Biotechnology and Bioengineering最新文献

{"title":"ETV2 Overexpression Promotes Efficient Differentiation of Pluripotent Stem Cells to Endothelial Cells","authors":"Yunfeng Ding, Soniya Tamhankar, Feifan Du, Tessa Christopherson, Nate Schlueter, Jenna R. Cohen, Eric V. Shusta, Sean P. Palecek","doi":"10.1002/bit.28979","DOIUrl":"https://doi.org/10.1002/bit.28979","url":null,"abstract":"Differentiating endothelial cells (ECs) from human pluripotent stem cells (hPSCs) typically takes 2 weeks and requires parameter optimization. Overexpression of cell type-specific transcription factors in hPSCs has shown efficient differentiation into various cell types. ETV2, a crucial transcription factor for endothelial fate, can be overexpressed in hPSCs to induce rapid and facile EC differentiation (iETV2-ECs). We developed a two-stage strategy which involves differentiating inducible <i>ETV2-</i>overexpressing hPSCs in a basal induction medium during stage I and expanding them in an endothelial medium during stage II. By optimizing seeding density and medium composition, we achieved 99% pure CD31+ CD144+ iETV2-ECs without cell sorting in 5 days. iETV2-ECs demonstrated in vitro angiogenesis potential, LDL uptake, and cytokine response. Transcriptomic comparisons revealed similar gene expression profiles between iETV2-ECs and traditionally differentiated ECs. Additionally, iETV2-ECs responded to Wnt signaling agonist and TGFβ inhibitor to acquire brain EC phenotypes, making them a scalable EC source for applications including blood-brain barrier modeling.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"57 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering Yarrowia lipolytica for Enhanced Gastrodin Production via High-Throughput Screening and Transcriptomics-Guided Optimization","authors":"Mengchen Hu, Yijin He, Yifan Ma, Na Wu, Zhongwen Gan, Jinhan Fu, Zhiliang Yang, Tianqiong Shi, Xiaoman Sun, Yang Gu, Peng Xu","doi":"10.1002/bit.28977","DOIUrl":"https://doi.org/10.1002/bit.28977","url":null,"abstract":"Gastrodin, the principal bioactive component of the renowned herb <i>Gastrodia elata</i>, is extensively utilized in medicinal drugs and nutraceuticals. This study seeks to enhance microbial production of gastrodin through high-throughput screening (HTS) and transcriptomics-guided optimization. Initially, atmospheric pressure and room temperature plasma (ARTP)-mediated mutagenesis were employed to develop a library of mutant strains. Furthermore, a transcription factor-based biosensor and a high-throughput solid-phase extraction mass spectrometry (HP-SPE-MS) were evaluated to establish an HTS method for gastrodin. Consequently, mutant strain MT8 was isolated, producing 9.8 g/L of gastrodin in YPD medium, which represents a 55.6% increase compared to the control strain. Next, key genes identified via transcriptomics were overexpressed in strain MT8, with the overexpression of gene <i>YALI2E01737g</i>, a gene involved in the synthesis of aromatic amino acids, significantly enhancing gastrodin production to reach 10.1 g/L. In addition, fermentation process optimization further improved gastrodin titer up to 13.1 g/L in shaking flasks. This study demonstrated the utility of HTS techniques to enhance gastrodin production and paved the way for its future industrial application.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"13 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143672623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering an Overflow-Responsive Regulation System for Balancing Cellular Redox and Optimizing Microbial Production","authors":"Jianli Zhang, Jian Wang, Tian Jiang, Xinyu Gong, Qi Gan, Yuxi Teng, Yusong Zou, Ainoor Anwar Dawadi, Yajun Yan","doi":"10.1002/bit.28976","DOIUrl":"https://doi.org/10.1002/bit.28976","url":null,"abstract":"<i>Escherichia coli</i> accumulates acetate as a byproduct in fast growth aerobic conditions when using glucose as carbon source. This phenomenon, known as overflow metabolism, has negative impacts on cell growth and protein expression, also causes carbon loss during biosynthesis in