adipose derived stem cells

脂肪干细胞
  • 文章类型: Journal Article
    背景:间充质干细胞(MSC)已被证明是伤口愈合治疗的可能候选者,因为它们的使用可以增强许多组织的再生能力。人脂肪干细胞(hADSCs)具有易获取、数量大,易于操作。它们可以完全应用于皮肤伤口的治疗。
    目的:在本研究中,我们旨在探讨hADSC在皮肤伤口愈合中的作用和潜在机制。
    方法:hADSC从人皮下脂肪获得。通过油红O和碱性磷酸酶(ALP)染色确定从hADSCs分化的脂肪细胞和骨细胞,分别。我们在小鼠损伤模型中评估了hADSC和hADSC条件培养基(CM)对伤口愈合的影响。然后,我们在体外研究了hADSCs对人角质形成细胞HaCAT细胞的生物学效应。
    结果:结果显示hADSCs可以成功分化为成骨和成脂细胞。hADSC和hADSC-CM在体内显著促进皮肤创伤愈合。hADSCs通过激活Notch信号通路和Rps6kb1激酶激活AKT信号通路显著促进HaCAT细胞增殖和迁移。此外,我们发现hADSC介导的Rps6kb1/AKT信号激活依赖于Notch信号通路。
    结论:我们证明hADSCs可以通过Notch途径促进皮肤细胞-HaCAT细胞的增殖和迁移,提示hADSC可能为皮肤损伤的治疗提供替代治疗方法。
    BACKGROUND: Mesenchymal stem cells (MSCs) have been documented as possible candidates for wound healing treatment because their use could reinforce the regenerative capacity of many tissues. Human adipose stem cells (hADSCs) have the advantages of easy access, large quantity and easy operation. They can be fully applied in the treatment of skin wounds.
    OBJECTIVE: In this study, we aim to explore the roles and potential mechanisms of hADSCs in cutaneous wound healing.
    METHODS: hADSCs were obtained from human subcutaneous fat. Adipocytes and osteocytes differentiated from hADSCs were determined by staining with Oil Red O and alkaline phosphatase (ALP), respectively. We assessed the effects of hADSCs and hADSC conditional medium (CM) on wound healing in an injury model of mice. Then, we investigated the biological effects of hADSCs on human keratinocytes HaCAT cells in vitro.
    RESULTS: The results showed that hADSCs could be successfully differentiated into osteogenic and lipogenic cells. hADSCs and hADSCs-CM significantly promote skin wound healing in vivo. hADSCs significantly promoted HaCAT cell proliferation and migration by activating the Notch signaling pathway and activated the AKT signaling pathway by Rps6kb1 kinase in HaCAT cells. In addition, we found that hADSCs-mediated activation of Rps6kb1/AKT signaling was dependent on the Notch signaling pathway.
    CONCLUSIONS: We demonstrated that hADSCs can promote skin cell-HaCAT cell proliferation and migration via the Notch pathway, suggesting that hADSCs may provide an alternative therapeutic approach for the treatment of skin injury.
