.19%. The abbreviation LE is used in the slide; the article uses EE for encapsulation efficiency. Release experiments in the article used dialysis with a 6-8 kDa MWCO cellulose membrane, 2.0 mL BER-loaded PS at 0.5 mg/mL, 3 L PBS release medium at 37 degrees C and 350 rpm, with and without 1 mM H2O2. Aliquots of 500 microL from the inner compartment were measured by UV-vis and returned to the dialysis tube. PrimaryData.pdf slide 1 contains a release curve for BER-loaded non-responsive PS in PBS and PBS containing 1 mM H2O2, with axes Time (h) and Cumulative release (%). The slide text states that the non-responsive PS were prepared using a PPPhA block and that BER release in the ROS-rich environment was about 50% higher than in PBS or in the comparison described on the slide. *Cryo-transmission electron microscopy (cryo-TEM) Instrument from the article: FEI Tecnai G2 Spirit TWIN microscope, bright-field mode, 120 kV accelerating voltage. Sample preparation from the article: 4 microL sample applied to holey or lacey carbon grids hydrophilized by glow discharge; excess sample blotted with Whatman no. 1 filter paper; grids plunged into liquid ethane at -182 degrees C and observed at -173 degrees C. Image analysis was performed using ImageJ. Note: no native cryo-TEM image files were supplied separately in the PowerPoint raw-data files, but the article contains cryo-TEM and size-distribution figures. *In vivo mouse study and serum chemistry Model: male Balb/C mice, 2-3 months old, 5 groups with 6 animals per group: control, PS, CCl4, CCl4 + BER, and CCl4 + BER-PS. Treatment: BER 6 mg/kg intraperitoneally, dissolved in DMSO and diluted with saline to 5% DMSO, administered 1 h before CCl4. CCl4 was 10% v/v in olive oil, 2 mL/kg, intraperitoneally. PS and BER-PS were diluted in distilled water and 5% DMSO. Mice were sacrificed 48 h later. Serum ALT, AST, and ALP were measured with a Bio-Tek EL808 Ultra Microplate Reader according to diagnostic kit instructions. Raw numerical serum-enzyme tables were not present in the attached PowerPoint files. *Histopathology, immunohistochemistry, immunofluorescence, and TUNEL Histopathology: paraffin liver sections were stained with hematoxylin and eosin (HE). Liver necrosis was evaluated as the percentage of liver with destroyed hepatocytes using ImageJ version 1.54 g according to the article. Immunohistochemistry: TNF-alpha and NF-kB p65 were detected in paraffin sections with DAKO EnVision+ Peroxidase/DAB kit and analyzed with ImageJ/IHC profiler according to the article. Immunofluorescence: 8-OHdG was detected in paraffin sections using an 8-OHdG antibody and mouse-IgGk BP-CFL 594, imaged with an Olympus BX51 microscope and quantified in ImageJ. TUNEL: DNA fragmentation was detected with a Proteintech TUNEL kit and imaged by fluorescence microscopy. PrimaryData.pdf does not include raw TUNEL images as separate files, but the article contains the TUNEL figure. MVshortResultsb.pdf slide 2 includes working presentation images: an HCT116 24 h XTT bar chart under H2O2-induced oxidative stress and histology images from Balb/C mice comparing control, CCl4, CCl4 + berberine, and CCl4 + berberine-NP. *Western blotting Liver samples were homogenized in RIPA buffer, proteins were separated by SDS-PAGE and transferred to PVDF membranes. The article reports use of antibodies against oxidative-stress, apoptosis, autophagy, MAPK, and Akt markers, chemiluminescent detection, C-DiGit Blot Scanner, and ImageJ densitometry. PrimaryData.pdf slides 2-18 contain full uncropped blot images for 4-HNE, HO-1, cleaved caspase-3/caspase-3, caspase-9/cleaved caspase-9, LC3, p62, p21, GAPDH, p-ERK, ERK, p-p38, p38, p-JNK, JNK, p-Akt, Akt, and JNK + GAPDH. Lanes are annotated as Control, PS, CCl4, BER + CCl4, and BER-PS + CCl4. Molecular-weight markers visible on the slides are in kDa. *Statistical