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Exosomes Derived from Rejuvenated Stem Cells Inactivate NLRP3 Inflammasome and Pyroptosis of Nucleus Pulposus Cells via the Transfer of Antioxidants. 从再生干细胞中提取的外泌体通过抗氧化剂的转移激活 NLRP3 炎症体和核浆细胞的嗜热症。
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-10-01 Epub Date: 2024-07-26 DOI: 10.1007/s13770-024-00663-z
Shuai Peng, Xiangyang Liu, Lei Chang, Bin Liu, Mingyan Zhang, Yan Mao, Xiongjie Shen

Background: Accumulating evidence supports the potential of exosomes as a promising therapeutic approach for intervertebral disc degeneration (IDD). Nevertheless, enhancing the efficiency of exosome treatment remains an urgent concern. This study investigated the impact of quercetin on the characteristics of mesenchymal stem cells (MSCs) and their released exosomes.

Methods: Exosomes were obtained from quercetin pre-treated MSCs and quantified for the production based on nanoparticle tracking and western blot analysis. The molecules involved in the secretion and cargo sorting of exosomes were investigated using western blot and immunofluorescence analysis. Based on the in vitro biological analysis and in vivo histological analysis, the effects of exosomes derived from conventional or quercetin-treated MSCs on nucleus pulposus (NP) cells were compared.

Results: A significant enhancement in the production and transportation efficiency of exosomes was observed in quercetin-treated MSCs. Moreover, the exosomes derived from quercetin-treated MSCs exhibited a greater abundance of antioxidant proteins, specifically superoxide dismutase 1 (SOD1), which inhibit the activation of NOD-like receptor thermal protein domain associated protein 3 (NLRP3) inflammasome in NP cells. Through in vitro and in vivo experiments, it was elucidated that exosomes derived from quercetin-treated MSCs possessed enhanced anti-inflammatory and antioxidant properties.

Conclusion: Collectively, our research underscores an optimized therapeutic strategy for IDD utilizing MSC-derived exosomes, thereby augmenting the efficacy of exosomes in intervertebral disc regeneration.

背景:越来越多的证据表明,外泌体有望成为治疗椎间盘退变(IDD)的一种有效方法。然而,提高外泌体治疗的效率仍是一个亟待解决的问题。本研究探讨了槲皮素对间充质干细胞(MSCs)及其释放的外泌体特性的影响:方法:从经槲皮素预处理的间充质干细胞中获取外泌体,并根据纳米颗粒追踪和Western印迹分析对外泌体的产生进行定量。利用 Western 印迹和免疫荧光分析研究了参与外泌体分泌和货物分拣的分子。根据体外生物学分析和体内组织学分析,比较了传统间充质干细胞或槲皮素处理的间充质干细胞产生的外泌体对髓核细胞的影响:结果:在槲皮素处理的间充质干细胞中观察到外泌体的产生和运输效率明显提高。此外,经槲皮素处理的间充质干细胞产生的外泌体含有更多的抗氧化蛋白,特别是超氧化物歧化酶1(SOD1),它能抑制NP细胞中NOD样受体热蛋白域相关蛋白3(NLRP3)炎性体的激活。通过体外和体内实验,我们发现槲皮素处理的间充质干细胞产生的外泌体具有更强的抗炎和抗氧化特性:总之,我们的研究强调了利用间充质干细胞衍生的外泌体治疗IDD的优化策略,从而增强了外泌体在椎间盘再生中的功效。
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引用次数: 0
Rapamycin Attenuates H2O2-Induced Oxidative Stress-Related Senescence in Human Skin Fibroblasts. 雷帕霉素可减轻H2O2诱导的人皮肤成纤维细胞氧化应激导致的衰老
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-10-01 Epub Date: 2024-08-02 DOI: 10.1007/s13770-024-00660-2
Yuyang Tang, Sen Yang, Zhen Qiu, Li Guan, Yigui Wang, Guixin Li, Yuanyu Tu, Lijuan Guo

Background: Oxidative stress plays an important role in the skin aging process. Rapamycin has been shown to have anti-aging effects, but its role in oxidative senescence of skin cells remains unclear. The aim of this study was to explore the effect of rapamycin on oxidative stress-induced skin cell senescence and to illustrate the mechanism.

Methods: Primary human skin fibroblasts (HSFs) were extracted and a model of H2O2-induced oxidative senescence was constructed, and the effects of rapamycin on their value-added and migratory capacities were detected by CCK-8 and scratch assays. SA-β-gal was utilized to detect senescence, oxidatively closely related factors were also assessed. Gene and protein expressions of senescence, oxidative, and autophagy were detected by western blotting and quantitative-PCR. The data were analyzed by one-way analysis of variance.

