Tissue engineering. Part C, Methods最新文献

{"title":"Reproducible Preparation of Primary Rat Hepatocyte Sheets Using a Thermoresponsive Culture Dish.","authors":"Jun Kobayashi,&nbsp;Teruo Okano","doi":"10.1089/ten.TEC.2023.0099","DOIUrl":"10.1089/ten.TEC.2023.0099","url":null,"abstract":"<p><p>Hepatocyte transplantation has been utilized as a therapy for congenital metabolic liver diseases such as hemophilia and for liver function support in acute liver failure. Hepatocyte sheet technology using a thermoresponsive poly(<i>N</i>-isopropylacrylamide) (PIPAAm)-grafted dish is expected to provide an efficient cell transplantation method because the resulting hepatocyte sheet possesses extracellular matrix (ECM) on the basal surface, which enhances attachment to the target sites. However, the cultured hepatocytes consume large amounts of oxygen, leading to the loss of a few hepatocytes within the confluent culture sheet owing to a lack of oxygen. To circumvent this problem, this work demonstrates the shortening of diffusion distance, that is, the medium depth, to accelerate oxygen supply from the gas phase/medium interface to the cultured hepatocytes, allowing them to form a monolayer hepatocyte sheet. Incubation of hepatocytes with medium at a depth of 1.3 mm facilitates confluent culture of hepatocytes for 72 h, whereas viable hepatocytes decreased at 2.6 mm depth. Hepatocyte sheets are formed on a 0.5 μg/cm² fibronectin-physisorbed PIPAAm-grafted dish during 72 h incubation at 37°C. Detachment of the cultured hepatocyte sheet from the PIPAAm-grafted dish where the surface becomes hydrophilic at 20°C is accomplished by scraping the periphery of the sheet using a cell scraper. Furthermore, the apical side of the hepatocyte sheet can be physically grabbed using a gelatin-coated membrane, and the sheet with ECM on the basal surface can be readily transferred to the target site after melting the coated gelatin at 37°C. Both methods are beneficial for creating tissue models by layering with another type of cell sheets, and for quick transplantation, such as into the subcutaneous space and orthotopic transplantation on the surface of the liver.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"479-491"},"PeriodicalIF":3.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9974047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Harvest of Cell-Only Muscle Fibers Using Thermally Expandable Hydrogels with Adhesive Patterns.","authors":"Yu Bin Lee, Se-Jeong Kim, Eun Mi Kim, Hayeon Byun, Heungsoo Shin","doi":"10.1089/ten.TEC.2023.0096","DOIUrl":"10.1089/ten.TEC.2023.0096","url":null,"abstract":"<p><p>Muscle tissue engineering has been the focus of extensive research due to its potential for numerous medical applications, including <i>ex vivo</i> actuator development and clinical treatments. In this study, we developed a method for harvesting muscle fiber in a floatable and translocatable manner utilizing thermally expandable hydrogels with a chemically patterned polydopamine (PD) layer generated by microcontact printing (μCP). The μCP of PD on the hydrogel facilitated the formation of stripe patterns with varying widths of printed/nonprinted area (50/50, 100/100, and 200/200 μm). The spatially controlled adhesion of C2C12 myoblasts on the PD patterns produced clearly distinguishable muscle fibers, and translocated muscle fibers exhibited preserved extracellular matrix and junction proteins. Furthermore, the development of anisotropic arrangements and mature myotubes within the fibers suggests the potential for functional control of engineered muscle tissues. Overall, the muscle fiber harvesting method developed herein is suitable for both translocation and floating and is a promising technique for muscle tissue engineering as it mimics the structure-function relationship of natural tissue.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"447-458"},"PeriodicalIF":3.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9924577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Correction to:</i> Implanted Tissue-Engineered Vascular Graft Cell Isolation with Single-Cell RNA Sequencing Analysis, by Mirhaidari et al. Tissue Engineering Part C: Methods 2023;29(2):72-84; doi: 10.1089/ten.tec.2022.0189.","authors":"","doi":"10.1089/ten.tec.2022.0189.correx","DOIUrl":"10.1089/ten.tec.2022.0189.correx","url":null,"abstract":"","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":" ","pages":"492"},"PeriodicalIF":3.