Enhanced elastin synthesis and maturation in human vascular smooth muscle tissue derived from induced-pluripotent stem cells (original) (raw)
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Experimental Cell Research, 2016
In vitro research on vascular tissue engineering has extensively used isolated primary human or animal smooth muscle cells (SMC). Research programs that lack such facilities tend towards commercially available primary cells sources. Here, we aim to evaluate the capacity of commercially available human SMC to maintain their contractile phenotype, and determine if dedifferentiation towards the synthetic phenotype occurs in response to conventional cell culture and passaging without any external biochemical or mechanical stimuli. Lower passage SMC adopted a contractile phenotype marked by a relatively slower proliferation rate, higher expression of proteins of the contractile apparatus and smoothelin, elongated morphology, and reduced deposition of collagen types I and III. As the passage number increased, migratory capacity was enhanced, average cell speed, total distance and net distance travelled increased up to passage 8. Through the various assays, corroborative evidence pinpoints SMC at passage 7 as the transition point between the contractile and synthetic phenotypes, while passage 8 distinctly and consistently exhibited characteristics of synthetic phenotype. This knowledge is particularly useful in selecting SMC of appropriate passage number for the target vascular tissue engineering application, for example, a homeostatic vascular graft for blood vessel replacement versus recreating atherosclerotic blood vessel model in vitro.
Elastin biosynthesis: The missing link in tissue-engineered blood vessels
Cardiovascular Research, 2006
Nearly 20 years have passed since Weinberg and Bell attempted to make the first tissue-engineered blood vessels. Following this early attempt, vascular tissue engineering has emerged as one of the most promising approaches to fabricate orderly and mechanically competent vascular substitutes. In elastic and muscular arteries, elastin is a critical structural and regulatory matrix protein and plays an important and dominant role by conferring elasticity to the vessel wall. Elastin also regulates vascular smooth muscle cells activity and phenotype. Despite the great promise that tissue-engineered blood vessels have to offer, little research in the last two decades has addressed the importance of elastin incorporation into these vessels. Although cardiovascular tissue engineering has been reviewed in the past, very little attention has been given to elastin. Thus, this review focuses on the recent advances made towards elastogenesis and the challenges we face in the quest for appropriate functional vascular substitutes.
Proceedings of the National Academy of Sciences, 2010
Biodegradable scaffolds seeded with bone marrow mononuclear cells (BMCs) are the earliest tissue-engineered vascular grafts (TEVGs) to be used clinically. These TEVGs transform into living blood vessels in vivo, with an endothelial cell (EC) lining invested by smooth muscle cells (SMCs); however, the process by which this occurs is unclear. To test if the seeded BMCs differentiate into the mature vascular cells of the neovessel, we implanted an immunodeficient mouse recipient with human BMC (hBMC)-seeded scaffolds. As in humans, TEVGs implanted in a mouse host as venous interposition grafts gradually transformed into living blood vessels over a 6-month time course. Seeded hBMCs, however, were no longer detectable within a few days of implantation. Instead, scaffolds were initially repopulated by mouse monocytes and subsequently repopulated by mouse SMCs and ECs. Seeded BMCs secreted significant amounts of monocyte chemoattractant protein-1 and increased early monocyte recruitment. These findings suggest TEVGs transform into functional neovessels via an inflammatory process of vascular remodeling.
Tissue Engineering in the Vasculature
The Anatomical Record, 2013
Tissue engineering holds great promise to address complications and limitations encountered with the use of traditional prosthetic materials, such as thrombogenicity, infection, and future degeneration which represent the major morbidity and mortality after device implant surgery. The general concept of tissue engineering consists of three main components: a scaffold material, a cell type for seeding the scaffold, and biochemical, physio-chemical signaling and remodeling process. This remodeling process is guided by cell signals derived from both seeded cells and host inflammatory cells that infiltrate the scaffold and deposit extracellular matrix, forming the neotissue. Vascular tissue engineering is at the forefront in the translation of this technology to clinical practice, as tissue engineered vascular grafts (TEVGs) have now been successfully implanted in children with congenital heart disease. In this report, we review the history, advances, and state of the art in TEVGs. Anat Rec, 00:000-000, Abbreviations used: BMC 5 bone marrow cells; BMMNC 5 bone marrow mononuclear cell; CST 5 cell sheet technology; ECM 5 extracellular matrix; EB 5 embryonic bodies; ESC 5 embryonic stem cells; EC 5 endothelial cells; IVC 5 inferior vena cava; iPS cells 5 induced pluriopotent stem cells; PIPAAm 5 poly(N-isopropylacrylamide); SMC 5 smooth muscle cell.