Results: Rapamycin (0.1 nmol/L for 48 h) promoted the proliferative and migration of H2O2-treated HSFs (p < 0.05), decreased senescent phenotypes SA-β-gal staining and the expression of P53, and MMP-1 proteins, and increased the expression level of COL1A-1 (p < 0.001). Rapamycin also enhanced the activities of SOD and HO-1, and effectively removed intracellular ROS, MDA levels (p < 0.05), in addition, autophagy-related proteins and genes were significantly elevated after rapamycin pretreatment (p < 0.001). Rapamycin upregulated the autophagy pathway to exert its protective effects.

Conclusion: Our findings indicate that rapamycin shields HSFs from H2O2-induced oxidative damage, the mechanism is related to the reduction of intracellular peroxidation and upregulation of autophagy pathway. Therefore, rapamycin has the potential to be useful in the investigation and prevention of signs of aging and oxidative stress.

背景:氧化应激在皮肤老化过程中起着重要作用。雷帕霉素已被证明具有抗衰老作用,但其在皮肤细胞氧化衰老中的作用仍不清楚。本研究旨在探讨雷帕霉素对氧化应激诱导的皮肤细胞衰老的影响,并说明其机制:方法:提取原代人皮肤成纤维细胞(HSFs),构建H2O2诱导的氧化衰老模型,通过CCK-8和划痕试验检测雷帕霉素对其增值和迁移能力的影响。利用SA-β-gal检测衰老,还评估了与氧化密切相关的因子。通过 Western 印迹和定量 PCR 检测衰老、氧化和自噬的基因和蛋白质表达。数据采用单因素方差分析:雷帕霉素(0.1 nmol/L,48 h)促进了经 H2O2 处理的 HSFs 的增殖和迁移(p 结论:雷帕霉素(0.1 nmol/L,48 h)促进了经 H2O2 处理的 HSFs 的增殖和迁移:我们的研究结果表明,雷帕霉素能保护HSFs免受H2O2诱导的氧化损伤,其机制与细胞内过氧化物的减少和自噬途径的上调有关。因此,雷帕霉素有望用于研究和预防衰老迹象和氧化应激。
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引用次数: 0
Platelet-Rich Plasma-Embedded Porous Polycaprolactone Film with a Large Surface Area for Effective Hemostasis. 具有大表面积的富血小板血浆包覆多孔聚己内酯薄膜可实现有效止血。
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-10-01 Epub Date: 2024-06-19 DOI: 10.1007/s13770-024-00656-y
Min Ji Kim, Ye Jin Song, Tae Gyun Kwon, Jin Ho Lee, So Young Chun, Se Heang Oh

Background: Uncontrollable and widespread bleeding caused by surgery or sudden accidents can lead to death if not treated with appropriate hemostasis. To prevent excessive life-threatening bleeding, various hemostatic agents based on polymeric biomaterials with various additives for accelerated blood coagulation have been adopted in clinical fields. In particular, platelet-rich plasma (PRP), which contains many blood coagulation factors that can accelerate blood clot formation, is considered as one of the most effective hemostatic additives.

Methods: We investigated a PRP-embedded porous film using discarded (expired) PRP and a film with a leaf-stacked structure (FLSS), as a hemostatic agent to induce rapid hemostasis. The film, which contained an LSS on one side (PCL-FLSS), was fabricated by a simple heating-cooling technique using tetraglycol and polycaprolactone (PCL) film. Activated PRP was obtained by the thawing of frozen PRP at the end of its expiration date (the platelet cell membrane is disrupted during the freezing and thawing of PRP, thus releasing various coagulation factors) and embedded in the PCL-FLSS (PRP-FLSS).

Results: From in vitro and in vivo experiments using a rat hepatic bleeding model, it was recognized that PRP-FLSS is not only biocompatible but also significantly accelerates blood clotting and thus prevents rapid bleeding, probably due to a synergistic effect of the sufficient supply of various blood coagulants from activated PRP embedded in the LSS layer and the large surface area of the LSS itself.

Conclusion: The study suggests that PRP-FLSS, a combination of a porous polymer matrix with a unique morphology and discarded biofunctional resources, can be an advanced hemostatic agent as well as an upcycling platform to avoid the waste of biofunctional resources.

