Frontiers in Bioengineering and Biotechnology最新文献

{"title":"High-throughput platform for label-free sorting of 3D spheroids using deep learning.","authors":"Claudia Sampaio da Silva, Julia Alicia Boos, Jonas Goldowsky, Manon Blache, Noa Schmid, Tim Heinemann, Christoph Netsch, Francesca Luongo, Stéphanie Boder-Pasche, Gilles Weder, Alba Pueyo Moliner, Roos-Anne Samsom, Ary Marsee, Kerstin Schneeberger, Ali Mirsaidi, Bart Spee, Thomas Valentin, Andreas Hierlemann, Vincent Revol","doi":"10.3389/fbioe.2024.1432737","DOIUrl":"10.3389/fbioe.2024.1432737","url":null,"abstract":"<p><p>End-stage liver diseases have an increasing impact worldwide, exacerbated by the shortage of transplantable organs. Recognized as one of the promising solutions, tissue engineering aims at recreating functional tissues and organs <i>in vitro</i>. The integration of bioprinting technologies with biological 3D models, such as multi-cellular spheroids, has enabled the fabrication of tissue constructs that better mimic complex structures and <i>in vivo</i> functionality of organs. However, the lack of methods for large-scale production of homogeneous spheroids has hindered the upscaling of tissue fabrication. In this work, we introduce a fully automated platform, designed for high-throughput sorting of 3D spheroids based on label-free analysis of brightfield images. The compact platform is compatible with standard biosafety cabinets and includes a custom-made microscope and two fluidic systems that optimize single spheroid handling to enhance sorting speed. We use machine learning to classify spheroids based on their bioprinting compatibility. This approach enables complex morphological analysis, including assessing spheroid viability, without relying on invasive fluorescent labels. Furthermore, we demonstrate the efficacy of transfer learning for biological applications, for which acquiring large datasets remains challenging. Utilizing this platform, we efficiently sort mono-cellular and multi-cellular liver spheroids, the latter being used in bioprinting applications, and confirm that the sorting process preserves viability and functionality of the spheroids. By ensuring spheroid homogeneity, our sorting platform paves the way for standardized and scalable tissue fabrication, advancing regenerative medicine applications.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1432737"},"PeriodicalIF":4.3,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11663632/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142881790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Significant differences in knee kinematics of healthy subjects with high and low anterior tibial laxity.","authors":"Shiyang Chen, Shaohua Chen, Qingyang Kang, Fangzheng Lin, Shuting Zheng, Xixi Liu, Chunhong Guo, Yongjin Li, Dingkun Lin, Xiaolong Zeng","doi":"10.3389/fbioe.2024.1514516","DOIUrl":"10.3389/fbioe.2024.1514516","url":null,"abstract":"<p><strong>Background: </strong>Anterior tibial laxity is considered to be a risk factor for knee injuries, including anterior cruciate ligament ruptures. The anterior cruciate ligament reconstruction also aims to restore anterior tibial laxity. While anterior tibial laxity is considered to be linked to dynamic knee stability, the mechanisms connecting anterior tibial laxity to these stability issues are not fully understood. The purpose of this study was to investigate the kinematic alterations between different anterior tibial laxity in healthy subjects. We hypothesized that anterior tibial laxity affects the anteroposterior tibial displacement during dynamic movements.</p><p><strong>Methods: </strong>This study involved thirty-five healthy subjects. There were twenty males and fifteen females with an average age of 18.91 ± 0.78 years. Their knees were categorized into \"Tight\" (the smallest 50%) and \"Lax\" (the largest 50%) groups based on anterior tibial laxity measurements using a Kneelax3 arthrometer. Kinematic data were collected using a three-dimensional motion capture system when they performed level walking, upslope walking, and vertical jumping. The knee kinematics were recorded for statistical analysis. We used independent sample t-tests to analyze key kinematic differences between groups.</p><p><strong>Results: </strong>The \"Lax\" group exhibited increased posterior tibial translation during upslope walking (5.4 ± 2.22 mm at swing max flexion, <i>p</i> = 0.018) and vertical jumping (8.5 ± 2.78 mm at propulsion max flexion, <i>p</i> = 0.003; 7.6 ± 3.17 mm at landing max flexion, <i>p</i> = 0.019) than the \"Tight\" group. Significant differences in tibial internal rotation were observed during initial contact of the gait cycle of level walking (1.9° ± 0.95°, <i>p</i> = 0.049) and upslope walking (2.1° ± 1.03°, <i>p</i> = 0.041) in the \"Lax\" group compared to the \"Tight\" group. No significant differences in adduction/abduction or medial/lateral tibial translation were found between groups.