Scoping And Clinical Appling Of Debridement Surgery In Suppurative Osteomyelitis

Objective:To establish a model of Staphylococcus aureus biofilm on surfaces of titanium alloy disc and s-equestrum in vitro; To scoping the debridement surgery and guide the treatment of chronic osteomy-elitis via exploring whether the established model can be used in the research of biofilm-forming ab-ility of the same species of bacteria on surfaces of various foreign bodies; To obtain images of Staph-ylococcus aureus biofilm, and observe the structure of it; To make 3-d reconstruction images of Stap-hylococcus aureus biofilm, and observe the structure of it; To Compare biofilm-forming ability of St-aphylococcus aureus on the surface of titanium alloy disc with that of sequestrum.Methods:The culture method used in this experiment was modified from Unnanuntana's setting piece cul-ture method.A few cant breeds staphylococcus aureus were inoculated in trypticase soy broth,and got purebred bacterium fluid after 3 times purification.The bacterium fluid concentration was adjusted to reach 106CFU/ml,and then prepared for experimentation. Before the experimentation,10 titanium al-loy discs and sequestrums which were sterilized were soaked with phosphate buffer for 4h,and then were randomly divided into 10 groups.There were 10 Sterile cell culture flasks,each of which was a-dded with 20ml of tryptone soy broth and 5μl of bacterial suspension and a titanium alloy disc and a sequestrum, were cultivated in incubator for 24h to form a stable biofilm,and then all titanium alloy discs and sequestrums were removed from 10 flasks and analysed in a confocal laser scanning micr-oscope. All titanium alloy discs and sequestrums,which were stained followed kit instructionns, were analysed in a confocal laser scanning immediately to calculate the biofilm thickness and the percent-age of vital bacteria of each layer.From 2009.2 to 2010.2, using debridement surgery and tuo li xiao du san to treat 68 patient who getting suppurative osteomyelitis.Results:1. Bacterial biofilm is a highly organized structure of multi-cell population.Observed in a conf-ocal laser scanning microscope,bacterial biofilm is composed of red, green and yellow fluorescence. The red fluorescence is dead bacteria;the green,vital;the yellow, dead superposes vital. The underlyi-ng vital bacteria is less than the middle. 2. biofilm is homopolymers of mutual gathered conglobation lump.The spatial arrangement of the microorganisms in the biofilm showed voids outlined by layers of vital bacteria, which themselv-es were packed in layers of dead material.3. When Staphylococcus aureus is cultivated for 24h,the bacteria biofilm become stable.The m-ean BBF thickness was (39.20±4.26)μm on surfaces of titanium alloy disc,(44.70±4.22)μm on surfa-ces of sequestrum (P<0.05).The mean vital bacteria percentage was (52.20±5.45)% on surfaces of titanium alloy disc,(57.30±4.92)% on surfaces of sequestrum (P<0.05).4. On the titanium alloy disc,the mean vital bacteria percentage was (81.50±10.46)% in the mi-ddle layer,and (52.20±5.45)% in the underlying(P<0.01).On the sequestrum,the mean vital bacteria percentage was (85.50±6.42)% in the middle layer,and (57.30±4.92)% in the underlying(P<0.01).5.The long-term effect of observation group whose sequestrum is cleared is better than control group whose sequestrum is reserved.Conclusion:1. The model established in this experiment can be used to study biofilm-forming ability of the same species of bacteria on surfaces of various foreign bodies in this experiment.2. Biofilm-forming ability of Staphylococcus aureus on the surface of sequestrum is higher than that of titanium alloy disc. As far as the chronic osteomyelitis concerned,the titanium alloy disc not only should be removed, but also the sequestrum should be paid attention to the clean.3. In the biofilm, the vital bacteria percentage of middle-layer is higher than that of underlying layer.The metabolism of the underlying vital bacteria may be slower than the middle.Most of the vit-al bacteria were packed by the dead. The organic structure of these biofilms may one of the reasons that cause the chronic osteomyelitis difficult to cure.4. The shortcoming of the model established in this experiment is that it can't entirely reflect the complex environment in vivo.5.The long-term effect of debridement surgery whose sequestrum is cleared is better than that whose sequestrum is reserved.