Sinoviyal Eklem Sıvısında Fosfolipid Zenginleştirme için Monodispers-Mezoporoz SiO2 Metal Oksit Afinite Kromatografisi (MOAC) Sorbentinin Geliştirilmesi

Abstract

Diagnosing diseases, monitoring their course and observing the effectiveness of treatment have been among the leading focuses of medical science. In this respect, traditional methods may be insufficient in today's conditions in terms of early intervention opportunities. Developments in laboratory technologies and chromatographic methods have enabled the examination of clinical pictures at the molecular level at the meantime paved the way for time, labor and cost effective alternatives. Diagnosis of arthropathies, including osteoarthritis (OA), monitoring their course and evaluating treatment effectiveness are usually done through qualitative parameter-based surveys conducted with the patient. Developing an analytical method where quantitative comparisons can be made will benefit clinical processes, especially treatment planning. Synovial joint fluid (SJF) is a friction-reducing lubricant for the joint region, but also a transfer medium for the cartilage extracellular matrix and molecules released during vascular circulation. Structures such as synovium, meniscus, joint ligament and joint capsule express various markers through secretion into the SJF. Anabolites and catabolites formed as a result of regeneration and degeneration events for bone and cartilage homeostasis also diffuse into the SJF. Therefore, an abnormal situation that may occur in the tissues and systems with which SJF interacts is also reflected in the synovial joint fluid at the molecular level. Within the scope of the thesis, it was aimed to develop a microextraction sorbent for the specific and selective detection of phospholipids (PLs) in SJF. For this purpose, SiO2, SiO2@PEI and SiO2@PEI@Ti(IV) were determined as candidate sorbents. As a result of preliminary studies, experimental studies were carried out with monodisperse-mesoporous SiO2 microspheres, since the results obtained with SiO2 were more meaningful. Monodipers-mesoporous SiO2 microspheres were synthesized by multi-step hydrolysis condensation method. Characteristic structures of microspheres were examined by SEM, XPS, XRD, ICP-OES and BET methods. Since synovial joint fluid contains biomarkers associated with articular cartilage, studies were carried out on this biological material. The lipid fraction extracted from SJF by liquid-liquid extraction (LLE) method was examined in terms of phospholipid enrichment performance by interacting with monodisperse-mesoporous SiO2 microspheres. The lipid fraction extracted from SJF by LLE method was examined in terms of PL enrichment performance by interacting with monodisperse-mesoporous SiO2 microspheres. While 141 PLs were characterized in the fraction obtained with the solid phase extraction (SPE) protocol after LLE, the number of PLs detected in the fraction obtained after LLE from the concentrated extract was found to be around 164. These numbers are close to each other shows that the enrichment is successful and selective in terms of PLs. Extraction efficiency was evaluated based on 44 common peaks detected by both methods. In conclusion, PL enrichment was successfully achieved with the proposed SPE protocol. As a result of the PL enrichment process carried out in SJF with the SPE protocol, it was demonstrated that the sorbent was successful in PL enrichment. Monodisperse-mesoporous SiO2 microspheres were found suitable for selective enrichment of PLs in SJF samples. It is concluded that these microspheres can be used to identify PL changes in physiological and pathological conditions, including OA researches.