</p><p><strong>Conclusion: </strong>The study revealed that high anterior tibial laxity resulted in increased posterior tibial translation and tibial internal rotation. High anterior tibial laxity resulted in dynamic instability of knees during motions, especially in high-demanding activities like upslope or vertical jumping. However, further research is needed to explore the clinical functional effects of knee laxity.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1514516"},"PeriodicalIF":4.3,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11664222/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142880660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Computational modeling of superparamagnetic nanoparticle-based (affinity) diagnostics.","authors":"Loïc Van Dieren, Vlad Tereshenko, Haïzam Oubari, Yanis Berkane, Jonathan Cornacchini, Filip Thiessen Ef, Curtis L Cetrulo, Korkut Uygun, Alexandre G Lellouch","doi":"10.3389/fbioe.2024.1500756","DOIUrl":"10.3389/fbioe.2024.1500756","url":null,"abstract":"<p><strong>Introduction: </strong>Magnetic nanoparticles (MNPs), particularly iron oxide nanoparticles (IONPs), are renowned for their superparamagnetic behavior, allowing precise control under external magnetic fields. This characteristic makes them ideal for biomedical applications, including diagnostics and drug delivery. Superparamagnetic IONPs, which exhibit magnetization only in the presence of an external field, can be functionalized with ligands for targeted affinity diagnostics. This study presents a computational model to explore the induced voltage in a search coil when MNPs pass through a simulated blood vessel, aiming to improve non-invasive diagnostic methods for disease detection and monitoring.</p><p><strong>Methods: </strong>A finite element model was constructed using COMSOL Multiphysics to simulate the behavior of IONPs within a dynamic blood vessel environment. Governing equations such as Ampère's law and Faraday's law of induction were incorporated to simulate the induced voltage in a copper coil as MNPs of various sizes flowed through the vessel. Rheological parameters, including blood viscosity and flow rates, were factored into the model using a non-Newtonian fluid approach.</p><p><strong>Results: </strong>The amount of MNPs required for detection varies significantly based on the sensitivity of the detection equipment and the size of the nanoparticles themselves. For highly sensitive devices like a SQUID voltmeter, with a coil sensitivity approximately 10^-12 V, very low MNP concentrations-approximately 10^-4 μg/mL-are sufficient for detection, staying well within the safe range. As coil sensitivity decreases, such as with standard voltmeters at 10^-8 V or 10^-6 V, the MNP concentration required for detection rises, approaching or even exceeding potentially toxic levels. Additionally, the physical size of MNPs plays a role; larger nanoparticles (e.g., 50 nm radius) require fewer total particles for detection at the same sensitivity than smaller particles like those with a 2.5 nm radius. For instance, at a coil sensitivity of 10^-10 V, a 2.5 nm particle requires approximately 10¹² particles, whereas a 50-nm particle only needs 10⁸. This highlights the importance of optimizing both detection sensitivity and particle size to balance effective detection with safety.</p><p><strong>Conclusion: </strong>This computational model demonstrates the feasibility of using superparamagnetic nanoparticles in real-time, non-invasive diagnostic systems.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1500756"},"PeriodicalIF":4.3,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11658970/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876426","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of the effects of <i>Bacillus velezensis</i> HJ-16 inoculation on tobacco leaves based on multi-omics methods.","authors":"Qing Zhou, Jinchu Yang, Yingjie Feng, Zongcan Yang, Yixuan Wang, Zhan Zhang, Tingting Zhang, Wenzhao Liu, YongMing Xu, Yongfeng Yang, Jihong Huang","doi":"10.3389/fbioe.2024.1493766","DOIUrl":"10.3389/fbioe.2024.1493766","url":null,"abstract":"<p><p>In this study, a strain isolated from the surface of flue-cured tobacco leaves, identified as <i>Bacillus velezensis</i> HJ-16, was applied in the solid-state fermentation of tobacco leaves. This strain, known for producing thermally stable enzymes, including amylase, cellulase, and protease, significantly improved the sensory qualities of tobacco, enhancing aromatic intensity, density, and softness, while reducing irritation. Whole-genome sequencing and functional annotation revealed that <i>B. velezensis</i> HJ-16 