TY - JOUR TI - Witness Box Protocol: Automatic machine identification and authentication in industry 4.0 AU - Mehdi, Nabeel AU - Starly, Binil T2 - COMPUTERS IN INDUSTRY AB - The current wave of Industrial Internet of Things (IIoT) is reshaping the manufacturing sector with system interoperability, remote real-time process monitoring and advanced analytics. As digitally enabled manufacturing machines continue to grow exponentially, it becomes imperative to uniquely and securely identify them in the cyber-physical world, particularly in defense, biomedical, energy and aerospace manufacturing. Research about threats originating from internal adversaries’ i.e the machine/organization owner within a tiered digitally connected supply chain is scarce. This paper introduces a machine fingerprinting scheme named as the ‘Witness Box Protocol’ (WBP) that exploits the physical properties of manufacturing machines (legacy or smart) and their surroundings to create a unique biometric like fingerprint. WBP provides both machine registration and authentication on a digital network through a low cost, non-invasive approach. The fingerprint is generated by a Locality Sensitive Hashing (LSH) technique that accommodates small variations in physical signature data and can corroborate data provenance from machines by verifying machine identity through authentication. Additionally, this fingerprint hash simplifies asset management within a large enterprise or distributed network comprising of thousands of machines in a Manufacturing-as-a-Service (MaaS) paradigm. In this research, fingerprints were randomly generated from the statistical features of signals from 3D printers and CNC machine in a production-like lab environment. Using k-means clustering and Jaccard similarity index, these fingerprints are shown to identify the source equipment with 95 % accuracy. DA - 2020/12// PY - 2020/12// DO - 10.1016/j.compind.2020.103340 VL - 123 SP - SN - 1872-6194 KW - Cybersecurity KW - Smart manufacturing KW - Manufacturing fingerprinting KW - Cyber physical systems KW - Digital supply Chain traceability ER - TY - JOUR TI - Automating the Search and Discovery of Manufacturing Service Providers to Enable a Digital Supply Chain Network AU - Starly, Binil AU - Cohen, Paul AU - Raman, Shivakumar T2 - SMART AND SUSTAINABLE MANUFACTURING SYSTEMS AB - Uncertainty in manufacturing networks has created barriers to closing the gap between design enterprises and the American industrial base. Uncertainty arises from the lack of transparent access to manufacturer capabilities, the inability to auto-discover service providers who are best capable for a given job request, and the dependence on human word-of-mouth trust network relationships that exist in the manufacturing supply chain. This uncertainty slows down the pace of product development lifecycles from a viewpoint of inefficient forms of supplier assessment, vetting, selection, and compliance, leading to a trust tax tacked onto the final price of products. In times of global crisis such as the coronavirus disease pandemic, this uncertainty also leads to inefficient forms of gathering information on manufacturing capability, available capacity, and registered licenses and assessing compliance. This technical note outlines solution pathways that can help ease the search and discovery process of connecting clients and manufacturing service providers through digitally enabled technologies. DA - 2020/12// PY - 2020/12// DO - 10.1520/SSMS20200061 VL - 4 IS - 3 SP - 276-280 SN - 2572-3928 KW - coronavirus disease KW - manufacturing search engine optimization KW - schema.org vocabulary extensions KW - blockchain ER - TY - JOUR TI - Recent Development in Therapeutic Cardiac Patches AU - Mei, Xuan AU - Cheng, Ke T2 - FRONTIERS IN CARDIOVASCULAR MEDICINE AB - For the past decades, heart diseases remain the leading cause of death worldwide. In the adult mammalian heart, damaged cardiomyocytes will be replaced by non-contractile fibrotic scar tissues due to the poor regenerative ability of heart, causing heart failure subsequently. The development of tissue engineering has launched a new medical innovation for heart regeneration. As one of the most outstanding technology, cardiac patches hold the potential to restore cardiac function clinically. Consisted of two components: therapeutic ingredients and substrate scaffolds, the fabrication of cardiac patches requires both advanced bioactive molecules and biomaterials. In this review, we will present the most state-of-the-art cardiac patches and analysis their compositional details. The therapeutic ingredients will be discussed from cell sources to bioactive molecules. In the meanwhile, the recent advances to obtain scaffold biomaterials will be highlighted, including synthetic and natural materials. Also, we have focused on the challenges and potential strategies to fabricate clinically applicable cardiac patches. DA - 2020/11/27/ PY - 2020/11/27/ DO - 10.3389/fcvm.2020.610364 VL - 7 SP - SN - 2297-055X KW - biomaterials KW - cardiac patch KW - myocardial infarction KW - cell therapy KW - cardiac tissue regeneration ER - TY - JOUR TI - Three-dimensional imaging of intact porcine cochlea using tissue clearing and custom-built light-sheet microscopy AU - Moatti, Adele AU - Cai, Yuheng AU - Li, Chen AU - Sattler, Tyler AU - Edwards, Laura AU - Piedrahita, Jorge AU - Ligler, Frances S. AU - Greenbaum, Alon T2 - BIOMEDICAL OPTICS EXPRESS AB - Hearing loss is a prevalent disorder that affects people of all ages. On top of the existing hearing aids and cochlear implants, there is a growing effort to regenerate functional tissues and restore hearing. However, studying and evaluating these regenerative medicine approaches in a big animal model (e.g. pigs) whose anatomy, physiology, and organ size are similar to a human is challenging. In big animal models, the cochlea is bulky, intricate, and veiled in a dense and craggy otic capsule. These facts complicate 3D microscopic analysis that is vital in the cochlea, where structure-function relation is time and again manifested. To allow 3D imaging of an intact cochlea of newborn and juvenile pigs with a volume up to ∼ 250 mm 3 , we adapted the BoneClear tissue clearing technique, which renders the bone transparent. The transparent cochleae were then imaged with cellular resolution and in a timely fashion, which prevented bubble formation and tissue degradation, using an adaptive custom-built light-sheet fluorescence microscope. The adaptive light-sheet microscope compensated for deflections of the illumination beam by changing the angles of the beam and translating the detection objective while acquiring images. Using this combination of techniques, macroscopic and microscopic properties of the cochlea were extracted, including the density of hair cells, frequency maps, and lower frequency limits. Consequently, the proposed platform could support the growing effort to regenerate cochlear tissues and assist with basic research to advance cures for hearing impairments. DA - 2020/11/1/ PY - 2020/11/1/ DO - 10.1364/BOE.402991 VL - 11 IS - 11 SP - 6181-6196 SN - 2156-7085 ER - TY - JOUR TI - Cardiac Stromal Cell Patch Integrated with Engineered Microvessels Improves Recovery from Myocardial Infarction in Rats and Pigs AU - Su, Teng AU - Huang, Ke AU - Mathews, Kyle G. AU - Scharf, Valery F. AU - Hu, Shiqi AU - Li, Zhenhua AU - Frame, Brianna N. AU - Cores, Jhon AU - Dinh, Phuong-Uyen AU - Daniele, Michael A. AU - Ligler, Frances S. AU - Cheng, Ke T2 - ACS BIOMATERIALS