most microbial production scenarios. In this study, we regarded the “waste” metabolite as a useful metabolism indicator, constructed an overflow biosensor to monitor the change of acetate concentration and converted the signal into various regulation outputs. Phloroglucinol is a phenolic compound with several derivatives that exhibit various pharmacological activities. By applying the bifunctional dynamic regulation system on the phloroglucinol production, we released the cellular redox pressure in real-time and reduced the waste of carbon flux on overflow metabolism. Finally, carbon flux was redirected more favorably towards the desired product, resulting in a boosted phloroglucinol titer of 1.30 g/L, increased by 2.04-fold. Overall, this study explored the use of a central byproduct-responsive biosensor system on improving cellular metabolic status, providing a general approach for enhancing bioproduction.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"22 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143672625","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Single Cell Protein Production From Ethanol: Model-Based Bioreactor Operation at Industrial Scale","authors":"Eduardo Almeida Benalcázar, Wouter A. van Winden, Lars Puiman, John A. Posada, Mickel L. A. Jansen, Henk Noorman, Adrie J. J. Straathof","doi":"10.1002/bit.28969","DOIUrl":"https://doi.org/10.1002/bit.28969","url":null,"abstract":"Alternative fermentation feedstocks such as ethanol can be produced from CO&lt;sub&gt;2&lt;/sub&gt; via electrocatalytic processes that coproduce O&lt;sub&gt;2&lt;/sub&gt;. In this study, industrial-scale fermentation of ethanol with pure O&lt;sub&gt;2&lt;/sub&gt; for single cell protein (SCP) production was studied using a modeling approach. This approach considered (i) microbial kinetics, (ii) gas–liquid transfer, and (iii) an exploration of potential operational constraints. The technical feasibility for producing up to 58 kt/y of SCP in a 600 m&lt;sup&gt;3&lt;/sup&gt; bubble column operating in continuous mode was assessed and attributed mainly to a high O&lt;sub&gt;2&lt;/sub&gt; transfer rate of 1.1 mol/(kg h) through the use of pure O&lt;sub&gt;2&lt;/sub&gt;. However, most of the pure O&lt;sub&gt;2&lt;/sub&gt; fed to the fermenter remains unconsumed due to the large gas flows needed to maximize mass transfer. In addition, biomass production may be hampered by high dissolved CO&lt;sub&gt;2&lt;/sub&gt; concentrations and by large heat production. The model estimates a microbial biomass concentration of 114 g/kg, with a yield on ethanol of 0.61 g&lt;sub&gt;x&lt;/sub&gt;/g&lt;sub&gt;ethanol&lt;/sub&gt; (&gt; 95% &lt;span data-altimg=\"/cms/asset/cdee15e5-a923-4a44-97ff-c9692f5252ae/bit28969-math-0001.png\"&gt;&lt;/span&gt;&lt;mjx-container ctxtmenu_counter=\"403\" ctxtmenu_oldtabindex=\"1\" jax=\"CHTML\" role=\"application\" sre-explorer- style=\"font-size: 103%; position: relative;\" tabindex=\"0\"&gt;&lt;mjx-math aria-hidden=\"true\" location=\"graphic/bit28969-math-0001.png\"&gt;&lt;mjx-semantics&gt;&lt;mjx-mrow&gt;&lt;mjx-mrow&gt;&lt;mjx-msubsup data-semantic-children=\"0,4,5\" data-semantic-collapsed=\"(7 (6 0 4) 5)\" data-semantic- data-semantic-role=\"latinletter\" data-semantic-speech=\"upper Y Subscript x divided by s Superscript max\" data-semantic-type=\"subsup\"&gt;&lt;mjx-mi data-semantic-annotation=\"clearspeak:simple\" data-semantic-font=\"italic\" data-semantic- data-semantic-parent=\"7\" data-semantic-role=\"latinletter\" data-semantic-type=\"identifier\"&gt;&lt;mjx-c&gt;&lt;/mjx-c&gt;&lt;/mjx-mi&gt;&lt;mjx-script style=\"vertical-align: -0.355em; margin-left: -0.182em;\"&gt;&lt;mjx-mi data-semantic-font=\"normal\" data-semantic- data-semantic-parent=\"7\" data-semantic-role=\"limit function\" data-semantic-type=\"function\" size=\"s\" style=\"margin-left: 0.341em;\"&gt;&lt;mjx-c&gt;&lt;/mjx-c&gt;&lt;mjx-c&gt;&lt;/mjx-c&gt;&lt;mjx-c&gt;&lt;/mjx-c&gt;&lt;/mjx-mi&gt;&lt;mjx-spacer style=\"margin-top: 0.18em;\"&gt;&lt;/mjx-spacer&gt;&lt;mjx-mrow