possesses a single circular chromosome containing genes associated with enzyme production and metabolic activities, particularly in carbohydrate metabolism and amino acid metabolism. Untargeted metabolomics analysis identified significant changes in non-volatile metabolites induced by fermentation. These metabolites were enriched in pathways related to flavonoid biosynthesis, alkaloid biosynthesis, aromatic amino acid metabolism, lipid metabolism, and carbon metabolism. Metagenomic analysis showed that <i>Bacillus</i> became the dominant genus on the tobacco leaf surface following inoculation with <i>B. velezensis</i> HJ-16, altering the microbial community composition, reducing diversity and evenness, and enhancing microbial metabolic activity. These findings underscore the potential of <i>B. velezensis HJ-16</i> as a biotechnological tool to improve tobacco leaf quality.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1493766"},"PeriodicalIF":4.3,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11659759/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876314","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Collagen scaffold-seeded iTenocytes accelerate the healing and functional recovery of Achilles tendon defects in a rat model.","authors":"Thomas Später, Patricia Del Rio, Oksana Shelest, Jacob T Wechsler, Giselle Kaneda, Melissa Chavez, Julia Sheyn, Victoria Yu, Wolfgang Metzger, Dave Huang, Melodie Metzger, Wafa Tawackoli, Dmitriy Sheyn","doi":"10.3389/fbioe.2024.1407729","DOIUrl":"10.3389/fbioe.2024.1407729","url":null,"abstract":"<p><strong>Introduction: </strong>Tendon injuries represent an ongoing challenge in clinical practice due to poor regenerative capacity, structure, and biomechanical function recovery of ruptured tendons. This study is focused on the assessment of a novel strategy to repair ruptured Achilles tendons in a Nude rat model using stem cell-seeded biomaterial.</p><p><strong>Methods: </strong>Specifically, we have used induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (iMSCs) overexpressing the early tendon marker Scleraxis (SCX, iMSC^SCX+, iTenocytes) in combination with an elastic collagen scaffold. Achilles tendon defects in Nude rat models were created by isolating the tendon and excising 3 mm of the midsection. The Achilles tendon defects were then repaired with iTenocyte-seeded scaffolds, unseeded scaffolds, or suture only and compared to native Nude rat tendon tissue using gait analyses, biomechanical testing, histology, and immunohistochemistry.</p><p><strong>Results: </strong>The results show faster functional recovery of gait in iTenocyte-seeded scaffold group comparing to scaffold only and suture only groups. Both iTenocyte-seeded scaffold and scaffold only treatment groups had improved biomechanical properties when compared to suture only treatment group, however no statistically significant difference was found in comparing the cell seeding scaffold an scaffold only group in terms of biomechanical properties. Immunohistochemistry staining further demonstrated that iTenocytes successfully populated the collagen scaffolds and survived 9 weeks after implantation <i>in vivo</i>. Additionally, the repaired tissue of iTenocyte-treated injuries exhibited a more organized structure when compared to tendon defects that were repaired only with suturing or unseeded scaffolds.</p><p><strong>Conclusion: </strong>We suggest that iTenocyte-seeded DuRepair™ collagen scaffold can be used as potential treatment to regenerate the tendon tissue biomechanically and functionally.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1407729"},"PeriodicalIF":4.3,"publicationDate":"2024-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11658981/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142876372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimizing collagen-based biomaterials for periodontal regeneration: clinical opportunities and challenges.","authors":"Ye Feng, Hong-Peng Li","doi":"10.3389/fbioe.2024.1469733","DOIUrl":"10.3389/fbioe.2024.1469733","url":null,"abstract":"<p><p>Periodontal disease (PD) is a chronic inflammatory condition that affects the teeth and their supporting tissues, ultimately culminating in tooth loss. Currently, treatment modalities, such as systemic and local administration of antibiotics, serve to mitigate the progression of inflammation yet fall short in restoring the original anatomical structure and physiological function of periodontal tissues. Biocompatible material-based tissue engineering seems to be a promising therapeutic strategy for treating PD. Collagen, a component of the extracellular matrix commonly used for tissue engineering, has been regarded as a promising biogenic material for tissue regeneration owing to its high cell-activating and biocompatible properties. The structural and chemical similarities between collagen and components of the oral tissue extracellular matrix render it a promising candidate for dental regeneration. This review explored the properties of collagen and its current applications in periodontal regeneration. We also discussed the recent progression in collagen therapies and preparation techniques. The review also scrutinizes the pros and cons associated with the application of collagen-based biomaterials in PD treatment, aiming to pave the way for future applications of collagen-based biomaterials in the management of PD.