Tissue Engineering Part A最新文献

{"title":"Modular, Vascularized Hypertrophic Cartilage Constructs for Bone Tissue Engineering Applications.","authors":"Nicholas G Schott, Gurcharan Kaur, Rhima M Coleman, Jan P Stegemann","doi":"10.1089/ten.tea.2024.0367","DOIUrl":"https://doi.org/10.1089/ten.tea.2024.0367","url":null,"abstract":"<p><p>Insufficient vascularization is the main barrier to creating engineered bone grafts for treating large and ischemic defects. Modular tissue engineering approaches have promise in this application because of the ability to combine tissue types and localize microenvironmental cues to drive desired cell function. In direct bone formation approaches, it is challenging to maintain sustained osteogenic activity, since vasculogenic cues can inhibit tissue mineralization. This study harnessed the physiological process of endochondral ossification to create multiphase tissues that allowed concomitant mineralization and vessel formation. Mesenchymal stromal cells in pellet culture were differentiated toward a cartilage phenotype, followed by induction to chondrocyte hypertrophy. Hypertrophic pellets (HPs) exhibited increased alkaline phosphatase activity, calcium deposition, and osteogenic gene expression relative to chondrogenic pellets. In addition, HPs secreted and sequestered angiogenic factors, and supported new blood vessel formation by cocultured endothelial cells and undifferentiated stromal cells. Multiphase constructs created by combining HPs and vascularizing microtissues and maintained in an unsupplemented basal culture medium were shown to support robust vascularization and sustained tissue mineralization. These results demonstrate a promising <i>in vitro</i> strategy to produce multiphase-engineered constructs that concomitantly support the generation of mineralized and vascularized tissue in the absence of exogenous osteogenic or vasculogenic medium supplements.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144053523","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":"https://doi.org/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.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-25","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}

{"title":"Surface-Patterned Silicon Oxynitride for Aligned Myotubes and Neurite Outgrowth <i>In Vitro</i>.","authors":"Kamal Awad, Matthew Fiedler, Ahmed S Yacoub, Leticia Brotto, Pranesh B Aswath, Marco Brotto, Venu Varanasi","doi":"10.1089/ten.tea.2024.0358","DOIUrl":"https://doi.org/10.1089/ten.tea.2024.0358","url":null,"abstract":"<p><p>Traumatic injuries lead to volumetric muscle loss (VML) and nerve damage that cause chronic functional deficits. Due to the inability of mammalian skeletal muscle to regenerate after VML damage, engineered scaffolds have been explored to address this challenge, but with limited success in functional restoration. We introduce novel bioactive amorphous silicon oxynitride (SiONx) biomaterials with surface properties and Si ion release to accelerate muscle and nerve cell differentiation for functional tissue regeneration. Micropatterned scaffolds were designed and developed on Si-wafer to test the effect of SiONx on myogenesis and neurogenesis. The scaffolds were created using UV photolithography to first pattern their surface, followed by the deposition of SiONx through plasma enhanced chemical vapor deposition (PECVD). X-ray diffraction (XRD) and energy dispersive spectroscopy (EDS) confirmed the uniform chemical structure of an amorphous SiONx film on the patterned surfaces. Atomic force microscopy and scanning electron microscopy (SEM) elucidated the surface morphology with a uniform 2 μm grating microstructure. The 2 µm pattern size is within the range of cellular dimensions, allowing for effective cell-surface interactions. Further, 2 µm features provide sufficient contact points for cell adhesion without overwhelming the cell's ability to interact with the surface. Two separate studies were conducted with SiONx biomaterials and Si ions alone. This was done to understand how Si ions impact cell response separate from the surfaces. C2C12 mouse myoblasts and NG108 neuronal cells were cultured on SiONx biomaterials. In separate studies, we tested the effect of Si ion treatments with these cells (cultured on tissue culture plastic). Cell culture studies demonstrated enhanced C2C12 myoblast attachment and proliferation on SiONx surfaces. High-resolution SEM and fluorescence images revealed highly aligned myotubes (from C2C12 cells) and axons (from NG108 cells) in a parallel direction to the micropatterned SiONx scaffolds. GAP43 expression, neurite outgrowth, and alignment were significantly increased with the Si-ions and SiONx biomaterials. These findings suggest that SiONx scaffolds enhance muscle and nerve cell adhesion and growth and promote the formation of aligned myotubes and axons on the pattern surfaces.