Tissue Engineering Part A最新文献

{"title":"Development of a 3D <i>in Vitro</i> Wound Healing Model to Assess the Effect of ADSC-EVs on Vascularization.","authors":"Emma K C Symonds, Alfonso J Schmidt, Alexander W Brown, Margaret J Currie, Patries M Herst, Kathryn E Hally, Kirsty M Danielson","doi":"10.1177/19373341251383899","DOIUrl":"https://doi.org/10.1177/19373341251383899","url":null,"abstract":"<p><p>Angiogenesis is critical for effective wound healing and relies on the successful coordination of various cell types, including endothelial cells, macrophages, and fibroblasts. Adipose-derived stem cell extracellular vesicles (ADSC-EVs) have demonstrated proangiogenic properties and have been posited as a novel therapeutic to aid wound healing; however, their functional impact within human-derived multicellular models remains largely uncharacterized. This study explores the development and application of a 3D multicellular <i>in vitro</i> model to assess the effects of ADSC-EVs on vascularization in the context of wound healing. 3D multicellular <i>in vitro</i> models were developed by coculturing human umbilical vein endothelial cells (HUVECs), monocyte-derived macrophages, and fibroblasts within Matrigel to recapitulate the <i>in vivo</i> wound healing microenvironment. A five-color confocal microscopy panel was developed to visualize each cell type and EVs within the models. The optimized models were then treated with ADSC-EVs or control to determine their impact on angiogenesis and cell colocalization. We determined that vessel formation was significantly enhanced when HUVECs were cocultured in multicellular models compared with monocultures, with the greatest effect observed in the full three-cell-type model. This effect was even more pronounced with the addition of ADSC-EVs. ADSC-EV treatment also enhanced macrophage colocalization within endothelial structures. This study developed a multicellular model that can be used for future work assessing wound healing <i>in vitro</i> and will be additive to currently used single-cell and <i>in vivo</i> models. We have applied these models to demonstrate that ADSC-EVs significantly enhance tube formation in HUVECs and the development of tissue-like structures in multicell systems, highlighting their potential as a promising therapeutic approach for improving wound healing.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145214501","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}

{"title":"LINC00687 Regulates PRDX2 Expression in High Glucose-Induced Nonunion after Digital Replantation.","authors":"Xiangying Wang, Xin He","doi":"10.1177/19373341251381372","DOIUrl":"https://doi.org/10.1177/19373341251381372","url":null,"abstract":"<p><p>Diabetic nonunion is a major clinical challenge with unclear molecular mechanisms. This study systematically investigated the key genes and molecular mechanisms of bone nonunion after finger replantation induced by high glucose using Gene Expression Omnibus (GEO), bioinformatics, and experimental analyses. In total, 179 differentially expressed mRNAs and one lncRNA (DElncRNA) were identified using the GEO dataset. Functional enrichment analysis showed that these genes were mainly involved in the regulation of autophagy and metabolism. Protein-protein interaction network analysis identified five core genes (Peroxiredoxin 2 [PRDX2], FK506 binding protein 8 [FKBP8], SHANK-associated RH domain interactor [SHARPIN], WD repeat domain 45 [WDR45], and gamma-aminobutyric acid type A receptor-associated protein like 2 [GABARAPL2]), three of which exhibited good binding affinities for potential therapeutic agents. Immune infiltration analysis revealed significant differences in the CD8+ T cell proportions between nonunion and healthy samples. We constructed a competitive endogenous RNA network (long intergenic non-protein coding RNA 687 [LINC00687]-miR-4443-PRDX2) and verified its direct regulatory interaction using a dual-luciferase reporter assay. FKBP8, PRDX2, SHARPIN, WDR45, and GABARAPL2 were overexpressed in tissue samples from patients with type 2 diabetes mellitus fracture nonunion. Animal experiments further confirmed that LINC00687 upregulated PRDX2 expression by sponging miR-4443 in a hyperglycemic environment, thereby inhibiting bone healing. This study not only identified PRDX2 and other genes as potential biomarkers of diabetic nonunion but also clarified the regulatory role of the LINC00687/miR-4443/PRDX2 axis in hyperglycemia-induced nonunion, providing a new molecular target for clinical prevention and treatment. Impact Statement 1. PRDX2, KBP8, SHARPIN, WDR45, and GABARAPL2 were potential biomarkers for this study. 2. LINC00687-miR-4443-PRDX2 participated in high glucose-induced nonunion in this study. 3. Autophagy process and metabolic pathways contribute to the progression in this study.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145202175","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}