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1469733"},"PeriodicalIF":4.3,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11655217/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142863879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Repeated harvest enables efficient production of VSV-GP.","authors":"Rebecca Habisch, Peter Neubauer, Jorge Soza-Ried, Eva Puschmann","doi":"10.3389/fbioe.2024.1505338","DOIUrl":"10.3389/fbioe.2024.1505338","url":null,"abstract":"<p><p>Viral products keep gaining importance in multiple therapeutic fields. Considering the scale and production slot limitations, optimizing the outcome of every manufacturing batch is essential to minimize costs and make this therapeutic modality broadly available to patients. Most manufacturing processes for oncolytic viruses currently in clinical studies are based on a batch process. Here, we evaluated the benefits in terms of titer increase of a repeated harvest approach and compared it to the classical batch production process. While no effect on cell density was observed, the cumulated infectious titer following repeated harvest was over 400 times higher than the evaluated batch process yield. This shows that repeated harvests or perfusion have the potential to boost viral yields and should be considered when deciding on a process format for production.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1505338"},"PeriodicalIF":4.3,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11656157/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142863844","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent nanoengineered therapeutic advancements in sepsis management.","authors":"Li Liu, Li Li, Ting Wang, Zheyu Li, Bingpeng Yan, Ruirong Tan, Anqi Zeng, Wenbo Ma, Xin Zhu, Zhujun Yin, Chunhua Ma","doi":"10.3389/fbioe.2024.1495277","DOIUrl":"10.3389/fbioe.2024.1495277","url":null,"abstract":"<p><p>Sepsis (defined as sepsis 3.0) is a life-threatening organ dysfunction caused by a dysregulated host response to a variety of pathogenic microorganisms. Characterized by high morbidity and mortality, sepsis has become a global public health problem. However, there is a lack of appropriate diagnostic and therapeutic strategies for sepsis and current management rely on the limited treatment strategies. Recently, nanomedicines targeting and controlling the release of bio-active agents have shown excellent potency in sepsis management, with improved therapeutic efficacy and reduced adverse effects. In this review, we have summarized the advantages of nanomaterials. Also, the preparation and efficacy of the main categories of anti-sepsis nanomedicines applied in sepsis management are described in detail, including antibiotic-coated nanomaterials, antimicrobial peptides-coated nanomaterials, biomimetic nanomaterials, nanomaterials targeting macrophages and natural products loaded nanomaterials. These advances in nanomedicines establish the huge potential for nanomaterials-based sepsis management, especially in the improved pharmaceutical and pharmacological properties, enhanced therapeutic efficacy, controllable drug-targeting and reduced side effects. To further facilitate clinical translation of anti-sepsis nanomedicines, we propose that the issues involving safety, regulatory laws and cost-effectiveness should receive much more attention in the future.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1495277"},"PeriodicalIF":4.3,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11655211/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142863882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biomechanical assessment of Kirschner wires integrated with a novel external fixation device for treatment of pediatric supracondylar humeral fracture: a finite element analysis.","authors":"Yu-Hsin Lu, Ching-Lung Tai, Wei-Chun Lee, Si-Yao Wang, Chi-Yu Mao, Wen-E Yang, Chia-Hsieh Chang, Hsuan-Kai Kao","doi":"10.3389/fbioe.2024.1480298","DOIUrl":"10.3389/fbioe.2024.1480298","url":null,"abstract":"<p><strong>Background: </strong>Pediatric supracondylar humeral fractures present considerable surgical challenges due to the difficulty of achieving proper fracture alignment and stable fixation while avoiding injury to the ulnar nerve. This study assesses the biomechanical performance of a novel Kirschner wire (K-wire) fixation device (KFD), designed to enhance stability and reduce complications linked to traditional K-wire configurations.