注射PRP与透明质酸治疗膝骨关节炎的研究

通过对膝关节骨性关节炎(OA)患者临床试验,研究关节内注射血小板的血浆PRP和透明质酸HA对膝关节OA患者的临床结果和生活质量的长期影响。该非安慰剂对照的随机临床试验包括160例受膝关节OA影响的患者、1～4级的凯尔格伦-劳伦斯量表。在PRP组(n=87)中,在4周间隔内进行2次关节内注射,HA组(n=73)在1周间隔内进行了3次关节内注射。问卷调查之后,对所有患者在治疗前和治疗后12个月进行了前瞻性评估。使用SPSS 16.1软件对结果进行分析。在12个月的随访中,两组WOMAC疼痛评分和身体疼痛均有显著改善;然而,与HA组相比,PRP组的结果较好(p0.001)。其他WOMAC和SF-36参数仅在PRP组中得到改善。两组患者2级OA患者均得到更多改善。本研究表明PRP注射液比HA注射液在减少症状和改善生活质量方面更有效,是选择对常规治疗没有反应的膝关节OA患者的治疗方案。     

外周动脉疾病治疗进展

外周动脉疾病(PAD)与心血管疾病(CV)的发病率和死亡率有着密切的联系。ACCF/AHA指南建议无症状和症状性PAD患者戒烟并应用抗血小板/抗凝药物。对于存在严重肢体缺血(CLI)的PAD患者应考虑接受腔内与开放保肢手术治疗。即便存在CLI的PAD患者接受如上治疗,有时仍无法为患肢提供足够的血流灌注以消除症状。为建立有效血供,许多研究已经深入细胞治疗层面。内皮干细胞、单核细胞和骨髓间充质干细胞在临床应用中得到了很好的研究。血管内皮生长因子、成纤维细胞生长因子和肝细胞生长因子(HGF)也被应用于PAD患者,以诱导血管生成。其中,HGF最有优势,因为它可诱导血管生成却不伴有反应性血管炎及血管通透性增高。同时,血管腔内治疗器械及技术,如药物涂层球囊等也获得较快发展。本文将PAD治疗进展综述如下。     

临床级干细胞库及干细胞制剂

干细胞为解决重大医学难题带来了希望,现已开展多项干细胞治疗临床研究,且获得了一定的疗效。干细胞制剂作为干细胞治疗产品,在制备和应用过程中均需严格的操作和管理,我国还处于干细胞临床转化起步阶段,相关政策法规有待进一步细化。利用干细胞库进行干细胞制剂的标准化制备和应用已成为国际主流趋势。北京干细胞库在中国科学院"干细胞与再生医学研究"战略性先导科技专项支持下,建立了中国首家临床级干细胞库和临床级胚胎干细胞系,获得了一系列中国食品药品检定研究院的检验报告,并开展了一些干细胞的临床研究。此外,北京干细胞库参与了多项干细胞相关政策法规的制定。现将结合北京干细胞库的研究进展,对临床级干细胞制剂的管理和临床级干细胞库的建设进行综述,以期为我国干细胞研究领域的健康发展贡献一己之力。     

羟乙基淀粉沉淀(HES)法分离脐血造血干细胞的临床优势分析

目的:探讨羟乙基淀粉沉淀(HES)法分离脐血单个核细胞(MNCs)的效果。方法:采集脐血31份,应用羟乙基淀粉(HES)沉淀飞和密度离心(Ficoll)法分离脐血MNCs,、纯化CD3+细胞、CD14+细胞、CD34+细胞,以台盼蓝拒染法检测细胞活力。结果:HES法MNCs回收率、CFU-GM计数高于Ficoll法(85.29±3.79 VS 47.06±4.61,t=35.67,P0.05;67.31±11.57/l×105MNCs VS28.54±6.39/l×105MNCs,t=16.33,P0.05);HES法分离的MNCs中CD3+、CD14+、CD34+细胞数均高于Ficoll法(t=5.76,t=2.51,t=6.67,P0.05)。结论:HES法分离MNCs可获得较好的回收率,是人脐血干细胞理想的分离方法。     

