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Method Article

Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1

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DOI:

10.3791/55642

July 11th, 2017

* These authors contributed equally

In This Article

Summary

In this two-part study, a biological actuator was developed using highly flexible polydimethylsiloxane (PDMS) cantilevers and living muscle cells (cardiomyocytes), and characterized. The biological actuator was incorporated with a base made of modified PDMS materials to build a self-stabilizing, swimming biorobot.

Abstract

Biological machines often referred to as biorobots, are living cell- or tissue-based devices that are powered solely by the contractile activity of living components. Due to their inherent advantages, biorobots are gaining interest as alternatives to traditional fully artificial robots. Various studies have focused on harnessing the power of biological actuators, but only recently studies have quantitatively characterized the performance of biorobots and studied their geometry to enhance functionality and efficiency. Here, we demonstrate the development of a self-stabilizing swimming biorobot that can maintain its pitch, depth, and roll without external intervention. The design and fabrication of the PDMS scaffold for the biological actuator and biorobot followed by the functionalization with fibronectin is described in this first part. In the second part of this two-part article, we detail the incorporation of cardiomyocytes and characterize the biological actuator and biorobot function. Both incorporate a base and tail (cantilever) which produce fin-based propulsion. The tail is constructed with soft lithography techniques using PDMS and laser engraving. After incorporating the tail with the device base, it is functionalized with a cell adhesive protein and seeded confluently with cardiomyocytes. The base of the biological actuator consists of a solid PDMS block with a central glass bead (acts as a weight). The base of the biorobot consists of two composite PDMS materials, Ni-PDMS and microballoon-PDMS (MB-PDMS). The nickel powder (in Ni-PDMS) allows magnetic control of the biorobot during cells seeding and stability during locomotion. Microballoons (in MB-PDMS) decrease the density of MB-PDMS, and enable the biorobot to float and swim steadily. The use of these two materials with different mass densities, enabled precise control over the weight distribution to ensure a positive restoration force at any angle of the biorobot. This technique produces a magnetically controlled self-stabilizing swimming biorobot.

Introduction

Biological actuators and biorobots are being actively studied to provide an alternative to conventional robotics for numerous applications. Biorobots that walk5,6,7,8, swim1,2,3,4, pump9,10, or grip11,12,13 have already been developed. Similarly, muscle ....

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Protocol

1. Calculate Mass of PDMS and Additives

  1. Use the following equation to find the mass of PDMS needed for specific heights in the following procedures,
    M = ϱ*V = ϱ * Height*Area          (1),
    where 'Height' is the height of the layer, 'Area' is the area of a container that the PDMS will be cured in, 'ϱ' is the density of the mixture and 'V' is the volume.
    NOTE: Densities for height calculations are PDMS = 0.965 g/mL, Ni-PDMS = 1.639 g/mL, MB-PDMS = 0.648 g/mL.
  2. Use equation (1) to estimate the mass of PDMS needed, for a given container, to obtain a specific height (5 mm) for the base of the biological actuator. The resulting density of....

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Results

The biological actuator and biorobot have very similar fabrication processes, as the biorobot is a natural extension of the biological actuator (Figure 1). The biological actuator was developed first to establish techniques required for the biorobot, to analyze the force generated by the cells, and to characterize the cell maturation mechanically and biochemically, both of which are described in detail in Part 2 of this two-part article as well as in our recently publishe.......

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Discussion

Various locomotion mechanisms can be found among aquatic swimmers16. The locomotion mechanism of the biorobot in this study uses fin-based locomotion, specifically ostraciiform locomotion. Ostraciiform swimmers propel themselves by wagging a tail (cantilever) and having a rigid body (layered base)16. Fish such as the boxfish and cowfish use this type of locomotion. Ostraciiform swimmers are typically slow and have inefficient body dimensions. Although ostraciiform swimming .......

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Disclosures

The authors have nothing to disclose

Acknowledgements

M. T. Holley is supported by the Graduate Fellows program of the Louisiana Board of Regents and C. Danielson is supported by Howard Hughes Medical Institute Professors Program. This study is supported by NSF Grant No: 1530884. The authors would like to thank the support of the cleanroom at the Center for Advanced Microstructures and Devices (CAMD).

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
Polydimethylsiloxane (PDMS)Dow Corning184 sil elast kit  0.5kgSylgard 184
Nickel PowderSigma-Aldrich266981-100G
Phenolic microballoonsUS CompositesBJO-0930
Silicon wafers4 inch diameter
PWM101 light-duty spinnerSpin- coater
Positive photoresist (S1808)Dow CorningDEM-10018197
Hotplate
Vacuum chamber
M206 mechanical convection ovenConvection oven
Laser engraverUniversal Laser SystemVLS2.30Utilizes a 10 W, 10.6 µm wavelength, CO2 Laser
Universal Laser Systems ApplicationUniversal Laser SystemApplication for running the VLS 2.30
MatlabMathWorksNumerical analysis program
Scotch TapeScotch Brand
Solid-glass beadsSigma-AldrichZ265926-1EASoda-lime glass, diameter 3 mm
ScaleMettler ToledoEL303
BD-20AC Laboratory Corona TreaterElectrotechnic Products12051ACorona Discharger
Ultrasonic Bath 1.9 LFisher Scientific15-337-40240 kHz industrial transducer
Fibronectin from bovine plasmaSigma-AldrichF1141
Dulbecco’s Phosphate Buffer (PBS)Sigma-AldrichD1408-100ML
Dulbecco’s Modified Eagle Medium (DMEM)Hyclone Laboratories16750-074With 4500 mg/L glucose, 4.0 mM L-glutamine, and 110 mg/L sodium pyruvate.
Fetalclone III serumHyclone Industries, GE16777-240Fetal bovine serum
Penicillin-G sodium saltSigma-AldrichP3032

References

  1. Williams, B., Anand, S., Rajagopalan, J., Saif, M. A self-propelled biohybrid swimmer at low Reynolds number. Nat commun. 5, (2014).
  2. Nawroth, J., et al. A tissue-engineered jellyfish with biomimetic propulsion. Nat Biotechnol. 30 (8), 729-797 (2012).
  3. Huge ....

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Tags

Cardiac Muscle CellsBiological ActuatorPDMS ScaffoldSoft LithographyLaser EngravingMagnetic ControlFibronectin FunctionalizationCantilever FabricationBuoyancy Control