Microfluidic processing of synovial fluid for cytological analysis

Abstract

Cytological analysis of synovial fluid is widely used in the clinic to assess joint health and disease. However, in general practice, only the total number of white blood cells (WBCs) are available for cytologic evaluation of the joint. Moreover, sufficient volume of synovial aspirates is critical to run conventional analyses, despite limited volume of aspiration that can normally be obtained from a joint. Therefore, there is a lack of consistent and standardized synovial fluid cytological tests in the clinic. To address these shortcomings, we developed a microfluidic platform (Synovial Chip), for the first time in the literature, to achieve repeatable, cost- and time-efficient, and standardized synovial fluid cytological analysis based on specific cell surface markers. Microfluidic channels functionalized with antibodies against specific cell surface antigens are connected in series to capture WBC subpopulations, including CD4+, CD8+, and CD66b+ cells, simultaneously from miniscule volumes (100 μL) of synovial fluid aspirates. Cell capture specificity was evaluated by fluorescent labeling of isolated cells in microchannels and was around 90% for all three WBC subpopulations. Furthermore, we investigated the effect of synovial fluid viscosity on capture efficiency in the microfluidic channels and utilized hyaluronidase enzyme treatment to reduce viscosity and to improve cell capture efficiency (>60%) from synovial fluid samples. Synovial Chip allows efficient and standardized point-of-care isolation and analysis of WBC subpopulations in miniscule volumes of patient synovial fluid samples in the clinic.