data-semantic-children=\"1,3\" data-semantic-content=\"2\" data-semantic- data-semantic-parent=\"7\" data-semantic-role=\"division\" data-semantic-type=\"infixop\" size=\"s\"&gt;&lt;mjx-mi data-semantic-annotation=\"clearspeak:simple\" data-semantic-font=\"italic\" data-semantic- data-semantic-parent=\"4\" data-semantic-role=\"latinletter\" data-semantic-type=\"identifier\"&gt;&lt;mjx-c&gt;&lt;/mjx-c&gt;&lt;/mjx-mi&gt;&lt;mjx-mo data-semantic- data-semantic-operator=\"infixop,/\" data-semantic-parent=\"4\" data-semantic-role=\"division\" data-semantic-type=\"operator\" rspace=\"1\" space=\"1\"&gt;&lt;mjx-c&gt;&lt;/mjx-c&gt;&lt;/mjx-mo&gt;&lt;mjx-mi data-semantic-annotation=\"clearspeak:simple\" data-semantic-font=\"italic\" data-semantic- data-semantic-parent=\"4\" data-semantic-role=\"latin","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"24 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143672624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of Miniature AsCas12f1 Variants for Gene Editing and Activation","authors":"Chuanhong Ren, Zehua Bao","doi":"10.1002/bit.28978","DOIUrl":"https://doi.org/10.1002/bit.28978","url":null,"abstract":"Miniature CRISPR/Cas systems possess delivery advantages for gene therapy. The type V-F Cas12f1 from <i>Acidibacillus sulfuroxidans</i> is exceptionally compact (422 amino acids) and has been engineered by several studies as compact genome editing tools through protein and single guide RNA (sgRNA) engineering. However, a comparative evaluation of gene editing and activation efficiencies mediated by different AsCas12f1 variants and sgRNA scaffolds is lacking. This study tested combinations of four AsCas12f1 protein variants and six sgRNA scaffolds for their gene editing and transcription activation efficiencies. The protein variant AsCas12f1-HKRA performed the best in gene editing and activation when paired with sgRNA-en_v2.1 scaffold. Furthermore, we validated a super miniature gene activator by fusing a small activation domain to AsCas12f1-HKRA. Our findings recommend using AsCas12f1-HKRA and sgRNA-en_v2.1 for gene editing and activation applications.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"29 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143661119","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic Engineering and Adaptive Evolution of Escherichia coli for Enhanced Conversion of D‑Xylose to D-Glucaric Acid Mediated by Methanol","authors":"Wei-Xiang Chen, Ling-Jie Zheng, Xuan Luo, Shang-He Zheng, Hui-Dong Zheng, Li-Hai Fan, Qiang Guo","doi":"10.1002/bit.28974","DOIUrl":"https://doi.org/10.1002/bit.28974","url":null,"abstract":"<span>d</span>-Glucaric acid is a value-added dicarboxylic acid that can be utilized in the chemical, food, and pharmaceutical industries. Due to the complex process and environmental pollution associated with the chemical production of <span>d</span>-glucaric acid, bioconversion for its synthesis has garnered increasing attention in recent years. In this study, a novel cell factory was developed for the efficient production of <span>d</span>-glucaric acid using <span>d</span>-xylose and methanol. Mdh, Hps, Phi, Miox, Ino1, Suhb, and Udh were first co-expressed in <i>E. coli</i> JM109 to construct the <span>d</span>-glucaric acid synthesis pathway. The deletion of FrmRAB, RpiA, PfkA, and PfkB was then performed to block or weaken the endogenous competitive pathways. Next, adaptive evolution was carried out to improve cell growth and substrate utilization. With the purpose of further increasing the product titer, the NusA tag and myo-inositol biosensor were introduced into engineered <i>E. coli</i> to enhance Miox expression. After medium optimization and fermentation process control, 3.0 g/L of <span>d</span>-glucaric acid was finally obtained in the fed-batch fermentation using modified Terrific Broth medium.