In vitro analysis of skeletal muscles and Neuro-Muscolar

In vitro analysis of skeletal muscles and Neuro-Muscolar
Junction (NMJ) biomechanical properties
S. Carosio1, E. Rizzuto2, M. Faraldi3, S. Pisu3, Z. Del Prete2,1
1 Center for Life NanoScience – IIT@Sapienza, Istituto Italiano di Tecnologia, Italy
2 Dipartimento di Ingegneria Meccanica ed Aerospaziale – Sapienza Università di Roma, Roma, Italy
3 SAIMLAL – Sapienza Università di Roma, Roma, Italy
Biomechanics Laboratory
In the IIT Biomechanics Laboratory we have designed and realized novel experimental protocols and instrumentation to
perform a morphological and functional analyses on different neuromuscular junction (NMJ) districts both for in vitro and
ex vivo models [1-2]. Tissue surface strain field is measured contactless by a high-resolution and a high speed “Digital
Image Correlation” system realized ad-hoc [3].
In vitro analysis
y of NMJ functionalityy
Measuring
g X-MET biomechanical
properties
High frequency camera
X-MET
Micro force
transducer
NMJ functionality is measured by comparing muscle
contractile response elicited by nerve electrical
stimulation with that of the same muscle evoked by
direct stimulation of the membrane.
Platinum electrodes
3D co-culture : X-MET and embryonal spinal cord
Musarò A., Dobrowolny G., IIT, SAIMLAL, Sapienza Università di Roma
Ventral spinal cord
IF for neurofilament 45 days
Nerve-muscle
interaction
Study of NMJ functionality of ALS animal models
Musarò A
A., Dobrowolny G.,
G IIT,
IIT SAIMLAL,
SAIMLAL Sapienza Università di Roma
20x
40x
Variational methods for movie analisys
Pedone M
M.,Ruocco
Ruocco G.,
G InfoSapienza,
InfoSapienza Sapienza Università di Roma
The instrumentation and experimental protocols here illustrated are appropriate tools to detect early subtle abnormalities
of neuro-motor functionality in transgenic mouse models of neuro-muscular diseases. In particular, our systems allow to
discriminate the NMJ alterations induced directly by the skeletal muscle from those due to the motor neuron activity, both
in ex-vivo muscle and in 3D colture systems.
[1] Rizzuto E., Pisu S., Musarò A., Del Prete Z., Measuring neuromuscular junction functionality in the SOD1G93A animal
model of Amyotrophic Lateral Sclerosis, Ann Biomed Eng. 2015, doi: 10.1007/s10439-015-1259-x.
[2] Carosio S., Barberi L., Rizzuto E., Nicoletti C., Del Prete Z., Musarò A., Generation of eX vivo-vascularized Muscle
Engineered Tissue (X-MET), Sci Rep. 2013; doi: 10.1038/srep01420.
[3] Rizzuto E., Carosio S., Del Prete Z., Characterization of a Digital Image Correlation System for Dynamic Strain
Measurements of Small Biological Tissues, ExpTechniques. 2014, doi: 10.1111/ext.12115.