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  • 文章类型: Journal Article
    在欧洲,每年进行大约100000到500000个肌腱修复。这些手术与术后并发症的发生率相当高(从6%到11%)。自体微移植物(AAMG)和基质血管分数(SVF)已显示可改善60%至70%的治疗啮齿动物的肌腱愈合。这项研究的目的是评估AAMG在患有肌腱病的绵羊模型中的作用。我们使用绵羊模型是因为,作为一种大型动物,他们更像人类。假设与对照组相比,注射SVF可以改善肌腱愈合,减少炎症和基质降解,同时增加肌腱损伤早期的抗炎表达和胶原合成。16只2至5岁的亚平宁羊接受了500UII型胶原酶注射到两个常见的跟骨肌腱(CCT)中以诱发肌腱病。15天后(T0),每只绵羊随机接受1例CCT至2.5mL通过机械破碎获得的AAMG,对侧CCT未接受治疗.临床,生态,在4周(T1)和8周(T2)后进行超声检查。组织学,免疫组织化学,实时聚合酶链反应(RT-PCR),在T2进行生物力学评估。在T2时,治疗组的最终肌腱直径(9.1±1.4mm)和硬度表达(62%)与原始健康肌腱(8.1±1.1mm;100%)相似,与对照组相比显着恢复(9.5±1.7mm;39%)。此外,治疗组的组织学分析显示纤维取向评分有所改善,纤维水肿评分,浸润性炎症过程,坏死评分(4.3±3.3)与对照组(8.8±2.9)比较。免疫组织化学,治疗组显示胶原1、因子VIII的高表达和胶原3的显著低表达。通过RT-PCR分析证实了这些数据。研究结果表明,通过机械破坏获得的AAMG具有安全性,高效,和可靠的技术,增强肌腱愈合。
    In Europe, approximatively 100 000 to 500 000 tendon repairs are performed every year. These procedures are associated with a considerable rate of postoperative complications (from 6% to 11%). Autologous micro-grafts (AAMG) and stromal vascular fraction (SVF) have been shown to improve tendon healing in 60% to 70% of treated rodents. The purpose of this study was to evaluate the effects of AAMG in a sheep model with tendinopathy. We used sheep models because, as a large animal, they are more comparable to humans. The hypothesis was that SVF injection would improve tendon healing compared with the control group, reducing inflammatory and matrix degrading, while increasing anti-inflammatory expression and collagen synthesis in the early stage of tendon injury. Sixteen Apennine sheep aged 2 to 5 years underwent 500 UI type I collagenase injection into both common calcaneal tendons (CCT) to induce tendinopathy. After 15 days (T0), one CCT in every ovine underwent randomly to 2.5 mL of AAMG obtained by mechanical disruption and the contralateral CCTs received no treatment. Clinical, ecographic, and sonographic evaluations were performed after 4 weeks (T1) and 8 weeks (T2). Histological, immunohistochemical, real-time polymerase chain reaction (RT-PCR), and biomechanical evaluations were performed at T2. At T2, the treated group showed a final tendon diameter (9.1 ± 1.4 mm) and a hardness expression (62%) that were similar to the original healthy tendon (8.1 ± 1.1 mm; 100%), with a significant recovery compared with the control group (9.5 ± 1.7 mm; 39%). Moreover, histological analysis of the treated group revealed an improvement in the fiber orientation score, fiber edema score, infiltrative-inflammatory process, and necrosis score (4.3 ± 3.3) compared with control group (8.8 ± 2.9). Immunohistochemically, the treated group showed high expression of collagen 1, Factor VIII and significantly low expression of collagen 3. These data were confirmed by RT-PCR analysis. The study findings suggested that AAMGs obtained through mechanical disruption present a safe, efficient, and reliable technique, enhancing tendon healing.
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  • 文章类型: Journal Article
    BACKGROUND: A key moderator of wound healing is oxygen. Wound healing is a dynamic and carefully orchestrated process involving blood cells, cytokines, parenchymal cells (i.e. fibroblasts and mesenchymal stem cells) and extracellular matrix reorganization. Human adipose derived stem cells as well as human fibroblasts produce soluble factors, exhibit diverse effects on inflammation and anti inflammation response and are involved in wound healing processes.Hyperbaric oxygen therapy is an effective adjunct treatment for ischemic disorders such as chronic infection or chronic wounds. In vitro effects of hyperbaric oxygen therapy on human cells were presented in many studies except for those on mono- and co-cultures of human adipose derived stem cells and fibroblasts.
    OBJECTIVE: The aim of this study was to investigate the effects of hyperbaric oxygen therapy on mono- and co-cultures of human adipose derived stem cells and fibroblasts.
    METHODS: Mono- and co-cultures from human adipose derived stem cells and fibroblasts were established. These cultures were exposed to hyperbaric oxygen therapy every 24 h for five consecutive days. Measuring experiments were performed on the first, third and fifth day. Therapy effects on the expression of VEGF, IL 6 and reactive oxygen species were investigated.
    RESULTS: After exposure to hyperbaric oxygen, cell culturess showed a significant increase in the expression of VEGF after 3 and 5 days. All cultures showed significantly reduced formation of reactive oxygen species throughout the experiments. The expression of IL-6 decreased during the experiment in mono-cultures of human adipose derived stem cells and co-cultures. In contrast, mono-cultures of human skin fibroblasts showed an overall significantly increased expression of IL-6.
