The impacts of e-beam and X-ray irradiation on medical-grade high-density polyethylene (HDPE) are compared with that of gamma irradiation to evaluate their potential effects on pharmaceutical applications. An extensive suite of analytical techniques was employed to evaluate the chemical and physical transformations of irradiated HDPE. Key findings indicated that irradiation did not significantly alter the mechanical properties, as tensile strength and elongation at break remained stable across all irradiation types and doses. Thermal analysis via Differential Scanning Calorimetry (DSC) revealed a slight decrease in melting temperature at higher doses, with an equivalent melting temperature peak observed for all three irradiation technologies. Electron Spin Resonance (ESR) detected alkyl and allyl radicals, which decreased over time, showing no significant differences attributable to irradiation type or dose. Colorimetric analysis indicated yellowing in the samples, linked to specific additives. The quantification of methionine oxidized byproducts by High-Performance Liquid Chromatography (HPLC) demonstrated that the oxidation potential was equivalent for the three irradiation technologies for HDPE. The study concluded that there was a lack of significant impact of irradiation technologies on several physical, chemical and mechanical properties of HDPE.
{"title":"Comparative analysis of irradiation technologies on high-density polyethylene (HDPE) for biopharmaceutical applications","authors":"Blanche Krieguer , Fabien Girard , Samuel Dorey , Nathalie Dupuy , Sylvain R.A. Marque","doi":"10.1016/j.ejps.2025.107394","DOIUrl":"10.1016/j.ejps.2025.107394","url":null,"abstract":"<div><div>The impacts of e-beam and X-ray irradiation on medical-grade high-density polyethylene (HDPE) are compared with that of gamma irradiation to evaluate their potential effects on pharmaceutical applications. An extensive suite of analytical techniques was employed to evaluate the chemical and physical transformations of irradiated HDPE. Key findings indicated that irradiation did not significantly alter the mechanical properties, as tensile strength and elongation at break remained stable across all irradiation types and doses. Thermal analysis via Differential Scanning Calorimetry (DSC) revealed a slight decrease in melting temperature at higher doses, with an equivalent melting temperature peak observed for all three irradiation technologies. Electron Spin Resonance (ESR) detected alkyl and allyl radicals, which decreased over time, showing no significant differences attributable to irradiation type or dose. Colorimetric analysis indicated yellowing in the samples, linked to specific additives. The quantification of methionine oxidized byproducts by High-Performance Liquid Chromatography (HPLC) demonstrated that the oxidation potential was equivalent for the three irradiation technologies for HDPE. The study concluded that there was a lack of significant impact of irradiation technologies on several physical, chemical and mechanical properties of HDPE.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107394"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145676725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-12-10DOI: 10.1016/j.ejps.2025.107406
Yi Liu , Guiyun Song , Daniel Banov , Jennifer Denison , Courtaney Davis , Kendice Ip
This study aims to compare the single-dose pharmacokinetic profiles of semaglutide administered via sublingual, oral, and injectable routes in Sprague–Dawley rats. Semaglutide was delivered sublingually in a proprietary anhydrous suspension vehicle. Rats were randomized into five groups and received the following treatments: subcutaneous injection (0.011 mg/kg), sublingual suspension (1 mg/kg, prepared from either commercial tablets or peptide powder), and oral tablets (1 mg/kg and 20 mg/kg). Semaglutide was detectable in plasma within 2 minutes post-dosing in all groups except the oral 1 mg/kg group. Sublingual administration demonstrated lower variability in plasma concentrations compared to oral dosing. At 1 mg/kg, the sublingual route achieved a significantly higher area under the curve (AUC) than oral (82.53 vs.15.08 ng*h/ml, p=0.004), indicating improved bioavailability. The maximum plasma concentration (Cmax) was reached within 30 minutes for oral and sublingual routes, and at 8 hours for subcutaneous injection. The relative bioavailability was 0.06% for oral 1 mg/kg, 0.16% for oral 20 mg/kg, and 0.34% and 0.29% for sublingual 1 mg/kg using tablets or powder, respectively. No significant difference in AUC was observed between sublingual semaglutide prepared from oral tablets versus powder. These results highlight the potential of sublingual delivery of semaglutide and suggest this route may improve absorption while reducing variability. This proof-of-concept study supports further development of sublingual semaglutide formulations and pharmacokinetics research in humans.