背景:手术或突发事故造成的无法控制的大面积出血,如果不采取适当的止血措施,可能会导致死亡。为防止过量出血危及生命,临床上采用了各种基于高分子生物材料的止血剂,并添加了各种加速血液凝固的添加剂。其中,富血小板血浆(PRP)含有多种血液凝固因子,可加速血凝块的形成,被认为是最有效的止血添加剂之一:我们使用废弃(过期)的 PRP 和具有叶片叠层结构(FLSS)的薄膜研究了一种 PRP 嵌入多孔薄膜,作为止血剂诱导快速止血。这种一面含有 LSS 的薄膜(PCL-FLSS)是利用四甘醇和聚己内酯(PCL)薄膜通过简单的加热-冷却技术制成的。通过解冻过期的冷冻 PRP(在冷冻和解冻 PRP 的过程中血小板细胞膜被破坏,从而释放出各种凝血因子)来获得活性 PRP,并将其嵌入 PCL-FLSS 中(PRP-FLSS):利用大鼠肝脏出血模型进行的体外和体内实验表明,PRP-FLSS 不仅具有生物相容性,还能显著加速血液凝固,从而防止快速出血,这可能是由于嵌入 LSS 层的活化 PRP 提供了充足的各种凝血因子,而 LSS 本身又具有较大的表面积,二者产生了协同效应:该研究表明,PRP-FLSS 是一种具有独特形态的多孔聚合物基质与废弃生物功能资源的结合体,既可作为一种先进的止血剂,也可作为一种避免生物功能资源浪费的再循环平台。
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引用次数: 0
High-Speed Clearing and High-Resolution Staining for Analysis of Various Markers for Neurons and Vessels. 高速清除和高分辨率染色,用于分析神经元和血管的各种标记。
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-10-01 Epub Date: 2024-07-02 DOI: 10.1007/s13770-024-00658-w
Jung Min Park, Seock Hwan Choi, Eun-Shil Lee, Sang-Il Gum, Sungkuk Hong, Dong Sun Kim, Man-Hoon Han, Soung-Hoon Lee, Ji Won Oh

Background: Tissue clearing enables deep imaging in various tissues by increasing the transparency of tissues, but there were limitations of immunostaining of the large-volume tissues such as the whole brain.

Methods: Here, we cleared and immune-stained whole mouse brain tissues using a novel clearing technique termed high-speed clearing and high-resolution staining (HCHS). We observed neural structures within the cleared brains using both a confocal microscope and a light-sheet fluorescence microscope (LSFM). The reconstructed 3D images were analyzed using a computational reconstruction algorithm.

Results: Various neural structures were well observed in three-dimensional (3D) images of the cleared brains from Gad-green fluorescent protein (GFP) mice and Thy 1-yellow fluorescent protein (YFP) mice. The intrinsic fluorescence signals of both transgenic mice were preserved after HCHS. In addition, large-scale 3D imaging of brains, immune-stained by the HCHS method using a mild detergent-based solution, allowed for the global topological analysis of several neuronal markers such as c-Fos, neuronal nuclear protein (NeuN), Microtubule-associated protein 2 (Map2), Tuj1, glial fibrillary acidic protein (GFAP), and tyrosine hydroxylase (TH) in various anatomical regions in the whole mouse brain tissues. Finally, through comparisons with various existing tissue clearing methodologies such as CUBIC, Visikol, and 3DISCO, it was confirmed that the HCHS methodology results in relatively less tissue deformation and higher fluorescence retention.

Conclusion: In conclusion, the development of 3D imaging based on novel tissue-clearing techniques (HCHS) will enable detailed spatial analysis of neural and vascular networks present within the brain.

背景:方法:在此,我们使用一种称为高速清除和高分辨率染色(HCHS)的新型清除技术对小鼠全脑组织进行了清除和免疫染色。我们使用共聚焦显微镜和光片荧光显微镜(LSFM)观察了清除后大脑中的神经结构。我们使用计算重建算法对重建的三维图像进行了分析:结果:在Gad-绿色荧光蛋白(GFP)小鼠和Thy 1-黄色荧光蛋白(YFP)小鼠清除后大脑的三维图像中,可以很好地观察到各种神经结构。两种转基因小鼠的固有荧光信号在 HCHS 后都得到了保留。此外,通过使用温和去污剂溶液进行免疫染色的 HCHS 方法对大脑进行大规模三维成像,可以对小鼠全脑组织中不同解剖区域的多个神经元标记物(如 c-Fos、神经元核蛋白(NeuN)、微管相关蛋白 2(Map2)、Tuj1、胶质纤维酸性蛋白(GFAP)和酪氨酸羟化酶(TH))进行全局拓扑分析。最后,通过与 CUBIC、Visikol 和 3DISCO 等现有的各种组织清除方法进行比较,证实 HCHS 方法的组织变形相对较小,荧光保留率较高:总之,基于新型组织清除技术(HCHS)的三维成像技术的发展将有助于对大脑内的神经和血管网络进行详细的空间分析。
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引用次数: 0
Tissue Engineering and Regenerative Medicine in the Field of Otorhinolaryngology. 耳鼻喉科领域的组织工程和再生医学。
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-10-01 Epub Date: 2024-07-17 DOI: 10.1007/s13770-024-00661-1
Se-Young Oh, Ha Yeong Kim, Soo Yeon Jung, Han Su Kim

Background: Otorhinolaryngology is a medical specialty that focuses on the clinical study and treatments of diseases within head and neck regions, specifically including the ear, nose, and throat (ENT), but excluding eyes and brain. These anatomical structures play significant roles in a person's daily life, including eating, speaking as well as facial appearance and expression, thus greatly impacting one's overall satisfaction and quality of life. Consequently, injuries to these regions can significantly impact a person's well-being, leading to extensive research in the field of tissue engineering and regenerative medicine over many years.