{"title":"Potential of Low-Dose Carbon Monoxide in Promoting Osseointegration.","authors":"Jiahe Li, Liang Zhou, Mingxiao Liu, Tianyu Huang, Xian He","doi":"10.1177/19373341251383864","DOIUrl":"https://doi.org/10.1177/19373341251383864","url":null,"abstract":"<p><p>Successful osseointegration is crucial for dental implant stability, yet it remains challenging due to adverse local microenvironments, particularly infection and inflammation. While carbon monoxide (CO) has been recognized as a promising gaseous signaling molecule with diverse therapeutic properties, its clinical application faces significant limitations due to dose control challenges. To address this issue, we developed a polyetheretherketone (PEEK)-based photo-responsive implant system with surface-immobilized manganese carbonyl nanocrystals, enabling precisely controlled near-infrared light-triggered CO release. The system demonstrated efficient photoresponsiveness, achieving 13.83 ± 1.16 μM CO release within 10 min under optimal illumination conditions. <i>In vitro</i> studies revealed that low-dose CO significantly enhanced bone marrow mesenchymal stem cell osteogenic differentiation with upregulated expression of key markers, including Runx2, ALP, and OCN. In a rat femoral defect model, implants with controlled CO release exhibited significantly improved osseointegration. Comprehensive biosafety assessments confirmed the system's excellent biocompatibility without detectable organ toxicity. This research provides compelling evidence for controlled low-dose CO as an innovative strategy to enhance osseointegration, offering new possibilities for dental and orthopedic implant development, particularly for challenging clinical scenarios with compromised bone healing. Impact Statement This study introduces a novel approach for improving implant osseointegration through controlled carbon monoxide delivery, potentially offering a new strategy for enhancing the success rate of dental implant procedures.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145194056","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}

{"title":"Developing a Functional Osteoarthritis Model Using Human Osteochondral-Synovial Explants.","authors":"Luminita Labusca, Camelia-Mihaela Zara-Danceanu, Anca Emanuela Minuti, Cristina Stavila, Adriana Petrovici, Petru Plamadeala, Iuliu Ivanov, Florin Zugun-Eloae, Dragos Anita, Adriana Anita, Nicoleta Lupu","doi":"10.1177/19373341251377645","DOIUrl":"https://doi.org/10.1177/19373341251377645","url":null,"abstract":"<p><p>Osteochondral explants can serve as valuable <i>ex vivo</i> models for investigating joint development and testing therapeutic interventions in osteoarthritis (OA). The incorporation of synovial tissue in coculture settings more closely reproduces the inflammatory milieu characteristic of OA joints; however, no report exists regarding the culture media that can support such <i>ex vivo</i> systems. We investigated the reactivity of osteochondral explants using two media types: Dulbecco's modified essential medium (DMEM) and chondrogenic medium (CHONDRO). Additionally, we tested the potential therapeutic effect of serum-free conditioned media (CM) derived from allogeneic adipose-derived stem cells (ADSCs) in the context of OA. Osteochondral fragments with or without homologous synovium were cultured in DMEM and CHONDRO for up to 30 days. A subset of explants received treatment with CM. Explant reactivity was assessed by cytokine release, synovial cellularity, and osteochondral protein content using Western blot and immunohistochemistry. Explants kept in DMEM displayed diminished levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFα), together with increased Collagen II (Col II) expression. Notably, consistent suppression of TNFα was observed following CM treatment. Conversely, the CHONDRO-kept samples exhibited an increased prevalence of chondrocyte clusters; heightened Perlecan presence as well as IL-1β levels in response to CM treatment and synovial tissue-dependent fluctuations in Col II levels. Remarkably, significantly increased β-galactosidase levels could be detected in osteochondral tissues treated with CM, regardless of the culture media type. In the experimental conditions created, DMEM provided a neutral milieu and was less prone to confounding experimental outcomes, rendering it suitable for evaluating potential therapies. CHONDRO apparently increased chondrocyte clusters and facilitated extracellular matrix synthesis; however, its usage requires caution due to potential interference with experimental readouts. CM could exert an antisenescence effect, an effect that warrants further investigation.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145132869","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}

