explant

Explant
  • 文章类型: Journal Article
    肌肉骨骼损伤,包括肌腱病,为老龄化人群带来了巨大的临床负担。虽然与年龄相关的肌腱功能下降的生物学驱动因素知之甚少,众所周知,细胞外基质(ECM)重塑的失调在慢性肌腱变性中起作用。衰老细胞,与肌肉骨骼组织中的多种退行性病变有关,分泌具有促进ECM分解潜力的高度促炎衰老相关分泌表型(SASP)。然而,衰老细胞在肌腱ECM稳态失调中的作用尚不清楚.为了直接评估这一点,我们建立了小鼠肌腱外植体诱导细胞衰老的体外模型。这项新技术使我们能够研究衰老细胞及其天然ECM的分离相互作用,而不受年龄相关的系统性变化的干扰。我们记录了诱导肌腱外植体细胞衰老的多种生物标志物,包括细胞周期停滞,凋亡抗性,和持续的炎症反应。然后,我们利用这个体外衰老模型来比较幼体的ECM重塑反应,自然衰老,和诱导衰老的肌腱改变了机械刺激。我们发现衰老和衰老独立地导致ECM相关基因表达的改变,减少蛋白质合成,和组织组成变化。此外,MMP活性持续,从而使衰老和诱导衰老组织的重塑平衡向降解过度生产转移。一起,这表明细胞衰老在衰老肌腱的机械反应改变中起作用,并可能导致衰老人群的不良临床结局.
    Musculoskeletal injuries, including tendinopathies, present a significant clinical burden for aging populations. While the biological drivers of age-related declines in tendon function are poorly understood, it is well accepted that dysregulation of extracellular matrix (ECM) remodeling plays a role in chronic tendon degeneration. Senescent cells, which have been associated with multiple degenerative pathologies in musculoskeletal tissues, secrete a highly pro-inflammatory senescence-associated secretory phenotype (SASP) that has potential to promote ECM breakdown. However, the role of senescent cells in the dysregulation of tendon ECM homeostasis is largely unknown. To assess this directly, we developed an in vitro model of induced cellular senescence in murine tendon explants. This novel technique enables us to study the isolated interactions of senescent cells and their native ECM without interference from age-related systemic changes. We document multiple biomarkers of cellular senescence in induced tendon explants including cell cycle arrest, apoptosis resistance, and sustained inflammatory responses. We then utilize this in vitro senescence model to compare the ECM remodeling response of young, naturally aged, and induced-senescent tendons to an altered mechanical stimulus. We found that both senescence and aging independently led to alterations in ECM-related gene expression, reductions in protein synthesis, and tissue compositional changes. Furthermore, MMP activity was sustained, thus shifting the remodeling balance of aged and induced-senescent tissues towards degradation over production. Together, this demonstrates that cellular senescence plays a role in the altered mechano-response of aged tendons and likely contributes to poor clinical outcomes in aging populations.