SCIENCE & ENGINEERING AB - The vascularized cardiac patch strategy is promising for ischemic heart repair after myocardial infarction (MI), but current fabrication processes are quite complicated. Vascularized cardiac patches that can promote concurrent restoration of both the myocardium and vasculature at the injured site in a large animal model remain elusive. The safety and therapeutic benefits of a cardiac stromal cell patch integrated with engineered biomimetic microvessels (BMVs) were determined for treating MI. By leveraging a microfluidic method employing hydrodynamic focusing, we constructed the endothelialized microvessels and then encapsulated them together with therapeutic cardiosphere-derived stromal cells (CSCs) in a fibrin gel to generate a prevascularized cardiac stromal cell patch (BMV-CSC patch). We showed that BMV-CSC patch transplantation significantly promoted cardiac function, reduced scar size, increased viable myocardial tissue, promoted neovascularization, and suppressed inflammation in rat and porcine MI models, demonstrating enhanced therapeutic efficacy compared to conventional cardiac stromal cell patches. BMV-CSC patches did not increase renal and hepatic toxicity or exhibit immunogenicity. We noted a significant increase in endogenous progenitor cell recruitment to the peri-infarct region of the porcine hearts treated with BMV-CSC patch as compared to those that received control treatments. These findings establish the BMV-CSC patch as a novel engineered-tissue therapeutic for ischemic tissue repair. DA - 2020/11// PY - 2020/11// DO - 10.1021/acsbiomaterials.0c00942 VL - 6 IS - 11 SP - 6309-6320 SN - 2373-9878 KW - microfluidics KW - cardiac patch KW - cardiac stromal cells KW - myocardial infarction KW - porcine model ER - TY - JOUR TI - Biomechanical evaluation of an autologous flexor digitorum lateralis graft to augment the surgical repair of gastrocnemius tendon laceration in a canine ex vivo model AU - Duffy, Daniel J. AU - Curcillo, Chiara P. AU - Chang, Yi-Jen AU - Gaffney, Lewis AU - Fisher, Matthew B. AU - Moore, George E. T2 - VETERINARY SURGERY AB - Abstract Objective To evaluate the effect of an autologous flexor digitorum lateralis (FDL) graft to augment a three‐loop pulley (3LP) core repair in a canine cadaveric gastrocnemius tendon (GT) laceration model. Study design Ex vivo, biomechanical study. Sample population Twenty‐six canine cadaveric hind limbs. Methods Tendons were divided into two groups (n = 13). After sharp transection, paired GT were repaired with 3LP or 3LP + FDL tendon augmentation. Yield, peak and failure loads, tensile loads required to create 1 and 3‐mm gapping, and failure modes were analyzed. Significance was set at P < .05. Results Yield and failure force (mean ± SD) for 3LP + FDL were 134.9 ± 44.1 N and 205.4 ± 46.4 N, respectively, which were greater than for 3LP alone (67.9 ± 12.2 N and 91.8 ± 9.9 N, respectively, P < .0001). No constructs (0%) formed 1 or 3‐mm gaps in the 3LP + FDL graft group compared with 84% and 39% for 3LP, respectively ( P < .0001). Failure modes were different between groups ( P < .001), with 85% of 3LP + FDL constructs failing by tissue rupture at the myotendinous junction, distant to the repair site. Conclusion Addition of an autologous FDL graft to a core 3LP tendon repair increased yield, peak, and failure forces by twofold, 2.3‐fold, and 2.2‐fold, respectively, compared with core 3LP alone while preventing the occurrence of gap formation. Clinical significance Use of FDL tendon augmentation for GT laceration may increase repair site strength and resist gap formation better than 3LP core suture use alone. Additional studies are required in vivo to determine the effect of FDL graft augmentation on clinical function. DA - 2020/12// PY - 2020/12// DO - 10.1111/vsu.13453 VL - 49 IS - 8 SP - 1545-1554 SN - 1532-950X ER - TY - JOUR TI - In Vitro Validation of Transgene Expression in Gene-Edited Pias Using CRISPR Transcriptional Activators AU - Polkoff, Kathryn M. AU - Chung, Jaewook AU - Simpson, Sean G. AU - Gleason, Katherine AU - Piedrahita, Jorge A. T2 - CRISPR JOURNAL AB - The use of CRISPR-Cas and RNA-guided endonucleases has drastically changed research strategies for understanding and exploiting gene function, particularly for the generation of gene-edited animal models. This has resulted in an explosion in the number of gene-edited species, including highly biomedically relevant pig models. However, even with error-free DNA insertion or deletion, edited genes are occasionally not expressed and/or translated as expected. Therefore, there is a need to validate the expression outcomes gene modifications in vitro before investing in the costly generation of a gene-edited animal. Unfortunately, many gene targets are tissue specific and/or not expressed in cultured primary cells, making validation difficult without generating an animal. In this study, using pigs as a proof of concept, we show that CRISPR-dCas9 transcriptional activators can be used to validate functional transgene insertion in nonexpressing easily cultured cells such as fibroblasts. This is a tool that can be used across disciplines and animal species to save time and resources by verifying expected outcomes of gene edits before generating live animals. DA - 2020/10// PY - 2020/10// DO - 10.1089/crispr.2020.0037 VL - 3 IS - 5 SP - 409-418 SN - 2573-1602 ER - TY - JOUR TI - Decentralized cloud manufacturing-as-a-service (CMaaS) platform architecture with configurable digital assets AU - Hasan, Mahmud AU - Starly, Binil T2 - JOURNAL OF MANUFACTURING SYSTEMS AB - Contemporary Cloud Manufacturing-as-a-Service (CMaaS) platforms now promise customers instant pricing and access to a large capacity of manufacturing nodes. However, many of the CMaaS platforms are centralized with data flowing through an intermediary agent connecting clients with service providers. This paper reports the design, implementation and validation of middleware software architectures which aim to directly connect client users with manufacturing service providers while improving transparency, data integrity, data provenance and retaining data ownership to its creators. In the first middleware, clients have the ability to directly customize and configure parts parametrically, leading to an instant generation of downstream manufacturing process plan codes. In the second middleware, clients can track the data provenance generated in a blockchain based decentralized architecture across a manufacturing system. The design of digital assets across a distributed manufacturing system infrastructure controlled by autonomous smart contracts through Ethereum based ERC-721 non-fungible tokens is proposed to enable communication and collaboration across decentralized CMaaS platform architectures. The performance of the smart contracts was evaluated on three different global Ethereum blockchain test networks with the centrality and dispersion statistics on their performance provided as a reference benchmark for future smart contract implementations. DA - 2020/7// PY - 2020/7// DO - 10.1016/j.jmsy.2020.05.017 VL - 56 SP - 157-174 SN - 1878-6642 KW - Cybermanufacturing KW - Blockchain KW - Smart contracts KW - Distributed digital thread KW - Cloud manufacturing ER - TY - JOUR TI - Influence of barbed epitendinous suture combined with a core locking-loop sutures to repair experimental flexor tendon lacerations AU - Eby, Adam C. AU - Duffy, Daniel J. AU - Chang, Yi-Jen AU - Gaffney, Lewis AU - Fisher, Matthew B. AU - Moore, George E. T2 - VETERINARY SURGERY AB - Abstract Objective To determine the influence of barbed epitendinous sutures (ES) on the biomechanical properties and gap formation of repaired canine tendons. Study design Ex vivo, experimental study. Sample population Eighty (n = 16/group) canine superficial digital flexor tendons (SDFT). Methods After transection, SDFT were repaired with a locking‐loop (LL) pattern alone (group 1), an LL + smooth ES with monofilament suture (group 2), an LL + V‐loc‐ES (group 3), an LL + Quill‐ES (group 4), or an LL + Stratafix‐ES (group 5). All core LL repairs were performed with 0 USP polypropylene, and all ES were placed with 2‐0 USP equivalent. Constructs were preloaded and tested to failure. Yield, peak, and failure loads; occurrence of gap formation; and failure modes were compared. Results Yield loads were greater for groups 2 and 5 ( P < .0001). Peak and failure loads were greater when an ES was used ( P < .005), especially for groups 2 and 5 ( P < .0001). Groups with an ES required higher loads to generate 1‐ and 3‐mm gaps compared with specimens without an ES ( P < .002). Force to create 1‐ and 3‐mm gaps was greater for group 5 ( P < .0001) and groups 2 and 5 ( P < .0001), respectively. Failure mechanism did not differ ( P = .092) between ES groups, consisting of suture breakage in 51 of 64 constructs compared with pull‐through in seven of 16 group 1 constructs. Conclusion Epitendinous suture placement improved the biomechanical properties of repaired tendons. Stratafix barbed suture performed better as an ES compared with other barbed sutures and similarly to monofilament suture. Clinical significance Stratafix barbed suture eliminates the requirement for knot tying and seems to be equivalent to smooth monofilament suture when used as an ES in this pattern. DA - 2020/12// PY - 2020/12// DO - 10.1111/vsu.13496 VL - 49 IS - 8 SP - 1590-1599 SN - 1532-950X ER - TY - JOUR TI - Effect of partial vs complete circumferential epitendinous suture placement on the biomechanical properties and gap formation of canine cadaveric tendons AU - Duffy, Daniel J. AU - Chang, Yi-Jen AU - Fisher, Matthew B. AU - Moore, George E. T2 - VETERINARY SURGERY AB - Abstract Objective To determine the effect of partial vs complete circumferential epitendinous suture (ES) placement in addition to a core suture on the biomechanical strength and gapping characteristics of repaired canine tendinous constructs. Study design Ex vivo, biomechanical study. Sample population Thirty‐six canine superficial digital flexor tendons. Methods Superficial digital flexor tendons were randomly assigned to three groups (n = 12), sharply transected and repaired with a core locking‐loop suture with Group 1 a partial circumferential ES, 180° on the palmar side; Group 2 a complete circumferential ES, 360° and double knotting technique; or Group 3 a complete circumferential ES, 360° and single knotting technique. After preloading, constructs were distracted to monotonic failure. Failure mode, gap formation, yield, peak, and failure forces were analyzed. Results Mean yield (group 1 = 68.6 N, group 2 = 106.5 N, group 3 = 114 N, P < .013), peak (group 1 = 92.8 N, group 2 = 134.6 N, group 3 = 147.3 N; P < .001), and failure (group 1 = 88.7 N, group 2 = 133.0 N, group 3 = 145.5 N, P < .001) loads differed between groups. No difference in yield ( P = .874), peak ( P = .434), or failure load ( P = .434) was detected between complete circumferential ES groups. Force to create 1‐mm ( P < .001) and 3‐mm ( P < .038) gap formation was greater in specimens with complete vs partial circumferential ES placement. Complete circumferential ES repairs failed primarily by suture pull‐through compared with suture breakage in most partial circumferential ES constructs. Conclusion Addition of a complete circumferential ES with a single or double knotting technique increased the biomechanical strength of normal tendon repairs while reducing gap formation compared with partial ES placement alone. Clinical significance Complete circumferential ES is recommended over partial ES placement. DA - 2020/12// PY - 2020/12// DO - 10.1111/vsu.13494 VL - 49 IS - 8 SP - 1571-1579 SN - 1532-950X ER - TY - JOUR TI - Advances of exosome isolation techniques in lung cancer AU - Mahgoub, Elham O. AU - Razmara, Ehsan AU - Bitaraf, Amirreza AU - Norouzi, Fahimeh-Sadat AU - Montazeri, Maryam AU - Behzadi-Andouhjerdi, Roudabeh AU - Falahati, Mojtaba AU - Cheng, Ke AU - Haik, Yousif AU - Hasan, Anwarul AU - Babashah, Sadegh T2 - MOLECULAR BIOLOGY REPORTS DA - 2020/9// PY - 2020/9// DO - 10.1007/s11033-020-05715-w VL - 47 IS - 9 SP - 7229-7251 SN - 1573-4978 KW - Lung cancer KW - Exosome KW - Multi-vesicular bodies KW - Extracellular vesicles KW - MicroRNAs ER - TY - JOUR TI - Clinical xenotransplantation of the heart: At the watershed AU - Platt, Jeffrey L. AU - Piedrahita, Jorge A. AU - Cascalho, Marilia T2 - JOURNAL OF HEART AND LUNG TRANSPLANTATION AB - See Related Article, page 751 See Related Article, page 751 Transplantation has been the preferred treatment for severe failure of the heart and other organs for more than 3 decades. But, for these decades and longer transplantation could be offered to few who could benefit because donated human organs were scarce. The most obvious solution to the scarcity of human hearts and other organs today, 30 years ago and decades before has been to use animals instead of humans as the source of organs, that is xenotransplantation. Yet, if xenotransplantation has been the most obvious solution, it also has seemed the most frustrating, a proverbial carrot on a stick, held just beyond reach by 3 barriers—immunity, physiologic incompatibility, and infection—of which immunity appeared most daunting.1Cascalho M Platt JL The immunological barrier to xenotransplantation.Immunity. 2001; 14: 437-446Abstract Full Text Full Text PDF PubMed Scopus (154) Google Scholar The past 3 decades brought some dramatic advances in concept and technologies that might address the barriers to xenotransplantation. Yet, these advances were countered at every turn by discovery of new antigens, more incompatibilities and longer lists of zoonotic organisms. But in the current issue of Journal of Heart and Lung Transplantation, and in a previous report in another journal,2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar,3Längin M Mayr T Reichart B et al.Consistent success in life-supporting porcine cardiac xenotransplantation.Nature. 2018; 564: 430-433Crossref PubMed Scopus (142) Google Scholar Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar report > 3-month survival and function of orthotopic pig cardiac xenografts in baboons given clinically acceptable immunosuppression. The results “fulfill for the first time the preclinical efficacy” standard an International Society for Heart and Lung Transplantation (ISHLT) committee proposed as a premise for launching clinical trials in cardiac xenotransplantation.2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar,4Cooper DK Keogh AM Brink J et al.Report of the Xenotransplantation Advisory Committee of the International Society for Heart and Lung Transplantation: the present status of xenotransplantation and its potential role in the treatment of end-stage cardiac and pulmonary diseases.J Heart Lung Transplant. 