{"title":"Single-dose pharmacokinetics of sublingual semaglutide in rats","authors":"Yi Liu , Guiyun Song , Daniel Banov , Jennifer Denison , Courtaney Davis , Kendice Ip","doi":"10.1016/j.ejps.2025.107406","DOIUrl":"10.1016/j.ejps.2025.107406","url":null,"abstract":"<div><div>This study aims to compare the single-dose pharmacokinetic profiles of semaglutide administered via sublingual, oral, and injectable routes in Sprague–Dawley rats. Semaglutide was delivered sublingually in a proprietary anhydrous suspension vehicle. Rats were randomized into five groups and received the following treatments: subcutaneous injection (0.011 mg/kg), sublingual suspension (1 mg/kg, prepared from either commercial tablets or peptide powder), and oral tablets (1 mg/kg and 20 mg/kg). Semaglutide was detectable in plasma within 2 minutes post-dosing in all groups except the oral 1 mg/kg group. Sublingual administration demonstrated lower variability in plasma concentrations compared to oral dosing. At 1 mg/kg, the sublingual route achieved a significantly higher area under the curve (AUC) than oral (82.53 <em>vs.</em>15.08 ng*h/ml, <em>p</em>=0.004), indicating improved bioavailability. The maximum plasma concentration (C<sub>max</sub>) was reached within 30 minutes for oral and sublingual routes, and at 8 hours for subcutaneous injection. The relative bioavailability was 0.06% for oral 1 mg/kg, 0.16% for oral 20 mg/kg, and 0.34% and 0.29% for sublingual 1 mg/kg using tablets or powder, respectively. No significant difference in AUC was observed between sublingual semaglutide prepared from oral tablets <em>versus</em> powder. These results highlight the potential of sublingual delivery of semaglutide and suggest this route may improve absorption while reducing variability. This proof-of-concept study supports further development of sublingual semaglutide formulations and pharmacokinetics research in humans.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107406"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145735623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-12-17DOI: 10.1016/j.ejps.2025.107419
Nienke Johanna Maria Klaassen , Milou Boswinkel , Nino Chiron Morsink , Alexandra Gil Arranja , Guillaume Cornelis Maria Grinwis , Janneke Molkenboer-Kuenen , Jan Willem Hesselink , Sebastiaan Alexander van Nimwegen , Johannes Frank Wilhelmus Nijsen
Introduction
Holmium-166 microspheres are suitable for micro-brachytherapy for various solid tumours, due to their favourable characteristics regarding treatment and imaging. The purpose of this study was to investigate acute- to long-term toxicity, 3 days to 12 months, biodegradation and in vivo holmium release of two types of holmium-based microspheres, containing poly(L-lactic acid) (Ho-PLLA-MS) or hydroxide (Ho-(OH)3−MS), homogeneously suspended in an injection fluid comprised by colloidal microcrystalline cellulose (MCC) and phosphate buffer in healthy mice.
Methods
250 mice were included in the study. Non-radioactive holmium-165 microsphere suspensions, were injected subcutaneously on the flank of 150 mice, animals were intensively monitored. CT imaging was performed to visualise the location of the injection site, the distribution of the microspheres and the injection fluid and their possible translocation. Injection site and other relevant tissues were assessed histologically and possible holmium leakage or microsphere migration was examined by analysing the holmium content in the injection site and relevant tissues via inductively coupled plasma analysis (ICP).
Results
The injection of holmium-165 microspheres suspended in MCC-based injection fluid caused local inflammation ranging from slight to severe. No relevant deviations in haematological or biochemical parameters indicative for acute or chronic systemic toxicity were observed. ICP analysis and CT imaging showed no signs of microsphere translocation from the injection site to other organs.
Conclusion
These findings indicate that the injection fluid (MCC in phosphate buffer) in combination with Ho-(OH)3−MS or Ho-PLLA-MS can potentially be used as administration fluid for controlled localized microsphere delivery. However, prior to clinical application, clinically relevant doses have to be tested in sensitive anatomical regions.
{"title":"Preclinical safety assessment of holmium-based microspheres for micro-brachytherapy in mice","authors":"Nienke Johanna Maria Klaassen , Milou Boswinkel , Nino Chiron Morsink , Alexandra Gil Arranja , Guillaume Cornelis Maria Grinwis , Janneke Molkenboer-Kuenen , Jan Willem Hesselink , Sebastiaan Alexander van Nimwegen , Johannes Frank Wilhelmus Nijsen","doi":"10.1016/j.ejps.2025.107419","DOIUrl":"10.1016/j.ejps.2025.107419","url":null,"abstract":"<div><h3>Introduction</h3><div>Holmium-166 microspheres are suitable for micro-brachytherapy for various solid tumours, due to their favourable characteristics regarding treatment and imaging. The purpose of this study was to investigate acute- to long-term toxicity, 3 days to 12 months, biodegradation and <em>in vivo</em> holmium release of two types of holmium-based microspheres, containing poly(L-lactic acid) (Ho-PLLA-MS) or hydroxide (Ho-(OH)<sub>3</sub>−MS), homogeneously suspended in an injection fluid comprised by colloidal microcrystalline cellulose (MCC) and phosphate buffer in healthy mice.</div></div><div><h3>Methods</h3><div>250 mice were included in the study. Non-radioactive holmium-165 microsphere suspensions, were injected subcutaneously on the flank of 150 mice, animals were intensively monitored. CT imaging was performed to visualise the location of the injection site, the distribution of the microspheres and the injection fluid and their possible translocation. Injection site and other relevant tissues were assessed histologically and possible holmium leakage or microsphere migration was examined by analysing the holmium content in the injection site and relevant tissues via inductively coupled plasma analysis (ICP).</div></div><div><h3>Results</h3><div>The injection of holmium-165 microspheres suspended in MCC-based injection fluid caused local inflammation ranging from slight to severe. No relevant deviations in haematological or biochemical parameters indicative for acute or chronic systemic toxicity were observed. ICP analysis and CT imaging showed no signs of microsphere translocation from the injection site to other organs.</div></div><div><h3>Conclusion</h3><div>These findings indicate that the injection fluid (MCC in phosphate buffer) in combination with Ho-(OH)<sub>3</sub>−MS or Ho-PLLA-MS can potentially be used as administration fluid for controlled localized microsphere delivery. However, prior to clinical application, clinically relevant doses have to be tested in sensitive anatomical regions.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107419"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145793553","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-11-29DOI: 10.1016/j.ejps.2025.107392