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  • 文章类型: Journal Article
    已使用基于双重腺相关病毒(AAV)的方法在临床前解决了耳聋引起的耳聋缺陷(OTOF)。然而,转导的时机,mRNA的重组,和用双杂交AAV方法的蛋白质表达方法以前没有被表征。这里,我们已经建立了一种离体测定法来确定双AAV介导的OTOF在小鼠胞囊毛细胞中表达的动力学。我们利用了两种不同的重组载体,它们包含DB-OTO,一个包含在毛细胞特异性Myo15启动子控制下的OTOF的5'部分,另一个是OTOF的3'部分。我们探索了Myo15启动子在小鼠胞囊毛细胞中的特异性,在OTOF缺陷小鼠模型中建立的DB-OTO离体剂量反应特征,并证明了腺毛细胞中AAV1的耐受性。此外,我们确定了与5'至3'向量的一对一比率的偏差,对重组OTOF的影响很小。最后,我们在体外14至21天建立了重组OTOFmRNA和蛋白质表达量的平台,恢复时间与体内模型相当。这些发现证明了离体模型系统用于探索表达动力学并建立双重AAV介导的OTOF表达的体内和离体恢复时机的实用性。
    Deafness-causing deficiencies in otoferlin (OTOF) have been addressed preclinically using dual adeno-associated virus (AAV)-based approaches. However, timing of transduction, recombination of mRNA, and protein expression with dual hybrid AAV methods methods have not previously been characterized. Here, we have established an ex vivo assay to determine the kinetics of dual-AAV mediated expression of OTOF in hair cells of the mouse utricle. We utilized two different recombinant vectors that comprise DB-OTO, one containing the 5\' portion of OTOF under the control of the hair cell-specific Myo15 promoter, and the other the 3\' portion of OTOF. We explored specificity of the Myo15 promoter in hair cells of the mouse utricle, established dose response characteristics of DB-OTO ex vivo in an OTOF-deficient mouse model, and demonstrated tolerability of AAV1 in utricular hair cells. Furthermore, we established deviations from a one-to-one ratio of 5\' to 3\' vectors with little impact on recombined OTOF. Finally, we established a plateau in quantity of recombined OTOF mRNA and protein expression by 14 to 21 days ex vivo with comparable recovery timing to that in vivo model. These findings demonstrate the utility of an ex vivo model system for exploring expression kinetics and establish in vivo and ex vivo recovery timing of dual AAV-mediated OTOF expression.
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  • 文章类型: Journal Article
    背景:乳房植入物移除和囊切除术的数量继续迅速增加。外植体手术的目的应该是从美学和功能的角度优化患者的结果。
    目的:确认无引流全囊切除术的安全性,并确定肌肉修复在去除胸肌下或双平面美容乳房植入物后的外植体结局中的作用。
    方法:我们在2021年1月至2023年11月之间对我们的技术进行了回顾性评估。我们报告了一个单外科医生系列,包括140例从双平面或胸膜下口袋中取出美容乳房植入物的连续病例,全部进行全囊切除术。在每种情况下,囊切除术后对胸大肌进行了细致的修复。在任何情况下都不使用排水沟。所有患者均随访至少3个月。术后至少6个月评估患者满意度。
    结果:通过执行所描述的无排水技术,没有血清肿的病例,血肿,气胸或美容乳房变形在这个系列。83%的患者被视为日间病例,患者对结果的满意度很高。
    结论:不使用引流管的全囊切除术是一种新颖而安全的方法,精心修复胸大肌。血清肿的风险没有增加。肌肉修复可能有助于防止乳房外植体后美容畸形。
    方法:本期刊要求作者为每篇文章分配一定程度的证据。对于这些循证医学评级的完整描述,请参阅目录或在线作者说明www。springer.com/00266.
    BACKGROUND: The number of breast implant removal and capsulectomy procedures continues to increase rapidly. The aim of explant surgery should be to optimise patient outcomes from both an aesthetic and functional perspective.
    OBJECTIVE: To confirm the safety of drainless total capsulectomy and to determine the role of muscle repair in explant outcomes following the removal of sub-pectoral or dual-plane cosmetic breast implants.
    METHODS: We conducted a retrospective evaluation of our technique between January 2021 and November 2023. We report a single surgeon series of 140 consecutive cases of cosmetic breast implant removal from dual-plane or sub-pectoral pockets, all performed with total capsulectomy. In each case, meticulous repair of the Pectoralis major muscle was performed following capsulectomy. Drains were not used in any case. All patients were followed up for a minimum of 3 months. Patient satisfaction was assessed a minimum of 6 months post-operatively.
    RESULTS: By performing the described drainless technique, there were no cases of seroma, haematoma, pneumothorax or cosmetic breast distortion in this series. 83% of patients were treated as day cases and patient satisfaction with outcomes was high.