Methods: This chapter provides an overview of the anatomical characteristics of otorhinolaryngologic tissues and explores the tissue engineering and regenerative medicine research in otology (ear), rhinology (nose), facial bone, larynx, and trachea.

Results and conclusion: The integration of tissue engineering and regenerative medicine in otorhinolaryngology holds the promise of broadening the therapeutic choices for a wide range of conditions, ultimately improving quality of a patient's life.

背景:耳鼻咽喉科是一门医学专科,专注于头颈部疾病的临床研究和治疗,具体包括耳、鼻、喉(ENT),但不包括眼和脑。这些解剖结构在一个人的日常生活中起着重要作用,包括进食、说话以及面部外观和表情,因此极大地影响着一个人的整体满意度和生活质量。因此,这些区域的损伤会严重影响一个人的幸福感,从而导致多年来组织工程和再生医学领域的广泛研究:本章概述了耳鼻喉科组织的解剖学特征,并探讨了耳科(耳)、鼻科(鼻)、面骨、喉和气管的组织工程和再生医学研究:结果和结论:组织工程和再生医学在耳鼻咽喉科的结合有望拓宽多种疾病的治疗选择,最终提高患者的生活质量。
{"title":"Tissue Engineering and Regenerative Medicine in the Field of Otorhinolaryngology.","authors":"Se-Young Oh, Ha Yeong Kim, Soo Yeon Jung, Han Su Kim","doi":"10.1007/s13770-024-00661-1","DOIUrl":"10.1007/s13770-024-00661-1","url":null,"abstract":"<p><strong>Background: </strong>Otorhinolaryngology is a medical specialty that focuses on the clinical study and treatments of diseases within head and neck regions, specifically including the ear, nose, and throat (ENT), but excluding eyes and brain. These anatomical structures play significant roles in a person's daily life, including eating, speaking as well as facial appearance and expression, thus greatly impacting one's overall satisfaction and quality of life. Consequently, injuries to these regions can significantly impact a person's well-being, leading to extensive research in the field of tissue engineering and regenerative medicine over many years.</p><p><strong>Methods: </strong>This chapter provides an overview of the anatomical characteristics of otorhinolaryngologic tissues and explores the tissue engineering and regenerative medicine research in otology (ear), rhinology (nose), facial bone, larynx, and trachea.</p><p><strong>Results and conclusion: </strong>The integration of tissue engineering and regenerative medicine in otorhinolaryngology holds the promise of broadening the therapeutic choices for a wide range of conditions, ultimately improving quality of a patient's life.</p>","PeriodicalId":23126,"journal":{"name":"Tissue engineering and regenerative medicine","volume":" ","pages":"969-984"},"PeriodicalIF":4.4,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11416456/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141627741","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}
引用次数: 0
Transplantation of Neural Progenitor Cells Derived from Stem Cells from Apical Papilla Through Small-Molecule Induction in a Rat Model of Sciatic Nerve Injury. 通过小分子诱导在坐骨神经损伤大鼠模型中移植从顶端乳头干细胞中提取的神经祖细胞
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-08-01 Epub Date: 2024-06-21 DOI: 10.1007/s13770-024-00648-y
Junhao Koh, Junqing Liu, Chi Him Poon, Jun Kang, Mohammed S Basabrain, Lee Wei Lim, Chengfei Zhang

Background: Stem cell-based transplantation therapy holds promise for peripheral nerve injury treatment, but adult availability is limited. A cell culture protocol utilizing a small-molecule cocktail effectively reprogrammed stem cells from apical papilla (SCAPs) into neural progenitor cells, subsequently differentiating into neuron-like cells. This study aims to evaluate neural-induced SCAPs, with and without small-molecule cocktail, for sciatic nerve repair potential.

Methods: A scaffold-free cell sheet technique was used to construct a three-dimensional cell sheet. Subsequently, this cell sheet was carefully rolled into a tube and seamlessly inserted into a collagen conduit, which was then transplanted into a 5 mm sciatic nerve injury rat model. Functional sciatic nerve regeneration was evaluated via toe spread test, walking track analysis and gastrocnemius muscle weight. Additionally, degree of sciatic nerve regeneration was determined based on total amount of myelinated fibers.