    CONCLUSIONS: Hyperbaric oxygen therapy leads to immunmodulatory and proangiogenetic effects in a wound-like enviroment of adipose derived stem cells and fibroblasts.
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  • 文章类型: Journal Article
    UNASSIGNED: To explored the effect of stromal cell-derived factor 1α (SDF-1α) on promoting the migration ability of rat adipose derived stem cells (rADSCs) by constructed the rADSCs overexpression SDF-1α via adenovirus transfection.
    UNASSIGNED: rADSCs were isolated from adipose tissue of 6-week-old SPF Sprague Dawley rats. Morphological observation, multi-directional differentiations (osteogenic, adipogenic, and chondrogenic inductions), and flow cytometry identification were performed. Transwell cell migration experiment was used to observe and screen the optimal concentration of exogenous SDF-1α to optimize the migration ability of rADSCs; the optimal multiplicity of infection (MOI) of rADSCs was screened by observing the cell status and fluorescence expression after transfection. Then the third generation of rADSCs were divided into 4 groups: group A was pure rADSCs; group B was rADSCs co-cultured with SDF-1α at the best concentration; group C was rADSCs infected with recombinant adenovirus-mediated green fluorescent protein (Adv-GFP) with the best MOI; group D was rADSCs infected with Adv-GFP-SDF-1α overexpression adenovirus with the best MOI. Cell counting kit 8 (CCK-8) and Transwell cell migration experiment were preformed to detect and compare the effect of exogenous SDF-1α and SDF-1α overexpression on the proliferation and migration ability of rADSCs.
    UNASSIGNED: The cell morphology, multi-directional differentiations, and flow cytometry identification showed that the cultured cells were rADSCs. After screening, the optimal stimulating concentration of exogenous SDF-1α was 12.5 nmol/L; the optimal MOI of Adv-GFP adenovirus was 200; the optimal MOI of Adv-GFP-SDF-1α overexpression adenovirus was 400. CCK-8 method and Transwell cell migration experiment showed that compared with groups A and C, groups B and D could significantly improve the proliferation and migration of rADSCs ( P<0.05); the effect of group D on enhancing the migration of rADSCs was weaker than that of group B, but the effect of promoting the proliferation of rADSCs was stronger than that of group D ( P<0.05).
    UNASSIGNED: SDF-1α overexpression modification on rADSCs can significantly promote the proliferation and migration ability, which may be a potential method to optimize the application of ADSCs in tissue regeneration and wound repair.
    UNASSIGNED: 通过使用腺病毒构建基质细胞衍生因子 1α(stromal cell-derived factor 1α,SDF-1α)过表达的大鼠脂肪干细胞(rat adipose derived stem cells,rADSCs),研究 SDF-1α 修饰促进 rADSCs 迁移能力的效果。.
    UNASSIGNED: 取 6 周龄 SPF 级 SD 大鼠脂肪组织分离培养 rADSCs,行细胞形态观察、多向分化(成骨、成脂、成软骨)及流式细胞仪鉴定。取第 3 代 rADSCs,采用 Transwell 细胞迁移实验观察和筛选出优化 rADSCs 迁移能力的外源性 SDF-1α 最佳刺激浓度;通过观察转染后细胞状态和荧光表达情况筛选出 rADSCs 的最佳感染复数(multiplicity of infection,MOI)。然后取第 3 代 rADSCs 分为 4 组,A 组为单纯 rADSCs;B 组于 rADSCs 中加入最佳浓度 SDF-1α 共培养;C 组为携带绿色荧光蛋白的腺病毒载体(recombinant adenovirus-mediated green fluorescent protein,Adv-GFP)以最佳 MOI 感染 rADSCs;D 组采用 Adv-GFP-SDF-1α 过表达腺病毒以最佳 MOI 感染 rADSCs。通过细胞计数试剂盒 8(cell counting kit 8,CCK-8)法、Transwell 细胞迁移实验检测内外源性 SDF-1α 优化 rADSCs 增殖和迁移的能力。.
