Extracellular vesicles:Emerged as a promising strategy for regenerative medicine

Cell transplantation therapy has certain limitations including immune rejection and limited cell viability,which seriously hinder the transformation of stem cellbased tissue regeneration into clinical practice.Extracellular vesicles(EVs)not only possess the advantages of its derived cells,but also can avoid the risks of cell transplantation.EVs are intelligent and controllable biomaterials that can participate in a variety of physiological and pathological activities,tissue repair and regeneration by transmitting a variety of biological signals,showing great potential in cell-free tissue regeneration.In this review,we summarized the origins and characteristics of EVs,introduced the pivotal role of EVs in diverse tissues regeneration,discussed the underlying mechanisms,prospects,and challenges of EVs.We also pointed out the problems that need to be solved,application directions,and prospects of EVs in the future and shed new light on the novel cell-free strategy for using EVs in the field of regenerative medicine.

Extracellular vesicles in nanomedicine and regenerative medicine:A review over the last decade

Small extracellular vesicles(sEVs)are known to be secreted by a vast majority of cells.These sEVs,specifically exosomes,induce specific cell-to-cell interactions and can activate signaling pathways in recipient cells through fusion or interaction.These nanovesicles possess several desirable properties,making them ideal for regenerative medicine and nanomedicine applications.These properties include exceptional stability,biocompatibility,wide biodistribution,and minimal immunogenicity.However,the practical utilization of sEVs,particularly in clinical settings and at a large scale,is hindered by the expensive procedures required for their isolation,limited circulation lifetime,and suboptimal targeting capacity.Despite these challenges,sEVs have demonstrated a remarkable ability to accommodate various cargoes and have found extensive applications in the biomedical sciences.To overcome the limitations of sEVs and broaden their potential applications,researchers should strive to deepen their understanding of current isolation,loading,and characterization techniques.Additionally,acquiring fundamental knowledge about sEVs origins and employing state-of-the-art methodologies in nanomedicine and regenerative medicine can expand the sEVs research scope.This review provides a comprehensive overview of state-of-the-art exosome-based strategies in diverse nanomedicine domains,encompassing cancer therapy,immunotherapy,and biomarker applications.Furthermore,we emphasize the immense potential of exosomes in regenerative medicine.

Consensus statement on research and application of Chinese herbal medicine derived extracellular vesicles-like particles(2023 edition)

To promote the development of extracellular vesicles of herbal medicine especially the establishment of standardization,led by the National Expert Committee on Research and Application of Chinese Herbal Vesicles,research experts in the field of herbal medicine and extracellular vesicles were invited nationwide with the support of the Expert Committee on Research and Application of Chinese Herbal Vesicles,Professional Committee on Extracellular Vesicle Research and Application,Chinese Society of Research Hospitals and the Guangdong Engineering Research Center of Chinese Herbal Vesicles.Based on the collation of relevant literature,we have adopted the Delphi method,the consensus meeting method combined with the nominal group method to form a discussion draft of“Consensus statement on research and application of Chinese herbal medicine derived extracellular vesicles-like particles(2023)”.The first draft was discussed in online and offline meetings on October 12,14,November 2,2022 and April and May 2023 on the current status of research,nomenclature,isolation methods,quality standards and research applications of extracellular vesicles of Chinese herbal medicines,and 13 consensus opinions were finally formed.At the Third Academic Conference on Research and Application of Chinese Herbal Vesicles,held on May 26,2023,Kewei Zhao,convenor of the consensus,presented and read the consensus to the experts of the Expert Committee on Research and Application of Chinese Herbal Vesicles.The consensus highlights the characteristics and advantages of Chinese medicine,inherits the essence,and keeps the righteousness and innovation,aiming to provide a reference for colleagues engaged in research and application of Chinese herbal vesicles at home and abroad,decode the mystery behind Chinese herbal vesicles together,establish a safe,effective and controllable accurate Chinese herbal vesicle prevention and treatment system,and build a bridge for Chinese medicine to the world.