Xinhao Wan , Ming Yang , Zhijian Zhong , Xiaoqing Zhou , Xuecheng Wang , Qing Tao , Zhenfeng Wu
With the modernization of the pharmaceutical industry and increasingly stringent regulatory requirements, ensuring the quality and process stability of pharmaceutical tablets has become a critical challenge. In particular, chewable tablets, as a unique type of oral solid dosage form designed to be chewed before swallowing, require special attention to mechanical properties and sensory attributes such as texture, mouthfeel, and taste acceptability. This study aims to implement a forward design-based multi-attribute decision-making (MADM) framework for systematic and accurate multi-criteria quality assessment and prediction of pharmaceutical chewable tablets. Tablets were produced using batch-based tableting technology, and critical quality attributes—including weight variation, friability, porosity, and hardness—were systematically evaluated under varying process conditions. Tensile strength was employed as an external validation index. To overcome the limitations of traditional evaluation methods, the proposed MADM framework incorporates a game theory (GT)-based combinatorial weighting strategy, effectively integrating subjective experience-based judgment with objective data-driven metrics. In addition, a backpropagation neural network (BPNN) model was developed to capture the nonlinear relationships between process parameters and the comprehensive quality index. Compared to conventional weighting approaches such as the analytic hierarchy process (AHP), entropy method (EM), and mean-based weighting, the GT-based strategy demonstrated superior performance in terms of evaluation robustness and accuracy. The MADM results showed strong agreement with the tensile strength data, confirming the reliability and consistency of the evaluation framework. Furthermore, the BPNN model achieved high prediction accuracy, providing solid quantitative support for process optimization. Overall, the proposed method offers a robust and generalizable solution for multi-criteria quality assessment and prediction of pharmaceutical chewable tablets. When integrated with industrial information systems, it enables data-driven optimization, intelligent quality control, and automation, thereby contributing to the advancement of smart pharmaceutical manufacturing.
{"title":"Application of a forward design-based multi-attribute decision-making method in quality assessment in pharmaceutical tablet manufacturing","authors":"Xinhao Wan , Ming Yang , Zhijian Zhong , Xiaoqing Zhou , Xuecheng Wang , Qing Tao , Zhenfeng Wu","doi":"10.1016/j.ejps.2025.107392","DOIUrl":"10.1016/j.ejps.2025.107392","url":null,"abstract":"<div><div>With the modernization of the pharmaceutical industry and increasingly stringent regulatory requirements, ensuring the quality and process stability of pharmaceutical tablets has become a critical challenge. In particular, chewable tablets, as a unique type of oral solid dosage form designed to be chewed before swallowing, require special attention to mechanical properties and sensory attributes such as texture, mouthfeel, and taste acceptability. This study aims to implement a forward design-based multi-attribute decision-making (MADM) framework for systematic and accurate multi-criteria quality assessment and prediction of pharmaceutical chewable tablets. Tablets were produced using batch-based tableting technology, and critical quality attributes—including weight variation, friability, porosity, and hardness—were systematically evaluated under varying process conditions. Tensile strength was employed as an external validation index. To overcome the limitations of traditional evaluation methods, the proposed MADM framework incorporates a game theory (GT)-based combinatorial weighting strategy, effectively integrating subjective experience-based judgment with objective data-driven metrics. In addition, a backpropagation neural network (BPNN) model was developed to capture the nonlinear relationships between process parameters and the comprehensive quality index. Compared to conventional weighting approaches such as the analytic hierarchy process (AHP), entropy method (EM), and mean-based weighting, the GT-based strategy demonstrated superior performance in terms of evaluation robustness and accuracy. The MADM results showed strong agreement with the tensile strength data, confirming the reliability and consistency of the evaluation framework. Furthermore, the BPNN model achieved high prediction accuracy, providing solid quantitative support for process optimization. Overall, the proposed method offers a robust and generalizable solution for multi-criteria quality assessment and prediction of pharmaceutical chewable tablets. When integrated with industrial information systems, it enables data-driven optimization, intelligent quality control, and automation, thereby contributing to the advancement of smart pharmaceutical manufacturing.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107392"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145654019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-12-06DOI: 10.1016/j.ejps.2025.107402
Xiaoqing Zhuang , Jenni Virta , Heidi Liljenbäck , Lauri Paasonen , Anu J. Airaksinen , Anne Roivainen , Xiang-Guo Li
Nanofibrillar cellulose (NFC) hydrogel has emerged as a promising implantable material for therapeutic applications. In this study, the relatively longer-lived positron-emitting radionuclide zirconium-89 was chelated to the octadentate deferoxamine (DFO*)-conjugated NFC hydrogel ([89Zr]Zr-DFO*-NFC) to enable longitudinal monitoring of its in vivo fate using positron emission tomography techniques. Following subcutaneous implantation in healthy mice, [89Zr]Zr-DFO*-NFC retained radioactivity at the implant site for at least 14 days, with minimal signal detected in the kidneys, urinary bladder, and overlying skin. In contrast, mice receiving a control formulation of [89Zr]Zr-oxalate mixed with NFC hydrogel showed progressive accumulation of radioactivity in the bones, consistent with known [89Zr]Zr-oxalate distribution patterns, and only limited retention at the implant site by Day 7. These findings demonstrate that [89Zr]Zr-DFO*-NFC hydrogel implants exhibit high in vivo stability with negligible systemic release following subcutaneous implantation, supporting their potential use as safe and traceable biomaterial platforms.