    CONCLUSIONS: Total capsulectomy without the use of drains is a novel and safe approach, aided by careful repair of the Pectoralis major muscle. There is no increased risk of seroma. The muscle repair may help to prevent post-explant cosmetic deformity of the breast.
    METHODS: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .
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  • 文章类型: Journal Article
    脊髓刺激(SCS)是一种用于治疗各种慢性疼痛状况的治疗方式,在过去的50年中迅速发展。不幸的是,随着时间的推移,植入SCS的患者经历了导致疼痛缓解减少的习惯现象。因此,新的刺激波形和SCS应用的发现已被证明可以延长疗效并降低外植率。本文探讨了各种SCS波形,他们的应用,并提出了一种分级的习惯缓解方法。我们怀疑神经习惯现象与药理学相似。因此,我们敦促进一步探索早期引入这些刺激策略以减轻脊髓刺激习惯。
    Spinal cord stimulation (SCS) is a therapeutic modality for the treatment of various chronic pain conditions that has rapidly evolved over the past 50 years. Unfortunately, over time, patients implanted with SCS undergo a habituation phenomenon leading to decreased pain relief. Consequently, the discovery of new stimulation waveforms and SCS applications has been shown to prolong efficacy and reduce explantation rates. This article explores various SCS waveforms, their applications, and proposes a graded approach to habituation mitigation. We suspect the neural habituation phenomenon parallels that seen in pharmacology. Consequently, we urge further exploration of the early introduction of these stimulation strategies to abate spinal cord stimulation habituation.
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  • 文章类型: Journal Article
    左心室辅助装置(LVAD)是晚期心力衰竭患者的出色疗法,可用于移植或终身使用。LVAD还允许通常与功能改善相关的衰竭心脏的逆向重塑。的确,越来越多的人对更好地了解这类患者有热情,其中LVAD被用作介导心肌恢复的辅助手段.当患者达到表明他们不再需要LVAD支持的基准时,与终止LVAD治疗相关的问题成为首要问题。这篇综述的目的是为围绕LVAD停用的实际和技术问题提供外科观点。
    Left ventricular assist devices (LVADs) are excellent therapies for advanced heart failure patients either bridged to transplant or for lifetime use. LVADs also allow for reverse remodeling of the failing heart that is often associated with functional improvement. Indeed, growing enthusiasm exists to better understand this population of patients, whereby the LVAD is used as an adjunct to mediate myocardial recovery. When patients achieve benchmarks suggesting that they no longer need LVAD support, questions related to the discontinuation of LVAD therapy become front and center. The purpose of this review is to provide a surgical perspective on the practical and technical issues surrounding LVAD deactivation.
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  • 文章类型: Journal Article
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  • 文章类型: Journal Article
    肥大细胞通过多种不同的受体被激活以释放从头合成的预形成的颗粒和介质。然而,肥大细胞的生理和功能尚未完全了解。人类中肥大细胞活化的传统研究已经利用了组织来源的肥大细胞的培养物,包括CD34+祖细胞或充分表征的市售细胞系。这些方法的一个限制是肥大细胞不再处于自然状态。因此,它们对人类皮肤病的适用性可能是有限的。人类皮肤外植体模型已用于研究细胞介质的短期影响,毒品,和皮肤上的刺激物,同时避免了围绕未经批准的药物进行体内刺激研究的伦理问题。尽管如此,很少有研究利用完整的人体组织来研究肥大细胞脱颗粒。这篇“方法”论文描述了完整皮肤外植体模型在研究人类肥大细胞活化中的开发和应用。在这份手稿中,我们分享了建立离体人皮肤外植体的方案,并描述了刺激实验和技术的结果,以最大程度地减少创伤诱导的组胺释放。皮肤外植体是使用去识别,全厚度,来自整形和重建手术的非患病皮肤标本。结果是可重复的,并证明了使用BTK抑制剂和QWF可以抑制FcºRI-和MRGPRX2诱导的介质释放,分别。因此,这种外植体模型提供了一个快速和容易的方法来评估人类皮肤肥大细胞的活化和抑制。