《干细胞临床研究管理办法(试行)》发布

<正>国家卫生和计划生育委员会、国家食品药品监督管理总局近日联合发布《干细胞临床研究管理办法(试行)》(以下简称《管理办法》)。这是我国首个针对干细胞临床研究进行管理的规范性文件,旨在规范干细胞临床研究行为,保障受试者权益,促进干细胞研究健康发展。《管理办法》明确,从事干细胞临床研究的医疗机构     

人间充质干细胞的无异源培养体系研究进展

人间充质干细胞具有自我更新和多向分化的潜能,越来越多的应用于细胞再生医学领域。临床应用细胞制剂的制备,需要在GMP指导下对人间充质干细胞进行体外扩增。传统的方法人间充质干细胞扩增方法含FBS,但由于存在批次间质量差异、异源污染和免疫排斥风险,无异源培养方法显得尤为必要。多种替代FBS的人源性血液成分已经在研究之中,如血清、血浆和血小板衍生物等。化学成分限定性培养基是标准化的最终目标,目前已有商业化的培养基上市。无异源培养基是临床细胞分离和扩增的前提,可保证临床细胞治疗应用的安全性和有效性,是该领域走向产业化的必然趋势。本文对人间充质干细胞的无异源培养体系研究进展做一综述。     

富血小板血浆注射治疗在膝骨关节炎中的应用进展

富血小板血浆(platelet-rich plasma，PRP)由于富含多种活性生长因子，能够刺激软骨细胞增殖，促进软骨前体细胞增殖、迁移、向软骨细胞分化，促进胶原蛋白合成以及抑制软骨的炎性反应和退变，提供有利于组织修复的内环境，延缓病情进展。近年来PRP注射治疗已成为治疗与骨关节炎(osteoarthritis，OA)相关疾病的新型选择，并且疗效显著。为了进一步提高其效用，PRP注射治疗不仅在关节腔内进行，还可在软骨下骨内进行注射。软骨下骨的病变会加速软骨损耗，故有必要将软骨下骨也当作OA众多发病机制和病理过程的关键因素之一。根据PRP的生物特效以及PRP注射治疗在膝骨关节炎(knee osteoarthritis，KOA)中应用的研究进展进行了综述，同时对软骨下骨内PRP注射治疗KOA的研究进行了展望，以期为KOA的治疗提供更加有效的方法。     

康柏西普与雷珠单抗对年龄相关性黄斑变性患者血清CRP、VEGF、眼压及视力的影响

目的:探讨康柏西普与雷珠单抗对年龄相关性黄斑变性(AMD)患者血清中C反应蛋白(CRP)、血管内皮生长因子(VEGF)水平、眼压(IOP)及视力的影响。方法:选取2015年4月到2016年5月在我院接受治疗的AMD患者70例作为研究对象,采用随机数字表法将所有患者分为观察组和对照组各35例,所有患者给予内眼手术标准玻璃体注射治疗,观察组给予康柏西普注射液1.5mg腔内注射,对照组给予雷珠单抗注射液0.5 mL腔内注射,对比两组患者治疗前、治疗后3d血清CRP、VEGF水平;记录并对比两组患者视力、IOP水平及并发症情况。结果:两组患者治疗后3d血清CRP、VEGF水平较治疗前下降,且观察组患者CRP、VEGF水平低于对照组(P0.05);两组患者治疗后3d视力最小视角对数(logMAR)及IOP较治疗前降低,且观察组患者视力logMAR及IOP水平明显低于对照组,差异均有统计学意义(P0.05)。治疗后3d,两组各出现1例玻璃体出血,并发症发生率比较差异无统计学意义(P0.05)。结论:相对于雷珠单抗,康柏西普更能有效降低AMD患者血清CRP、VEGF水平和IOP,提高患者视力,改善病情,值得临床推广应用。     

脐血干细胞移植治疗肝硬化的临床研究

目的:观察脐血干细胞移植治疗失代偿期肝硬化的临床疗效。方法:对60例失代偿期肝硬化患者进行自身对照的临床研究。在无菌条件下取健康产妇足月生产的脐带血,分离纯化脐血干细胞,通过肝动脉途径,将纯化的脐血干细胞移植入患者肝脏内,于移植后2周、4周、8周进行肝功能、凝血指标检测,并于4周及8周行腹部B超及胃镜检查,观察患者移植后不同时间症状改善情况及术后不良反应的发生情况。结果:60例接受脐血干细胞移植的肝硬化患者术中术后无明显不良反应发生。移植后,患者临床症状改善明显,食欲不振、乏力、腹水等减轻甚至消失;血清学检测:白蛋白水平较术前明显升高,凝血酶原时间、总胆红素较术前明显下降;术后8周复查胃镜,食道静脉曲张没有明显变化。结论:脐血干细胞移植治疗失代偿期肝硬化是一种安全、有效的方法,尤其是在提高白蛋白水平及改善凝血功能方面有很好的疗效,可作为肝移植治疗的过渡或补充治疗。     

