In this study of Candida albicans biofilm-associated peri-prosthetic joint, Candida albicans infection model in mice was constructed using titanium-nickel wire, commonly used to simulate joint implants. This PJI model is easy to operate, has a high success rate, and is clinically relevant. It will help understanding the prevention and treatment of Candida albicans biofilm-related PJI.
Traditional animal models cannot accurately simulate the actual situation of PJI, due to the complex environment surrounding the processes. The Candida albicans biofilm-related mouse PJI model can address the research gap. The establishment of PJI animal model is very important for the research and the development of new drugs or therapeutics for PJI.
The Candida albicans biofilm-associated mouse PJI model established in this paper is expected to be an important model for the prevention and treatment of Candida albicans biofilm-associated PJI. To begin, take an inoculating loop to pick a monoclonal colony of Candida albicans from a YPD plate. Transfer the colony to five milliliters of YPD liquid medium supplemented with 50 micrograms per milliliter of carbenicillin.
Shake the cells at a constant speed of 220 RPM at 30 degrees Celsius overnight. Then centrifuge the cell suspension at 400G for five minutes at room temperature. After discarding the supernatant, resuspend the cells in normal saline solution and adjust the cell concentration by matching the turbidity of the cell suspension to a 0.5 McFarland standard.
To create 0.9%normal saline solution, weigh 0.9 grams of sodium chloride and dissolve it in 100 milliliters of deionized water. Next, autoclave the surgical instruments and titanium nickel alloy wire at 121 degrees Celsius for 30 minutes prior to use. Then randomly divide 30 male C57BL/6 mice into control, blank implant, and PGI group.
After anesthetizing the mice, remove hair from the left hind limb and use iodine to disinfect the shaved area. Verify the depth of anesthesia by observing the loss of the righting reflex. For the mice in the control group, do not administer any treatment.
For the mice in the blank implant and PGI group, use a number 10 blade to create a five millimeter longitudinal incision on the knee of the left hind limb to expose the joints for further procedures. Now, insert a sterile syringe needle to create a five millimeter long hole in the femoral intramedullary canal. Carefully insert a nickel titanium alloy wire into the hole before cutting it with scissors.
Then administer two microliters of YPD medium drop by drop along the nickel titanium alloy wire for the mice in the blank implant group. Using a nylon suture, close the wound layer by layer. Next, inoculate two microliters of Candida albicans cells into the joint space of mice along the nickel titanium alloy wire in the PJI group.
Using a nylon suture, close the wound layer by layer. House the mice with free access to water and food for 14 days. To begin, create a Candida albicans biofilm-associated PJI mouse model.
After 14 days of the model establishment, euthanize the mouse and harvest the kidneys, liver, and spleen. For each organ, introduce 500 microliters of sterile normal saline. Grind the tissues on a homogenizer at four degrees Celsius.
Then add 100 microliters of the prepared homogenate onto a YPD plate and use a bent rod to spread it evenly. Place the YPD plates inverted in a 37 degree Celsius incubator for 48 hours. At the end of the incubation, visually examine the plates, counting the visible colonies.
