N= 29 Wistar rats
Weight mean : 421g

Origin of animals : Biotério Central of Campus of Ribeirão Preto of São Paulo University
Maintenance : Experimental Pathology Lab of Pathology Department on Medicine School of Ribeirão Preto – USP
Animals keeping : polypropylen boxes
Animals food : basic diet of lab and water
Light conditions : natural light/dark cycles under controlled basic conditions
Experimentators : A.Tenenbaum,R.Mené,Kim

The mice were euthanized in CO2 chambers.

• Incision of the skin with a blade nº 23
• Finding and isolation of the pretibial muscle using Metzenbaum scissors
• Section of tendons with a Mayo scissors.
• Fixation of Portions of the muscles on sections of cork oak
• Embedding using 50% formaldehyde solution - 10% buffered and 50% of pure paraffin for conventional optical microscopic study.
• Staining Sections as routine histological process.
• Microtome used : Leika RM 2155
• Coloration of muscle slices with

-hematoxilin-eosin
-Masson´s trichrome
-and red picro-sirius.

• Histologic examination was realized with a conventional microscope (Olympus BH-2).
• Observation under polarized lampe
• Photographic documentation was realized with a Leika DMR microscope joined to a digital camera Leika CD 300F and compatible PC.

• Animals were divided in 6 boxes, so that boxes from number 1 to 5 had 5 rats and box number 6 had 4 rats 
• All animals were submitted to a walking evaluation by footprint impression on special paper , and previous immersion of back legs on a soap solution. 
• Box 1 had the control group, that received the application of 0,1 ml saline solution ( NaCl 0,9%) on the right pretibial muscle. 

• The mice of boxes 2 to 5 received 0,1 ml of Endopeels Original Main Product on the same muscle as control group.

• Box 6 was constituted of 4 rats, and each one received 0,5 ml of Endopeels original main product injected on the subcutaneous layer. 

• All the animals were submitted to walking evaluation before and after their respective injections.

1 month after Endopeel Injection 0.1ml in the right pretibial muscle.
What is seen in black on the pictures is not a necrosis like could imagine some scientifics !
In fact, 4 conclusions have to be taken in consideration

• an artefact of coloration
• an absence of necrosis
• an apoptosis
• a bioregenerative process 

3 months (D90)after Endopeel Injection 0.1ml in the right pretibial muscle.

7 months (D210)after Endopeel IM Injection 0.1ml in the right pretibial muscle.

Complete Restitutio ad integrum after 7 months

0.5 ml ( 5x 0.1ml)  Endopeel SC Injection in the right subcutaneous pretibial area.