鞘内注射脐带间充质干细胞治疗神经系统疾病的安全性和疗效研究（英文）

目的:探讨腰穿鞘内注射脐带间充质干细胞治疗神经系统疾病的安全性和疗效并且分析腰穿治疗过程中遇到的各种技术难题。方法:2008年12月至2011年5月88例伴有神经系统疾病的患者给予鞘内注射干细胞治疗,并且评价其在治疗过程中的技术难题,采用HAI评分系统对这些患者神经功能的恢复情况进行评估,定期观察患者的临床症状、生物学指标和影像学检查观察其治疗前后的变化。结果:88例患者中具有技术难题的患者有20例,主要表现为在鞘内注射脐带间充质干细胞的过程中需要行全麻补充、腰穿定位困难等,18例患者在治疗过程中出现副作用(头痛、低热、腰痛和下肢痛),但这些副作用在48小时内经过系统治疗后完全缓解。随访1年,50例患者神经功能得到一定改善,包括15例脊髓损伤患者、10例脑瘫患者、10例脑外伤后综合征患者、5例脑梗死后综合征患者、5例脊髓小脑共济失调患者和5例运动神经元病患者。结论:鞘内注射脐带间充质干细胞治疗神经系统疾病是安全和有效的,由于随访时间较短,有必要对这种治疗模式进行扩大的、双盲、安慰剂对照研究来进一步推广其临床应用。     

Production of canine mesenchymal stem cells from adipose tissue and their application in dogs with chronic osteoarthritis of the humeroradial joints

Autologous AD-MSC [adipose-derived MSC (mesenchymal stem cell)] therapy involves harvesting fat from the patient by isolating the stem and regenerative cells and administering the cells back to the patient. This study evaluated the production of canine AD-MSCs and their possible application in cellular therapy for dogs. To assess whether cellular therapy can replace drug therapy, the clinical effect of a single intra-articular injection of AD-MSCs was evaluated on 4 dogs with lameness associated with OA (osteoarthritis) of the humeroradial joints. MSCs were readily isolated from adult dog adipose tissue, and their ability to form colony and differentiate into various phenotypes was confirmed. AD-MSCs expressed OCT4, NANOG and SOX2 at the mRNA level, pluripotency markers usually ascribed to embryonic stem cells. The results suggest the stemness of the cells isolated from canine fat, and good quality control made them available for both experimental and clinical use. Follow-up studies to evaluate the effects of AD-MSC therapy showed that OA of the elbow joints improved with time, indicating significant potential for clinical use in the treatment of lameness, particularly when administered before the injury becomes severe.     

Therapeutic applications of adipose-derived stromal vascular fractions in osteoarthritis

Osteoarthritis (OA) is considered to be a highly heterogeneous disease with progressive cartilage loss, subchondral bone remodeling, and low-grade inflammation. It is one of the world's leading causes of disability. Most conventional clinical treatments for OA are palliative drugs, which cannot fundamentally cure this disease. The stromal vascular fraction (SVF) from adipose tissues is a heterogeneous cell population. According to previous studies, it contains a large number of mesenchymal stem cells, which have been used to treat OA with good therapeutic results. This safe, simple, and effective therapy is expected to be applied and promoted in the future. In this paper, the detailed pathogenesis, diagnosis, and current clinical treatments for OA are introduced. Then, clinical studies and the therapeutic mechanism of SVF for the treatment of OA are summarized.     