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11075514/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10168900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improved Production of Induced Pluripotent Stem Cells Using Dot Pattern Culture Plates.","authors":"Yoshiki Nakashima,&nbsp;Hiroki Iguchi,&nbsp;Eiko Shimizu,&nbsp;Minh N T Le,&nbsp;Kenta Takakura,&nbsp;Yuta Nakamura,&nbsp;Teruhiko Yanagisawa,&nbsp;Rutvi Sanghavi,&nbsp;Satoshi Haneda,&nbsp;Masayoshi Tsukahara","doi":"10.1089/ten.TEC.2023.0068","DOIUrl":"10.1089/ten.TEC.2023.0068","url":null,"abstract":"<p><p>The rate of cell proliferation is a crucial factor in cell production under good manufacturing practice (GMP) control. In this study, we identified a culture system for induced pluripotent cells (iPSCs) that supports cell proliferation and viability and maintains the cells in an undifferentiated state even at 8 days after seeding. This system involves the use of dot pattern culture plates that have been coated with a chemically defined scaffold which has high biocompatibility. Under cell starvation conditions, where medium exchange was not performed for 7 days or where the amount of medium exchange was reduced to half or a quarter, iPSC viability and lack of differentiation were maintained. The rate of cell viability in this culture system was greater than generally obtained by standard culture methods. The cells in this compartmentalized culture system could be induced to differentiate in a controlled and consistent manner: differentiation of endoderm occurred in a controlled and consistent manner: endoderm, mesoderm, and ectoderm could be consistently induced to differentiate in the cultures. In conclusion, we have developed a culture system that supports high viability in iPSCs and allows their controlled differentiation. This system has the potential for use in GMP-based production of iPSCs for clinical purposes.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 9","pages":"410-423"},"PeriodicalIF":3.0,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10517333/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10299523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effective Technical Protocol for Producing a Mono-Iodoacetate-Induced Temporomandibular Joint Osteoarthritis in a Rat Model.","authors":"So-Yeon Yun,&nbsp;Yerin Kim,&nbsp;Hyunjeong Kim,&nbsp;Bu-Kyu Lee","doi":"10.1089/ten.TEC.2023.0066","DOIUrl":"10.1089/ten.TEC.2023.0066","url":null,"abstract":"<p><p>An animal model of osteoarthritis (OA) induced by monosodium iodoacetate (MIA) can be effectively adjusted based on the concentration of MIA to control the onset, progression, and severity of OA as required. The rat temporomandibular joint osteoarthritis (TMJOA) model using MIA is a useful tool for studying the effectiveness of disease-modifying OA drugs in TMJOA research. However, the intricate and complex anatomy of the rat TMJ often poses challenges in achieving consistent TMJOA induction during experiments. In the previous article, a reference point was established by drawing parallel lines based on the line connecting the external ear and the zygomatic arch. However, this is not suitable for the anatomical characteristics of the rat. We used the zygomatic arch as a reference, which is a technical protocol that considers it. In our protocol, we designated a point ∼1 mm away from the point where the zygomatic arch bends toward the ear as the injection site. To ensure precise injection of MIA and increase the likelihood of inducing OA, it is recommended to insert the needle at a 45° angle so that the needle tip contacts the joint projection. To confirm TMJOA induction, we identified changes in the condyle using <i>in vivo</i> microcomputed tomography (CT) in a rat model of MIA-induced OA and measured the degree of pain-related inflammation using head withdrawal threshold (HWT) measurements. Micro-CT scanning revealed typical OA-like lesions, including degenerative changes and subchondral bone remodeling induced by MIA in the TMJ. Pain, a major clinical feature of OA, showed an appropriate response corresponding to the structural changes shown in micro-CT scanning. In addition, the MIA concentration suitable for long-term observation of lesions was determined through <i>ex vivo</i> micro-CT imaging and HWT measurements. The 8 mg concentration exhibited a significant difference compared with others, confirming the sustained presence of lesions, particularly through changes in subchondral bone over an extended period. Consequently, we have successfully established a reliable rat TMJOA induction model and identified the MIA concentration suitable for long-term observation of subchondral bone research, which will greatly contribute to the study of TMJOA-an incurable disease lacking specific treatment options. The Clinical Trial Registration number is 2021-12-208.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 9","pages":"438-445"},"PeriodicalIF":3.0,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10298512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Upscale Manufacture of Chondrocytes for Allogeneic Cartilage Therapies.","authors":"Charlotte H Hulme, John K Garcia, Claire Mennan, Jade Perry, Sally Roberts, Kevin Norris, Duncan Baird, Larissa Rix, Robin Banerjee, Carl Meyer, Karina T Wright","doi":"10.1089/ten.TEC.2023.0037","DOIUrl":"10.1089/ten.TEC.2023.0037","url":null,"abstract":"<p><p>Allogeneic chondrocyte therapies need to be developed to allow more individuals to be treated with a cell therapy for cartilage repair and to reduce the burden and cost of the current two-stage autologous procedures. Upscale manufacture of chondrocytes using a