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"40 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143653802","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Perfusion-Based Production of rAAV via an Intensified Transient Transfection Process","authors":"Tam N. T. Nguyen, Damdae Park, Christopher T. Canova, Jose Sangerman, Prasanna Srinivasan, Rui Wen Ou, Paul W. Barone, Caleb Neufeld, Jacqueline M. Wolfrum, Stacy L. Springs, Anthony J. Sinskey, Richard D. Braatz","doi":"10.1002/bit.28967","DOIUrl":"https://doi.org/10.1002/bit.28967","url":null,"abstract":"Increasing demand for recombinant adeno-associated virus (rAAV)-based gene therapies necessitates increased manufacturing production. Transient transfection of mammalian cells remains the most commonly used method to produce clinical-grade rAAVs due to its ease of implementation. However, transient transfection processes are often characterized by suboptimal yields and low fractions of full-to-total capsids, both of which contribute to the high cost of goods of many rAAV-based gene therapies. Our previously developed mechanistic model for rAAV2/5 production indicated that the inadequate capsid filling is due to a temporal misalignment between viral DNA replication and capsid synthesis within the cells and the repression of later phase capsid formation by Rep proteins. We experimentally validated this prediction and showed that performing multiple, time-separated doses of plasmid increases the production of rAAV. In this study, we use the insights generated by our mechanistic model to develop an intensified process for rAAV production that combines perfusion with high cell density re-transfection. We demonstrate that performing multiple, time-separated doses at high cell density boosts both cell-specific and volumetric productivity and improves plasmid utilization when compared to a single bolus at standard operating conditions. Our results establish a new paradigm for continuously manufacturing rAAV via transient transfection that improves productivity and reduces manufacturing costs.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"14 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143653799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioengineered Tumor-Stroma Prostate Cancer In Vitro Models for Screening Therapeutics","authors":"Maria V. Monteiro, Filipa Moreira-Silva, Matilde Lagarto, Luís P. Ferreira, Carlota Ramalhinho, Iola F. Duarte, Carmen Jerónimo, Vítor M. Gaspar, João F. Mano","doi":"10.1002/bit.28971","DOIUrl":"https://doi.org/10.1002/bit.28971","url":null,"abstract":"Cancer-associated fibroblasts are increasingly recognized to have a high impact on prostate tumor growth and drug resistance. Here, we bioengineered organotypic prostate cancer 3D in vitro models to better understand tumor-stroma interplay, the metabolomic profile underlying such interactions, and their impact on standard-of-care therapeutics performance. The assembly of robust and uniform spheroids was evaluated and compared in monotypic PC-3 and heterotypic microtumors comprised of either a healthy or malignant stroma and prostate cancer cells. Our findings demonstrate that the precise inclusion of prostate cancer stromal elements is crucial to generating robust PC-3 prostate cancer spheroids with reproducible morphology and size. The inclusion of cancer-associated fibroblasts promoted the establishment of more compact microtumors exhibiting characteristic expression of major proteins. Exometabolomic profile analysis also highlighted the impact of stromal cells on tumor models metabolism. The optimized heterotypic spheroids were additionally exploited for screening standard-of-care therapeutics, exhibiting a higher resistance when compared to their monotypic counterparts. Our findings demonstrate that including stromal elements in PC-3 prostate cancer models is crucial for their use as increasingly organotypic testing platforms, being relevant for screening candidate anti-cancer therapeutics and for the discovery of potential combinations with emerging anti-stroma therapies.