{"title":"Longitudinal monitoring of nanofibrillar cellulose hydrogel medical implants in mice using positron emission tomography","authors":"Xiaoqing Zhuang , Jenni Virta , Heidi Liljenbäck , Lauri Paasonen , Anu J. Airaksinen , Anne Roivainen , Xiang-Guo Li","doi":"10.1016/j.ejps.2025.107402","DOIUrl":"10.1016/j.ejps.2025.107402","url":null,"abstract":"<div><div>Nanofibrillar cellulose (NFC) hydrogel has emerged as a promising implantable material for therapeutic applications. In this study, the relatively longer-lived positron-emitting radionuclide zirconium-89 was chelated to the octadentate deferoxamine (DFO*)-conjugated NFC hydrogel ([<sup>89</sup>Zr]Zr-DFO*-NFC) to enable longitudinal monitoring of its <em>in vivo</em> fate using positron emission tomography techniques. Following subcutaneous implantation in healthy mice, [<sup>89</sup>Zr]Zr-DFO*-NFC retained radioactivity at the implant site for at least 14 days, with minimal signal detected in the kidneys, urinary bladder, and overlying skin. In contrast, mice receiving a control formulation of [<sup>89</sup>Zr]Zr-oxalate mixed with NFC hydrogel showed progressive accumulation of radioactivity in the bones, consistent with known [<sup>89</sup>Zr]Zr-oxalate distribution patterns, and only limited retention at the implant site by Day 7. These findings demonstrate that [<sup>89</sup>Zr]Zr-DFO*-NFC hydrogel implants exhibit high <em>in vivo</em> stability with negligible systemic release following subcutaneous implantation, supporting their potential use as safe and traceable biomaterial platforms.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107402"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145707293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The putative proton/organic cation (H+/OC) antiporter has been shown to mediate transport of CNS drug compounds like oxycodone and pyrilamine across the blood-brain barrier (BBB). This transporter has a broad substrate profile and is able to transport substrates against their concentration gradient, making it an interesting target for brain drug delivery. However, the molecular identity of this transporter remains unknown. Recent studies have indicated that the two proteins TM7SF3 and LHFPL6 might be components of this transporter. The present study aimed to investigate the roles of TM7SF3 and LHFPL6 in the H+/OC antiporter function to advance understanding of its molecular identity and potential in CNS drug delivery.
Methods
CRISPR-Cas9 gene-editing was used to generate three hCMEC/D3 knockout (KO) cell lines: TM7SF3 KO (TM-KO), LHFPL6 KO (LH-KO), and a double KO of TM7SF3 and LHFPL6 (TMLH-KO). The uptake of pyrilamine analogue (EDMPG) and [3H]-pyrilamine was assessed in wild type (WT) and KO lines. Quantitative Realtime Polymerase Chain Reaction (qRT-PCR) confirmed successful gene knockouts. Passive diffusion properties and the expression and functionality of known BBB transporters, including LAT1 (SLC7A5), GLUT1 (SLC2A1), and MCT1 (SLC16A1), were also examined.
Results
The EDMPG uptake was significantly reduced in TM-, LH-, and TMLH-KO cells, suggesting that TM7SF3 and LHFPL6 contribute to the H+/OC antiporter function. However, [3H]-pyrilamine uptake remained unchanged across all KOs, indicating a TM7SF3- and LHFPL6-independent transport mechanism. This was further supported by the persistent inhibition of [3H]-pyrilamine uptake in the presence of known H+/OC antiporter substrates. While passive diffusion and GLUT1- and MCT1-mediated transport were unaffected, LAT1-mediated uptake of [3H]L-leucine and gabapentin (Neurontin) was significantly reduced in LH- and TMLH-KO cells, correlating with decreased LAT1 mRNA expression in these cells.
Conclusions
This study suggests that the H+/OC antiporter operates via two distinct mechanisms: a high-capacity, TM7SF3- and LHFPL6-independent pathway and a low-capacity, TM7SF3- and LHFPL6-dependent pathway. These findings underscore the complexity of the H+/OC antiporter molecular composition and highlight the need for further research to fully elucidate its identity.