Results: Small-molecule cocktail induced SCAPs enhanced motor function recovery, evident in improved sciatic function index and gastrocnemius muscle retention. We also observed better host myelinated fiber retention than undifferentiated SCAPs or neural-induced SCAPs without small-molecule cocktail. However, clusters of neuron-like cell bodies (surrounded by sparse myelinated fibers) were found in all cell sheet-implanted groups in the implantation region. This suggests that while the implanted cells likely survived transplantation, integration was poor and would likely hinder long-term recovery by occupying the space needed for host nerve fibers to project through.

Conclusion: Neural-induced SCAPs with small-molecule cocktail demonstrated promising benefits for nerve repair; further research is needed to improve its integration and optimize its potential for long-term recovery.

背景:以干细胞为基础的移植疗法有望治疗周围神经损伤,但成人可用性有限。一种利用小分子鸡尾酒的细胞培养方案能有效地将来自顶端乳头的干细胞(SCAPs)重编程为神经祖细胞,随后分化为神经元样细胞。本研究旨在评估使用或不使用小分子鸡尾酒的神经诱导SCAPs修复坐骨神经的潜力:方法:采用无支架细胞片技术构建三维细胞片。方法:采用无支架细胞片技术构建三维细胞片,然后将细胞片小心卷成管状,无缝插入胶原导管,再移植到 5 毫米坐骨神经损伤大鼠模型中。通过脚趾伸展试验、行走轨迹分析和腓肠肌重量来评估坐骨神经的功能性再生。此外,根据有髓鞘纤维的总量确定坐骨神经再生的程度:结果:小分子鸡尾酒诱导的 SCAPs 促进了运动功能的恢复,坐骨神经功能指数和腓肠肌保持力的改善就是明证。与未分化的 SCAPs 或未使用小分子鸡尾酒的神经诱导 SCAPs 相比,我们还观察到宿主髓鞘纤维的保留更好。然而,在植入区域的所有细胞片植入组中都发现了神经元样细胞体集群(周围有稀疏的髓鞘纤维)。这表明,虽然植入的细胞很可能在移植后存活下来,但整合能力很差,很可能会占据宿主神经纤维投射所需的空间,从而阻碍长期恢复:结论:使用小分子鸡尾酒的神经诱导 SCAPs 对神经修复大有益处;需要进一步研究以提高其整合性并优化其长期恢复的潜力。
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引用次数: 0
Extracellular Vesicles from Adipose Tissue-Derived Stromal Cells Stimulate Angiogenesis in a Scaffold-Dependent Fashion. 脂肪组织来源基质细胞的细胞外小泡以支架依赖的方式刺激血管生成
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-08-01 Epub Date: 2024-07-08 DOI: 10.1007/s13770-024-00650-4
V E Getova, E Orozco-García, S Palmers, G Krenning, R Narvaez-Sanchez, M C Harmsen

Background: The extracellular vesicles (EVs) secreted by adipose tissue-derived stromal cells (ASC) are microenvironment modulators in tissue regeneration by releasing their molecular cargo, including miRNAs. However, the influence of ASC-derived extracellular vesicles (ASC-EVs) on endothelial cells (ECs) and vascularisation is poorly understood. The present study aimed to determine the pro-angiogenic effects of ASC-EVs and explore their miRNA profile.

Methods: EVs were isolated from normoxic and hypoxic cultured ASC conditioned culture medium. The miRNA expression profile was determined by miRseq, and EV markers were determined by Western blot and immunofluorescence staining. The uptake dynamics of fluorescently labelled EVs were monitored for 24 h. ASC-EVs' pro-angiogenic effect was assessed by sprouting ex vivo rat aorta rings in left ventricular-decellularized extracellular matrix (LV dECM) hydrogel or basement membrane hydrogel (Geltrex®).

Results: ASC-EVs augmented vascular network formation by aorta rings. The vascular network topology and stability were influenced in a hydrogel scaffold-dependent fashion. The ASC-EVs were enriched for several miRNA families/clusters, including Let-7 and miR-23/27/24. The miRNA-1290 was the highest enriched non-clustered miRNA, accounting for almost 20% of all reads in hypoxia EVs.

Conclusion: Our study revealed that ASC-EVs augment in vitro and ex vivo vascularisation, likely due to the enriched pro-angiogenic miRNAs in EVs, particularly miR-1290. Our results show promise for regenerative and revascularisation therapies based on ASC-EV-loaded ECM hydrogels.