    Mast cells are activated through a variety of different receptors to release preformed granules and mediators synthesized de novo. However, the physiology and function of mast cells are not fully understood. Traditional studies of mast cell activation in humans have utilized cultures of tissue-derived mast cells including CD34+ progenitor cells or well-characterized commercially available cell lines. One limitation of these methods is that mast cells are no longer in a natural state. Therefore, their applicability to human skin disorders may be limited. Human skin explant models have been utilized to investigate the short-term effects of cell mediators, drugs, and irritants on skin while avoiding the ethical concerns surrounding in vivo stimulation studies with non-approved agents. Nonetheless, few studies have utilized intact human tissue to study mast cell degranulation. This \"Methods\" paper describes the development and application of an intact skin explant model to study human mast cell activation. In this manuscript, we share our protocol for setting up ex vivo human skin explants and describe the results of stimulation experiments and techniques to minimize trauma-induced histamine release. Skin explants were generated using de-identified, full-thickness, non-diseased skin specimens from plastic and reconstructive surgeries. Results were reproducible and demonstrated FcɛRI- and MRGPRX2-induced mediator release which was inhibited with the use of a BTK inhibitor and QWF, respectively. Thus, this explant model provides a quick and accessible method of assessing human skin mast cell activation and inhibition.
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  • 文章类型: Journal Article
    衰老是退行性肌腱损伤的主要危险因素,然而,这种变性的病因和进展知之甚少。虽然老化的肌腱具有先天的细胞差异,支持降低的能力,以维持机械组织稳态,尚未研究老化的肌腱对机械载荷水平变化的响应。为了解决这个问题,我们对年轻和年老的鼠屈肌腱外植体进行不同水平的体外拉伸应变。我们首先比较了静态和循环应变对年轻肌腱基质重塑的影响,发现循环应变是研究体外重塑的最佳选择。然后,我们研究了在7天的各种机械刺激(应力剥夺,1%,3%,5%,或7%循环应变)通过评估组织组成,生物合成能力,和降解配置文件。我们假设老化的肌腱会对拉伸应变的变化表现出柔和的自适应响应,并表现出改变的机械设定值。在这种情况下,重塑平衡是最佳的。有趣的是,我们发现1%的循环菌株最好地维持天然生理,同时促进两个年龄组的ECM更新。然而,老化的肌腱显示较少的应变相关变化,表明适应机械载荷水平变化的能力降低。这项工作对理解衰老肌腱组织稳态的调节具有重大影响,这可以为治疗老年患者提供临床康复策略。
    Aging is a primary risk factor for degenerative tendon injuries, yet the etiology and progression of this degeneration are poorly understood. While aged tendons have innate cellular differences that support a reduced ability to maintain mechanical tissue homeostasis, the response of aged tendons to altered levels of mechanical loading has not yet been studied. To address this question, we subjected young and aged murine flexor tendon explants to various levels of in vitro tensile strain. We first compared the effect of static and cyclic strain on matrix remodeling in young tendons, finding that cyclic strain is optimal for studying remodeling in vitro. We then investigated the remodeling response of young and aged tendon explants after 7 days of varied mechanical stimulus (stress deprivation, 1%, 3%, 5%, or 7% cyclic strain) via assessment of tissue composition, biosynthetic capacity, and degradation profiles. We hypothesized that aged tendons would show muted adaptive responses to changes in tensile strain and exhibit a shifted mechanical setpoint, at which the remodeling balance is optimal. Interestingly, we found that 1% cyclic strain best maintains native physiology while promoting extracellular matrix (ECM) turnover for both age groups. However, aged tendons display fewer strain-dependent changes, suggesting a reduced ability to adapt to altered levels of mechanical loading. This work has a significant impact on understanding the regulation of tissue homeostasis in aged tendons, which can inform clinical rehabilitation strategies for treating elderly patients.
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  • 文章类型: Letter
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  • 文章类型: Journal Article
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