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144054089","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":"Three-Dimensional Bioprinting of Astrocytes and Endothelial Cells to Direct Retinal Axon Growth and Vascularization.","authors":"Fatima E Abukunna, Afnan M Aladdad, Kiran J McLoughlin, Khyathi Thallapureddy, Michael Vierra, Zoya Siddiqui, Karl E Kador","doi":"10.1089/ten.tea.2024.0326","DOIUrl":"https://doi.org/10.1089/ten.tea.2024.0326","url":null,"abstract":"<p><p>Retinal organoids (ROs) are currently used to study retinal development and diseases but cannot model glaucoma because they fail to form a nerve fiber layer (NFL) and optic nerve (ON). Utilizing three-dimensional bioprinting, ON head astrocytes (ONHAs) and vascular endothelial cells, both of which contribute to NFL development <i>in vivo</i> but are absent in ROs, were positioned at the center of scaffolds seeded with retinal ganglion cells (RGCs). In experiments using ONHAs isolated from developing retinas, polarization of RGC neurite growth increased by 43% while ONHA from adult retinas or astrocytes from the developing peripheral retina or developing cortex did not increase polarization above controls. Furthermore, RGC-seeded scaffolds increased both the number and rate of ONHAs migrating out from the printed center compared to scaffolds lacking RGCs, mimicking the migration pattern observed during retinal development. Finally, in scaffolds containing both ONHAs and endothelial cells, the endothelial cells preferentially migrate on and only form vascular tube structures on scaffolds also containing RGCs. These results suggest that recreating the developmental organization of the retina can recapitulate the mechanism of NFL development and retinal vascularization <i>in vitro.</i> This step is not only necessary for the development of retinal models of glaucoma but has the potential for translation to other parts of the central nervous system.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144058614","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":"Microencapsulation of Liver Spheroids with Poly(Vinyl Alcohol) Enhances Function Compared with Alginate.","authors":"Stephen Harrington, Edward Larson, Aldyn Wildey, Vincent Ling, Lisa Stehno-Bittel, Francis Karanu","doi":"10.1089/ten.tea.2024.0312","DOIUrl":"https://doi.org/10.1089/ten.tea.2024.0312","url":null,"abstract":"<p><p><b><i>Background and Aims:</i></b> Cell therapy approaches to treating chronic liver disease provide only transient improvements, mainly due to loss of hepatocytes after infusion. Microencapsulation in alginate has been shown to protect transplanted cells from physical stress and rejection, but the poor biocompatibility of alginate can lead to graft failure. This study aimed to evaluate a biocompatible poly(vinyl alcohol) (PVA)-based microcapsule against standard alginate for improved transplantation outcome of liver spheroids. <b><i>Materials and Methods:</i></b> Human hepatocyte spheroids were microencapsulated in alginate or PVA hydrogel microspheres. Viability and function (albumin secretion and CYP activity) of the encapsulated spheroids were assessed <i>in vitro</i> at 3, 10, and 30 days postencapsulation and compared with unencapsulated spheroids. Spheroids were implanted intraperitoneally into immunodeficient mice, and human albumin levels in serum were monitored over 30 days. Cell-free microspheres were implanted in immune-competent mice to assess material biocompatibility. <b><i>Results:</i></b> Unencapsulated spheroids aggregated extensively beyond 10 days, precluding day 30 assessment. At day 30, PVA spheroids showed significantly higher CYP1A1 induction, albumin secretion, and metabolic activity compared with alginate. Mice receiving PVA spheroids had significantly higher serum albumin after 30 days compared with alginate and unencapsulated spheroids. Empty PVA microspheres showed less evidence of foreign body response <i>in vivo</i>, whereas thicker regions of inflamed tissue were observed in the alginate group. <b><i>Conclusions:</i></b> PVA-encapsulated hepatocyte spheroids maintained better overall viability, metabolic activity, and function compared with alginate-encapsulated cells both <i>in vitro</i> and <i>in vivo</i>. Both encapsulated groups demonstrated substantially improved outcomes compared with unencapsulated cells.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144044608","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":"Osteoblast-Mesenchymal Stem Cell Coculture Drives <i>In Vitro</i> Osteogenesis in 3D Bioprinted Periosteum.","authors":"Shannon T McLoughlin, Paige Wilcox, John F Caccamese, John P Fisher","doi":"10.1089/ten.tea.2025.0038","DOIUrl":"https://doi.org/10.1089/ten.tea.2025.0038","url":null,"abstract":"<p><p>The periosteum serves as a local source of osteoprogenitor cells and vasculature, therefore influencing the key processes of osteogenesis and neovascularization during bone healing. However, it is often not considered in traditional bone tissue engineering strategies. The periosteum consists of two stratified cell layers, including an inner cambium layer, which serves as a local source of osteoblasts (OBs) and osteoprogenitor cells, and an outer fibrous layer, which hosts vasculature, collagen fibers, and support cells. While several studies have investigated