analysis The article reports analysis using StatSoft STATISTICA version 13.1. Differences between groups were assessed by one-way ANOVA followed by Tukey's post hoc test. Values were expressed as mean +/- SD, and P < 0.05 was considered statistically significant. -------------------------------------------------------------------- DATASET STRUCTURE -------------------------------------------------------------------- BER_PS_raw_data.zip (estimated archive name; update to the final repository archive name if different) BER_PS_raw_data/ ├── 000_ReadMe_filled_BER_PS_tracked_changes.docx - this ReadMe document ├── PrimaryData.pdf - primary/source data slides, including release data for BER-loaded non-responsive PS and full uncropped western-blot images └── MVshortResults.pdf - short results slides containing DLS reports, BER loading/encapsulation calculations, preliminary HCT116 XTT chart, and histology images PrimaryData.pdf slide-level contents: Slide 1: BER release from BER-loaded non-responsive PS in PBS and PBS + 1 mM H2O2; PPPhA non-responsive polymer description; time versus cumulative release plot. Slide 2: full uncropped 4-HNE blot image; lanes Control, PS, CCl4, BER + CCl4, BER-PS + CCl4; molecular-weight markers 60, 45, and 35 kDa. Slide 3: full uncropped HO-1 blot image; markers 45, 35, and 25 kDa. Slide 4: full uncropped caspase-3 blot image; markers 25, 20, and 15 kDa. Slide 5: full uncropped caspase-9 and cleaved caspase-9 blot images; markers 60, 45, and 35 kDa. Slide 6: full uncropped LC3 blot image; markers 25, 20, and 15 kDa. Slide 7: full uncropped p62 blot image; markers 75, 60, and 45 kDa. Slide 8: full uncropped p21 blot image; markers 25, 20, and 15 kDa. Slide 9: full uncropped GAPDH blot image; markers 45, 35, and 25 kDa. Slide 10: full uncropped p-ERK blot image; markers 60, 45, and 35 kDa. Slide 11: full uncropped ERK blot image; markers 60, 45, and 35 kDa. Slide 12: full uncropped p-p38 blot image; markers 45, 35, and 25 kDa. Slide 13: full uncropped p38 blot image; markers 45, 35, and 25 kDa. Slide 14: full uncropped p-JNK blot image; markers 75, 60, 45, and 35 kDa. Slide 15: full uncropped JNK blot image; markers 75, 60, 45, and 35 kDa. Slide 16: full uncropped p-Akt blot image; markers 75, 60, and 45 kDa. Slide 17: full uncropped Akt blot image; markers 75, 60, and 45 kDa. Slide 18: JNK + GAPDH full uncropped blot image or combined control image; markers 75, 60, 45, and 35 kDa are visible in the slide text. MVshortResults.pdf slide-level contents: Slide 1: DLS size-distribution by intensity reports and loading calculations for NR-PSs Berberine and ROS-PSs Berberine. Visible values include NR-PSs Z-average 96.57 nm, PDI 0.163, peak 116.9 nm, BER concentration 902.45 microg/mL, volume 4.8 mL, LC 18.8%, LE/EE 30.08%; ROS-PSs Z-average 85.73 nm, PDI 0.070, peak 93.35 nm, BER concentration 562.753 microg/mL, volume 4.8 mL, LC 11.73%, LE/EE 34.19%. Slide 2: preliminary working images: HCT116 colon cancer cell XTT bar chart at 24 h under H2O2-induced oxidative stress; Balb/C mouse liver histology panels labeled Control, CCl4, CCl4 + berberine, and CCl4 + berberine-NP. -------------------------------------------------------------------- FILENAME STRUCTURE -------------------------------------------------------------------- The raw-data package does not use a formal project/WP naming convention. It retains working presentation names assigned during manuscript preparation. The following interpretation should be used: * PrimaryData.pdf = primary/source-data slide deck. It contains the supporting-information release plot for non-responsive PS and full uncropped western-blot images corresponding mainly to article Fig. 6 and Fig. 9 marker panels. * MVshortResults.pdf = short results/working-summary slide deck. It contains DLS report screenshots, BER concentration/loading calculations, and preliminary