质子/有机阳离子（H+/OC）反转运体已被证明可介导中枢神经系统药物化合物如羟考酮和吡啶胺通过血脑屏障（BBB）的转运。这种转运体具有广泛的底物特征，能够不顾底物的浓度梯度运输底物，使其成为脑药物输送的有趣靶点。然而，这种转运体的分子特性仍然未知。最近的研究表明TM7SF3和LHFPL6两种蛋白可能是该转运体的组分。本研究旨在探讨TM7SF3和LHFPL6在H+/OC反转运蛋白功能中的作用，以进一步了解其在中枢神经系统药物传递中的分子特性和潜力。方法：采用CRISPR-Cas9基因编辑技术，生成3株hCMEC/D3敲除（KO）细胞系：TM7SF3 KO （TM-KO）、LHFPL6 KO （LH-KO）和TM7SF3和LHFPL6双KO （TMLH-KO）。测定了野生型（WT）和KO系对吡咯胺类似物（EDMPG）和[3H]-吡咯胺的摄取。定量实时聚合酶链反应（qRT-PCR）证实基因敲除成功。我们还检测了已知血脑屏障转运蛋白LAT1 （SLC7A5）、GLUT1 （SLC2A1）和MCT1 （SLC16A1）的被动扩散特性以及表达和功能。结果：TM-、LH-和TMLH-KO细胞的EDMPG摄取明显减少，提示TM7SF3和LHFPL6参与了H+/OC反转运蛋白功能。然而，在所有ko中，[3H]-吡啶胺摄取保持不变，表明其转运机制不依赖于TM7SF3-和lhfpl6。在已知的H+/OC反转运底物存在下，[3H]-吡咯胺摄取的持续抑制进一步支持了这一点。虽然被动扩散和GLUT1-和mct1介导的转运不受影响，但LAT1介导的[3H] l -亮氨酸和加巴喷丁的摄取在LH-和TMLH-KO细胞中显著减少，这与这些细胞中LAT1 mRNA表达减少有关。结论：本研究表明，H+/OC反向转运蛋白通过两种不同的机制起作用：高容量的、不依赖TM7SF3-和lhfpl6的途径和低容量的、依赖TM7SF3-和lhfpl6的途径。这些发现强调了H+/OC反转运分子组成的复杂性，并强调了进一步研究以充分阐明其身份的必要性。
{"title":"Elucidating the roles of TM7SF3 and LHFPL6 in the putative H+/OC antiporter function in the human brain capillary endothelial cell line, hCMEC/D3","authors":"Nana Svane , Toshiki Kurosawa , Benjamin Schmid , Lasse Saaby , Mie Kristensen , Hidetsugu Tabata , Yoshiyuki Kubo , Tetsuya Terasaki , Birger Brodin , Yoshiharu Deguchi","doi":"10.1016/j.ejps.2025.107409","DOIUrl":"10.1016/j.ejps.2025.107409","url":null,"abstract":"<div><h3>Introduction</h3><div>The putative proton/organic cation (H<sup>+</sup>/OC) antiporter has been shown to mediate transport of CNS drug compounds like oxycodone and pyrilamine across the blood-brain barrier (BBB). This transporter has a broad substrate profile and is able to transport substrates against their concentration gradient, making it an interesting target for brain drug delivery. However, the molecular identity of this transporter remains unknown. Recent studies have indicated that the two proteins TM7SF3 and LHFPL6 might be components of this transporter. The present study aimed to investigate the roles of TM7SF3 and LHFPL6 in the H<sup>+</sup>/OC antiporter function to advance understanding of its molecular identity and potential in CNS drug delivery.</div></div><div><h3>Methods</h3><div>CRISPR-Cas9 gene-editing was used to generate three hCMEC/D3 knockout (KO) cell lines: TM7SF3 KO (TM-KO), LHFPL6 KO (LH-KO), and a double KO of TM7SF3 and LHFPL6 (TMLH-KO). The uptake of pyrilamine analogue (EDMPG) and [<sup>3</sup>H]-pyrilamine was assessed in wild type (WT) and KO lines. Quantitative Realtime Polymerase Chain Reaction (qRT-PCR) confirmed successful gene knockouts. Passive diffusion properties and the expression and functionality of known BBB transporters, including LAT1 (<em>SLC7A5</em>), GLUT1 (<em>SLC2A1</em>), and MCT1 (<em>SLC16A1</em>), were also examined.</div></div><div><h3>Results</h3><div>The EDMPG uptake was significantly reduced in TM-, LH-, and TMLH-KO cells, suggesting that TM7SF3 and LHFPL6 contribute to the H<sup>+</sup>/OC antiporter function. However, [<sup>3</sup>H]-pyrilamine uptake remained unchanged across all KOs, indicating a TM7SF3- and LHFPL6-independent transport mechanism. This was further supported by the persistent inhibition of [<sup>3</sup>H]-pyrilamine uptake in the presence of known H<sup>+</sup>/OC antiporter substrates. While passive diffusion and GLUT1- and MCT1-mediated transport were unaffected, LAT1-mediated uptake of [<sup>3</sup>H]L-leucine and gabapentin (Neurontin) was significantly reduced in LH- and TMLH-KO cells, correlating with decreased LAT1 mRNA expression in these cells.</div></div><div><h3>Conclusions</h3><div>This study suggests that the <em>H</em>+/OC antiporter operates via two distinct mechanisms: a high-capacity, TM7SF3- and LHFPL6-independent pathway and a low-capacity, TM7SF3- and LHFPL6-dependent pathway. These findings underscore the complexity of the H<sup>+</sup>/OC antiporter molecular composition and highlight the need for further research to fully elucidate its identity.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107409"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145741596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-10-20DOI: 10.1016/j.ejps.2025.107335
Dariusz Wawrzyniak , Klaudia Zielinkiewicz , Ewa Mijowska , Katarzyna Rolle , Elżbieta Czarniewska
The medical application of borophene in fields such as in vivo diagnosis, drug delivery, and photochemical therapy of cancer requires detailed assessment. There is no knowledge about its effects on cells, organs, and the whole organism of humans and animals. It can be assumed that the impact of borophene on cells will be correlated with the aspect ratio of the 2D flakes and their thickness. This study sought to determine if these nanomaterials are biocompatible and, if not, whether their cytotoxicity is caused by their thickness, dosage, or exposure duration in the model cells employed in the study. The influence of 3-, 4- or 6-layer borophene in the concentration range from 0.2 to 1 mg/ml of the medium on the morphology, integrity of the cell membrane, induction of apoptosis and reactive oxygen species and the level of reducing power in MRC-5 cells was investigated. The study showed no effect of borophene on the