    UNASSIGNED: 经细胞形态观察、多向分化检测以及流式细胞仪检测显示所培养细胞为 rADSCs。经筛选,优化 rADSCs 迁移能力的外源性 SDF-1α 最佳刺激浓度为 12.5 nmol/L;Adv-GFP 腺病毒最佳 MOI 为 200,Adv-GFP-SDF-1α 过表达腺病毒最佳 MOI 为 400。CCK-8 法和 Transwell 细胞迁移实验检测示,与 A、C 组比较,B、D 组均可显著提高 rADSCs 的增殖、迁移能力( P<0.05);D 组提高 rADSCs 迁移能力的作用弱于 B 组,但促进 rADSCs 增殖能力的作用强于 D 组,差异均有统计学意义( P<0.05)。.
    UNASSIGNED: SDF-1α 修饰 rADSCs 可明显促进其增殖、趋化性迁移能力,可优化 ADSCs 在组织再生、创伤修复治疗中的疗效。.
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  • 文章类型: Journal Article
    关节软骨是无血管的,淋膜,和各向异性组织,这些特征导致软骨组织损伤的显著愈合问题。为了克服这个问题,各种技术已经被开发和广泛使用,但是成本效益和由此产生的组织再生从未实现具有最佳生物力学特性的透明样软骨。该实验的想法是使用可生物降解的多孔海绵软骨(BPSC)支架,结合微骨折技术和脂肪来源的干细胞(ASC)或分泌组在动物模型上增强透明样软骨的再生。
    在新西兰白兔的股骨滑车上制作了模型缺损。比较四组骨软骨缺损的不同治疗方法。各组分别为:(1)对照组;(2)支架组;(3)支架+ASCs组;(4)支架+Secretome组。12周后,我们终止动物模型,然后使用国际软骨研究协会(ICRS)评分系统和Oswestry关节镜评分(OAS)进行宏观评估,然后使用O\'Driscoll评分系统将标本切片进行显微镜评估。
    在总体和组织学上,所有治疗组的平均评分均优于对照组。宏观和微观评估的最佳平均评分是给予支架+ASCs的组。
    BPSC支架的应用增强了较大骨软骨缺损中的软骨再生。此外,在支架植入的同时添加ASCs或分泌组进一步增强软骨再生,其中ASC显示更好的结果。
    Articular cartilage is an avascular, alymphatic, and anisotropic tissue, these characteristics cause significant healing problems to injuries to the cartilage tissue. To overcome this problem, various techniques have been developed and widely used, but the cost-effectiveness and resulting tissue regeneration have never achieved hyaline-like cartilage that has the best biomechanical properties. The idea of this experiment is to use a Biodegradable Porous Sponge Cartilage (BPSC) Scaffold to enhance the regeneration of hyaline-like cartilage combined with microfracture technique and Adipose Derived Stem Cells (ASCs) or secretome on an animal model.
    A model defect was made on the femoral trochlea of a New Zealand white rabbit. Four groups were made to compare different treatment methods for osteochondral defects. The groups were: (1) Control group; (2) Scaffold Group; (3) Scaffold + ASCs Group; (4) Scaffold + Secretome Group. After 12 weeks, we terminate the animal models, then a macroscopic evaluation using the International Cartilage Research Society (ICRS) scoring system and Oswestry Arthroscopy Score (OAS) was done, followed by sectioning the specimen for microscopic evaluation using the O\'Driscoll scoring system.
    The mean score for all treatment group were better compared to the control group grossly and histologically. The best mean score for macroscopic and microscopic evaluation was the group given Scaffold + ASCs.
    The application of BPSC scaffold enhances cartilage regeneration in larger osteochondral defects. Furthermore, the addition of ASCs or secretome along with the scaffold implantation further enhances the cartilage regeneration, in which ASCs shows better results.
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  • 文章类型: Journal Article
    Since the discovery of adipose-derived mesenchymal stem cell (ADSC) in more than ten years, a great progress has been made from its basic research to clinical application. Compared with bone marrow mesenchymal stem cells, ADSCs are more abundant in reserve, easier to obtain with fewer injuries and less complications. These cells have multiple differentiation potential and can differentiate into adipocytes, chondrocytes and osteoblasts with the influence of different inducing factors. Early studies of ADSCs mainly focused on the ability of multi-directional differentiation, espe-cially on the regeneration of bone defects and cartilage tissue. At present, the researches mainly focus on immunoregulation and paracrine function of ADSCs. Although ADSCs have made a great progress in clinical application, the cell preparation, use pattern, and mechanisms in clinical treatment are not clear. This paper elaborates on these issues.