背景:脂肪组织源性基质细胞(ASC)分泌的细胞外囊泡(EVs)通过释放包括 miRNAs 在内的分子载体,成为组织再生过程中的微环境调节剂。然而,ASC源性细胞外囊泡(ASC-EVs)对内皮细胞(ECs)和血管生成的影响却鲜为人知。本研究旨在确定ASC-EVs的促血管生成作用,并探索其miRNA谱:方法:从常氧和缺氧培养的 ASC 条件培养液中分离出 EVs。方法:从正常缺氧和缺氧培养的 ASC 条件培养液中分离出 EVs,通过 miRseq 测定其 miRNA 表达谱,并通过 Western 印迹和免疫荧光染色确定 EV 标记。通过在左心室脱细胞细胞外基质(LV dECM)水凝胶或基底膜水凝胶(Geltrex®)中萌发体外大鼠主动脉环,评估了ASC-EVs的促血管生成作用:结果:ASC-EVs增强了主动脉环的血管网络形成。血管网络的拓扑结构和稳定性受到水凝胶支架依赖性的影响。ASC-EV富集了多个miRNA家族/集群,包括Let-7和miR-23/27/24。miRNA-1290是富集度最高的非成簇miRNA,占缺氧EVs中所有读数的近20%:我们的研究表明,ASC-EVs能增强体外和体内血管生成,这可能是由于EVs中富集了促血管生成的miRNA,尤其是miR-1290。我们的研究结果表明,基于 ASC-EV 负载 ECM 水凝胶的再生和血管再通疗法大有可为。
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引用次数: 0
The Porous SilMA Hydrogel Scaffolds Carrying Dual-Sensitive Paclitaxel Nanoparticles Promote Neuronal Differentiation for Spinal Cord Injury Repair. 携带双敏感紫杉醇纳米颗粒的多孔 SilMA 水凝胶支架促进脊髓损伤修复中的神经元分化
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-08-01 Epub Date: 2024-07-15 DOI: 10.1007/s13770-024-00659-9
Zhixiang Li, Tao Zhou, Zhengqi Bao, Min Wu, Yingji Mao

Background: In the intricate pathological milieu post-spinal cord injury (SCI), neural stem cells (NSCs) frequently differentiate into astrocytes rather than neurons, significantly limiting nerve repair. Hence, the utilization of biocompatible hydrogel scaffolds in conjunction with exogenous factors to foster the differentiation of NSCs into neurons has the potential for SCI repair.

Methods: In this study, we engineered a 3D-printed porous SilMA hydrogel scaffold (SM) supplemented with pH-/temperature-responsive paclitaxel nanoparticles (PTX-NPs). We analyzed the biocompatibility of a specific concentration of PTX-NPs and its effect on NSC differentiation. We also established an SCI model to explore the ability of composite scaffolds for in vivo nerve repair.

Results: The physical adsorption of an optimal PTX-NPs dosage can simultaneously achieve pH/temperature-responsive release and commendable biocompatibility, primarily reflected in cell viability, morphology, and proliferation. An appropriate PTX-NPs concentration can steer NSC differentiation towards neurons over astrocytes, a phenomenon that is also efficacious in simulated injury settings. Immunoblotting analysis confirmed that PTX-NPs-induced NSC differentiation occurred via the MAPK/ERK signaling cascade. The repair of hemisected SCI in rats demonstrated that the composite scaffold augmented neuronal regeneration at the injury site, curtailed astrocyte and fibrotic scar production, and enhanced motor function recovery in rat hind limbs.

Conclusion: The scaffold's porous architecture serves as a cellular and drug carrier, providing a favorable microenvironment for nerve regeneration. These findings corroborate that this strategy amplifies neuronal expression within the injury milieu, significantly aiding in SCI repair.

背景:在脊髓损伤(SCI)后错综复杂的病理环境中,神经干细胞(NSCs)经常分化为星形胶质细胞而非神经元,这极大地限制了神经的修复。因此,利用生物相容性水凝胶支架结合外源因子促进神经干细胞分化为神经元,有可能实现脊髓损伤的修复:在这项研究中,我们设计了一种三维打印多孔 SilMA 水凝胶支架(SM),并在其中添加了 pH/ 温度响应型紫杉醇纳米颗粒(PTX-NPs)。我们分析了特定浓度的 PTX-NPs 的生物相容性及其对 NSC 分化的影响。我们还建立了一个 SCI 模型,以探索复合支架在体内修复神经的能力:结果:最佳剂量的 PTX-NPs 物理吸附可同时实现 pH 值/温度响应释放和良好的生物相容性,这主要体现在细胞活力、形态和增殖上。适当的 PTX-NPs 浓度能引导 NSC 向神经元分化,而不是向星形胶质细胞分化,这种现象在模拟损伤环境中也很有效。免疫印迹分析证实,PTX-NPs 通过 MAPK/ERK 信号级联诱导 NSC 分化。对大鼠半损伤性脊髓损伤的修复表明,复合支架促进了损伤部位的神经元再生,减少了星形胶质细胞和纤维化瘢痕的生成,并增强了大鼠后肢的运动功能恢复:结论:支架的多孔结构可作为细胞和药物载体,为神经再生提供有利的微环境。这些研究结果证实,这种策略能在损伤环境中扩大神经元的表达,大大有助于 SCI 的修复。
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引用次数: 0
Implantation of Culture-Expanded Bone Marrow Derived Mesenchymal Stromal Cells for Treatment of Osteonecrosis of the Femoral Head. 植入培养扩增的骨髓间充质基质细胞治疗股骨头骨坏死。
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-08-01 Epub Date: 2024-06-14 DOI: 10.1007/s13770-024-00647-z
Seong-Dae Yoon, Bum-Jin Shim, Seung-Hoon Baek, Shin-Yoon Kim