Role of hypoxia preconditioning in therapeutic potential of mesenchymal stem-cell-derived extracellular vesicles

The use of mesenchymal stem-cells(MSC)in cell therapy has received considerable attention because of their properties.These properties include high expansion and differentiation in vitro,low immunogenicity,and modulation of biological processes,such as inflammation,angiogenesis and hematopoiesis.Curiously,the regenerative effect of MSC is partly due to their paracrine activity.This has prompted numerous studies,to investigate the therapeutic potential of their secretome in general,and specifically their extracellular vesicles(EV).The latter contain proteins,lipids,nucleic acids,and other metabolites,which can cause physiological changes when released into recipient cells.Interestingly,contents of EV can be modulated by preconditioning MSC under different culture conditions.Among them,exposure to hypoxia stands out;these cells respond by activating hypoxia-inducible factor(HIF)at low O_(2) concentrations.HIF has direct and indirect pleiotropic effects,modulating expression of hundreds of genes involved in processes such as inflammation,migration,proliferation,differentiation,angiogenesis,metabolism,and cell apoptosis.Expression of these genes is reflected in the contents of secreted EV.Interestingly,numerous studies show that MSC-derived EV conditioned under hypoxia have a higher regenerative capacity than those obtained under normoxia.In this review,we show the implications of hypoxia responses in relation to tissue regeneration.In addition,hypoxia preconditioning of MSC is being evaluated as a very attractive strategy for isolation of EV,with a high potential for clinical use in regenerative medicine that can be applied to different pathologies.

Bronchoalveolar lavage fluid assessment facilitates precision medicine for lung cancer

Lung cancer is the most common and fatal malignant disease worldwide and has the highest mortality rate among tumor-related causes of death.Early diagnosis and precision medicine can significantly improve the survival rate and prognosis of lung cancer patients.At present,the clinical diagnosis of lung cancer is challenging due to a lack of effective non-invasive detection methods and biomarkers,and treatment is primarily hindered by drug resistance and high tumor heterogeneity.Liquid biopsy is a method for detecting circulating biomarkers in the blood and other body fluids containing genetic information from primary tumor tissues.Bronchoalveolar lavage fluid(BALF)is a potential liquid biopsy medium that is rich in a variety of bioactive substances and cell components.BALF contains information on the key characteristics of tumors,including the tumor subtype,gene mutation type,and tumor environment,thus BALF may be used as a diagnostic supplement to lung biopsy.In this review,the current research on BALF in the diagnosis,treatment,and prognosis of lung cancer is summarized.The advantages and disadvantages of different components of BALF,including cells,cell-free DNA,extracellular vesicles,and micro RNA are introduced.In particular,the great potential of extracellular vesicles in precision diagnosis and detection of drug-resistant for lung cancer is highlighted.In addition,the performance of liquid biopsies with different body fluid sources in lung cancer detection are compared to facilitate more selective studies involving BALF,thereby promoting the application of BALF for precision medicine in lung cancer patients in the future.

New paradigms in regenerative engineering:Emerging role of extracellular vesicles paired with instructive biomaterials

Mesenchymal stem cells(MSCs)have long been regarded as critical components of regenerative medicine strategies,given their multipotency and persistence in a variety of tissues.Recently,the specific role of MSCs in mediating regenerative outcomes has been attributed(in part)to secreted factors from transplanted cells,namely extracellular vesicles.This viewpoint manuscript highlights the promise of cell-derived extracellular vesicles as agents of regeneration,enhanced by synergy with appropriate biomaterials platforms.Extracellular vesicles are a potentially interesting regenerative tool to enhance the synergy between MSCs and biomaterials.As a result,we believe these technologies will improve patient outcomes through efficient therapeutic strategies resulting in predictable patient outcomes.

Plant-derived nanovesicles: Current understanding and applications for cancer therapy

Plant-derived vesicles(PDVs)are membranous structures that originate from plant cells and are responsible for multiple physiological and pathological functions.In the last decade,PDVs have gained much attention for their involvement in different biological processes,including intercellular communication and defense response,and recent scientific evidence has opened a new avenue for their applications in cancer treatment.Nevertheless,much remains unknown about these vesicles,and current research remains inconsistent.This review aims to provide a comprehensive introduction to PDVs,from their biological characteristics to purification methods,and to summarize the status of their potential development for cancer therapy.

Plant-derived nanovesicles: Further exploration of biomedical function and application potential

Extracellular vesicles(EVs)are phospholipid bilayer vesicles actively secreted by cells,that contain a variety of functional nucleic acids,proteins,and lipids,and are important mediums of intercellular communication.Based on their natural properties,EVs can not only retain the pharmacological effects of their source cells but also serve as natural delivery carriers.Among them,plant-derived nanovesicles(PNVs)are characterized as natural disease therapeutics with many advantages such as simplicity,safety,eco-friendliness,low cost,and low toxicity due to their abundant resources,large yield,and low risk of immunogenicity in vivo.This review systematically introduces the biogenesis,isolation methods,physical characterization,and components of PNVs,and describes their administration and cellular uptake as therapeutic agents.We highlight the therapeutic potential of PNVs as therapeutic agents and drug delivery carriers,including anti-inflammatory,anticancer,wound healing,regeneration,and antiaging properties as well as their potential use in the treatment of liver disease and COVID-19.Finally,the toxicity and immunogenicity,the current clinical application,and the possible challenges in the future development of PNVs were analyzed.We expect the functions of PNVs to be further explored to promote clinical translation,thereby facilitating the development of a new framework for the treatment of human diseases.

