RESUMEN
BACKGROUND: Fibrin glue has proven to be a good delivery system for cell transplantation but the factors that influence the fibrin-cell relationships are not well understood. The purpose of this study was to assess the effect of different concentrations of fibrin glue components (thrombin and fibrinogen) on the function of pancreatic islets. METHOD: Islets were isolated from rat pancreata and combined with 6 different fibrin glue formulations. Each islet sample was incubated sequentially with RPMI containing low and high glucose, and culture supernatants were harvested for insulin determination using enzyme-linked immunosorbent assay (ELISA). RESULTS: The control group (no fibrin glue) and group 3 (with thrombin 50 U/mL and fibrinogen 10 mg/mL) had the highest insulin secretion in response to glucose stimulation. These were followed by groups 5 and 4 with 2.6 and 1.8 stimulation indexes, respectively. Group 2 (with thrombin 50 U/mL and fibrinogen 5 mg/mL) and group 6 (commercial kit with thrombin 250 U/mL and fibrinogen 75-115 mg/mL) had the lowest insulin response after glucose stimulation. CONCLUSION: This study demonstrates that different fibrin glue formulations significantly impact pancreatic islets function. In the future, when using fibrin glue as a carrier for pancreatic islet transplantation, lower concentrations of fibrinogen and thrombin are recommended to obtain more viable and functional grafts.
Asunto(s)
Adhesivo de Tejido de Fibrina/farmacología , Fibrinógeno/farmacología , Insulina/metabolismo , Islotes Pancreáticos/metabolismo , Trombina/farmacología , Animales , Ensayo de Inmunoadsorción Enzimática , Glucosa/farmacología , Secreción de Insulina , Islotes Pancreáticos/efectos de los fármacos , Cinética , RatasRESUMEN
Whether T cells circulating peripherally express changes at a clonal level after renal transplantation is uncertain. To clarify this issue, we analyzed T-cell clonality of peripheral blood lymphocytes (PBLs) in 12 renal transplant recipients by a novel polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method that can discriminate T-cell clones with different T-cell receptor (TCR) Vbeta motifs. The PCR-SSCP study showed that after transplantation, only a few distinct T-cell clonotypes accumulated in the absence of clinical episodes, irrespective of the compatibility of HLA antigens. In contrast, various T-cell clones appeared in cases of acute rejection (AR) and infection. These subsided immediately after the AR was resolved; however, they remained long after the resolution of the infection. In a case of AR followed by an infectious episode, distinct T-cell clones appeared concomitantly with each episode. Several of them disappeared or remained thereafter. In one case, significant numbers of accumulating bands were observed by in-vitro stimulation by mixed lymphocyte reaction (MLR); several were identical to those found in vivo. However, some of those that did not appear in vitro were apparent in vivo. In conclusion, the appearance of T-cell clonotypes at a peripheral level indicates the existence of immunologically activated T-cell clones, which were significantly affected by immunosuppressive therapy. It was also determined that the T-cell immune system is much more complicated in vivo than in vitro.