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

Candida albicans Biyofilm ile İlgili PJI için Bir Fare Modeli Oluşturma

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.

Yazar Spotlight: Candida albicans Biyofilm İlişkili Protez Eklem Enfeksiyonları Üzerine İleri Araştırmalar 

Farede Periprostetik Eklem Candida albicans Enfeksiyon Modeli

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  

Candida albicans Biyofilm İlişkili Periprostetik Eklem Enfeksiyonu (PJI) Fare Modelinin Oluşturulması

Başlamak için, bir YPD plakasından monoklonal bir candida albicans kolonisi seçmek için bir aşılama döngüsü alın. Koloniyi, mililitre başına 50 mikrogram karbenisilin ile takviye edilmiş beş mililitre YPD sıvı ortamına aktarın. Hücreleri gece boyunca 30 santigrat derecede 220 RPM sabit hızda sallayın.Daha sonra hücre süspansiyonunu oda sıcaklığında beş dakika boyunca 400 g'da santrifüjleyin. Süpernatanı attıktan sonra, hücreleri normal salin çözeltisinde yeniden süspanse edin ve hücre süspansiyonunun bulanıklığını 0.5 McFarland standardıyla eşleştirerek hücre konsantrasyonunu ayarlayın. % 0.9 normal tuzlu su çözeltisi oluşturmak için, 0.9 gram sodyum klorürü tartın ve 100 mililitre deiyonize suda çözün.Daha sonra, cerrahi aletleri kullanmadan önce 30 dakika boyunca titanyum nikel alaşımlı tel içinde 121 santigrat derecede otoklavlayın. Daha sonra rastgele 30 erkek C57BL / 6 fareyi kontrol boş implantına ve PJI grubuna bölün. Fareleri uyuşturduktan sonra, sol arka bacaktaki tüyleri alın ve traş edilen bölgeyi dezenfekte etmek için iyot kullanın.Doğrultma refleksinin kaybını gözlemleyerek anestezi derinliğini doğrulayın. Kontrol grubundaki fareler için herhangi bir tedavi uygulamayın. Boş implant ve PJI grubundaki fareler için, daha sonraki prosedürler için eklemleri ortaya çıkarmak için sol arka ekstremitenin dizinde beş milimetre uzunlamasına bir kesi oluşturmak için 10 numaralı bir bıçak kullanın.Şimdi, femoral intramedüller kanalda beş milimetre uzunluğunda bir delik oluşturmak için steril bir şırınga iğnesi yerleştirin. Makasla kesmeden önce deliğe nikel titanyum alaşımlı bir tel dikkatlice yerleştirin. Ardından, boş implant grubundaki fareler için nikel titanyum alaşımlı tel boyunca damla damla iki mikrolitre YPD ortamı uygulayın.Naylon bir sütür kullanarak, yarayı katman katman kapatın. Daha sonra, iki mikrolitre candida albicans hücresini, PJI grubundaki nikel titanyum alaşımlı tel boyunca farelerin eklem boşluğuna aşılayın. Naylon bir sütür kullanarak, yarayı katman katman kapatın.Fareleri 14 gün boyunca suya ve yiyeceğe ücretsiz erişimi olan bir yerde barındırın.

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 Görüntüleme.  Xi’an Jiaotong University. Başlamak için, bir YPD plakasından monoklonal bir candida albicans kolonisi seçmek için bir aşılama döngüsü alın. Koloniyi, mililitre başına 50 mikrogram karbenisilin ile takviye edilmiş beş mililitre YPD sıvı ortamına aktarın. Hücreleri gece boyunca 30 santigrat derecede 220 RPM sabit hızda sallayın.Daha sonra hücre süspansiyonunu oda sıcaklığında beş dakika boyunca 400 g'da santrifüjleyin. Süpernatanı attıktan sonra, hücreleri normal salin çözeltis...

Video: 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 ...