间充质干细胞分泌组发挥作用的分子机制

背景:大量研究证实间充质干细胞分泌组的治疗有效性与间充质干细胞相当,但对于其发挥作用的机制尚不清楚。目的:总结近年来关于间充质干细胞分泌组的研究进展,探究其发挥治疗作用的分子机制,分析目前面临的问题并展望未来发展方向。方法:以“exosomes,mesenchymal stem cells secrete,extracellular vesicles,mesenchymal stem cells,mechanism”为英文检索词检索PubMed数据库相关文献,排除与文章研究目的无关及重复性文章,同时结合文献追踪的方法,最终纳入符合要求的109篇文献进行综述。结果与结论:(1)间充质干细胞分泌组通过递送遗传物质、免疫调节因子和生长因子等到靶细胞,通过激活靶细胞抗凋亡和促生存途径,调节血管生成,调节纤维化以及调节免疫微环境,促进组织修复和再生。(2)间充质干细胞分泌组在疾病治疗方面的潜力也得到证实,大量研究结果表明,间充质干细胞分泌组可以作为炎症疾病和退行性疾病的新型无细胞替代治疗方法。(3)近年来,经过工程化改造间充质干细胞分泌组发挥的治疗作用更加高效,但是由于间充质干细胞分泌组的异质性以及成分的复杂性,其发挥治疗作用的具体机制尚不清楚。(4)目前,仍需要进一步研究来明确间充质干细胞分泌组发挥治疗作用的关键靶点,并且未来应结合工程化和基因工程技术等,研发出具有创新性的特异型、增强型间充质干细胞分泌组。

Disease-derived circulating extracellular vesicle preconditioning:A promising strategy for precision mesenchymal stem cell therapy

Mesenchymal stem cell(MSC)-based therapies have emerged as promising methods for regenerative medicine;however,how to precisely enhance their tissue repair effects is still a major question in the field.Circulating extracellular vesicles(EVs)from diseased states carry diverse pathological information and affect the functions of recipient cells.Based on this unique property,we report that disease-derived circulating EV(disease-EV)preconditioning is a potent strategy for precisely enhancing the tissue repair potency of MSCs in diverse disease models.Briefly,plasma EVs from lung or kidney tissue injuries were shown to contain distinctly enriched molecules and were shown to induce tissue injury-specific gene expression responses in cultured MSCs.Disease-EV preconditioning improved the performance(including proliferation,migration,and growth factor production)of MSCs through metabolic reprogramming(such as via enhanced oxidative phosphorylation and lipid metabolism)without inducing an adverse immune response.Consequently,compared with normal MSCs,disease-EV-preconditioned MSCs exhibited superior tissue repair effects(including anti-inflammatory and antiapoptotic effects)in diverse types of tissue injury(such as acute lung or kidney injury).Disease-derived EVs may serve as a type of“off-the-shelf”product due to multiple advantages,such as flexibility,stability,long-term storage,and ease of shipment and use.This study highlights the idea that disease-EV preconditioning is a robust strategy for precisely enhancing the regenerative capacity of MSC-based therapies.

基于水凝胶的细胞外囊泡递送系统在再生医学中的研究进展

再生医学是通过替换和修复受损的组织、器官使其恢复正常结构与功能的科学,其常用技术包括组织工程、器官移植、无细胞治疗等。其中,无细胞治疗降低了免疫排斥的风险,减少了致瘤性,成为再生医学的研究热点。近年来发现的细胞外囊泡(extracellular vesicles,EVs)为实现无细胞治疗拓宽了道路。EVs是由几乎所有细胞释放到胞外的膜性囊泡,是细胞间通信的重要媒介,它们作为治疗药物和药物递送载体具有广阔应用前景。然而,基于EVs的治疗存在半衰期短、缺乏靶向性及滞留性等问题。生物材料水凝胶可一定程度提高EVs在体内的稳定性和滞留性,基于此,研究者们探究了基于水凝胶的EVs递送策略。本文系统综述了近年来基于水凝胶的EVs递送系统在再生医学中的研究进展,探讨了其在临床应用中的前景和挑战。

Mesenchymal stem cell-derived exosomes: Toward cell-free therapeutic strategies in regenerative medicine

Mesenchymal stem cells(MSCs)are multipotent stem cells with marked potential for regenerative medicine because of their strong immunosuppressive and regenerative abilities.The therapeutic effects of MSCs are based in part on their secretion of biologically active factors in extracellular vesicles known as exosomes.Exosomes have a diameter of 30-100 nm and mediate intercellular communication and material exchange.MSC-derived exosomes(MSC-Exos)have potential for cell-free therapy for diseases of,for instance,the kidney,liver,heart,nervous system,and musculoskeletal system.Hence,MSC-Exos are an alternative to MSCbased therapy for regenerative medicine.We review MSC-Exos and their therapeutic potential for a variety of diseases and injuries.