2000; 19: 1125-1165Abstract Full Text Full Text PDF PubMed Scopus (136) Google Scholar Did Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar finally capture the carrot and take, what they call “the final step toward clinical xenotransplantation” or did they just advance the stick, with unenumerated barriers keeping clinical application beyond reach? In 2000, an advisory committee of the ISHLT reported on the status of research into the barriers to clinical cardiac and pulmonary xenotransplantation and recommended that “a clinical trial should be considered when approximately 60% survival of life-supporting pig organs in non-human primates for a minimum of 3 months [has been achieved in] at least 10 animals.”4Cooper DK Keogh AM Brink J et al.Report of the Xenotransplantation Advisory Committee of the International Society for Heart and Lung Transplantation: the present status of xenotransplantation and its potential role in the treatment of end-stage cardiac and pulmonary diseases.J Heart Lung Transplant. 2000; 19: 1125-1165Abstract Full Text Full Text PDF PubMed Scopus (136) Google Scholar The report culminated a decade of high profile discovery and debate. Key immune, physiologic, and infectious barriers to xenotransplantation had been identified and dramatic advances had been reported in genetic engineering, reproductive cloning, and development of biologic technologies for suppressing immunity and inducing tolerance (see Cascalho and Platt1Cascalho M Platt JL The immunological barrier to xenotransplantation.Immunity. 2001; 14: 437-446Abstract Full Text Full Text PDF PubMed Scopus (154) Google Scholar for review). But, the exciting advances that made clinical application seem so close also fueled concern about a long-known but little-understood endogenous retrovirus of pigs (porcine endogenous retrovirus) and about novel organisms that might develop in and spread beyond the immunosuppressed recipient. Although the need for organs for transplantation was urgent and increasing the ISHLT committee suggested only "consideration" of clinical trials because the proposed standard was still far from being achieved. Only 1 of numerous heterotopic cardiac xenografts in non-human primates had survived 3 months, and this graft suffered severe immune and inflammatory injury. Thus, the ISHLT committee correctly asserted that as of 2000, the “immune barriers [had] not yet been overcome.”4Cooper DK Keogh AM Brink J et al.Report of the Xenotransplantation Advisory Committee of the International Society for Heart and Lung Transplantation: the present status of xenotransplantation and its potential role in the treatment of end-stage cardiac and pulmonary diseases.J Heart Lung Transplant. 2000; 19: 1125-1165Abstract Full Text Full Text PDF PubMed Scopus (136) Google Scholar Equally important was that only 1 orthotopic porcine cardiac xenograft had as yet supported the life of a non-human primate treated with clinically acceptable immunosuppression regimen and that for 39 days.5Vial CM Ostlie DJ Bhatti FN et al.Life supporting function for over one month of a transgenic porcine heart in a baboon.J Heart Lung Transplant. 2000; 19: 224-229Abstract Full Text Full Text PDF PubMed Scopus (97) Google Scholar In 2016, the prospects for clinical application of xenotransplantation changed dramatically. Mohiuddin et al6Mohiuddin MM Singh AK Corcoran PC et al.Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft.Nat Commun. 2016; 7: 11138Crossref PubMed Scopus (190) Google Scholar reported that heterotopic porcine cardiac xenografts expressing a human complement regulator (CD46) and human thrombomodulin and lacking Galα1-3Gal (owing targeting of α1,3-galactosyltransferase) could survive more than a year (up to 945 days) in baboons treated with a clinically-acceptable regimen that included anti-rhesus monkey CD40. Because the xenografts were heterotopic, the potential for providing physiologic support remained uncertain but this success in overcoming immune barriers reinvigorated discussion about whether and how xenotransplantation might be applied in patients. In 2018, Längin et al3Längin M Mayr T Reichart B et al.Consistent success in life-supporting porcine cardiac xenotransplantation.Nature. 2018; 564: 430-433Crossref PubMed Scopus (142) Google Scholar reported 4 successful orthotopic porcine xenografts in baboons that functioned until the recipients were euthanized, per protocol on days 90, 90, 182, and 195. The xenografts were from pigs with the same or similar genetic modifications as those used by Mohiuddin et al,6Mohiuddin MM Singh AK Corcoran PC et al.Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft.Nat Commun. 2016; 7: 11138Crossref PubMed Scopus (190) Google Scholar and the baboons were treated with immunosuppression similar to that of Mohiuddin et al6Mohiuddin MM Singh AK Corcoran PC et al.Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft.Nat Commun. 2016; 7: 11138Crossref PubMed Scopus (190) Google Scholar (plus an mammalian target of rapamycin inhibitor to limit myocardial remodeling7Kushwaha SS Raichlin E Sheinin Y et al.Sirolimus affects cardiomyocytes to reduce left ventricular mass in heart transplant recipients.Eur Heart J. 2008; 29: 2742-2750Crossref PubMed Scopus (41) Google Scholar). The present report2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar adds 4 orthotopic xenografts to the series. The xenografts in 2 recipients exhibited no evidence of significant rejection but systemic complications in the baboons with grafts harboring Cytomegalovirus (porcine cytomegalovirus [pCMV]) necessitated termination on Days 15 and 27. Two other xenografts functioned at 90 days, when the recipients were euthanized per protocol. Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar portray the 6 of 8 orthotopic xenografts functioning 3 months or longer as “well exceeding the key requirement of the ISHLT of the ISHLT International Advisory Board.” Some might questionwhether 6 of 8 xenografts functioning 3 months or more well exceeds or meets the ISHLT standard (60% of at least 10 life-supporting xenografts functioning 3 months4Cooper DK Keogh AM Brink J et al.Report of the Xenotransplantation Advisory Committee of the International Society for Heart and Lung Transplantation: the present status of xenotransplantation and its potential role in the treatment of end-stage cardiac and pulmonary diseases.J Heart Lung Transplant. 2000; 19: 1125-1165Abstract Full Text Full Text PDF PubMed Scopus (136) Google Scholar). We think giving more than passing mention to this standard formulated more than 15 years before the immunologic barriers had been overcome, obscures the significance of Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar’s accomplishment. If a porcine cardiac xenograft can support a non-human primate treated with a clinically acceptable regimen of immunosuppression; then, we must urgently consider what more, if anything, is needed to offer a cardiac xenograft to a dying human patient. And, we must explain why those facing imminent death for want of a transplant cannot be permitted xenotransplantation instead. Yet, after repeatedly claiming to have met or exceeded the standard for clinical application of xenotransplantation,2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar,3Längin M Mayr T Reichart B et al.Consistent success in life-supporting porcine cardiac xenotransplantation.Nature. 2018; 564: 430-433Crossref PubMed Scopus (142) Google Scholar Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar now demurs, saying only that “more experiments will be necessary.” But, the justification for more experiments is unpersuasive. The authors assert more experiments would “address the efficacy of humanized anti-CD40/CD40L,” but orthotopic cardiac xenografts are extraordinarily inefficient, complex, and insensitive system for testing the efficacy of immune modifiers; indeed, the efficacy of anti-CD40/CD40L was established using heterotopic xenografts.6Mohiuddin MM Singh AK Corcoran PC et al.Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft.Nat Commun. 2016; 7: 11138Crossref PubMed Scopus (190) Google Scholar Eight orthotopic xenografts are certainly too few to firmly establish the frequency of success and durability of function porcine cardiac xenografts in baboons. But, larger numbers of xenografts in baboons will reveal little more about the efficacy and durability xenografts might have in humans. The ISHLT committee wisely avoided stipulating a standard error for 60% survival or an end-point beyond 90 days because experimental transplants in non-human primates do not model and potentially misrepresent some key variables in clinical cardiac transplantation.8Platt JL Cascalho M Piedrahita JA Xenotransplantation: progress along paths uncertain from models to application.ILAR J. 2018; 59: 286-308Crossref PubMed Scopus (5) Google Scholar That is not to say experimentation in this model or in others should cease. Clinical cardiac xenotransplantation is likely to raise questions and challenges not foreseen today, and an experimental model could prove invaluable for addressing these. There is another perhaps more pressing reason to question what more experiments can accomplish. We suspect porcine cardiac xenografts in non-human primates might overestimate or misrepresent the barriers to clinical xenotransplantation.8Platt JL Cascalho M Piedrahita JA Xenotransplantation: progress along paths uncertain from models to application.ILAR J. 2018; 59: 286-308Crossref PubMed Scopus (5) Google Scholar The xenografts reported by Mohiuddin et al6Mohiuddin MM Singh AK Corcoran PC et al.Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft.Nat Commun. 2016; 7: 11138Crossref PubMed Scopus (190) Google Scholar and Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar were from pigs engineered to express human CD46 and human thrombomodulin to better regulate complement and coagulation in recipients. Yet, these and other proteins interact inefficiently with cognatepartners in baboons.9Lawson JH Platt JL Molecular barriers to xenotransplantation.Transplantation. 1996; 62: 303-310Crossref PubMed Scopus (162) Google Scholar Still more important is the possibility that the human proteins might elicit cellular and/or humoral immunity in baboons. Such immunity developing over time, perhaps when immunosuppression decreases to maintenance, might block regulatory functions or initiate injury to the grafts. Perhaps the anti-CD40 antibodies Mohiuddin et al6Mohiuddin MM Singh AK Corcoran PC et al.Chimeric 2C10R4 anti-CD40 antibody therapy is critical for long-term survival of GTKO.hCD46.hTBM pig-to-primate cardiac xenograft.Nat Commun. 2016; 7: 11138Crossref PubMed Scopus (190) Google Scholar considers essential are needed to disrupt immunity to the human proteins. Since intensity of immunosuppression could limit clinical application of xenotransplantation, it would be ironic if humans were found to need less intense regimens than baboons because humans are tolerant to the products of these transgenes. Of course, clinical trials might reveal higher or different barriers to xenotransplantation in humans than in baboons. But, that possibility has eluded detection in Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar’s model, and it would seem unlikely that more experiments would overcome the deficiency. Nor will the conduct of more porcine xenografts in baboons reveal much about the barrier posed by infection. The microorganisms of pigs known to be capable of infecting humans have been identified almost entirely by investigation of humans and pigs. Indeed, concerns about porcine endogenous retrovirus arose from investigation of human and pig cells in culture and the waning of those concerns from investigation of human contact with pig organs and xenografts.10Paradis K Langford G Long Z et al.Search for cross-species transmission of porcine endogenous retrovirus in patients treated with living pig tissue. The XEN 111 Study Group.Science. 1999; 285: 1236-1241Crossref PubMed Scopus (631) Google Scholar Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar associate the demise of 2 baboon recipients with the presence of pCMV in in cardiac xenografts, but the paper provides no evidence that pCMV was transmitted to the recipients or directly caused multiorgan failure that forced termination of the experiments. Perhaps a yet unidentified organism harbored by the baboons compromised innate immune control of pCMV. Regardless, pCMV will be excluded from potential sources of xenografts and since pCMV has not been shown to infect humans, Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar’s experience might reflect an instance in which non-human primates misrepresent a biologic barrier. In 2018, Längin et al3Längin M Mayr T Reichart B et al.Consistent success in life-supporting porcine cardiac xenotransplantation.Nature. 2018; 564: 430-433Crossref PubMed Scopus (142) Google Scholar called the first 4 successful orthotopic cardiac xenografts in baboons a milestone. If those xenografts and the 2 successful xenografts now reported2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar are a milestone, it is because they erase serious doubt about whether a porcine heart can survive and function for months in a non-human primate recipient receiving a clinically acceptable (if not yet optimal) immunosuppression regimen. But, the experiments do not prove clinical xenotransplantation will succeed—only clinical xenotransplants can prove that. If this regimen were used successfully in clinical xenotransplants, the clinical xenotransplants would be appropriately called a milestone, if the regimen failed some other term might be used. Reichart et al2Reichart B, Längin M, Radan J, et al. Pig-to-non-human primate heart transplantation: the final step toward clinical xenotransplantation [e-pub ahead of print]?J Heart Lung Transplant, https://doi.org/10.1016/j.healun.2020.05.004. Accessed July 6, 2020.Google Scholar’s report brings into focus an impending watershed in management heart failure. When clinical trials in cardiac xenotransplantation do begin, whether soon or after many more experiments, interest in the multitude of pre-clinical experiments and committee deliberations concerning the feasibility of xenotransplantation will fade. And, the central question will be whether the essential but intrusive immune modifiers and/or the less than optimal genetic background of engineered pigs8Platt JL Cascalho M Piedrahita JA Xenotransplantation: progress along paths uncertain from models to application.ILAR J. 2018; 59: 286-308Crossref PubMed Scopus (5) Google Scholar can be omitted or changed so that xenotransplantation can approach or even surpass the safety and efficacy of allotransplantation and become the preferred treatment for organ failure. That shift in focus as difficult to imagine as it may be will fuel identification and surmounting of barriers unapparent in today's pre-clinical models. If this impending change in interest in xenotransplantation seems fanciful, it just recapitulates how interest in allotransplantation changed 60 years ago. The authors have no conflicts of interest to declare. The work be the authors pertinent to this communication was supported by the National Institutes of Health (AI122369; OD023138) Pig-to-non-human primate heart transplantation: The final step toward clinical xenotransplantation?The Journal of Heart and Lung TransplantationVol. 