bioreactor could help provide an off-the-shelf allogeneic chondrocyte therapy with many doses being produced in a single manufacturing run. In this study, we assess a good manufacturing practice-compliant hollow-fiber bioreactor (Quantum^®) for adult chondrocyte manufacture. Chondrocytes were isolated from knee arthroplasty-derived cartilage (<i>n</i> = 5) and expanded in media supplemented with 10% fetal bovine serum (FBS) or 5% human platelet lysate (hPL) on tissue culture plastic (TCP) for a single passage. hPL-supplemented cultures were then expanded in the Quantum bioreactor for a further passage. Matched, parallel cultures in hPL or FBS were maintained on TCP. Chondrocytes from all culture conditions were characterized in terms of growth kinetics, morphology, immunoprofile, chondrogenic potential (chondrocyte pellet assays), and single telomere length analysis. Quantum expansion of chondrocytes resulted in 86.4 ± 38.5 × 10⁶ cells in 8.4 ± 1.5 days, following seeding of 10.2 ± 3.6 × 10⁶ cells. This related to 3.0 ± 1.0 population doublings in the Quantum bioreactor, compared with 2.1 ± 0.6 and 1.3 ± 1.0 on TCP in hPL- and FBS-supplemented media, respectively. Quantum- and TCP-expanded cultures retained equivalent chondropotency and mesenchymal stromal cell marker immunoprofiles, with only the integrin marker, CD49a, decreasing following Quantum expansion. Quantum-expanded chondrocytes demonstrated equivalent chondrogenic potential (as assessed by ability to form and maintain chondrogenic pellets) with matched hPL TCP populations. hPL manufacture, however, led to reduced chondrogenic potential and increased cell surface positivity of integrins CD49b, CD49c, and CD51/61 compared with FBS cultures. Quantum expansion of chondrocytes did not result in shortened 17p telomere length when compared with matched TCP cultures. This study demonstrates that large numbers of adult chondrocytes can be manufactured in the Quantum hollow-fiber bioreactor. This rapid, upscale expansion does not alter chondrocyte phenotype when compared with matched TCP expansion. Therefore, the Quantum provides an attractive method of manufacturing chondrocytes for clinical use. Media supplementation with hPL for chondrocyte expansion may, however, be unfavorable in terms of retaining chondrogenic capacity.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 9","pages":"424-437"},"PeriodicalIF":3.0,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10517319/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10299041","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Bibliometric Analysis of Research on Decellularized Matrix for Two Decades.","authors":"Xueying Hou,&nbsp;Enchong Zhang,&nbsp;Yukun Mao,&nbsp;Jie Luan,&nbsp;Su Fu","doi":"10.1089/ten.TEC.2023.0013","DOIUrl":"10.1089/ten.TEC.2023.0013","url":null,"abstract":"<p><p>The articles and reviews in the field of decellularized extracellular matrix (dECM) from 2001 to 2021 were retrieved and extracted from the Web of Science Core Collection. The R package Bibliometrix, CiteSpace, VOSviewer, and the online BIBLIOMETRC platform were utilized for bibliometric analysis, including specific characteristics of annual publications, influential countries/regions, core journals, leading institutions, keywords, key references, cocited authors, journals and institutions, cooperation, and historical direct citations. Our study concluded core references that fueled the development of dECM and highlighted current research directions, hotpots, and trends. From 2001 to 2021, 3,046 publications were retrieved in total, including 2,700 articles and 349 reviews. The United States (<i>n</i> = 895) produced the majority of publications, and the University of Pittsburgh (<i>n</i> = 318) published most productions. <i>Biomaterials</i> were identified as the most productive and influential journal in the dECM field considering the number of publications (<i>n</i> = 194), and total citations (<i>n</i> = 15,694). Immunomodulation, bioreactors, aging, three-dimensional (3D) bioprinting, bone tissue engineering, bioink, hydrogel, biomaterials, and regeneration were the latest high-frequency keywords, indicating the emerging frontiers of dECM. In the field, decellularization techniques lay the foundation. Orthotopic transplantation of recellularized dECM and induction of specific cell differentiation promoted the bursts of research. The 3D bioprinting and hydrogel based on dECM were extensively studied in recent years. The present study provided developmental trajectories, current research status, global collaboration patterns, hotpots, and trending topics of dECM. Decellularization techniques, tissue engineering to regenerate organs, and improvements in application are the major themes over the past two decades. Impact Statement The review article is significant because decellularized extracellular matrix (dECM), which derived from biological tissues and removal of immunogenic cells, is characterized by safety, biocompatibility, and low in toxicity. Showing great application prospects, dECM has been applied in multiple scenarios of tissue repairment and reconstruction, among the most popular topics in tissue engineering. Thus, analyzing and concluding the development, current condition and future trends are of great significance. Comparing to conventional review, this review article systemically and comprehensively concluded the historical development, current status, and research trending topics. Thus, it allows scholars to get a rapid overview of the dECM field, and plan research directions.