establishing a candida albicans biofilm-associated periprosthetic joint infection (pji) mouse model

建立 白色念珠菌 生物膜相关 PJI 的小鼠模型

Candida albicans (C. albicans) commensally reside in many parts of the human body1, which is also the most common opportunistic pathogen that causes life-threatening invasive fungal infections, especially in immunocompromised patients2,3. C. albicans can transform between yeast and mycelium states as a polymorphic fungus. The mycelium state exhibits higher virulence, stronger adhesion, and invasion of cells and tissues4,5. Besides, C. albicans can form biofilms on the surfaces of biomedical materials such as dentures, catheters, and stents1,6,7. The dense three-dimensional structure of biofilms restricts the infiltration of antifungal drugs, expresses drug-resistant genes, and down-regulates the metabolism of fungal cells to resist immune system clearance6,7. Therefore, biofilms-related infections&#160;are quite challenging in clinics8.
Staphylococcus aureus, coagulase-negative staphylococcus, and enterobacter are the main pathogens causing PJI9. Although the incidence of fungal PJI is relatively low (about 1%)10, the treatment cost of fungal PJI is higher11, the treatment cycle is longer11, and the treatment success rate is lower10 than bacterial PJI. In recent years, the incidence of fungal PJI has been increasing year-by-year10. Candida PJI accounts for 77%-84% of fungal PJI10,12, and C. albicans is the most common in Candida (54%). Therefore, fungal PJI needs to be studied.
Currently, PJI is treated via revision surgery by (1) removing the infected implant, (2) debridement, (3) antimicrobial treatment, and (4) reimplantation. After thorough debridement, an antibiotic containing bone cement is placed, and the patient is treated with antibiotics systemically for more than 6 weeks to effectively control the infection before a new implant is placed13. However, this method cannot fully eliminate pathogens in tissues, and recurrent infections treated with long-term antimicrobial therapy are highly likely to develop in drug-resistant strains14,15,16.
Establishing animal models of PJI is important for the research and development of new drugs or therapeutics for PJI. In the development of PJI, large dead spaces are formed around the prosthesis, leading to the formation of hematomas, which further block the blood supply of the surrounding tissues and impair the effect of antibiotics11,15. Due to the difficulty in mimicking the surrounding environment of the prosthesis, traditional animal models cannot accurately simulate the actual situation of PJI17,18.
In this paper, a C. albicans biofilm-associated PJI model in mice was constructed by using a clinically widely used titanium-nickel wire to simulate joint implants19,20. This PJI model exhibits the advantages of easy operation, high successful rate, high repeatability, and high clinical correlation. It is expected to be an important model for studying the prevention and treatment of C. albicans biofilm-related PJI.

作者聚焦：推进白色念珠菌生物膜相关人工关节感染的研究 

小鼠假体周围关节 白色念珠菌 感染模型

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. 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.

Watch this Scientific Journal Video about Establishing a Candida albicans Biofilm-Associated Periprosthetic Joint Infection PJI Mouse Model at JoVE.com

Establishing a Candida albicans Biofilm-Associated Periprosthetic Joint Infection (PJI) Mouse Model  

建立 白色念珠菌 生物膜相关假体周围感染（PJI）小鼠模型

首先，采用接种环从 YPD 平板中挑选白色念珠菌的单克隆菌落。将菌落转移到5毫升YPD液体培养基中，补充每毫升50μg的羧苄青霉素。在 220 摄氏度下以 30 RPM 的恒定速度摇动电池过夜。然后在室温下以400g离心细胞悬浮液5分钟。弃去上清液后，将细胞重悬于生理盐水溶液中，并通过将细胞悬液的浊度与0.5McFarland标准品相匹配来调节细胞浓度。要制备 0.9% 生理盐水溶液，称取 0.9 克氯化钠并将其溶解在 100 毫升去离子水中。接下来，在使用前将手术器械在 121 摄氏度的钛镍合金丝中高压灭菌 30 分钟。然后将30只雄性C57BL/6小鼠随机分为对照空白植入物组和PJI组。麻醉小鼠后，去除左后肢的毛发，并用碘对剃光区域进行消毒。通过观察矫正反射的丧失来验证麻醉深度。对于对照组中的小鼠，不要进行任何治疗。对于空白植入物和PJI组中的小鼠，使用10号刀片在左后肢膝盖上创建一个5毫米的纵向切口，以暴露关节以进行进一步的操作。现在，插入无菌注射器针头，在股骨髓内管上开一个五毫米长的孔。小心地将镍钛合金线插入孔中，然后用剪刀切割。然后沿镍钛合金丝为空白植入组中的小鼠逐滴施用两微升YPD培养基。使用尼龙缝合线，逐层闭合伤口。接下来，将两微升白色念珠菌细胞沿着PJI组的镍钛合金丝接种到小鼠的关节空间中。使用尼龙缝合线，逐层闭合伤口。将小鼠饲养 14 天，免费获得水和食物。

establishing-candida-albicans-biofilm-associated-periprosthetic-joint

Research

JoVE Journal

Immunology and Infection

Establishing a Candida albicans Biofilm-Associated Periprosthetic Joint Infection (PJI) Mouse Model

92 次观看.  Xi’an Jiaotong University. 首先，采用接种环从 YPD 平板中挑选白色念珠菌的单克隆菌落。将菌落转移到5毫升YPD液体培养基中，补充每毫升50μg的羧苄青霉素。在 220 摄氏度下以 30 RPM 的恒定速度摇动电池过夜。然后在室温下以400g离心细胞悬浮液5分钟。弃去上清液后，将细胞重悬于生理盐水溶液中，并通过将细胞悬液的浊度与0.5McFarland标准品相匹配来调节细胞浓度。要制备 0.9% 生理盐水溶液，称?...

视频: Establishing a Candida albicans Biofilm-Associated Periprosthetic Joint Infection (PJI) Mouse Model  

A Periprosthetic Joint Candida albicans Infection Model in Mouse

Periprosthetic joint infection (PJI) is one of the common infections caused by Candida albicans (C. albicans), which increasingly concerns surgeons and scientists. Generally, biofilms that can shield C. albicans from antibiotics and immune clearance are formed at the infection site. Surgery involving the removal of the infected implant, debridement, antimicrobial treatment, and reimplantation is the gold standard for the treatment of PJI. Thus, establishing animal PJI models is of great significance for the research and development of new drugs or therapeutics for PJI. In this study, a smooth nickel-titanium alloy wire, a widely used implant in orthopedic clinics, was inserted into the femoral joint of a C57BL/6 mouse before the C. albicans were inoculated into the articular cavity along the wire. After 14 days, mature and thick biofilms were observed on the surface of implants under a scanning electronic microscope (SEM). A significantly reduced bone trabecula was found in the H&amp;E staining of the infected joint specimens. To sum up, a mouse PJI model with the advantages of easy operation, high successful rate, high repeatability, and high clinical correlation was established. This is expected to be an important model for clinical studies of C. albicans biofilm-related PJI prevention.

Periprosthetic joint infection (PJI) caused by dangerous pathogens is common in clinical orthopedics. Existing animal models cannot accurately simulate the actual situation of PJI. Here, we established a Candida albicans biofilm-associated PJI mouse model to research and develop new therapeutics for PJI.

Periprosthetic joint infection (PJI) is one of the common infections caused by Candida albicans (C. albicans), which increasingly concerns surgeons and ...