</p><p><strong>Methods: </strong>Using finite element analysis (FEA), we evaluated four fixation strategies for treatment of pediatric supracondylar humeral simple transverse fractures: crossed pin fixation, crossed pin fixation with KFD, two lateral pin fixation, and two lateral pin fixation with KFD, under various mechanical loads. The analysis focused on the stress and strain experienced by the K-wires at the fracture site during torsional and bending forces.</p><p><strong>Results: </strong>FEA revealed that the KFD significantly reduced the stress and strain on the K-wires in all configurations. In both crossed pin and two lateral pin fixation methods, the addition of the KFD showed lower stress and strain levels compared to setups without the KFD.</p><p><strong>Conclusion: </strong>This study demonstrates the potential of the KFD to enhance fracture stability and reduce mechanical stress at the fracture site, suggesting a promising improvement in the treatment of pediatric supracondylar humeral fractures. This innovation may contribute to safer and more reliable outcomes in pediatric orthopedic surgery.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1480298"},"PeriodicalIF":4.3,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11655205/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142863873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Modeling and optimization of culture media for recombinant <i>Helicobacter pylori</i> vaccine antigen HpaA.","authors":"Runqing Tan, Song Zhou, Min Sun, Yu Liu, Xiumei Ni, Jin He, Gang Guo, Kaiyun Liu","doi":"10.3389/fbioe.2024.1499940","DOIUrl":"10.3389/fbioe.2024.1499940","url":null,"abstract":"<p><strong>Introduction: </strong><i>H. pylori</i> (<i>Helicobacter pylori</i>) infection represents a significant global health concern, exacerbated by the emergence of drug-resistant strains resulting from conventional antibiotic treatments. Consequently, the development of vaccines with both preventive and therapeutic properties has become crucial in addressing <i>H. pylori</i> infections. The <i>H. pylori</i> adhesin protein HpaA has demonstrated strong immunogenicity across various adjuvants and dosage forms, positioning it as a key candidate antigen for recombinant subunit vaccines against <i>H. pylori</i>. Optimizing fermentation culture conditions is an effective strategy to enhance product yield and lower production costs. However, to date, there has been no systematic investigation into methods for improving the fermentation yield of HpaA. Enhancing the fermentation medium to increase HpaA yield holds significant potential for application and economic benefits in the prevention and detection of <i>H. pylori</i> infection.</p><p><strong>Methods: </strong>To achieve a stable and high-yielding <i>H. pylori</i> vaccine antigen HpaA, this study constructed recombinant <i>Escherichia coli</i> expressing HpaA. The impact of fermentation medium components on the rHpaA yield was assessed using a one-factor-at-a-time approach alongside Plackett-Burman factorial experiments. Optimal conditions were effectively identified through response surface methodology (RSM) and artificial neural network (ANN) statistical computational models. The antigenicity and immunogenicity of the purified rHpaA were validated through immunization of mice, followed by Western Blot analysis and serum IgG ELISA quantification.</p><p><strong>Results: </strong>Glucose, yeast extract, yeast peptone, NH₄Cl and CaCl₂ all contributed to the production of rHpaA, with glucose, yeast extract, and NH₄Cl demonstrating particularly significant effects. The artificial neural network linked genetic algorithm (ANN-GA) model exhibited superior predictive accuracy, achieving a rHpaA yield of 0.61 g/L, which represents a 93.2% increase compared to the initial medium. Animal immunization experiments confirmed that rHpaA possesses good antigenicity and immunogenicity.</p><p><strong>Discussion: </strong>This study pioneers the statistical optimization of culture media to enhance rHpaA production, thereby supporting its large-scale application in <i>H. pylori</i> vaccines. Additionally, it highlights the advantages of the ANN-GA approach in bioprocess optimization.</p>","PeriodicalId":12444,"journal":{"name":"Frontiers in Bioengineering and Biotechnology","volume":"12 ","pages":"1499940"},"PeriodicalIF":4.3,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11652157/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142853624","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}