Mitigating challenges and expanding the future of vascular tissue engineering—are we there yet?
Frontiers in Physiology
Atherosclerosis is still a significant cause of death in western societies. The leading cause of this cardiovascular disease is lipid accumulation and inflammation of the large arteries, which may lead to clinical complications such as arterial thrombosis, myocardial infarction, and ischemic stroke (Xenotransplantation, 1997). Drugs are usually the first treatment choice, even in the late stages of atherosclerosis. Sometimes, more aggressive treatment like Coronary artery bypass surgery (CABG) is needed. CABGs are performed by harvesting a vessel from the patient, but the patient undergoing two surgeries only added to their comorbidities. However, long-term results after CABG depend not only on the completeness of revascularization and the initial severity of coronary and myocardial lesions but also on comorbidities like diabetes mellitus, arterial hypertension, and pulmonary and renal disorders. Moreover, the limited availability of autografts was soon realized, and new options proposed by tissue engineering were started exploring to design synthetic grafts (Corridon et al., 2013). Vascular tissue engineering (VTE) is focused on constructing vessels using different biomaterials, cell sources, biomolecules, and mechanical stimuli that can function in physiological environments (Lovett et al., 2009; Pradeep et al., 2019; Wang et al., 2022a). Such vessels replace non-functional vascular compartments and generate networks within bio (artificial) scaffolds. Pioneering efforts in this field date back to 1950, when artificial vascular grafts made from synthetic polymer materials were used to replace occluded arterial vessels (Song et al., 2018). It was perceived that biomaterials could support microvascular function showing great potential in accelerating the transition away from xenogenic materials for clinical application. For example, Li et al. (2017) developed a hyaluronic acid-based hydrogel chemically modified with fibronectin motifs that promote EC binding of α3/α5 b1 integrins, resulting in better vascularization to a non-modified hydrogel in a mouse stroke model. Similarly, advances in nanotechnology can bring additional functionality to vascular scaffolds, optimize internal vascular graft surface, reduce early thrombosis and inflammatory responses, and even direct the differentiation of stem cells into the vascular cell phenotype (Mironov et al., 2008; Wang et al., 2022a).
PLoS ONE, 2013
Aims: Tissue engineering is an innovative method to restore cardiovascular tissue function by implanting either an in vitro cultured tissue or a degradable, mechanically functional scaffold that gradually transforms into a living neo-tissue by recruiting tissue forming cells at the site of implantation. Circulating endothelial colony forming cells (ECFCs) are capable of differentiating into endothelial cells as well as a mesenchymal ECM-producing phenotype, undergoing Endothelial-to-Mesenchymal-transition (EndoMT). We investigated the potential of ECFCs to produce and organize ECM under the influence of static and cyclic mechanical strain, as well as stimulation with transforming growth factor b1 (TGFb1). Methods and Results: A fibrin-based 3D tissue model was used to simulate neo-tissue formation. Extracellular matrix organization was monitored using confocal laser-scanning microscopy. ECFCs produced collagen and also elastin, but did not form an organized matrix, except when cultured with TGFb1 under static strain. Here, collagen was aligned more parallel to the strain direction, similar to Human Vena Saphena Cell-seeded controls. Priming ECFC with TGFb1 before exposing them to strain led to more homogenous matrix production. Conclusions: Biochemical and mechanical cues can induce extracellular matrix formation by ECFCs in tissue models that mimic early tissue formation. Our findings suggest that priming with bioactives may be required to optimize neo-tissue development with ECFCs and has important consequences for the timing of stimuli applied to scaffold designs for both in vitro and in situ cardiovascular tissue engineering. The results obtained with ECFCs differ from those obtained with other cell sources, such as vena saphena-derived myofibroblasts, underlining the need for experimental models like ours to test novel cell sources for cardiovascular tissue engineering.