The versatility of mesenchymal stem cells:From regenerative medicine to COVID,what is next?

Mesenchymal stem cells(MSCs)play key roles in regenerative medicine by promoting tissue healing.MSCs can be isolated from different adult tissues and they are able to differentiate into several lineages.Due to their antiinflammatory,angiogenic and immune-modulatory properties,MSCs are suitable for tissue engineering applications and,when associated with biomaterials,their benefits can be improved.Moreover,recently,MSCs have been studied for new clinical applications,such as in the treatment of patients with COVID-19.MSCs regenerative potential has been attributed to their secretome,which comprises extracellular matrix,soluble proteins and several elements,including the release of extracellular vesicles.Even though,in order to explore all their therapeutic potential,it is still necessary to advance in the investigation of their basic cell biology characteristics.

黄芪甲苷调控外泌体介导的JMJD3/H3K27me3/OPN通路抗结直肠癌肝转移机制研究

目的研究黄芪甲苷(ASⅣ)抑制结直肠癌肝转移的作用机制,探索ASⅣ对外泌体介导的Jumonji结构域包含蛋白3(JMJD3)/第27位的赖氨酸三甲基化的组蛋白H3(H3K27me3)/骨桥蛋白(OPN)信号通路的影响。方法取对数生长期的MC38细胞,超速离心法提取MC38细胞外泌体(MC38-EVs)及ASⅣ干预后MC38细胞外泌体(ASⅣ/MC38-EVs),粒径鉴定后用于动物实验。选取C57BL/6J小鼠15只,建立结直肠癌原位肝转移模型后,按照随机数字表法分为模型组、MC38-EVs组、ASⅣ/MC38-EVs组,每组5只。模型组造模前1周尾静脉注射磷酸盐缓冲液(PBS),100μL/次,隔日1次,连续3周;MC38-EVs组和ASⅣ/MC38-EVs组造模前1周尾静脉分别注射0.1 g/L的MC38-EVs、ASⅣ/MC38-EVs,100μL/次,隔日1次,连续3周。4周后处死小鼠,观察结直肠癌肝转移情况;苏木精-伊红(HE)染色法观察肝转移灶病理学改变;蛋白质免疫印迹(Western blot)法检测肝转移灶中JMJD3、H3K27me3、OPN蛋白表达情况;免疫荧光法检测肝转移灶中F4/80+、CD206+巨噬细胞的浸润和JMJD3、H3K27me3、OPN蛋白表达;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)检测瘤体中JMJD3、OPN mRNA表达。结果与模型组比较,MC38-EVs组肝转移灶数目和肝脏质量明显增加(P<0.05);与MC38-EVs组比较,ASⅣ/MC38-EVs组肝转移灶数目和肝脏质量明显减少(P<0.05);免疫荧光结果显示,与模型组比较,MC38-EVs组中肝组织F4/80+、CD206+巨噬细胞浸润程度明显升高(P<0.05);与MC38-EVs组比较,ASⅣ/MC38-EVs组中肝组织巨噬细胞浸润程度明显降低(P<0.05);Western blot、RT-qPCR和免疫荧光结果显示,与模型组比较,MC38-EVs组JMJD3、OPN蛋白和mRNA表达水平明显升高(P<0.05),H3K27me3蛋白表达水平明显降低(P<0.05);与MC38-EVs组相比,ASⅣ/MC38-EVs组JMJD3、OPN蛋白和mRNA表达水平明显降低(P<0.05),H3K27me3蛋白表达水平明显升高(P<0.05)。结论ASⅣ能够有效抑制结直肠癌肝转移,其机制与调控外泌体介导的JMJD3/H3K27me3/OPN通路激活相关。