Background: Although core decompression (CD) with stem cell for the treatment of osteonecrosis of the femoral head (ONFH) showed promising results in many reports, the efficacy remains uncertain. We aimed to evaluate the efficacy of CD with culture-expanded autologous bone marrow-derived mesenchymal stem cell (BM-MSC) implantation in early stage ONFH.

Methods: A total of 18 patients (22 hips) with ONFH who underwent CD with culture-expanded BM-MSC implantation from September 2013 to July 2020 were retrospectively reviewed. The median age was 35.0 years [interquartile range (IQR), 28.5-42.0], and the median follow-up period was 4.0 years (IQR, 2.0-5.3). The median number of MSCs was 1.06 × 108. To evaluate radiographic and clinical outcomes, Association Research Circulation Osseous (ARCO) classifications, Japanese Investigation Committee classification, combined necrotic angle (CNA) visual analogue scale (VAS) and Harris Hip Score (HHS) were checked at each follow-up.

Results: The preoperative stage of ONFH was ARCO 2 in 14 hips and ARCO 3a in 8 hips. The ARCO staging was maintained in 7 hips in ARCO 2 and 4 hips in ARCO 3a. The radiographic failure rate of ARCO 2 and 3a was 14.3 and 50%, respectively. Furthermore, CNA decreased to more than 20° in 6 hips (four were ARCO 2 and two were ARCO 3a).There was no significant difference in the VAS and HHS (P = 0.052 and P = 0.535, respectively). Total hip arthroplasty was performed in 4 hips.

Conclusion: CD with culture-expanded autologous BM-MSCs showed promising results for the treatment of early stage ONFH.

背景:尽管许多报道显示干细胞核心减压术(CD)治疗股骨头坏死(ONFH)效果良好,但疗效仍不确定。我们的目的是评估在早期股骨头坏死患者中植入培养扩增的自体骨髓间充质干细胞(BM-MSC)进行核心减压的疗效:回顾性分析2013年9月至2020年7月期间接受CD联合培养扩增自体骨髓间充质干细胞植入术的18例ONFH患者(22髋)。中位年龄为35.0岁[四分位距(IQR)为28.5-42.0],中位随访时间为4.0年(IQR为2.0-5.3)。间充质干细胞的中位数为 1.06 × 108。为了评估放射学和临床结果,每次随访时都检查了骨关节研究协会(ARCO)分类、日本调查委员会分类、联合坏死角(CNA)视觉模拟量表(VAS)和哈里斯髋关节评分(HHS):14个髋关节的ONFH术前分期为ARCO 2,8个为ARCO 3a。有 7 个处于 ARCO 2 期的髋关节和 4 个处于 ARCO 3a 期的髋关节维持了 ARCO 分期。ARCO 2 和 3a 的影像学失败率分别为 14.3% 和 50%。此外,6 个髋关节的 CNA 下降到 20° 以上(其中 4 个为 ARCO 2,2 个为 ARCO 3a),VAS 和 HHS 没有显著差异(分别为 P = 0.052 和 P = 0.535)。4个髋关节接受了全髋关节置换术:结论:使用培养扩增的自体骨髓间充质干细胞进行 CD 治疗早期 ONFH 有良好效果。
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引用次数: 0
Desktop-Stereolithography 3D Printing of a Decellularized Extracellular Matrix/Mesenchymal Stem Cell Exosome Bioink for Vaginal Reconstruction. 用于阴道重建的脱细胞细胞外基质/间充质干细胞外泌体生物墨水的桌面立体光刻三维打印。
IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING Pub Date : 2024-08-01 Epub Date: 2024-06-27 DOI: 10.1007/s13770-024-00649-x
Wenxin Shi, Jiahua Zheng, Jingkun Zhang, Xiaoli Dong, Zhongkang Li, Yanlai Xiao, Qian Li, Xianghua Huang, Yanfang Du

Background: 3D-printing is widely used in regenerative medicine and is expected to achieve vaginal morphological restoration and true functional reconstruction. Mesenchymal stem cells-derived exosomes (MSCs-Exos) were applyed in the regeneration of various tissues. The current study aimed to explore the effctive of MSCs-Exos in vaginal reconstruction.

