Drug-screening platform based on the contractility of tissue-engineered muscle

Department of Pathology, RISE Research Building, Brown Medical School/Miriam Hospital, Providence, RI 02906, USA.
Muscle & Nerve (Impact Factor: 2.28). 04/2008; 37(4):438-47. DOI: 10.1002/mus.20931
Source: PubMed


A tissue-based approach to in vitro drug screening allows for determination of the cumulative positive and negative effects of a drug at the tissue rather than the cellular or subcellular level. Skeletal muscle myoblasts were tissue-engineered into three-dimensional muscle with parallel myofibers generating directed forces. When grown attached to two flexible micro-posts (mu posts) acting as artificial tendons in a 96-well plate format, the miniature bioartificial muscles (mBAMs) generated tetanic (active) forces upon electrical stimulation measured with a novel image-based motion detection system. mBAM myofiber hypertrophy and active force increased in response to insulin-like growth factor 1. In contrast, mBAM deterioration and weakness was observed with a cholesterol-lowering statin. The results described in this study demonstrate the integration of tissue engineering and biomechanical testing into a single platform for the screening of compounds affecting muscle strength.

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    • "In vitro drug screening Flexible silicone posts Primary mouse myoblasts (0.2 × 10 6 cells/well) Collagen (1 mg/ml) and Matrigel (Ratio of 6:1 v/v) Fibrinogen–thrombin (0.5 mg/ml) and thrombin (1 U/ml) Vandenburgh et al., 2008 "
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    ABSTRACT: Bioengineering skeletal muscle often requires customized equipment and intricate casting techniques. One of the major hurdles when initially trying to establish in vitro tissue engineered muscle constructs is the lack of consistency across published methodology. Although this diversity allows for specialization according to specific research goals, lack of standardization hampers comparative efforts. Differences in cell type, number and density, variability in matrix and scaffold usage as well as inconsistency in the distance between and type of adhesion posts complicates initial establishment of the technique with confidence. We describe an inexpensive, but readily adaptable silicone chamber system for the generation of skeletal muscle constructs that can readily be standardized and used to elucidate myoblast behavior in a three-dimensional space. Muscle generation, regeneration and adaptation can also be investigated in this model, which is more advanced than differentiated myotubes.
    Frontiers in Physiology 11/2013; 4:349. DOI:10.3389/fphys.2013.00349 · 3.53 Impact Factor
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    • "The abundance of proteins within the sarcoplasm of skeletal muscle fibres is such that in mature fibres, the nuclei are distributed peripherally [5], and are numerous in order to transcriptionally regulate the often large fibre area [6]. Skeletal muscle is of high clinical importance due to its role in metabolism and movement, and as such, tissue engineered skeletal muscle has been highlighted as having important applications in transplantation in diseased, wasted or injured muscle [7], and as a model in its own right for physiological and clinical testing [8] [9]. However, in order for engineered muscle to have a significant impact in these areas, it must structurally resemble a native skeletal muscle. "
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    ABSTRACT: Tissue engineered skeletal muscle has great utility in experimental studies of physiology, clinical testing and its potential for transplantation to replace damaged tissue. Despite recent work in rodent tissue or cell lines, there is a paucity of literature concerned with the culture of human muscle derived cells (MDCs) in engineered constructs. Here we aimed to tissue engineer for the first time in the literature human skeletal muscle in self-assembling fibrin hydrogels and determine the effect of MDC seeding density and myogenic proportion on the structure and maturation of the constructs. Constructs seeded with 4 x 105 MDCs assembled to a greater extent than those at 1 x 105 or 2 x 105, and immunostaining revealed a higher fusion index and a higher density of myotubes within the constructs, showing greater structural semblance to in vivo tissue. These constructs primarily expressed perinatal and slow type I myosin heavy chain mRNA after 21 days in culture. In subsequent experiments MACS(R) technology was used to separate myogenic and non-myogenic cells from their heterogeneous parent population and these cells were seeded at varying myogenic (desmin +) proportions in fibrin based constructs. Only in the constructs seeded with 75% desmin + cells was there evidence of striations when immunostained for slow myosin heavy chain compared with constructs seeded with 10 or 50% desmin + cells. Overall, this work reveals the importance of cell number and myogenic proportions in tissue engineering human skeletal muscle with structural resemblance to in vivo tissue.
    Biomaterials 07/2013; 34(23). DOI:10.1016/j.biomaterials.2013.04.002 · 8.56 Impact Factor
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    • "This is currently a key obstacle for model system development, especially for a system that attempts to integrate a tissue mimetic (as opposed to two-dimensional monolayer culture) in a high-content and high-throughput manner. A few groups have utilized polymer-based cantilever systems to culture miniature tissues that simultaneously restrain the remodeling of tissue and report forces exerted [18,49,50,53]. It would be interesting to integrate electrical control with these types of systems to both stimulate and record electrical activity while maintaining appropriate force dynamics. "
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    ABSTRACT: Tissue engineering has developed many paradigms and techniques on how to best integrate cells and extracellular matrix to create in vitro structures that replicate native tissue. The strategy best suited for building these constructs depends mainly on the target cells, tissues, and organ of interest, and how readily their respective niches can be recapitulated in vitro with available technologies. In this review we examine engineered heart tissue and two techniques that can be used to induce tissue morphogenesis in artificial niches in vitro: engineered surface topology and electrical control of the system. For both the differentiation of stem cells into heart cells and further assembly of these cells into engineered tissues, these two techniques are effective in inducing in vivo like structure and function. Biophysical modulation through the control of topography and manipulation of the electrical microenvironment has been shown to have effects on cell growth and differentiation, expression of mature cardiac-related proteins and genes, cell alignment via cytoskeletal organization, and electrical and contractile properties. Lastly, we discuss the evolution and potential of these techniques, and bridges to regenerative therapies.
    Stem Cell Research & Therapy 02/2013; 4(1):14. DOI:10.1186/scrt162 · 3.37 Impact Factor
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