different methodologies to produce tissue-engineered periosteum (TEP) substitutes, few have evaluated the roles of specific cell types within the inner cambium layer and their patterning in 3D environments on underlying bone tissue development. Therefore, we sought to investigate whether mesenchymal stem cells (MSCs) alone, OBs alone, or a 1:1 mixture of the two would result in increased osteogenic differentiation of bone layer MSCs in a 3D bioprinted periosteum-bone coculture model <i>in vitro</i>. We first evaluated these effects in a 2D transwell model, demonstrating that OB-containing cultures, either alone or in a mixed population with MSCs, upregulated alkaline phosphatase activity and runt-related transcription factor 2 (<i>RUNX2</i>) expression. In the 3D bioprinted model, the mixed population showed higher levels of <i>RUNX2</i> expression and calcium deposition, indicating increased osteogenic differentiation within the bone layer. Results obtained from this study provide evidence that a mixed population of MSCs and OBs within the inner cambium layer of TEP can increase bone regeneration.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144058947","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":"Aging Impairs Implant Osseointegration Through a Novel Reactive Oxygen Species-Hypoxia-Inducible Factor 1α/p53 Axis.","authors":"Jingjing Shao, Shibo Liu, Chenfeng Chen, Wenchuan Chen, Zhimin Zhu, Lei Li","doi":"10.1089/ten.tea.2024.0355","DOIUrl":"https://doi.org/10.1089/ten.tea.2024.0355","url":null,"abstract":"<p><p>Enhancing bone-vessel coupling to form high-quality vascular-rich peri-implant bone is crucial for improving implant prognosis in elder patients. Notably, hypoxia-inducible factor 1α (HIF1α) is known to promote osteogenesis-angiogenesis coupling; however, this effect remains to be investigated in aged bone owing to the dual effect of HIF1α in different aged organs. In this study, HIF1α inhibitor or activator was applied to aged mice and their bone mesenchymal stem cells (BMSCs) to investigate the effects and inner mechanism of HIF1α on the peri-implant osteogenesis and angiogenesis in senescent status. Cell senescence, along with osteogenic and angiogenic abilities of aged BMSCs, was detected, respectively. Meanwhile, a femur implant implantation model was constructed on aged mice, and the bone-vessel coupling of peri-implant bone was observed. Mandibular bone morphology was also detected to further provide evidence for clinical oral implantation. Furthermore, p53 expression was examined <i>in vivo</i> and <i>in vitro</i> following HIF1α intervention. A reactive oxygen species (ROS) scavenger was also adopted to further investigate the roles of ROS in the HIF1α-p53 axis. Results showed that the suppression of HIF1α alleviated senescence and osteogenesis-angiogenesis coupling of aged BMSCs, while its activation aggravated these effects. The mandible phenotype and bone-vessel coupling in aged peri-implant bone also changed accordingly upon regulation of HIF1α. Mechanistically, p53 changed in the same direction as HIF1α <i>in vivo</i> and <i>in vitro</i>. Moreover, the ROS scavenger reversed the HIF1α-p53 relationship and weakened the effect of HIF1α inhibitor on peri-implant bone improvement. In conclusion, in aged mice, highly expressed HIF1α impaired peri-implant bone-vessel coupling and implant osseointegration through p53, and accumulated ROS was a prerequisite for HIF1α to positively regulate p53. These findings provide new insights into the role of HIF1α and the ROS-HIF1α/p53 signaling axis, offering potential therapeutic targets to improve implant outcomes in elderly patients.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143765905","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":"Enhancing Bone Healing Through Localized Cold Therapy in a Murine Femoral Fracture Model.","authors":"Matthew Zakaria, Jerome Allard, Jose Garcia, Justin Matta, Yazan Honjol, Drew Schupbach, Michael Grant, Fackson Mwale, Edward Harvey, Geraldine Merle","doi":"10.1089/ten.TEA.2024.0069","DOIUrl":"10.1089/ten.TEA.2024.0069","url":null,"abstract":"<p><p>Fracture healing, a critical and complex biological process, often presents challenges in clinical practice with the current standards failing to fully address the medical needs for rapid and effective recovery. In this work, a localized cold therapy is investigated as an alternative approach to expedite bone healing. We hypothesized that optimized cold application can enhance bone healing within a fracture model by inducing hypoxia, leading to accelerated angiogenesis along with improved osteogenesis. A short, localized cold exposure is directly applied to the fracture site over a 4-week period in a mouse fracture model, aiming to assess its impact on bone formation through mechanisms of angiogenesis and osteogenesis. Our results revealed a significantly greater volume of new bone tissue and enhanced vascularity at the fracture site in the cold-treated group compared with controls. Calcified tissue histology analysis showed that the accelerated callus maturation and development of the vascular network following cold exposure were associated with an activity increase of alkaline phosphatase and transient receptor potential vanilloid 1. These biological changes were accompanied by a hypoxic environment induced during cold therapy. The study provides compelling evidence supporting the efficacy of intermittent cold therapy in accelerating fracture healing. These promising results highlight the need for further research in larger-scale studies and diverse fracture models, underlining the potential of cold therapy as a novel, noninvasive treatment strategy in orthopedic care.