1. MyoBio: An Automated Bioreactor System Technology for Standardized Perfusion-Decellularization of Whole Skeletal Muscle.
- Author
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Ritter, Paul, Cai, Aijia, Reischl, Barbara, Fiedler, Maren, Prols, Gerhard, Fries, Benjamin, Kretzschmar, Elke, Michael, Mena, Hartmann, Kristin, Lesko, Christian, Salti, Haitham, Arkudas, Andreas, Horch, Raymund, Paulsen, Friedrich, Friedrich, Oliver, and Haug, Michael
- Subjects
SECOND harmonic generation ,SOLUTION (Chemistry) ,SKELETAL muscle ,REGENERATIVE medicine ,PLASTIC surgery ,ENVIRONMENTAL sampling - Abstract
Objective: Decellularizing solid organs is a promising top-down process to produce acellular bio-scaffolds for ‘de novo’ regrowth or application as tissue ‘patches’ that compensate, e.g., large volumetric muscle loss in reconstructive surgery. Therefore, generating standardized acellular muscle scaffolds marks a pressing area of need. Although animal muscle decellularization protocols were established, those are mostly manually performed and lack defined bioreactor environments and metrologies to assess decellularization quality in real-time. To close this gap, we engineered an automated bioreactor system to provide chemical decellularization solutions to immersed whole rat gastrocnemius medialis muscle through perfusion of the main feeding arteries. Results: Perfusion control is adjustable according to decellularization quality feedback. This was assessed both from (i) ex situ assessment of sarcomeres/nuclei through multiphoton fluorescence and label-free Second Harmonic Generation microscopy and DNA quantification, along with (ii) in situ within the bioreactor environment assessment of the sample’s passive mechanical elasticity. Conclusion: We find DNA and sarcomere-free constructs after 72 h of 0.1% SDS perfusion-decellularization. Furthermore, passive elasticity can be implemented as additional online decellularization quality measure, noting a threefold elasticity decrease in acellular constructs. Significance: Our MyoBio represents a novel and useful automated bioreactor environment for standardized and controlled generation of acellular whole muscle scaffolds as a valuable source for regenerative medicine. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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