Methods: In this work, hydrogel was designed using decellularized extracellular matrix (dECM) and gelatin methacrylate (GelMA) and silk fibroin (SF). The biological scaffolds were constructed using desktop-stereolithography. The physicochemical properties of the hydrogels were evaluated; Some experiments have been conducted to evaluate exosomes' effect of promotion vaginal reconstruction and to explore the mechanism in this process.

Results: It was observed that the sustained release property of exosomes in the hydrogel both in vitro and in vitro.The results revealed that 3D scaffold encapsulating exosomes expressed significant effects on the vascularization and musule regeneration of the regenerative vagina tissue. Also, MSCs-Exos strongly promoted vascularization in the vaginal reconstruction of rats, which may through the PI3K/AKT signaling pathway.

Conclusion: The use of exosome-hydrogel composites improved the epithelial regeneration of vaginal tissue, increased angiogenesis, and promoted smooth muscle tissue regeneration. 3D-printed, lumenal scaffold encapsulating exosomes might be used as a cell-free alternative treatment strategy for vaginal reconstruction.

背景:三维打印技术被广泛应用于再生医学领域,有望实现阴道形态恢复和真正的功能重建。间充质干细胞衍生的外泌体(MSCs-Exos)被应用于多种组织的再生。本研究旨在探索间充质干细胞外泌体在阴道重建中的功效:方法:本研究使用脱细胞细胞外基质(dECM)、甲基丙烯酸明胶(GelMA)和丝纤维素(SF)设计了水凝胶。生物支架采用台式立体光刻技术构建。对水凝胶的理化性质进行了评估;还进行了一些实验来评估外泌体促进阴道重建的效果,并探索这一过程的机制:结果表明,包裹外泌体的三维支架对再生阴道组织的血管化和肌肉再生有显著效果。此外,间充质干细胞-外泌体还能强烈促进大鼠阴道重建中的血管生成,这可能是通过 PI3K/AKT 信号通路实现的:结论:使用外泌体-水凝胶复合材料能改善阴道组织的上皮再生,增加血管生成,促进平滑肌组织再生。包裹外泌体的三维打印腔隙支架可作为阴道重建的无细胞替代治疗策略。
{"title":"Desktop-Stereolithography 3D Printing of a Decellularized Extracellular Matrix/Mesenchymal Stem Cell Exosome Bioink for Vaginal Reconstruction.","authors":"Wenxin Shi, Jiahua Zheng, Jingkun Zhang, Xiaoli Dong, Zhongkang Li, Yanlai Xiao, Qian Li, Xianghua Huang, Yanfang Du","doi":"10.1007/s13770-024-00649-x","DOIUrl":"10.1007/s13770-024-00649-x","url":null,"abstract":"<p><strong>Background: </strong>3D-printing is widely used in regenerative medicine and is expected to achieve vaginal morphological restoration and true functional reconstruction. Mesenchymal stem cells-derived exosomes (MSCs-Exos) were applyed in the regeneration of various tissues. The current study aimed to explore the effctive of MSCs-Exos in vaginal reconstruction.</p><p><strong>Methods: </strong>In this work, hydrogel was designed using decellularized extracellular matrix (dECM) and gelatin methacrylate (GelMA) and silk fibroin (SF). The biological scaffolds were constructed using desktop-stereolithography. The physicochemical properties of the hydrogels were evaluated; Some experiments have been conducted to evaluate exosomes' effect of promotion vaginal reconstruction and to explore the mechanism in this process.</p><p><strong>Results: </strong>It was observed that the sustained release property of exosomes in the hydrogel both in vitro and in vitro.The results revealed that 3D scaffold encapsulating exosomes expressed significant effects on the vascularization and musule regeneration of the regenerative vagina tissue. Also, MSCs-Exos strongly promoted vascularization in the vaginal reconstruction of rats, which may through the PI3K/AKT signaling pathway.</p><p><strong>Conclusion: </strong>The use of exosome-hydrogel composites improved the epithelial regeneration of vaginal tissue, increased angiogenesis, and promoted smooth muscle tissue regeneration. 3D-printed, lumenal scaffold encapsulating exosomes might be used as a cell-free alternative treatment strategy for vaginal reconstruction.</p>","PeriodicalId":23126,"journal":{"name":"Tissue engineering and regenerative medicine","volume":" ","pages":"943-957"},"PeriodicalIF":4.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11286906/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141470932","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}
引用次数: 0
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Tissue engineering and regenerative medicine
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