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":"303-314"},"PeriodicalIF":3.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141749809","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":"Oxidized Low-Density Lipoprotein Decreases the Survival of Bone Marrow Stem Cells via Inhibition of Bcl-2 Expression.","authors":"Xin Li, Yu Li, Hao Yu, Li-Li Men, Glenn Deng, Zhenguo Liu, Jian-Ling Du","doi":"10.1089/ten.TEA.2024.0025","DOIUrl":"10.1089/ten.TEA.2024.0025","url":null,"abstract":"<p><p>Therapy with mesenchymal stem cells (MSCs) is considered an attractive strategy for the repair or regeneration of damaged tissues. However, low survival of MSCs limits their applications clinically. Oxidized low-density lipoprotein (ox-LDL) is significantly increased in patients with hyperlipidemia and decreases the survival of MSCs. Bcl-2 is critically involved in important cell functions, including cell membrane integrity and cell survival. The present study was designed to test the hypothesis that ox-LDL attenuates the survival of MSCs through suppression of Bcl-2 expression. Bone marrow MSCs from C57BL/6 mice were cultured with ox-LDL at different concentrations (0-140 μg/mL) for 24 h with native LDL as control. Ox-LDL treatment substantially decreased the survival of MSCs dose-dependently and enhanced the release of intracellular lactate dehydrogenase (LDH) in association with a significant decrease in Bcl-2 protein level without change in BAX protein expression in MSCs. Bcl-2 overexpression effectively protected MSCs against ox-LDL-induced damages with preserved cell numbers without significant increase in LDH release. Treatment with <i>N</i>-acetylcysteine (NAC) (1 mM) effectively preserved Bcl-2 protein expression in MSCs and significantly attenuated ox-LDL-induced decrease of cell number and increase in the release of intracellular LDH. These data indicated that ox-LDL treatment resulted in a significant damage of cell membrane and dramatically decreased the survival of MSCs dose-dependently through inhibition of Bcl-2 expression. NAC treatment significantly protected MSCs against the damage of cell membrane by ox-LDL and promoted the survival of MSCs in association with preserved Bcl-2 expression.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":"325-333"},"PeriodicalIF":3.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141181618","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":"Impact of Passaging Primary Skeletal Muscle Cell Isolates on the Engineering of Skeletal Muscle.","authors":"Olga M Wroblewski, Christopher S Kennedy, Emmanuel E Vega-Soto, Celeste E Forester, Eileen Y Su, Matthew H Nguyen, Paul S Cederna, Lisa M Larkin","doi":"10.1089/ten.TEA.2024.0044","DOIUrl":"10.1089/ten.TEA.2024.0044","url":null,"abstract":"<p><p>Volumetric muscle loss (VML) is a clinical state that results in impaired skeletal muscle function. Engineered skeletal muscle can serve as a treatment for VML. Currently, large biopsies are required to achieve the cells necessary for the fabrication of engineered muscle, leading to donor-site morbidity. Amplification of cell numbers using cell passaging may increase the usefulness of a single muscle biopsy for engineering muscle tissue. In this study, we evaluated the impact of passaging cells obtained from donor muscle tissue by analyzing characteristics of <i>in vitro</i> cellular growth and tissue-engineered skeletal muscle unit (SMU) structure and function. Human skeletal muscle cell isolates from three separate donors (P0-Control) were compared with cells passaged once (P1), twice (P2), or three times (P3) by monitoring SMU force production and determining muscle content and structure using immunohistochemistry. Data indicated that passaging decreased the number of satellite cells and increased the population doubling time. P1 SMUs had slightly greater contractile force and P2 SMUs showed statistically significant greater force production compared with P0 SMUs with no change in SMU muscle content. In conclusion, human skeletal muscle cells can be passaged twice without negatively impacting SMU muscle content or contractile function, providing the opportunity to potentially create larger SMUs from smaller biopsies, thereby producing clinically relevant sized grafts to aid in VML repair.</p>","PeriodicalId":56375,"journal":{"name":"Tissue Engineering Part A","volume":" ","pages":"315-324"},"PeriodicalIF":3.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141319138","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}