plasma membrane, no cytotoxic effects leading to apoptosis or necrosis of MRC-5 cells even at high doses of tested nanomaterials and regardless of the number of layers. Borophene had a beneficial effect on human fibroblasts because it increased or did not change the level of reducing power, which balanced of oxidative stress in cells, if induced. These effects allowed maintaining cell viability and functionality. This study opens up research perspectives leading to understanding the mechanisms responsible for maintaining viability in the borophene-exposed cells. The question remains whether borophene is biocompatible in other models, e.g., non-phagocytic cells, and what other molecular mechanisms are involved in maintaining the viability of the borophene-exposed cells. If the biocompatibility of borophene with a wide range of cells is confirmed, it will find numerous applications in biomedicine with special emphasis on Boron Neutron Therapy.
{"title":"Influence of 3-, 4- and 6-layered borophene on biocompatibility with “non-professional” human phagocytes","authors":"Dariusz Wawrzyniak , Klaudia Zielinkiewicz , Ewa Mijowska , Katarzyna Rolle , Elżbieta Czarniewska","doi":"10.1016/j.ejps.2025.107335","DOIUrl":"10.1016/j.ejps.2025.107335","url":null,"abstract":"<div><div>The medical application of borophene in fields such as <em>in vivo</em> diagnosis, drug delivery, and photochemical therapy of cancer requires detailed assessment. There is no knowledge about its effects on cells, organs, and the whole organism of humans and animals. It can be assumed that the impact of borophene on cells will be correlated with the aspect ratio of the 2D flakes and their thickness. This study sought to determine if these nanomaterials are biocompatible and, if not, whether their cytotoxicity is caused by their thickness, dosage, or exposure duration in the model cells employed in the study. The influence of 3-, 4- or 6-layer borophene in the concentration range from 0.2 to 1 mg/ml of the medium on the morphology, integrity of the cell membrane, induction of apoptosis and reactive oxygen species and the level of reducing power in MRC-5 cells was investigated. The study showed no effect of borophene on the plasma membrane, no cytotoxic effects leading to apoptosis or necrosis of MRC-5 cells even at high doses of tested nanomaterials and regardless of the number of layers. Borophene had a beneficial effect on human fibroblasts because it increased or did not change the level of reducing power, which balanced of oxidative stress in cells, if induced. These effects allowed maintaining cell viability and functionality. This study opens up research perspectives leading to understanding the mechanisms responsible for maintaining viability in the borophene-exposed cells. The question remains whether borophene is biocompatible in other models, <em>e.g.</em>, non-phagocytic cells, and what other molecular mechanisms are involved in maintaining the viability of the borophene-exposed cells. If the biocompatibility of borophene with a wide range of cells is confirmed, it will find numerous applications in biomedicine with special emphasis on Boron Neutron Therapy.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107335"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145336438","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Polymeric micelles are promising nanocarriers for improving the solubility and therapeutic efficacy of poorly water-soluble drugs. In this study, bicalutamide (BIC)-loaded polymeric micelles were developed and optimized using central composite design (CCD) by varying two formulation factors: the Soluplus® percentage (%) and the Pluronic F127/Pluronic F68 ratio (w/w). The selected formulations exhibited favorable physicochemical properties with particle size (PS) below 100 nm, low polydispersity index (PDI) (≤ 0.066), and high encapsulation efficiencies (EE) (up to 90.6 %). Transmission electron microscopy (TEM) confirmed the spherical and monodisperse structure. The micelles exhibited near-neutral zeta potentials. Lyophilization with trehalose did not significantly alter particle size or uniformity. In vitro release studies demonstrated sustained drug release profiles for 72 h, and in vitro solubility measurements revealed a significant increase (∼161 to 335-fold) compared to free BIC. The formulations also remained colloidally stable upon dilution and were physically stable for up to 6 months at 4 °C, 25 °C/60 % Relative Humidity (RH), and 40 °C/75 % RH. Cellular uptake studies in the human prostate cancer (PC-3) cell line confirmed effective internalization of the micelles. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays demonstrated a concentration- and time-dependent cytotoxicity. F7 exhibited superior cytotoxicity among the tested formulations, compared to free BIC, while its blank formulation showed no significant toxicity, indicating favorable biocompatibility. These results suggest that the developed polymeric micelle systems have potential as stable and biocompatible delivery systems for BIC, warranting further investigation.