39Issue 8PreviewThe demand for donated human hearts far exceeds the number available. Xenotransplantation of genetically modified porcine organs provides an alternative. In 2000, an Advisory Board of the International Society for Heart and Lung Transplantation set the benchmark for commencing clinical cardiac xenotransplantation as consistent 60% survival of non-human primates after life-supporting porcine heart transplantations. Recently, we reported the stepwise optimization of pig-to-baboon orthotopic cardiac xenotransplantation finally resulting in consistent success, with 4 recipients surviving 90 (n = 2), 182, and 195 days. Full-Text PDF DA - 2020/8// PY - 2020/8// DO - 10.1016/j.healun.2020.06.002 VL - 39 IS - 8 SP - 758-760 SN - 1557-3117 ER - TY - JOUR TI - High-Throughput Manufacture of 3D Fiber Scaffolds for Regenerative Medicine AU - Shirwaiker, Rohan A. AU - Fisher, Matthew B. AU - Anderson, Bruce AU - Schuchard, Karl G. AU - Warren, Paul B. AU - Maze, Benoit AU - Grondin, Pierre AU - Ligler, Frances S. AU - Pourdeyhimi, Behnam T2 - TISSUE ENGINEERING PART C-METHODS AB - Engineered scaffolds used to regenerate mammalian tissues should recapitulate the underlying fibrous architecture of native tissue to achieve comparable function. Current fibrous scaffold fabrication processes, such as electrospinning and three-dimensional (3D) printing, possess application-specific advantages, but they are limited either by achievable fiber sizes and pore resolution, processing efficiency, or architectural control in three dimensions. As such, a gap exists in efficiently producing clinically relevant, anatomically sized scaffolds comprising fibers in the 1–100 μm range that are highly organized. This study introduces a new high-throughput, additive fibrous scaffold fabrication process, designated in this study as 3D melt blowing (3DMB). The 3DMB system described in this study is modified from larger nonwovens manufacturing machinery to accommodate the lower volume, high-cost polymers used for tissue engineering and implantable biomedical devices and has a fiber collection component that uses adaptable robotics to create scaffolds with predetermined geometries. The fundamental process principles, system design, and key parameters are described, and two examples of the capabilities to create scaffolds for biomedical engineering applications are demonstrated. Three-dimensional melt blowing (3DMB) is a new, high-throughput, additive manufacturing process to produce scaffolds composed of highly organized fibers in the anatomically relevant 1–100 μm range. Unlike conventional melt-blowing systems, the 3DMB process is configured for efficient use with the relatively expensive polymers necessary for biomedical applications, decreasing the required amounts of material for processing while achieving high throughputs compared with 3D printing or electrospinning. The 3DMB is demonstrated to make scaffolds composed of multiple fiber materials and organized into complex shapes, including those typical of human body parts. DA - 2020/7/1/ PY - 2020/7/1/ DO - 10.1089/ten.tec.2020.0098 VL - 26 IS - 7 SP - 364-374 SN - 1937-3392 KW - 3D printing KW - melt blowing KW - fibers KW - scaffolds ER - TY - JOUR TI - Dermal exosomes containing miR-218-5p promote hair regeneration by regulating beta-catenin signaling AU - Hu, Shiqi AU - Li, Zhenhua AU - Lutz, Halle AU - Huang, Ke AU - Su, Teng AU - Cores, Jhon AU - Dinh, Phuong-Uyen Cao AU - Cheng, Ke T2 - SCIENCE ADVANCES AB - The progression in the hair follicle cycle from the telogen to the anagen phase is the key to regulating hair regrowth. Dermal papilla (DP) cells support hair growth and regulate the hair cycle. However, they gradually lose key inductive properties upon culture. DP cells can partially restore their capacity to promote hair regrowth after being subjected to spheroid culture. In this study, results revealed that DP spheroids are effective at inducing the progression of the hair follicle cycle from telogen to anagen compared with just DP cell or minoxidil treatment. Because of the importance of paracrine signaling in this process, secretome and exosomes were isolated from DP cell culture, and their therapeutic efficacies were investigated. We demonstrated that miR-218-5p was notably up-regulated in DP spheroid-derived exosomes. Western blot and immunofluorescence imaging were used to demonstrate that DP spheroid-derived exosomes up-regulated β-catenin, promoting the development of hair follicles. DA - 2020/7// PY - 2020/7// DO - 10.1126/sciadv.aba1685 VL - 6 IS - 30 SP - SN - 2375-2548 ER - TY - JOUR TI - Exosome therapeutics for lung regenerative medicine AU - Popowski, Kristen AU - Lutz, Halle AU - Hu, Shiqi AU - George, Arianna AU - Dinh, Phuong-Uyen AU - Cheng, Ke T2 - JOURNAL OF EXTRACELLULAR VESICLES AB - ABSTRACT Exosomes are 30 to 100 nm extracellular vesicles that are secreted by many cell types. Initially viewed as cellular garbage with no biological functions, exosomes are now recognized for their therapeutic potential and used in regenerative medicine. Cell‐derived exosomes are released into almost all biological fluids, making them abundant and accessible vesicles for a variety of diseases. These naturally occurring nanoparticles have a wide range of applications including drug delivery and regenerative medicine. Exosomes sourced from a specific tissue have been proven to provide greater therapeutic effects to their native tissue, expanding exosome sources beyond traditional cell lines such as mesenchymal stem cells. However, standardizing production and passing regulations remain obstacles, due to variations in methods and quantification techniques across studies. Additionally, obtaining pure exosomes at sufficient quantities remains difficult due to the heterogeneity of exosomes. In this review, we will underline the uses of exosomes as a therapy and their roles in lung regenerative medicine, as well as current challenges in exosome therapies. DA - 2020/1/1/ PY - 2020/1/1/ DO - 10.1080/20013078.2020.1785161 VL - 9 IS - 1 SP - SN - 2001-3078 KW - Exosome KW - regenerative medicine KW - lung spheroid cell KW - stem cell ER - TY - JOUR TI - Light-triggered NO-releasing nanoparticles for treating mice with liver fibrosis AU - Liang, Hongxia AU - Li, Zhenhua AU - Ren, Zhigang AU - Jia, Qiaodi AU - Guo, Linna AU - Li, Shasha AU - Zhang, Hongyu AU - Hu, Shiqi AU - Zhu, Dashuai AU - Shen, Deliang AU - Yu, Zujiang AU - Cheng, Ke T2 - NANO RESEARCH DA - 2020/8// PY - 2020/8// DO - 10.1007/s12274-020-2833-6 VL - 13 IS - 8 SP - 2197-2202 SN - 1998-0000 KW - liver fibrosis KW - nitric oxide (NO) KW - hepatic stellate cells (HSC) KW - nanoparticles KW - near infrared light (NIR)-controlled release ER - TY - JOUR TI - The bioprinting roadmap AU - Sun, Wei AU - Starly, Binil AU - Daly, Andrew C. AU - Burdick, Jason A. AU - Groll, Juergen AU - Skeldon, Gregor AU - Shu, Wenmiao AU - Sakai, Yasuyuki AU - Shinohara, Marie AU - Nishikawa, Masaki AU - Jang, Jinah AU - Cho, Dong-Woo AU - Nie, Minghao AU - Takeuchi, Shoji AU - Ostrovidov, Serge AU - Khademhosseini, Ali AU - Kamm, Roger D. AU - Mironov, Vladimir AU - Moroni, Lorenzo AU - Ozbolat, Ibrahim T. T2 - BIOFABRICATION AB - This bioprinting roadmap features salient advances in selected applications of the technique and highlights the status of current developments and challenges, as well as envisioned advances in science and technology, to address the challenges