Treatment of osteoarthritis with mesenchymal stem cells

Osteoarthritis(OA)is one of the most prevalent joint diseases with prominent symptoms affecting the daily life of millions of middle aged and elderly people.Despite this,there are no successful medical interventions that can prevent the progressive destruction of OA joints.The onset of pathological changes in OA is associated with deviant activity of mesenchymal stem cells(MSCs),the multipotent precursors of connective tissue cells that reside in joints.Current therapies for OA have resulted in poor clinical outcomes without repairing the damaged cartilage.Intra-articular delivery of culture-expanded MSCs has opened new avenues of OA treatment.Pre-clinical and clinical trials demonstrated the feasibility,safety,and efficacy of MSC therapy.The Wnt/β-catenin,bone morphogenetic protein 2,Indian hedgehog,and Mitogen-activated protein kinase signaling pathways have been demonstrated to be involved in OA and the mechanism of action of MSC therapies.     

Application of mesenchymal stem cell therapy for the treatment of osteoarthritis of the knee: A concise review

Osteoarthritis(OA) refers to a chronic joint disease characterized by degenerative changes of articular cartilage and secondary bone hyperplasia. Since articular cartilage has a special structure, namely the absence of blood vessels as well as the low conversion rate of chondrocytes in the cartilage matrix, the treatment faces numerous clinical challenges. Traditional OA treatment(e.g., arthroscopic debridement, microfracture, autologous or allogeneic cartilage transplantation,chondrocyte transplantation) is primarily symptomatic treatment and pain management, which cannot contribute to regenerating degenerated cartilage or reducing joint inflammation. Also, the generated mixed fibrous cartilage tissue is not the same as natural hyaline cartilage. Mesenchymal stem cells(MSCs) have turned into the most extensively explored new therapeutic drugs in cell-based OA treatment as a result of their ability to differentiate into chondrocytes and their immunomodulatory properties. In this study, the preliminary results of preclinical(OA animal model)/clinical trials regarding the effects of MSCs on cartilage repair of knee joints are briefly summarized, which lay a solid application basis for more and deeper clinical studies on cell-based OA treatment.     

Progress in the use of mesenchymal stromal cells for osteoarthritis treatment

Literature review of MSCs in the treatment of osteoarthritis in the past five yearsOsteoarthritis (OA) is one of the most common chronic joint diseases, with prominent symptoms caused by many factors. However, current medical interventions for OA have resulted in poor clinical outcomes, demonstrating that there are huge unmet medical needs in this area. Cell therapy has opened new avenues of OA treatment. Different sources of mesenchymal stromal cells (MSCs) may have different phenotypes and cellular functions. Pre-clinical and clinical studies have demonstrated the feasibility, safety and efficacy of MSC therapy. Mitogen-activated protein kinase, Wnt and Notch signaling pathways are involved in the chondrogenesis of MSC-mediated treatments. MSCs may also exert effective immunoregulatory and paracrine effects to stimulate tissue repair. Therapy with extracellular vesicles containing cytokines, which are secreted by MSCs, might be a potential treatment for OA.     

Cellular Reprogramming of Human Peripheral Blood Cells

Breakthroughs in cell fate conversion have made it possible to generate large quantities of patient-specific cells for regenerative medicine. Due to multiple advantages of peripheral blood cells over fibroblasts from skin biopsy, the use of blood mononuclear cells （MNCs） instead of skin fibroblasts will expedite reprogramming research and broaden the application of reprogramming technology. This review discusses current progress and challenges of generating induced pluripotent stem cells （iPSCs） from peripheral blood MNCs and of in vitro and in vivo conversion of blood cells into cells of therapeutic value, such as mesenchymal stem cells, neural cells and hepatocytes. An optimized design of lentiviral vectors is necessary to achieve high reprogramming efficiency of peripheral blood cells. More recently, non-integrating vectors such as Sendai virus and episomal vectors have been successfully employed in generating integration-free iPSCs and somatic stem cells.     