间充质干细胞的细胞治疗策略研究进展

间充质干细胞(mesenchymal stem cells,MSCs)的细胞治疗是近年研究热点,包括传统意义上的以活细胞为基础的细胞治疗策略和新出现的以细胞外囊泡和其他可溶性蛋白质或生物活性分子为基础的去细胞治疗策略。目前,MSCs诱导获得的细胞已具备成熟的功能和特异性的结构,结合生物材料或类器官技术的原位移植在定植率及功能性方面有了较大提升。另一方面,随着大规模培养技术和细胞外囊泡分离技术的成熟,大量获得高纯度的细胞外囊泡难度降低,细胞外囊泡的研究也逐渐成为当前研究的热点。越来越多的研究支持MSCs的治疗效果不仅是通过植入和分化效应进行修复,MSCs旁分泌也表现出良好效果。笔者对MSCs的细胞治疗策略和去细胞治疗策略的最新进展进行综述。

Advances in extracellular vesicle functionalization strategies for tissue regeneration

Extracellular vesicles(EVs)are nano-scale vesicles derived by cell secretion with unique advantages such as promoting cell proliferation,anti-inflammation,promoting blood vessels and regulating cell differentiation,which benefit their wide applications in regenerative medicine.However,the in vivo therapeutic effect of EVs still greatly restricted by several obstacles,including the off-targetability,rapid blood clearance,and undesired release.To address these issues,biomedical engineering techniques are vastly explored.This review summarizes different strategies to enhance EV functions from the perspective of drug loading,modification,and combination of biomaterials,and emphatically introduces the latest developments of functionalized EV-loaded biomaterials in different diseases,including cardio-vascular system diseases,osteochondral disorders,wound healing,nerve injuries.Challenges and future directions of EVs are also discussed.

中药联合干细胞治疗多发性硬化研究进展

多发性硬化病机复杂,目前临床尚无治愈办法。中药和干细胞移植对多发性硬化均具有良好疗效,二者联合相较于单独应用具有更好的治疗效果。综述中药联合干细胞移植治疗多发性硬化作用及机制并探讨中药联合干细胞源性细胞外囊泡治疗多发性硬化的潜力,以期为多发性硬化的防治提供参考。

Extracellular vesicles:Emerging tools as therapeutic agent carriers

Extracellular vesicles(EVs)are secreted by both eukaryotes and prokaryotes,and are present in all biological fluids of vertebrates,where they transfer DNA,RNA,proteins,lipids,and metabolites from donor to recipient cells in cell-to-cell communication.Some EV components can also indicate the type and biological status of their parent cells and serve as diagnostic targets for liquid biopsy.EVs can also natively carry or be modified to contain therapeutic agents(e.g.,nucleic acids,proteins,polysaccharides,and small molecules)by physical,chemical,or bioengineering strategies.Due to their excellent biocompatibility and stability,EVs are ideal nanocarriers for bioactive ingredients to induce signal transduction,immunoregulation,or other therapeutic effects,which can be targeted to specific cell types.Herein,we review EV classification,intercellular communication,isolation,and characterization strategies as they apply to EV therapeutics.This review focuses on recent advances in EV applications as therapeutic carriers from in vitro research towards in vivo animal models and early clinical applications,using representative examples in the fields of cancer chemotherapeutic drug,cancer vaccine,infectious disease vaccines,regenerative medicine and gene therapy.Finally,we discuss current challenges for EV therapeutics and their future development.

Mesenchymal stem cell-derived extracellular vesicles:a possible therapeutic strategy for orthopaedic diseases:a narrative review

Accumulating evidence suggests that the therapeutic role of mesenchymal stem cells(MSCs)in bone diseases is closely related to paracrine-generated extracellular vesicles(EVs).MSC-derived EVs(MSC-EVs)carry proteins,nucleic acids,and lipids to the extracellular space and affect the bone microenvironment.They have similar biological functions to MSCs,such as the ability to repair organ and tissue damage.In addition,MSC-EVs also have the advantages of long half-life,low immunogenicity,attractive stability,ability to pass through the blood-brain barrier,and demonstrate excellent performance with potential practical applications in bone diseases.In this review,we summarise the current applications and mechanisms of MSC-EVs in osteoporosis,osteoarthritis,bone tumours,osteonecrosis of the femoral head,and fractures,as well as the development of MSC-EVs combined with materials science in the field of orthopaedics.Additionally,we explore the critical challenges involved in the clinical application of MSC-EVs in orthopaedic diseases.

特络细胞及其胞外囊泡与细胞间交流

特络细胞(telcocytes,TCs)是一种间质细胞,广泛存在于人和哺乳动物的许多器官和组织中。TCs是一种带有突起的细胞,通过同型连接、异型连接或释放细胞外囊泡直接参与细胞间交流。越来越多的证据表明,TCs可能是一种潜在的再生细胞。本文总结了TCs及其胞外囊泡参与细胞间交流的证据,并讨论了它们潜在的功能作用。TCs的研究为再生医学提供了新的见解,在未来可考虑利用TC_s作为新型靶细胞治疗相关疾病。