Study In Antimicrobial Activities Of The Supernatant Of HAMSCs To Propionibacterium Acnes In Vitro

Objective: To collect The Conditioned medium of Human amonion mesenchymal stem cells(h AMSCs-CS)which were isolated,cultured and identified,and investigate whether h AMSCs-CS could inhibit the growth of P.acnes.Methods: The h AMSCs were identified by flow cytometry and vimentin immunotaxation after isolating and extracting from human amniotic membrane by mechanical separation combined with trypsin-collagenase digestion,and h AMSCs-CS of all generations were collected at the same time.The smearing method was used to directly observe whether the h AMSCs-CS which were thawed at room temperature had the effect of inhibiting the growth of P.acnes.And then,the punching method was used to measure the size of the inhibition zone,and determine the drug sensitivity results according to the drug susceptibility test criteria.The results were judged using SPSS19.0 software for statistical analysis.Consults:The h AMSCs showed typical adherent growth.The cells were fibrous and long spindle type,with high expression of CD90,CD105,CD73,almost no expression of CD34,CD11 b,CD19,CD45 and HLA-DR,and positive expression of vimentin.Bacterial growth occurred in the primary-second-generation h AMSCs-CS groups by smearing method,but no growth was observed in the 3rd to 5th h AMSCs-CS groups.There was bacteriostatic rings in the 3rd to 5th h AMSCs-CS and normal saline groups by the punching method,but no bacteriostatic rings in the primary-second-generation h AMSCs-CS and clomatone groups.The mean/median of bacteriostatic rings from 3rd to 5th h AMSCs-CS was 29.50 mm,30.65 mm,and 30.00 mm.There was no statistically significant difference between the groups(P>0.05).The mean of bacteriostatic rings of the clomazone group was54.58 mm which had a statistically significant difference Compared with the 3rd to 5th generation h AMSCs(P<0.001).Conclusions:The h AMSCs can be successfully isolated in vitro by mechanical separation combined with trypsin-collagenase digestion,which is a stem cell growth mode,has stem cell characteristics and low immunogenicity,and expresses vimentin derived from the mesoderm.The primary to second generation h AMSCs-CS can not inhibit the the growth of P.acnes,the 3rd to 5th h AMSCs-CS had the effect of inhibiting the growth of P.acnes,and the antibacterial effect was stable,but weaker than the clomazone.