to the young and evolving technique. The topics covered in this roadmap encompass the broad spectrum of bioprinting; from cell expansion and novel bioink development to cell/stem cell printing, from organoid-based tissue organization to bioprinting of human-scale tissue structures, and from building cell/tissue/organ-on-a-chip to biomanufacturing of multicellular engineered living systems. The emerging application of printing-in-space and an overview of bioprinting technologies are also included in this roadmap. Due to the rapid pace of methodological advancements in bioprinting techniques and wide-ranging applications, the direction in which the field should advance is not immediately clear. This bioprinting roadmap addresses this unmet need by providing a comprehensive summary and recommendations useful to experienced researchers and newcomers to the field. DA - 2020/4// PY - 2020/4// DO - 10.1088/1758-5090/ab5158 VL - 12 IS - 2 SP - SN - 1758-5090 KW - biofabrication KW - bioprinting KW - cell printing KW - in vitro biological models KW - disease models KW - organoids KW - organ-on-a-chip ER - TY - JOUR TI - Engineering better stem cell therapies for treating heart diseases AU - Li, Junlang AU - Hu, Shiqi AU - Cheng, Ke T2 - ANNALS OF TRANSLATIONAL MEDICINE AB - : For decades, stem cells and their byproducts have shown efficacy in repairing tissues and organs in numerous pre-clinical studies and some clinical trials, providing hope for possible cures for many important diseases. However, the translation of stem cell therapy for heart diseases from bench to bed is still hampered by several limitations. The therapeutic benefits of stem cells are mediated by a combo of mechanisms. In this review, we will provide a brief summary of stem cell therapies for ischemic heart disease. Basically, we will talk about these barriers for the clinical application of stem cell-based therapies, the investigation of mechanisms behind stem-cell based cardiac regeneration and also, what bioengineers can do and have been doing on the translational stage of stem cell therapies for heart repair. DA - 2020/4// PY - 2020/4// DO - 10.21037/atm.2020.03.44 VL - 8 IS - 8 SP - SN - 2305-5847 KW - Stem cell therapy KW - ischemic heart diseases KW - bioengineering strategies KW - clinical translation ER - TY - JOUR TI - Tumor cell-derived exosomes home to their cells of origin and can be used as Trojan horses to deliver cancer drugs AU - Qiao, Li AU - Hu, Shiqi AU - Huang, Ke AU - Su, Teng AU - Li, Zhenhua AU - Vandergriff, Adam AU - Cores, Jhon AU - Dinh, Phuong-Uyen AU - Allen, Tyler AU - Shen, Deliang AU - Liang, Hongxia AU - Li, Yongjun AU - Cheng, Ke T2 - THERANOSTICS AB - Cancer is the second leading cause of death worldwide and patients are in urgent need of therapies that can effectively target cancer with minimal off-target side effects. Exosomes are extracellular nano-shuttles that facilitate intercellular communication between cells and organs. It has been established that tumor-derived exosomes contain a similar protein and lipid composition to that of the cells that secrete them, indicating that exosomes might be uniquely employed as carriers for anti-cancer therapeutics. Methods: We isolated exosomes from two cancer cell lines, then co-cultured each type of cancer cells with these two kinds of exosomes and quantified exosome. HT1080 or Hela exosomes were systemically injected to Nude mice bearing a subcutaneous HT1080 tumor to investigate their cancer-homing behavior. Moreover, cancer cell-derived exosomes were engineered to carry Doxil (a common chemotherapy drug), known as D-exo, were used to detect their target and therapeutic efficacy as anti-cancer drugs. Exosome proteome array analysis were used to reveal the mechanism underly this phenomenon. Results: Exosomes derived from cancer cells fuse preferentially with their parent cancer cells, in vitro. Systemically injected tumor-derived exosomes home to their original tumor tissues. Moreover, compared to Doxil alone, the drug-loaded exosomes showed enhanced therapeutic retention in tumor tissues and eradicated them more effectively in nude mice. Exosome proteome array analysis revealed distinct integrin expression patterns, which might shed light on the underlying mechanisms that explain the exosomal cancer-homing behavior. Conclusion: Here we demonstrate that the exosomes' ability to target the parent cancer is a phenomenon that opens up new ways to devise targeted therapies to deliver anti-tumor drugs. DA - 2020/// PY - 2020/// DO - 10.7150/thno.39434 VL - 10 IS - 8 SP - 3474-3487 SN - 1838-7640 KW - exosome KW - homing KW - integrin KW - doxorubicin KW - cancer therapy ER - TY - JOUR TI - Inhalation of lung spheroid cell secretome and exosomes promotes lung repair in pulmonary fibrosis AU - Dinh, Phuong-Uyen C. AU - Paudel, Dipti AU - Brochu, Hayden AU - Popowski, Kristen D. AU - Gracieux, M. Cyndell AU - Cores, Jhon AU - Huang, Ke AU - Hensley, M. Taylor AU - Harrell, Erin AU - Vandergriff, Adam C. AU - George, Arianna K. AU - Barrio, Raina T. AU - Hu, Shiqi AU - Allen, Tyler A. AU - Blackburn, Kevin AU - Caranasos, Thomas G. AU - Peng, Xinxia AU - Schnabel, Lauren V. AU - Adler, Kenneth B. AU - Lobo, Leonard J. AU - Goshe, Michael B. AU - Cheng, Ke T2 - NATURE COMMUNICATIONS AB - Abstract Idiopathic pulmonary fibrosis (IPF) is a fatal and incurable form of interstitial lung disease in which persistent injury results in scar tissue formation. As fibrosis thickens, the lung tissue loses the ability to facilitate gas exchange and provide cells with needed oxygen. Currently, IPF has few treatment options and no effective therapies, aside from lung transplant. Here we present a series of studies utilizing lung spheroid cell-secretome (LSC-Sec) and exosomes (LSC-Exo) by inhalation to treat different models of lung injury and fibrosis. Analysis reveals that LSC-Sec and LSC-Exo treatments could attenuate and resolve bleomycin- and silica-induced fibrosis by reestablishing normal alveolar structure and decreasing both collagen accumulation and myofibroblast proliferation. Additionally, LSC-Sec and LSC-Exo exhibit superior therapeutic benefits than their counterparts derived from mesenchymal stem cells in some measures. We showed that an inhalation treatment of secretome and exosome exhibited therapeutic potential for lung regeneration in two experimental models of pulmonary fibrosis. DA - 2020/2/28/ PY - 2020/2/28/ DO - 10.1038/s41467-020-14344-7 VL - 11 IS - 1 SP - SN - 2041-1723 ER - TY - JOUR TI - Joint laxity varies in response to partial and complete anterior cruciate ligament injuries throughout skeletal growth AU - Cone, Stephanie G. AU - Lambeth, Emily P. AU - Piedrahita, Jorge A. AU - Spang, Jeffrey T. AU - Fisher, Matthew B. T2 - JOURNAL OF BIOMECHANICS AB - Anterior cruciate ligament (ACL) injuries are increasingly common in the skeletally immature population. As such there is a need to increase our understanding of the biomechanical function of the joint following partial and complete ACL injury during skeletal growth. In this work, we aimed to assess changes in knee kinematics and loading of the remaining soft tissues following both partial and complete ACL injury in a porcine model. To do so, we applied anterior-posterior tibial loads and varus-valgus moments to stifle joints of female pigs ranging from early juvenile to late adolescent ages and assessed both kinematics and in-situ loads carried in the bundles of the ACL and other soft tissues including the collateral ligaments and the menisci. Partial