PURPOSE: Transglutaminase type 2 (TG2) is an extracellular matrix crosslinking enzyme with a pivotal role in kidney fibrosis. We tested whether quantification of urinary TG2 may represent a noninvasive method to estimate the severity of kidney allograft fibrosis. METHODS: We prospectively collected urine specimens from 18 deceased donor kidney transplant recipients at 1-day, 7-day, 1-month, 3-month, and 6-month posttransplant. In addition, kidney allograft tissue specimens at 0-day and 6-month posttransplant were sampled to analyze the correlation of urinary TG2 and kidney allograft fibrosis. RESULTS: Thirteen recipients had increased interstitial fibrosis and tubular atrophy (IFTA) scores at the 6-month protocol biopsy (IFTA group). The mean level of urinary TG2 in the IFTA group was higher compared to that of 5 other recipients without IFTA (no IFTA group). Conversely, the mean level of urinary syndecan-4 in the IFTA group was lower than levels in patients without IFTA. In the IFTA group, double immunofluorescent staining revealed that TG2 intensity was significantly upregulated and colocalizations of TG2/heparin sulfate proteoglycan and nuclear syndecan-4 were prominent, usually around tubular structures. CONCLUSION: Urinary TG2 in early posttransplant periods is a potent biomarker for kidney allograft inflammation or fibrosis.
Allografts ; Atrophy ; Biomarkers ; Biopsy ; Extracellular Matrix ; Fibrosis ; Humans ; Inflammation ; Kidney Transplantation ; Kidney ; Methods ; Prospective Studies ; Proteoglycans ; Syndecan-4 ; Tissue Donors ; Transplant Recipients

PURPOSE: Transplantation of microencapsulated islets is proposed as an ideal therapy for the treatment of type 1 diabetes mellitus without immunosuppression. This is based on the principle that foreign cells are protected from the host immune system by an artificial membrane. The aim of this study is to establish an ideal condition of microencapsulation by using an air-driven droplet generator and alginate in vitro. METHODS: Islets were prepared from Sprague Dawley rat and semi SPF-micro pig. Alginate concentrations were changed from 1.5% to 3.0%, and inflow rate of alginate was varied from 10 mL/hr to 40 mL/hr. CO2 flow rate was regulated from 2.0 L/min to 4.0 L/min. Viability was checked by dithizone and FDA/PI staining. Secretory function was tested with glucose challenge and insulin stimulation index was investigated. RESULTS: The optimal conditions for islet encapsulation were revealed with alginate inflow rate of 10 mL/hr, CO2 flow rate of 2.0 L/min in concentration of 2% alginate. In concentration of 2.5% alginate, alginate inflow rate of 20 mL/hr, CO2 flow rate 3.0 L/min was ideal, and alginate inflow rate of 40 mL/hr, CO2 flow rate of 4.0 L/min showed good conditions of microcapsules in concentration of 3% alginate. Viability of encapsulated islets was higher than 90% in both rat and porcine. In terms of insulin secretion, encapsulated islets secreted insulin in response to glucose in static culture medium. However there was no normal response to low and high glucose challenge with stimulation index of less than 2.0. CONCLUSION: Microencapsulation of islets in rat and pig was successful with air-driven droplet generator and alginate in vitro. Further studies about biocompatibility and glucose control in vivo should be followed to be a useful tool for treatment of diabetes mellitus patients in clinical setting.
Animals ; Capsules ; Diabetes Mellitus ; Diabetes Mellitus, Type 1 ; Dithizone ; Drug Compounding ; Glucose ; Humans ; Immune System ; Immunosuppression ; Insulin ; Islets of Langerhans* ; Membranes, Artificial ; Rats