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"86 Suppl 1 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143618151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Physiological Response of Penicillium chrysogenum to Mimicked Local and Global Perturbations of Substrate and Dissolved Oxygen Gradients at Industrial-Scale","authors":"Yining Chen, Cees Haringa, Zejian Wang, Yingping Zhuang, Guan Wang","doi":"10.1002/bit.28968","DOIUrl":"https://doi.org/10.1002/bit.28968","url":null,"abstract":"Industrial-scale microbial fermentation processes often face limitations in mixing and mass transfer, leading to the formation of environmental gradients within the bioreactor. These gradients expose microbes to heterogeneous conditions over time and space. In this study, we evaluated the effects of combined substrate and dissolved oxygen (DO) gradients on the metabolic response of <i>Penicillium chrysogenum</i> at an industrial scale. Three representative heterogeneous environments were simulated in scale-down systems: (1) feed inlet (high glucose, low oxygen (HGLO): C_S &gt; 20 mM, DO &lt; 0.012 mM), (2) aeration inlet (high oxygen, low glucose (HOLG): C_S &lt; 0.8 mM, DO &gt; 0.2 mM), and (3) global environment (periodic 360 s fluctuation cycle with 45 s of HGLO and 75 s of HOLG conditions). Results showed that prolonged exposure to feed inlet conditions led to a complete loss of penicillin production capacity, accompanied by significant excretion of intracellular metabolites, and this effect was largely irreversible. While, cells randomly walking under the top impeller zone did not lose production capacity but showed signs of premature degeneration due to increased energy demand. When exposed to the global environment, cells finely tuned their metabolism in a periodical manner, with nearly a 50% loss of penicillin productivity. In summary, substrate gradients alone did not cause irreversible effects, but large substrate gradients contributed to reduced productivity. Oxygen gradients, however, not only reduced production but also caused irreversible cellular damage. These findings provide valuable insights for developing scale-up criteria and strain engineering strategies aimed at improving large-scale culture performance.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"26 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"3D Printable Self-Healing Mineralized Hydrogels Loaded With Diclofenac Sodium: In Vitro and In Vivo Assessment","authors":"Nachiketa Palit, Purushottam Suryavanshi, Subham Banerjee","doi":"10.1002/bit.28973","DOIUrl":"https://doi.org/10.1002/bit.28973","url":null,"abstract":"The use of self-healing mineralized hydrogels in 3D printing has demonstrated significant advantages, including enhanced printing accuracy and the ability to maintain high shape fidelity throughout the printing process. After conducting an initial optimization study, we incorporated our self-healing mineralized hydrogel into semi-solid extrusion-based 3D printing to print diclofenac-loaded oral films. The dependence of the print speed on the nature of the material was established by varying the print speed. The process of optimizing the print speed was conducted using a blank hydrogel, which involved analyzing specific parameters, such as printing accuracy and the percentage of pore area under sizing. The results demonstrated that 2 mm/sec print speed showed a higher printing accuracy of 98.13% and pore area under-sizing value of 41.31%. Interestingly, the viscosity of the hydrogel increased from 5.30 to 133 PaS upon addition of the drug. The percentage pore area under sizing also decreased from 41.31% to 11.48% as the drug loading was increased from 0% to 3% <i>w</i>/<i>w</i>. The in vitro drug release study demonstrated that the 3% <i>w</i>/<i>w</i> diclofenac sodium-loaded oral films printed at 2 mm/sec exhibited a faster release profile. Furthermore, considerable bioavailability of diclofenac sodium (DS) was achieved from the 3D-printed oral films during the in vivo study. These results can be effectively used to develop a drug delivery system that can release medications accurately and consistently, either in a targeted area or systemically.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":"68 1","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143598861","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}