الفهرس 
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Abstract
This study aimed to study aimed to explore the role of stem cells in wound healing in animals by its antibacterial effects.
This study was depends upon the collection of MSC that collected by aspiration of bone marrow from the dogs which exposed to complete sterilization against any external parasites or any external diseases.
Antimicrobial activity of MSC CM and MSCs was tested by subculture from each dilution at different time intervals 1 ,6 , 24 & 48 hours and incubated for 16 - 24 hours at 37°C. Then CFUs were counted.
For Staphylococcus aureus and Escherichia coli, isolation and identification:
1-A total of 5 samples of different donkeys wounds were collected using sterilized cotton swabs and were incubated in nutrient broth and transferred to the laboratory to be incubated at 37 oC for 2 – 3 hours.
2-Cultivation of the samples from nutrient broth:-
-Broth tube of each wound sample was plated in parallel onto Manitol salt agar for isolation of Staphylococcus aureus and MacConkey agar for isolation of Escherichia coli.
-The two plates were incubated aerobically at 37 oC for 24 h – 48 h.
-Staphylococcus aureus proved to be coagulase possitive , while, E. coli isolates from Mackonkey agar were grown on MBE medium, E. coli were serological investigated also.
Bacterial Culture for antimicrobial Assays: Escherichia coli (E. coli) , and Staphylococcus aureus (SA) were used for these experiments.
The results indicated that:
1- Isolated MSCs had a fibroblastic-like appearance with adherent property to the culture plate. Differentiation function was proved with the formation of lipid drops and calcium oxalates on the differentiated MSCs and finally, purified MSCs from bone marrow were positive for cell surface markers, CD73, CD90, and CD105 while being negative for CD34 and CD45. The purified MSCs from bone marrow were positive for cell markers , CD44, CD90, CD105 which negative for CD34 and CD45.
2-Flow cytometry of the cells revealed positivity for CD44, CD90, and CD105 & lacked expression of CD45, and CD34.s
3-Identification of MSC-MVs by flow cytometry, showed the presence of several molecules such as CD44, CD29, CD105, but not CD34, CD45 (a, b, c, and d, respectively).
4-Canine bone marrow MSCs Inhibition of E. coli Bacterial Growth
-BM-derived MSCs significantly inhibited bacterial growth compared with control medium (RPMI) and that inhibition Is directly proportional to time of incubation ( inhibition was more at 48h culture )
a-E.Coli with MSCs
Incubation of E. coli alone resulted in an average of 2.86 × 107 CFU, E. coli with MSCs produced 2.87 × 106 CFU after 24h , and E. coli with MSCs for 48 h resulted in 1.14 × 103 CFU .
No inhibition was noticed at 1h & 6h incubation with MSCS
-BM-derived MSCs microvesicles significantly inhibited bacterial growth compared with control medium (RPMI) and that inhibition Is directly proportional to time of incubation ( inhibition was more at 48h culture ). No inhibition was noticed at 1h incubation with microvisicles
b-E.coli with Microvesicles :
Incubation of E. coli alone resulted in an average of 2.86 × 107 CFU, E. coli with MSCs microvesicles produced 2.40 × 106 CFU after 6th , E. coli with MSCs microvesicles for 24 h resulted in 1.90 × 104 CFU and , E. coli with MSCs microvesicles for 48 h resulted in 1.10 × 103 CFU. There were no inhibition was noticed at 1st h incubation with MSCs microvesicles
c- Staphylococcus aureus with MSCs:
Incubation of Staphylococcus aureus alone resulted in an average of 3.35 × 107 CFU, Staphylococcus aurous with MSCs produced 2.45 × 106 CFU after 24h , and Staphylococcus aurous with MSCs for 48 h resulted in 1.45× 102 CFU. No inhibition of Staphylococcus aurous was noticed at 1h & 6h incubation with MSCS.
d-Staphylococcus aurous with Microvesicles:
The incubation of Staphylococcus aureas alone resulted in an average of 3.35 × 107 CFU, Staphylococcus aureus with MSCs microvesicles produced 2.55 × 106 CFU after 6h , Staphylococcus aureus with MSCs microvesicles for 24 h resulted in 1.50 × 104 CFU and , Staphylococcus aureus with MSCs microvesicles for 48 h resulted in 1.10 × 102 CFU. This results indicated that, by increasing the time of exposure of Staphylococcus aureus to MSCs causes reduction of staphylococcus level .