Presentation of research projects. Bioibérica-FCB-Leitat Mario Marotta & G. Quintás 8th MuscleTech Network Workshop

1. Combined chondroitin sulfate and
glucosamine treatment improves skeletal
muscle recovery in a rat-injury model
Mario Marotta, Ph.D.

2. Chondroitin sulfate (CS) and Glucosamine (GLU) are constituents of proteoglycans and important
components of extracellular matrix in cartilage, skin, ligaments, tendons and skeletal muscle.
BACKGROUND
Chondroitin sulfate plus Glucosamine provides beneficial effects in Osteoarthritis
Clinical trials confirmed their efficacy for Osteoarthritis treatment.

3. Chondroitin sulfate (CS) and Glucosamine (GLU) are constituents of proteoglycans and important
components of extracellular matrix in cartilage, skin, ligaments, tendons and skeletal muscle.
BACKGROUND
Chondroitin sulfate plus Glucosamine provides beneficial effects in Osteoarthritis
Clinical trials confirmed their efficacy for Osteoarthritis treatment.
WHAT ABOUT OTHER APPLICATIONS? MUSCLE INJURY?
Could be also effective for the treatment of skeletal muscle injuries?

4. OBJECTIVES
Study of the effects of chondroitin sulfate + glucosamine on muscle healing after injury.
Collaborative research project MTN – Bioiberica S.A.
Muscle injury rat model.
Pre-clinical study
New treatments
Chondroitin sulfate
Glucosamine

5. IN VIVO pre-clinical study in rat model
Use of a recently developed skeletal muscle injury rat model which mimics the
muscle strain injuries detected in elite athletes.
STUDY DESIGN: Animal model

6. IN VIVO pre-clinical study in rat model
Contreras-Muñoz P, et al. Int J Sports Med. 2016; 37(3):183-90.
RAT MODEL PROFESSIONAL SOCCER P
A. 24h Post-injury in rat model. Coronal view,
Grade I-II lesion. Edema surrounding the
musculotendinous junction demonstrating a
classical feather-like pattern.
B. Coronal view. Proximal biceps, Grade
C. Coronal view. Distal biceps, Grade II
Characteristic feather-like pattern alon
fibers.
RAT MODEL PROFESSIONAL SOCCER PLAYERS
A. 24h Post-injury in rat model. Coronal view,
Grade I-II lesion. Edema surrounding the
musculotendinous junction demonstrating a
classical feather-like pattern.
B. Coronal view. Proximal biceps, Grade I lesion.
C. Coronal view. Distal biceps, Grade II lesion.
Characteristic feather-like pattern along muscle
fibers.
MRI imaging evaluationSurgically-induced skeletal muscle lesion
STUDY DESIGN: Animal model

7. STUDY DESIGN: treatments
0 3 weeks
Oral or intraperitoneal administration for 3-weeks of chondroitin sulfate + glucosamine
(CS+GLU) after skeletal muscle injury.
IN VIVO pre-clinical study in rat model
Gastrocnemius muscle injury

8. IN VIVO pre-clinical study in rat model
STUDY DESIGN: treatments
Oral gavage administration (140 and 175 mg/kg of CS and GLU, respectively)
Intraperitoneal injection (400 and 500 mg/kg of CS and GLU, respectively)
0 3 weeks
Daily CS+GLU administration after injury
Oral or intraperitoneal administration for 3-weeks of chondroitin sulfate + glucosamine
(CS+GLU) after skeletal muscle injury.

9. IN VIVO pre-clinical study in rat model
STUDY DESIGN: Data analysis
Muscle force measurement and sample collection for subsequent histological and
immunofluorescence analysis.
Muscle force analysis
Sample collection and histology
Data analysis
0 3 weeks

10. RESULTS: CS deposition
H&E
Merged
CS
DAPI
Healthy CS+GLU (i.p.)Untreated
***
* P<0.05, ** P<0.01 vs Untreated.
A B
CS+GLU (oral)
CS+GLU increased more than 30% the intramuscular Chondroitin sulfate levels

11. RESULTS: Muscle force
Improvement of gastrocnemius muscle force
*
*
*p<0.01 vs untreated
Oral and i.p. CS+GLU treatments for 3 weeks significantly increased muscle strength
1.22-fold
1.28-fold

12. RESULTS: Muscle regeneration
CS+GLU treatment promoted the growth of regenerating muscle fibers (cross sectional
area, CSA) and reduced intramuscular fibrosis (collagen-I deposition) vs untreated rats
+36%
-21%
-28%
+77%

13. CONCLUSIONS
Combined chondroitin sulfate and glucosamine treatment by oral or
intraperitoneal administration improves skeletal muscle recovery in a
rat-injury model
1. CS+GLU administration increase intramuscular Chondroitin sulfate deposition.
2. Both oral and i.p. treatments increase muscle force after injury
3. CS+GLU-based therapy stimulate the growth of regenerating myofibers and
prevent intramuscular fibrosis
Suggesting an important role of these products as potential new
therapies for the treatment of muscle injuries in sports medicine.

14. MTN team
Muscle Tech Network- Fundació leitat
Dr. Gil Rodas Font
FCB Medical Services
Dra. Meritxell de la Varga
Leitat Technological Center
Dr. Mario Marotta
Ms. Paola Contreras

15. Bioibérica team
Pharmascience Division, Bioibérica S.A.
Dr. Josep Vergés
Dra. Laia Montell
Ms. Anna M. Torrent
Mr. Roger Sabata

16. Thank you for
your attention!!!!