{"title":"Development and optimization of Soluplus®/Pluronic-based polymeric micelles for bicalutamide delivery: characterization, lyophilization, stability, and cellular studies","authors":"Nihal Tugce Ozaksun , Tugce Tayyar , Aysun Ozdemir , Mustafa Ark , Tuba Incecayir","doi":"10.1016/j.ejps.2025.107395","DOIUrl":"10.1016/j.ejps.2025.107395","url":null,"abstract":"<div><div>Polymeric micelles are promising nanocarriers for improving the solubility and therapeutic efficacy of poorly water-soluble drugs. In this study, bicalutamide (BIC)-loaded polymeric micelles were developed and optimized using central composite design (CCD) by varying two formulation factors: the Soluplus® percentage (%) and the Pluronic F127/Pluronic F68 ratio (w/w). The selected formulations exhibited favorable physicochemical properties with particle size (PS) below 100 nm, low polydispersity index (PDI) (≤ 0.066), and high encapsulation efficiencies (EE) (up to 90.6 %). Transmission electron microscopy (TEM) confirmed the spherical and monodisperse structure. The micelles exhibited near-neutral zeta potentials. Lyophilization with trehalose did not significantly alter particle size or uniformity<em>. In vitro</em> release studies demonstrated sustained drug release profiles for 72 h, and <em>in vitro</em> solubility measurements revealed a significant increase (∼161 to 335-fold) compared to free BIC. The formulations also remained colloidally stable upon dilution and were physically stable for up to 6 months at 4 °C, 25 °C/60 % Relative Humidity (RH), and 40 °C/75 % RH. Cellular uptake studies in the human prostate cancer (PC-3) cell line confirmed effective internalization of the micelles. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays demonstrated a concentration- and time-dependent cytotoxicity. F7 exhibited superior cytotoxicity among the tested formulations, compared to free BIC, while its blank formulation showed no significant toxicity, indicating favorable biocompatibility. These results suggest that the developed polymeric micelle systems have potential as stable and biocompatible delivery systems for BIC, warranting further investigation.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107395"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145700073","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-01Epub Date: 2025-12-12DOI: 10.1016/j.ejps.2025.107414
Joona Sorjonen , Luis Martin de Juan , Pirjo Tajarobi , Håkan Wikström , Jarkko Ketolainen
The appearance of a tablet is a critical quality attribute that can be directly evaluated by patients. During tablet manufacturing, mechanical stress may lead to visual defects, such as edge chipping, the prediction of which remains a major challenge, particularly during technology transfer or scale-up processes. This study aimed to evaluate whether the number and severity of defects could be predicted under two mechanical stress environments: pan coating and friabilator testing.
A breakage model was calibrated using drop tests with two placebo formulations: mannitol:microcrystalline cellulose (7:3) and microcrystalline cellulose:anhydrous dibasic calcium phosphate (7:3), both at target tensile strengths of 1.5–2.5 MPa. Discrete element method (DEM) simulations were used to estimate the collision velocities and frequencies in the coating pan, enabling defect prediction using the breakage model. One verification run was performed using the mannitol:MCC 7:3 formulation at 2.0 MPa, which was also used to predict the tablet appearance in the friabilator.
The model accurately predicted the severity of defects (Classes I–III, where I is the least and III the most severe defect) in the coating trial but underestimated the number of Class I defects. The numerical predictions made by the model are exponentially affected by inaccuracies in the calibration of model parameters, particularly for small damage. Visual differences between tablets with equal mass loss in the friabilator and pan coater also suggest that wear from low-force collisions contributed to damage during the coating process. In contrast, the number of appearance defects was overpredicted in the friabilator, highlighting the model’s limitations in systems with frequent collisions. Overall, with careful calibration and under conditions with few defect-causing events, the model can provide useful guidance for defect prediction.