{"title":"Tissue-Engineered Endothelial Keratoplasty with Controlled Cell Density: Toward Super TEEKs.","authors":"Inès Aouimeur, Louise Coulomb, Sofiane Fraine, Zhiguo He, Guillaume Bonnet, Tomy Sagnial, Gauthier Travers, Sédao Xxx, Cyril Mauclair, Anaick Moisan, Philippe Gain, Gilles Thuret, Corantin Maurin","doi":"10.1177/19373341251381346","DOIUrl":"https://doi.org/10.1177/19373341251381346","url":null,"abstract":"<p><p>Over the past 20 years, endothelial keratoplasty procedures have revolutionized the treatment of corneal endothelial disorders. These conditions have now become the leading indication for corneal transplantation in Western countries and account for half of all donor cornea usage. Despite their undeniable success, the global shortage of donor tissues and major disparities between nations justify the development of alternatives to donor grafts. Cell therapy using injections of suspended endothelial cells has proven effective, and tissue-engineered endothelial keratoplasty (TEEK), comprising a membrane coated with cultured endothelial cells, is under development to better mimic the native endothelial graft. Our team utilizes a femtosecond-laser-cut lens capsule disc as a bioengineering scaffold, taking advantage of this novel tissue's biocompatibility, transparency, curvature, and availability. In the present study, we provide proof of concept, in 12 TEEKs, that it is possible to control the final endothelial cell density (ECD) by varying the seeding density per mm². Cell characterization was performed through morphometric analysis of the endothelial mosaic stained with anti-NCAM (a lateral membrane marker used as a differentiation marker), using the CellPose artificial intelligence algorithm specifically trained for <i>in vitro</i> endothelium segmentation. Five criteria related to pleomorphism, polymorphism, and elongation were combined into a single endothelial quality score. The median cell viability at 28 days of culture, assessed by Hoechst 33342 and Calcein-AM staining, reached 98% (range: 83-99%). The median viable ECD (number of live cells per surface unit) in the highest-density group was 3.245 cells/mm² (range: 2.778-3.753), paving the way for the bioengineering of supra-physiological TEEKs, or \"super TEEKs\".</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145132886","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}

{"title":"Electrically Triggered Fluorescein and Dexamethasone Release from Conducting Polymer Hydrogels.","authors":"Matthew S Horrocks, Kirill E Zhurenkov, Matthew S Ting, Darren Svirskis, Jenny Malmström","doi":"10.1177/19373341251373100","DOIUrl":"https://doi.org/10.1177/19373341251373100","url":null,"abstract":"<p><p>Spatially and temporally controlled drug delivery is an important field to address the limitations of conventional pharmaceutical administration. While many effective controlled drug delivery systems exist, the repertoire of systems that additionally present a beneficial mechanical environment to cells remains scarce. To address this, a comprehensive release study of fluorescein as a model drug, and the corticosteroid dexamethasone, from poly(<i>N</i>-isopropylacrylamide)/polypyrrole (pNIPAM/PPy) conducting polymer hydrogels is presented within this study. Cyclic voltammetry and scanning electron microscopy (SEM) indicated that having the pNIPAM hydrogel phase present and doping with drugs reduced PPy thickness and shifted/suppressed redox peaks to some degree but not enough to prevent release. Fluorescein release was initiated by constant reduction, with a maximum of 54.5 ± 6.8 µg/cm² from PPy films and 6.3 ± 1.1 µg/cm² from pNIPAM/PPy. The quantity of fluorescein released was shown to be tunable by modulating the charge passed during PPy electropolymerization. Fluorescein-loaded pNIPAM/PPy samples were capable of multiple cycles of depletion and reloading via re-incorporation through re-oxidation in a fluorescein solution. The stability of pNIPAM/PPy regarding drug release was demonstrated, with no difference in release profiles and quantities after soaking samples for 1, 8, and 15 days. Interestingly, constant reduction did not elicit release of dexamethasone, while a biphasic pulsed potential of ±0.8 V at 0.5 Hz was effective. Minimal leaching of dexamethasone without stimulation was shown, alongside a multi-day, multi-triggerable release profile upon short stimulations. pNIPAM/PPy conducting polymer hydrogels are a promising platform for on/off drug delivery, with a nondegrading matrix, minimal passive drug-leaching, and where the drug payload can be reloaded, all while providing a suitable mechanical environment to interface with living cells.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145034775","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}