Circulation, 2003
Background-Materials commonly used to repair complex cardiac defects lack growth potential and have other unwanted side effects. We designed and tested a bone marrow cell (BMC)-seeded biodegradable scaffold that avoids these problems. Methods and Results-To demonstrate the contribution of the BMCs to histogenesis, we labeled them with green fluorescence, seeded them onto scaffolds, and implanted them in the inferior vena cava of dogs. The implanted grafts were analyzed immunohistochemically at 3 hours and subsequently at 2, 4, and 8 weeks after implantation using antibodies against endothelial cell lineage markers, endothelium, and smooth muscle cells. There was no stenosis or obstruction caused by the tissue-engineered vascular autografts (TEVAs) implanted into the dogs. Immunohistochemically, the seeded BMCs expressing endothelial cell lineage markers, such as CD34, CD31, Flk-1, and Tie-2, adhered to the scaffold. This was followed by proliferation and differentiation, resulting in expression of endothelial cells markers, such as CD146, factor VIII, and CD31, and smooth muscle cell markers, such as ␣-smooth muscle cell actin, SMemb, SM1, and SM2. Vascular endothelial growth factor and angiopoietin-1 were also produced by cells in TEVAs. Conclusions-These results provide direct evidence that the use of BMCs enables the establishment of TEVAs. These TEVAs are useful for cardiovascular surgery in humans and especially in children, who require biocompatible materials with growth potential, which might reduce the instance of complications caused by incompatible materials and lead to a reduced likelihood of further surgery.
Journal of Biomedical Materials Research Part A, 2009
Small-diameter blood vessel substitutes are urgently needed for patients requiring replacements of their coronary and below-the-knee vessels and for better arteriovenous dialysis shunts. Circulatory diseases, especially those arising from atherosclerosis, are the predominant cause of mortality and morbidity in the developed world. Current therapies include the use of autologous vessels or synthetic materials as vessel replacements. The limited availability of healthy vessels for use as bypass grafts and the failure of purely synthetic materials in small-diameter sites necessitate the development of a biological substitute. Tissue engineering is such an approach and has achieved promising results, but reconstruction of a functional vascular tunica media, with circumferentially oriented contractile smooth muscle cells (SMCs) and extracellular matrix, appropriate mechanical properties, and vasoactivity has yet to be demonstrated. This review focuses on strategies to effect the switch of SMC phenotype from synthetic to contractile, which is regarded as crucial for the engineering of a functional vascular media. The synthetic SMC phenotype is desired initially for cell proliferation and tissue remodeling, but the contractile phenotype is then necessary for sufficient vasoactivity and inhibition of neointima formation. The factors governing the switch to a more contractile phenotype with in vitro culture are reviewed. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009
Advanced Functional Materials, 2020
Formation of capillary blood vasculature is a critical requirement for native as well as engineered organs and can be induced in vitro by coculturing endothelial cells with fibroblasts. However, whether these fibroblasts are required only in the initial morphogenesis of endothelial cells or needed throughout is unknown, and the ability to remove these stromal cells after assembly can be useful for clinical translation. In this study, a technique termed CAMEO (Controlled Apoptosis in Multicellular Tissues for Engineered Organogenesis) is introduced, whereby fibroblasts are selectively ablated on demand, and it is utilized to probe the dispensability of fibroblasts in vascular morphogenesis. The presence of fibroblasts is shown to be necessary only during the first few days of endothelial cell morphogenesis, after which they can be ablated without significantly affecting the structural and functional features of the developed vasculature. Furthermore, the use of CAMEO to vascularize a...