{"title":"Early prediction of tablet defects during pan coating","authors":"Joona Sorjonen , Luis Martin de Juan , Pirjo Tajarobi , Håkan Wikström , Jarkko Ketolainen","doi":"10.1016/j.ejps.2025.107414","DOIUrl":"10.1016/j.ejps.2025.107414","url":null,"abstract":"<div><div>The appearance of a tablet is a critical quality attribute that can be directly evaluated by patients. During tablet manufacturing, mechanical stress may lead to visual defects, such as edge chipping, the prediction of which remains a major challenge, particularly during technology transfer or scale-up processes. This study aimed to evaluate whether the number and severity of defects could be predicted under two mechanical stress environments: pan coating and friabilator testing.</div><div>A breakage model was calibrated using drop tests with two placebo formulations: mannitol:microcrystalline cellulose (7:3) and microcrystalline cellulose:anhydrous dibasic calcium phosphate (7:3), both at target tensile strengths of 1.5–2.5 MPa. Discrete element method (DEM) simulations were used to estimate the collision velocities and frequencies in the coating pan, enabling defect prediction using the breakage model. One verification run was performed using the mannitol:MCC 7:3 formulation at 2.0 MPa, which was also used to predict the tablet appearance in the friabilator.</div><div>The model accurately predicted the severity of defects (Classes I–III, where I is the least and III the most severe defect) in the coating trial but underestimated the number of Class I defects. The numerical predictions made by the model are exponentially affected by inaccuracies in the calibration of model parameters, particularly for small damage. Visual differences between tablets with equal mass loss in the friabilator and pan coater also suggest that wear from low-force collisions contributed to damage during the coating process. In contrast, the number of appearance defects was overpredicted in the friabilator, highlighting the model’s limitations in systems with frequent collisions. Overall, with careful calibration and under conditions with few defect-causing events, the model can provide useful guidance for defect prediction.</div></div>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":"217 ","pages":"Article 107414"},"PeriodicalIF":4.7,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145755167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-22DOI: 10.1016/j.ejps.2026.107446
Josi Ann Steinke, Pierre-Louis Destruel, Benjamin Weiche, Stefan Dengl, Gregoire Schwach, Gerhard Pohlmann, Marlon J Hinner
The therapeutic potential of inhaled biologics is limited by protein instability during aerosolization, which can cause aggregation and increase immunogenicity risks. This study aims to identify key molecular and formulation parameters that minimize the aggregation of antibody Fab fragments during vibrating mesh nebulization. A set of 14 engineered Fabs with a broad range of melting temperatures (Tm 60-90°C) were nebulized using two different commercial vibrating mesh devices. We systematically assessed the impact of Fab thermostability, protein concentration (10-80 mg/mL), formulation excipients, and nebulizer device on the formation of high molecular weight species (HMWS) and subvisible particles (SVP). Aerosol characteristics, including fine particle fraction (FPF) and output rate, were also evaluated. High intrinsic thermostability and high protein concentration were identified as the two most critical factors for preventing aggregation. Fabs with a Tm above 80°C demonstrated exceptional stability with negligible aggregation. We attribute this effect to a correlation of high Tm with a higher resistance against unfolding and therefore a better tolerance against nebulization-induced stresses, in particular air/liquid interfacial stress. Counterintuitively, increasing the protein concentration from 10 mg/mL to 80 mg/mL suppressed aggregation for all Fabs, which had the highest benefit for Fabs with lower thermostability. This effect can at least in part be explained by a saturation of the air/liquid interface at higher Fab concentrations. While higher concentrations modestly reduced nebulizer output rates, the overall inhalable protein output (mg protein/min) was significantly enhanced. In comparison, full-length monoclonal antibodies showed poor aerosolization performance at high concentrations. Our findings provide clear guidance for developing inhaled Fab therapeutics. To ensure stability and minimize aggregation, priority should be given to selecting Fabs with high thermostability (Tm > 80°C) and formulating them at high concentrations (≥ 40 mg/mL). These strategies are expected to mitigate aggregation-induced immunogenicity and concomitant safety risks, facilitating the development of the next generation of inhaled protein drugs.
{"title":"Minimizing Aggregates when Nebulizing Antibody Fab Fragments: High Thermostability and High Concentration are Key.","authors":"Josi Ann Steinke, Pierre-Louis Destruel, Benjamin Weiche, Stefan Dengl, Gregoire Schwach, Gerhard Pohlmann, Marlon J Hinner","doi":"10.1016/j.ejps.2026.107446","DOIUrl":"https://doi.org/10.1016/j.ejps.2026.107446","url":null,"abstract":"<p><p>The therapeutic potential of inhaled biologics is limited by protein instability during aerosolization, which can cause aggregation and increase immunogenicity risks. This study aims to identify key molecular and formulation parameters that minimize the aggregation of antibody Fab fragments during vibrating mesh nebulization. A set of 14 engineered Fabs with a broad range of melting temperatures (T<sub>m</sub> 60-90°C) were nebulized using two different commercial vibrating mesh devices. We systematically assessed the impact of Fab thermostability, protein concentration (10-80 mg/mL), formulation excipients, and nebulizer device on the formation of high molecular weight species (HMWS) and subvisible particles (SVP). Aerosol characteristics, including fine particle fraction (FPF) and output rate, were also evaluated. High intrinsic thermostability and high protein concentration were identified as the two most critical factors for preventing aggregation. Fabs with a T<sub>m</sub> above 80°C demonstrated exceptional stability with negligible aggregation. We attribute this effect to a correlation of high T<sub>m</sub> with a higher resistance against unfolding and therefore a better tolerance against nebulization-induced stresses, in particular air/liquid interfacial stress. Counterintuitively, increasing the protein concentration from 10 mg/mL to 80 mg/mL suppressed aggregation for all Fabs, which had the highest benefit for Fabs with lower thermostability. This effect can at least in part be explained by a saturation of the air/liquid interface at higher Fab concentrations. While higher concentrations modestly reduced nebulizer output rates, the overall inhalable protein output (mg protein/min) was significantly enhanced. In comparison, full-length monoclonal antibodies showed poor aerosolization performance at high concentrations. Our findings provide clear guidance for developing inhaled Fab therapeutics. To ensure stability and minimize aggregation, priority should be given to selecting Fabs with high thermostability (T<sub>m</sub> > 80°C) and formulating them at high concentrations (≥ 40 mg/mL). These strategies are expected to mitigate aggregation-induced immunogenicity and concomitant safety risks, facilitating the development of the next generation of inhaled protein drugs.</p>","PeriodicalId":12018,"journal":{"name":"European Journal of Pharmaceutical Sciences","volume":" ","pages":"107446"},"PeriodicalIF":4.7,"publicationDate":"2026-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146044020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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