{"title":"A Human Progenitor Cell-Based Tissue Engineered Intervertebral Disc.","authors":"Sage S Frehner, Matthew Fainor, Galina Dulatov, Ryan Ringwood, Hannah Loftus, Cody Warner, Aira Bazaz, Harvey E Smith, Robert L Mauck, Isaac Erickson, Sarah E Gullbrand PhD","doi":"10.1177/19373341251373104","DOIUrl":"https://doi.org/10.1177/19373341251373104","url":null,"abstract":"<p><p>Cell and tissue engineering therapies provide promise for regenerating damaged intervertebral disc (IVD) tissue and resolving the low back pain that often accompanies it. However, these treatments remain experimental and unavailable for patients. Furthermore, the large body of work characterizing and utilizing mesenchymal stromal cells (MSCs) for these applications has, unfortunately, not resulted in any FDA-approved spinal therapies. Herein, we characterized DiscGenics's human cadaver-derived discogenic nucleus pulposus (NP) progenitor cells and, for the first time, their discogenic annulus fibrosus (AF) progenitor cells. We then used these discogenic NP and AF cells to create biomimetic human-sized total tissue-engineered IVD replacements, also known as endplate-modified angle ply structures (eDAPS), and compared these with eDAPS formulated with goat or human MSCs. Prior to eDAPS fabrication, discogenic cells were expanded using either two-dimensional attachment culture or three-dimensional suspension culture. Currently, no data exist as to how these discogenic progenitor cells deposit extracellular matrix in a 3D culture environment, nor do data exist characterizing whether the unique expansion environment influences subsequent discogenic cell behavior. Our data support that NP and AF discogenic cells occupy unique niches and serve distinct functions, both in the IVD and in an <i>in vitro</i> 3D culture environment. As a result, discogenic cells deposited more matrix overall than did MSCs. That matrix was distinct between the NP and AF analogs of the tissue-engineered IVDs while also being more homogeneous within each region. Most importantly, unlike both MSC groups, discogenic cells deposited little to no collagen X, suggesting that discogenic eDAPS possess a more stable regional phenotype that will be less susceptible to hypertrophy and downstream calcification. Overall, DiscGenics's discogenic NP and AF cells made compositionally and mechanically superior eDAPS when compared with both human and goat MSCs, with only minor differences between attachment- and suspension-derived discogenic cell eDAPS, supporting their use as a cell source for the creation of human-scale living whole disc replacements.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145002052","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}

{"title":"Chitosan Bioceramic Composites for Bone Regeneration: Insights from <i>In Vitro</i> and <i>In Vivo</i> Studies.","authors":"Jeffrey Huang, Jithendra Ratnayake, Maree Gould, George Dias","doi":"10.1177/19373341251372969","DOIUrl":"https://doi.org/10.1177/19373341251372969","url":null,"abstract":"<p><p>Bone-related pathologies due to injuries, trauma, and disease are a burden on the current health system that will only continue to grow as the population's life expectancy increases. The field of biomaterials aims to address these concerns by exploring, investigating, and optimizing bioregenerative grafts. In the context of bone regeneration, many biomaterials aim to achieve autograft-level regenerative properties, such as osteoconduction, osteoinduction, and low immunogenicity but also aim to address the disadvantages, such as the need for a secondary operation, donor site burden, and limited donor availability. Chitosan (CS) is a natural polymer well-studied in the field of biomaterials; it is known for its ease of fabrication, biocompatibility, antibacterial nature, and being a nonproteinaceous polysaccharide, which offers the advantage of low immunogenicity. However, CS lacks any osteogenic potential and is often combined with a bioceramic, creating a biocomposite scaffold. Bioceramics are ceramics specifically designed to aid bone regeneration due to their potential osteogenic properties. Although CS-bioceramic composites have been extensively studied, most research emphasizes their physicochemical properties, with limited attention to biological performance and <i>in vivo</i> outcomes. This review presents current findings on the regenerative potential of various CS-bioceramic composites, with a particular focus on <i>in vitro</i> and <i>in vivo</i> studies.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":"0"},"PeriodicalIF":2.9,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144994454","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}