ACL injury led to increased anterior tibial translation only in late adolescence and small increases in varus-valgus rotation at all ages. Complete ACL injury led to substantial increases in translation and rotation at all ages. At all ages, the medial collateral ligament and the medial meniscus combined to resist the majority of applied anterior tibial load following complete ACL transection. Across all ages and flexion angles, the contribution of the MCL ranged from 45 to 90% of the anterior load and the contribution of the medial meniscus ranged from 14 to 35% of the anterior load. These findings add to our current understanding of age-specific functional properties of both healthy and injured knees during skeletal growth. DA - 2020/3/5/ PY - 2020/3/5/ DO - 10.1016/j.jbiomech.2020.109636 VL - 101 SP - SN - 1873-2380 KW - Anterior cruciate ligament KW - Pediatric KW - Injury KW - Joint mechanics KW - Animal model ER - TY - JOUR TI - Atorvastatin enhances the therapeutic efficacy of mesenchymal stem cells-derived exosomes in acute myocardial infarction via up-regulating long non-coding RNA H19 AU - Huang, Peisen AU - Wang, Li AU - Li, Qing AU - Tian, Xiaqiu AU - Xu, Jun AU - Xu, Junyan AU - Xiong, Yuyan AU - Chen, Guihao AU - Qian, Haiyan AU - Jin, Chen AU - Yu, Yuan AU - Cheng, Ke AU - Qian, Li AU - Yang, Yuejin T2 - CARDIOVASCULAR RESEARCH AB - Abstract Aims Naturally secreted nanovesicles, known as exosomes, play important roles in stem cell-mediated cardioprotection. We have previously demonstrated that atorvastatin (ATV) pretreatment improved the cardioprotective effects of mesenchymal stem cells (MSCs) in a rat model of acute myocardial infarction (AMI). The aim of this study was to investigate if exosomes derived from ATV-pretreated MSCs exhibit more potent cardioprotective function in a rat model of AMI and if so to explore the underlying mechanisms. Methods and results Exosomes were isolated from control MSCs (MSC-Exo) and ATV-pretreated MSCs (MSCATV-Exo) and were then delivered to endothelial cells and cardiomyocytes in vitro under hypoxia and serum deprivation (H/SD) condition or in vivo in an acutely infarcted Sprague-Dawley rat heart. Regulatory genes and pathways activated by ATV pretreatment were explored using genomics approaches and functional studies. In vitro, MSCATV-Exo accelerated migration, tube-like structure formation, and increased survival of endothelial cells but not cardiomyocytes, whereas the exosomes derived from MSCATV-Exo-treated endothelial cells prevented cardiomyocytes from H/SD-induced apoptosis. In a rat AMI model, MSCATV-Exo resulted in improved recovery in cardiac function, further reduction in infarct size and reduced cardiomyocyte apoptosis compared to MSC-Exo. In addition, MSCATV-Exo promoted angiogenesis and inhibited the elevation of IL-6 and TNF-α in the peri-infarct region. Mechanistically, we identified lncRNA H19 as a mediator of the role of MSCATV-Exo in regulating expression of miR-675 and activation of proangiogenic factor VEGF and intercellular adhesion molecule-1. Consistently, the cardioprotective effects of MSCATV-Exo was abrogated when lncRNA H19 was depleted in the ATV-pretreated MSCs and was mimicked by overexpression of lncRNA H19. Conclusion Exosomes obtained from ATV-pretreated MSCs have significantly enhanced therapeutic efficacy for treatment of AMI possibly through promoting endothelial cell function. LncRNA H19 mediates, at least partially, the cardioprotective roles of MSCATV-Exo in promoting angiogenesis. DA - 2020/2/1/ PY - 2020/2/1/ DO - 10.1093/cvr/cvz139 VL - 116 IS - 2 SP - 353-367 SN - 1755-3245 KW - Exosomes KW - Atorvastatin KW - MSCs KW - lncRNA H19 KW - Myocardial infarction ER - TY - JOUR TI - Targeted anti-IL-1 beta platelet microparticles for cardiac detoxing and repair AU - Li, Zhenhua AU - Hu, Shiqi AU - Huang, Ke AU - Su, Teng AU - Cores, Jhon AU - Cheng, Ke T2 - SCIENCE ADVANCES AB - An acute myocardial infarction (AMI) induces a sterile inflammatory response that facilitates further heart injury and promotes adverse cardiac remodeling. Interleukin-1β (IL-1β) plays a central role in the sterile inflammatory response that results from AMI. Thus, IL-1β blockage is a promising strategy for treatment of AMI. However, conventional IL-1β blockers lack targeting specificity. This increases the risk of serious side effects. To address this problem herein, we fabricated platelet microparticles (PMs) armed with anti-IL-1β antibodies to neutralize IL-1β after AMI and to prevent adverse cardiac remodeling. Our results indicate that the infarct-targeting PMs could bind to the injured heart, increasing the number of anti-IL-1β antibodies therein. The anti-IL-1β platelet PMs (IL1-PMs) protect the cardiomyocytes from apoptosis by neutralizing IL-1β and decreasing IL-1β-driven caspase-3 activity. Our findings indicate that IL1-PM is a promising cardiac detoxification agent that removes cytotoxic IL-1β during AMI and induces therapeutic cardiac repair. DA - 2020/2// PY - 2020/2// DO - 10.1126/sciadv.aay0589 VL - 6 IS - 6 SP - SN - 2375-2548 ER - TY - JOUR TI - Mechanical properties of tissue formed in vivo are affected by 3D-bioplotted scaffold microarchitecture and correlate with ECM collagen fiber alignment AU - Huebner, Pedro AU - Warren, Paul B. AU - Chester, Daniel AU - Spang, Jeffrey T. AU - Brown, Ashley C. AU - Fisher, Matthew B. AU - Shirwaiker, Rohan A. T2 - CONNECTIVE TISSUE RESEARCH AB - Purpose: Musculoskeletal soft tissues possess highly aligned extracellular collagenous networks that provide structure and strength. Such an organization dictates tissue-specific mechanical properties but can be difficult to replicate by engineered biological substitutes. Nanofibrous electrospun scaffolds have demonstrated the ability to control cell-secreted collagen alignment, but concerns exist regarding their scalability for larger and anatomically relevant applications. Additive manufacturing processes, such as melt extrusion-based 3D-Bioplotting, allow fabrication of structurally relevant scaffolds featuring highly controllable porous microarchitectures.Materials and Methods: In this study, we investigate the effects of 3D-bioplotted scaffold design on the compressive elastic modulus of neotissue formed in vivo in a subcutaneous rat model and its correlation with the alignment of ECM collagen fibers. Polycaprolactone scaffolds featuring either 100 or 400 µm interstrand spacing were implanted for 4 or 12 weeks, harvested, cryosectioned, and characterized using atomic-force-microscopy-based force mapping.Results: The compressive elastic modulus of the neotissue formed within the 100 µm design was significantly higher at 4 weeks (p < 0.05), but no differences were observed at 12 weeks. In general, the tissue stiffness was within the same order of magnitude and range of values measured in native musculoskeletal soft tissues including the porcine meniscus and anterior cruciate ligament. Finally, a significant positive correlation was noted between tissue stiffness and the degree of ECM collagen fiber alignment (p < 0.05) resulting from contact guidance provided by scaffold strands.Conclusion: These findings demonstrate the significant effects of 3D-bioplotted scaffold microarchitectures in the organization and sub-tissue-level mechanical properties of ECM in vivo. DA - 2020/3/3/ PY - 2020/3/3/ DO - 10.1080/03008207.2019.1624733 VL - 61 IS - 2 SP - 190-204 SN - 1607-8438 KW - 3D printing KW - atomic force microscopy KW - elastic modulus KW - tissue engineering KW - tissue scaffolds ER -