间充质干细胞的来源及其对肝脏损伤及修复的研究进展

全球终末期肝病、肝衰竭的发病率和死亡率逐年升高,且目前肝移植是唯一疗效确切的治疗选择,但是,肝移植的使用受到肝源供体严重不足,长期存活率低,医疗费用昂贵等缺点使得原位肝移植的应用受限,绝大多数患者无法受益。为了克服肝脏器官短缺,干细胞替代治疗策略逐渐成为另一个肝病治疗的重要选择,干细胞治疗,特别是间充质干细胞（MSC）提供了一个新的肝病治疗选择。MSC是一群贴壁生长的成纤维细胞样细胞,由于MSC能够分化为多种类型的细胞,能够产生多种的细胞因子和生长因子,具有造血支持和免疫调节和抗炎功能,MSC被认为在再生医学领域具有重大的科学和实用价值。另外,由于MSC应用于治疗实验性肝损伤能明显提高动物存活率,明显改善肝功能。此外,一些临床前研究和临床研究也表明MSC对肝损伤性疾病具有显著地疗效。因此MSC在损伤性和退行性肝脏疾病的治疗具有广阔的应用前景。本文综述了MSC在肝损伤疾病治疗应用的进展,并对MSC在肝病治疗中的应用前景进行了展望。     

Doxycycline,salinomycin, monensin and ivermectin repositioned as cancer drugs

Chemotherapy is one of the standard methods for the treatment of malignant tumors. It aims to cause lethal damage to cellular structures, mainly DNA. Noteworthy, in recent years discoveries of novel anticancer agents from well-known antibiotics have opened up new treatment pathways for several cancer diseases. The aim of this review article is to describe new applications for the following antibiotics: doxycycline (DOX), salinomycin (SAL), monensin (MON) and ivermectin (IVR) as they are known to show anti-tumor activity, but have not yet been introduced into standard oncological therapy. To date, these agents have been used for the treatment of a broad-spectrum of bacterial and parasitic infectious diseases and are widely available, which is why they were selected. The data presented here clearly show that the antibiotics mentioned above should be recognised in the near future as novel agents able to eradicate cancer cells and cancer stem cells (CSCs) across several cancer types.     

Mesenchymal stem cell-based treatment for cartilage defects in osteoarthritis

Osteoarthritis (OA) is a common disorder and the restoration of the diseased articular cartilage in patients with OA is still a challenge for researchers and clinicians. Currently, a variety of experimental strategies have investigated whether mesenchymal stem cells (MSCs) instead of chondrocytes can be used for the regeneration and maintenance of articular cartilage in OA. MSCs can modulate the immune response of individuals and positively influence the microenvironment of the stem cells already present in the diseased tissue. Through direct cell–cell interaction or the secretion of various factors, MSCs can initiate endogenous regenerative activities in the OA joint. Targeted gene-modified MSC-based therapy might further enhance the cartilage regeneration in OA. Conventionally, delivery of MSCs was attained by graft of engineered constructs derived from cell-seeded scaffolds. However, intra-articular MSCs transplantation without scaffolds is a more attractive option for OA treatment. This article briefly summarizes the current knowledge about MSC-based therapy for prevention or treatment of OA, discussing the direct intra-articular injection of MSCs for the treatment of OA in animal models and in clinical applications, as well as potential future strategies for OA treatment.     

Teratomas from pluripotent stem cells: A clinical hurdle

Although basic research on human embryonic stem cells (hESCs) at the laboratory bench has progressed with enviable speed there has been little head way in terms of its clinical application. A look at the Internet however shows several stem cell clinics worldwide offering direct transplantation of undifferentiated hESCs to patients for the cure of a variety of diseases before bona fide evidence‐based results can be demonstrated from large controlled studies. This raises concern because reliable protocols have to be first developed to resolve the three major hurdles delaying clinical trials such as inadequate cell numbers, immunorejection and tumorigenesis. Cell expansion methods using bioreactors, rotary culture and mitotic agents have now been developed to generate stem cell derivatives in large numbers. The problem of immunorejection can now be overcome with the development of indirect and direct reprogramming protocols to personalize tissues to patients (human induced pluripotent stem cells, hiPSCs; nuclear transfer stem cells, NTSCs; induced neuronal cells, iN). However, hESC, hiPSC, and NTSCs being pluripotent have the disadvantage of teratoma formation in vivo which has to be carefully addressed so as to provide safe stem cell based therapies to the patient. This review addresses the issue of tumorigenesis and discusses approaches by which this concern may be overcome and at the same time emphasizes the need to concurrently explore alternative stem cell sources that do not confer the disadvantages of pluripotency but are widely multipotent so as to yield safe desirable tissues for clinical application as soon as possible. J. Cell. Biochem. 111: 769–781, 2010. © 2010 Wiley‐Liss, Inc.