{"title":"Engineering Vascularized Transplantable Soft Tissue Free Flaps in Sheep Using the Arteriovenous Loop Technique.","authors":"Florian Falkner, Simon Andreas Mayer, Benjamin Thomas, Arno Dimmler, Patrick Heimel, Karl Schneider, Annika Kengelbach-Weigand, Anne-Margarethe Kramer, Rebecca Luisa Schaefer, Adriana C Panayi, Jonathan P Sleeman, Wilko Thiele, Bruno Podesser, Helga Bergmeister, Ulrich Kneser, Volker J Schmidt, Amir K Bigdeli","doi":"10.1177/19373341251372950","DOIUrl":"10.1177/19373341251372950","url":null,"abstract":"<p><p>The aim of this study was to grow axially vascularized soft tissue flaps in sheep using the arteriovenous loop (AVL) technique to be transplanted for defect reconstruction. This technique may be a promising alternative to conventional free flaps to further reduce flap donor site morbidity. In this pilot study, AVLs (<i>n</i> = 12) were created in the groins of six sheep, placed into an isolation chamber, and embedded in Matriderm®. Tissue volume, vascularization, and cell proliferation were assessed on postoperative day (POD) 28 using immunohistochemical staining and microcomputed tomography (µCT). Four AVL free flaps were microsurgically anastomosed to the neck vessels in a standardized defect sheep model on POD 28. Defect closure and intrinsically vascularized scaffold-based bioengineered flaps (IVSBs) flap perfusion were studied by angiography and histology 10 days after transplantation. One IVSB flap was lost due to chamber infection. At POD 28, the remaining 11 IVSB flaps had filled the isolation chamber. Histological examination and µCT analysis of seven IVSB flaps verified homogeneous microvascular networks within the flaps. The mean number of microvessels, vessel volume, and the percentage of proliferating cells increased significantly over time. In the defect model, all four transplanted flaps showed macroscopically, angiographically, and histologically stable defect closure 10 days after transplantation, with homogeneous vascular integration into the surrounding tissue. This pilot study demonstrates that in a large animal model complex, defects can be reconstructed using free IVSB flaps with a clinically relevant tissue volume. These data provide the preclinical proof prior to human application.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144980484","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}

{"title":"Osteoclast Incorporation in an <i>In Vitro</i> 3D Model of Endochondral Ossification.","authors":"Amaia Garmendia Urdalleta, Janneke Witte-Bouma, Nicole Kops, Andrea Lolli, Eric Farrell","doi":"10.1089/ten.tea.2024.0281","DOIUrl":"10.1089/ten.tea.2024.0281","url":null,"abstract":"<p><p><i>In vitro</i> models aim to recapitulate human physiological processes, improving upon and replacing the need for animal-based models. Modeling bone formation via endochondral ossification <i>in vitro</i> is a very complex process due to the large number of cell types involved. Most current models are limited to mimicking the initial stages of the process (i.e., cartilage template formation and mineralization of the matrix), using a single cell type. Chondroclasts/osteoclasts are key players in cartilage resorption during endochondral ossification, but their introduction into <i>in vitro</i> models has thus far proven challenging. In this study, we aimed toward a new level of model complexity by introducing human monocyte-derived osteoclasts into 3D <i>in vitro-</i>cultured cartilage templates undergoing mineralization. Chondrogenic and mineralized chondrogenic pellets were formed from human pediatric bone marrow stromal cells and cultured in the presence of transforming growth factor-β3 (TGF-β) and TGF-β/β-glycerophosphate, respectively. These pellets have the capacity to form bone if implanted <i>in vivo.</i> To identify suitable <i>in vitro</i> co-culture conditions and investigate cell interactions, pellets were co-cultured with CD14+ monocytes in an indirect (transwell) or direct setting for up to 14 days, and osteoclastogenesis was assessed by means of histological stainings, osteoclast counting, and gene expression analysis. Upon direct co-culture, we achieved effective osteoclast formation <i>in situ</i> in regions of both mineralized and unmineralized cartilages. Notably, <i>in vitro</i>-generated osteoclasts showed the ability to form tunnels in the chondrogenic matrix and infiltrate the mineralized matrix. Addition of osteoclasts in human <i>in vitro</i> models of endochondral ossification increases the physiological relevance of these models. This will allow for the development of robust 3D human <i>in vitro</i> systems for the study of bone formation, disease modeling, and drug discovery, further reducing the need for animal models in the future. Impact Statement <i>In vitro</i> bone formation models of endochondral ossification are currently limited to the recapitulation of the initial stages of the process. In this article, we present a novel <i>in vitro</i> endochondral ossification model where osteoclasts were incorporated into mineralized hypertrophic cartilage templates, adding a new level of complexity toward the modeling of cartilage resorption during endochondral ossification.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":"1156-1170"},"PeriodicalIF":2.9,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144021318","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}