A Novel bioreactor for stimulating skeletal muscle in vitro

Kenneth Donnelly, Alastair Khodabukus, Andrew Philp, Louise Deldicque, Robert G. Dennis, Keith Baar

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5V input to outputs of up to 50V, with pulse widths from 0.05 to 4ms, and frequencies up to 100Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100Hz), this resulted in an increase in the rate of protein synthesis (at 12h, control [CTL]=5.0±0.16; 10Hz=5.0±0.07; and 100Hz=5.5±0.13fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1ms and one or two times rheobase, stimulation improved force production (CTL=0.07±0.009; 1.25V/mm=0.10±0.011; 2.5V/mm=0.14146±0.012; and 5V/mm=0.03756±0.008kN/mm2) and excitability (CTL=0.53±0.022; 1.25V/mm=0.44±0.025; 2.5V/mm=0.41±0.012; and 5V/mm=0.60±0.021V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio.

Original languageEnglish (US)
Pages (from-to)711-718
Number of pages8
JournalTissue Engineering - Part C: Methods
Volume16
Issue number4
DOIs
StatePublished - Aug 1 2010
Externally publishedYes

Fingerprint

Bioreactors
Muscle
Skeletal Muscle
Muscles
Electric Stimulation
Pulse circuits
Production control
Physiology
DC-DC converters
Microcontrollers
Monolayers
In Vitro Techniques
Tissue
Proteins
Engineers

ASJC Scopus subject areas

  • Biomedical Engineering
  • Bioengineering
  • Medicine (miscellaneous)
  • Medicine(all)

Cite this

Donnelly, K., Khodabukus, A., Philp, A., Deldicque, L., Dennis, R. G., & Baar, K. (2010). A Novel bioreactor for stimulating skeletal muscle in vitro. Tissue Engineering - Part C: Methods, 16(4), 711-718. https://doi.org/10.1089/ten.tec.2009.0125

A Novel bioreactor for stimulating skeletal muscle in vitro. / Donnelly, Kenneth; Khodabukus, Alastair; Philp, Andrew; Deldicque, Louise; Dennis, Robert G.; Baar, Keith.

In: Tissue Engineering - Part C: Methods, Vol. 16, No. 4, 01.08.2010, p. 711-718.

Research output: Contribution to journalArticle

Donnelly, K, Khodabukus, A, Philp, A, Deldicque, L, Dennis, RG & Baar, K 2010, 'A Novel bioreactor for stimulating skeletal muscle in vitro', Tissue Engineering - Part C: Methods, vol. 16, no. 4, pp. 711-718. https://doi.org/10.1089/ten.tec.2009.0125
Donnelly K, Khodabukus A, Philp A, Deldicque L, Dennis RG, Baar K. A Novel bioreactor for stimulating skeletal muscle in vitro. Tissue Engineering - Part C: Methods. 2010 Aug 1;16(4):711-718. https://doi.org/10.1089/ten.tec.2009.0125
Donnelly, Kenneth ; Khodabukus, Alastair ; Philp, Andrew ; Deldicque, Louise ; Dennis, Robert G. ; Baar, Keith. / A Novel bioreactor for stimulating skeletal muscle in vitro. In: Tissue Engineering - Part C: Methods. 2010 ; Vol. 16, No. 4. pp. 711-718.
@article{52c8ecd8f9574576815239c558bf2bb3,
title = "A Novel bioreactor for stimulating skeletal muscle in vitro",
abstract = "For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5V input to outputs of up to 50V, with pulse widths from 0.05 to 4ms, and frequencies up to 100Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100Hz), this resulted in an increase in the rate of protein synthesis (at 12h, control [CTL]=5.0±0.16; 10Hz=5.0±0.07; and 100Hz=5.5±0.13fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1ms and one or two times rheobase, stimulation improved force production (CTL=0.07±0.009; 1.25V/mm=0.10±0.011; 2.5V/mm=0.14146±0.012; and 5V/mm=0.03756±0.008kN/mm2) and excitability (CTL=0.53±0.022; 1.25V/mm=0.44±0.025; 2.5V/mm=0.41±0.012; and 5V/mm=0.60±0.021V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio.",
author = "Kenneth Donnelly and Alastair Khodabukus and Andrew Philp and Louise Deldicque and Dennis, {Robert G.} and Keith Baar",
year = "2010",
month = "8",
day = "1",
doi = "10.1089/ten.tec.2009.0125",
language = "English (US)",
volume = "16",
pages = "711--718",
journal = "Tissue Engineering - Part C: Methods",
issn = "1937-3384",
publisher = "Mary Ann Liebert Inc.",
number = "4",

}

TY - JOUR

T1 - A Novel bioreactor for stimulating skeletal muscle in vitro

AU - Donnelly, Kenneth

AU - Khodabukus, Alastair

AU - Philp, Andrew

AU - Deldicque, Louise

AU - Dennis, Robert G.

AU - Baar, Keith

PY - 2010/8/1

Y1 - 2010/8/1

N2 - For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5V input to outputs of up to 50V, with pulse widths from 0.05 to 4ms, and frequencies up to 100Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100Hz), this resulted in an increase in the rate of protein synthesis (at 12h, control [CTL]=5.0±0.16; 10Hz=5.0±0.07; and 100Hz=5.5±0.13fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1ms and one or two times rheobase, stimulation improved force production (CTL=0.07±0.009; 1.25V/mm=0.10±0.011; 2.5V/mm=0.14146±0.012; and 5V/mm=0.03756±0.008kN/mm2) and excitability (CTL=0.53±0.022; 1.25V/mm=0.44±0.025; 2.5V/mm=0.41±0.012; and 5V/mm=0.60±0.021V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio.

AB - For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5V input to outputs of up to 50V, with pulse widths from 0.05 to 4ms, and frequencies up to 100Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100Hz), this resulted in an increase in the rate of protein synthesis (at 12h, control [CTL]=5.0±0.16; 10Hz=5.0±0.07; and 100Hz=5.5±0.13fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1ms and one or two times rheobase, stimulation improved force production (CTL=0.07±0.009; 1.25V/mm=0.10±0.011; 2.5V/mm=0.14146±0.012; and 5V/mm=0.03756±0.008kN/mm2) and excitability (CTL=0.53±0.022; 1.25V/mm=0.44±0.025; 2.5V/mm=0.41±0.012; and 5V/mm=0.60±0.021V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio.

UR - http://www.scopus.com/inward/record.url?scp=77955006289&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955006289&partnerID=8YFLogxK

U2 - 10.1089/ten.tec.2009.0125

DO - 10.1089/ten.tec.2009.0125

M3 - Article

C2 - 19807268

AN - SCOPUS:77955006289

VL - 16

SP - 711

EP - 718

JO - Tissue Engineering - Part C: Methods

JF - Tissue Engineering - Part C: Methods

SN - 1937-3384

IS - 4

ER -