presentation panels for cell viability and mouse histology. NR-PSs = non-responsive polymersomes; ROS-PSs = ROS-sensitive polymersomes; BER = berberine; PS = polymersomes; NP = nanoparticles/polymersome nanoparticles. The slides sometimes use NP and PS interchangeably for the nanocarrier. Experimental group labels in blot and histology slides: Control; PS; CCl4; BER + CCl4 or CCl4 + BER; BER-PS + CCl4 or CCl4 + BER-PS. Molecular-weight labels on western blot slides are in kDa. DLS diameters are in nm. BER concentrations are in microg/mL. Loading values are in percent. -------------------------------------------------------------------- FILE TYPES & FORMATS, SW TO OPEN AND DIMENSIONS & UNITS -------------------------------------------------------------------- * Annotated Data (.PDF) original format: PPTX (Microsoft PowerPoint Office Open XML) converted format: PDF *DLS/ELS data represented in slides format in supplied files: Zetasizer report screenshots/images embedded in .pptx; no native .dts/.del files supplied SW to inspect values: PowerPoint/Impress for slide viewing; ImageJ/Fiji or OCR/digitization software only if numerical extraction from images is necessary horizontal axis: particle size, d.nm or nm, plotted on logarithmic scale in size-distribution reports vertical axis: intensity, percent summary values: Z-average diameter (d.nm or nm), PDI, intercept, peak size, peak % intensity, peak standard deviation, result quality *HPLC/UV-vis loading and release data represented in PowerPoint format in supplied files: calculation text boxes and graph image embedded in .pptx; no native chromatogram or spreadsheet files supplied SW: PowerPoint/Impress for viewing; original HPLC/UV-vis software is not identified in the supplied files loading dimensions/units: BER concentration (microg/mL), sample volume (mL), LC (%), LE/EE (%) release-plot dimensions/units: time (h) on the x-axis and cumulative release (%) on the y-axis *Western blot data represented in PowerPoint format in supplied files: full uncropped blot images embedded in .pptx; no separate original membrane-image files supplied SW: PowerPoint/Impress for viewing; ImageJ/Fiji for densitometry if required; LI-COR Image Studio or related scanner software may have been used originally, but this is not confirmed in the supplied files dimensions/units: molecular-weight markers in kDa; band intensity in arbitrary optical-density or chemiluminescence units if quantified from images lane labels: Control, PS, CCl4, BER + CCl4, BER-PS + CCl4 *Histology, immunostaining, cell-viability, and illustrative images represented in PowerPoint format in supplied files: microscope or chart images embedded in .pptx SW: PowerPoint/Impress for viewing; ImageJ/Fiji for image analysis if original images or calibrated exports are available dimensions/units: scale bars and magnifications as shown in article figures; MVshortresultsb.pptx slide 2 does not provide fully extractable numeric raw data for the histology images or XTT chart --------------------------------------------------------------------FUNDING--------------------------------------------------------------------* Ministry of Education, Youth and Sports of the Czech Republic: Advanced stimuli-responsive giant vesicles – towards the mimicking of essential cell functions (LUAUS24137) * Ministry of Education, Youth and Sports of the Czech Republic: “New Technologies for Translational Research in Pharmaceutical Sciences/NETPHARM” (CZ.02.01.01/00/22_008/0004607), co-funded by the European Union. * Faculty of Medicine in Rijeka (MT-ERK-100.24.0006) * University of Rijeka (uniri-biomed-18-30) * University of Rijeka (uniri-mladi-biomed-20-17) * University of Rijeka (uniri-iskusni-biomed-23-51) * University of Rijeka (uniri-mladi-biomed-23-18) * University of Rijeka (uniri-mzi-25-5) * University of Rijeka (uniri-iz-25-289)