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 9","pages":"395-409"},"PeriodicalIF":3.0,"publicationDate":"2023-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10663606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microfluidic Platform for Microparticle Fabrication and Release of a Cathepsin Inhibitor.","authors":"Elda A Treviño, Jimmy Shah, Joseph J Pearson, Manu O Platt, Younan Xia, Johnna S Temenoff","doi":"10.1089/ten.TEC.2023.0015","DOIUrl":"10.1089/ten.TEC.2023.0015","url":null,"abstract":"<p><p>Cathepsins are a family of cysteine proteases responsible for a variety of homeostatic functions throughout the body, including extracellular matrix remodeling, and have been implicated in a variety of degenerative diseases. However, clinical trials using systemic administration of cathepsin inhibitors have been abandoned due to side effects, so local delivery of cathepsin inhibitors may be advantageous. In these experiments, a novel microfluidic device platform was developed that can synthesize uniform, hydrolytically degradable microparticles from a combination of poly(ethylene glycol) diacrylate (PEGDA) and dithiothreitol (DTT). Of the formulations examined, the 10-polymer weight percentage 10 mM DTT formulation degraded after 77 days <i>in vitro</i>. A modified assay using the DQ Gelatin Fluorogenic Substrate was used to demonstrate sustained release and bioactivity of a cathepsin inhibitor (E-64) released from hydrogel microparticles over 2 weeks <i>in vitro</i> (up to ∼13 μg/mL released with up to ∼40% original level of inhibition remaining at day 14). Altogether, the technologies developed in this study will allow a small-molecule, broad cathepsin inhibitor E-64 to be released in a sustained manner for localized inhibition of cathepsins for a wide variety of diseases.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 8","pages":"361-370"},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10442676/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10431218","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"<i>Call for Special Issue Papers:</i> Artificial Intelligence in Tissue Engineering and Biology.","authors":"Jason L Guo,&nbsp;Michael Januszyk,&nbsp;Michael T Longaker","doi":"10.1089/ten.tec.2023.29040.cfp","DOIUrl":"https://doi.org/10.1089/ten.tec.2023.29040.cfp","url":null,"abstract":"","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 8","pages":"347-348"},"PeriodicalIF":3.0,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9971472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advanced Methodology for Rapid Isolation of Single Myofibers from Flexor Digitorum Brevis Muscle.","authors":"Kamal Awad, Logan Moore, Jian Huang, Lauren Gomez, Leticia Brotto, Venu Varanasi, Christopher Cardozo, Noah Weisleder, Zui Pan, Jingsong Zhou, Lynda Bonewald, Marco Brotto","doi":"10.1089/ten.TEC.2023.0012","DOIUrl":"10.1089/ten.TEC.2023.0012","url":null,"abstract":"<p><p>Isolated individual myofibers are valuable experimental models that can be used in various conditions to understand skeletal muscle physiology and pathophysiology at the tissue and cellular level. This report details a time- and cost-effective method for isolation of single myofibers from the flexor digitorum brevis (FDB) muscle in both young and aged mice. The FDB muscle was chosen for its documented history in single myofiber experiments. By modifying published methods for FDB myofiber isolation, we have optimized the protocol by first separating FDB muscle into individual bundles before the digestion, followed by optimizing the subsequent digestion medium conditions to ensure reproducibility. Morphological and functional assessments demonstrate a high yield of isolated FDB myofibers with sarcolemma integrity achieved in a shorter time frame than previous published procedures. This method could be also adapted to other types of skeletal muscle. Additionally, this highly reproducible method can greatly reduce the number of animals needed to yield adequate numbers of myofibers for experiments. Thus, this advanced method for myofiber isolation has the potential to accelerate research in skeletal muscle physiology and screening potential therapeutics \"<i>ex vivo</i>\" for muscle diseases and regeneration.</p>","PeriodicalId":23154,"journal":{"name":"Tissue engineering. Part C, Methods","volume":"29 8","pages":"349-360"},"PeriodicalIF":2.7,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10686193/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9972435","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}