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  • 文章类型: Journal Article
    背景:细胞辅助脂肪移植是一种新的乳房重建和隆胸技术。这项系统评价评估了疗效,在涉及细胞辅助脂肪转运的研究中,患者报告的结局指标的安全性和使用.我们还对学习质量进行客观评估,重点是招聘,随访并提供最新的临床试验景观分析。
    方法:根据PRISMA指南和预定义的纳入和排除标准检索关键电子数据库。两名独立审阅者检查了检索到的出版物并进行了数据提取。
    结果:确定了3980种出版物。筛选后,11项研究被纳入全面审查,代表总共336例患者,随访时间为6至42个月。在移植物保留和报告的满意度方面注意到一定程度的差异,尽管只有三项比较研究的结果相互矛盾.并发症发生率为37%。此外,在选定的研究中,客观结局评估(如3D评估模式或经过验证的患者报告结局指标)很少.
    结论:细胞辅助脂肪移植是一种外科技术,目前在整形和重建外科领域很少使用。目前,需要进一步的技术和成果标准化,除了严格的随机对照试验和支持长期随访数据以更好地确定手术安全性和有效性.常规使用更客观的结果衡量标准,特别是3D评估和验证患者报告的结果指标,还将有助于促进更广泛的临床采用和建立程序效用。
    BACKGROUND: Cell assisted lipotransfer serves as a novel technique for both breast reconstruction and breast augmentation. This systematic review assesses the efficacy, safety and use of patient reported outcome measures in studies involving cell assisted lipotransfer. We also carry out an objective assessment of study quality focussing on recruitment, follow-up and provide an up-to-date clinical trial landscaping analysis.
    METHODS: Key electronic databases were searched according to PRISMA guidelines and pre-defined inclusion and exclusion criteria. Two independent reviewers examined the retrieved publications and performed data extraction.
    RESULTS: 3980 publications were identified. Following screening, 11 studies were included for full review, representing a total of 336 patients with a follow-up time ranging from six to 42 months. A degree of variation was noted in graft retention and reported satisfaction levels, although there were only three comparative studies with conflicting results. Complications occurred at a rate of 37%. Additionally, there was a paucity of objective outcomes assessments (e.g. 3D assessment modalities or validated patient reported outcome measures) in the selected studies.
    CONCLUSIONS: Cell assisted lipotransfer is a surgical technique that is currently employed sparingly within the plastic & reconstructive surgery community. Presently, further technical and outcome standardization is required, in addition to rigorous randomized controlled trials and supporting long-term follow-up data to better determine procedural safety and efficacy. Routine use of more objective outcome measures, particularly 3D assessments and validated patient reported outcome measures, will also help facilitate wider clinical adoption and establish procedural utility.
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  • 文章类型: Journal Article
    The mechanical properties of highly porous (90% porosity) poly(l-lactide) (PLLA), poly(ε-caprolactone) (PCL) and poly(l-lactide/ε-caprolactone) (PLCL) were investigated. Young\'s modulus of non-porous PLLA, PCL and PLCL dropped from 2263.4, 183.7 and 5.7 MPa to 16.8, 1.0 and 1.0 MPa, respectively, for their ~90% porous counterparts. Elongation at break of PCL decreased noticeably with porosity fraction while PLCL maintained a highly elastomeric character and strain recovery capacity even in the presence of pores. Inorganic bioactive particles (hydroxyapatite or bioglass) were added to confer bioactivity to the aforementioned synthetic bioresorbable polymers, and their effect on the mechanical properties was also investigated. Addition of 15 vol.% of inorganic bioactive particles increased the Young\'s modulus of highly porous PLLA from 16.2 to ~30 MPa. On the contrary, the difference between Young\'s modulus of filled and unfilled PCL and PLCL scaffolds was not statistically significant. Finally, an in vitro study of the cytocompatibility and adhesion of adipose derived stem cells (ADSCs) was conducted. The observed viability and excellent adhesion of these cells to both porous and non-porous templates indicate that the employed materials can be good candidates for application in tissue engineering.
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