| Summary: | 碩士 === 國立臺灣大學 === 獸醫學研究所 === 104 === Kidney disease affects millions of people in the world because of increasing incidence and prevalence and the high costs of treatment. Renal injury is mainly caused by toxic and ischemic injury. Renal ischemia-reperfusion occurs in different clinical situations including kidney transplantation, cardiopulmonary bypass, aortic bypass surgery, accidental trauma, sepsis, and hydronephrosis. Post ischemia-reperfusion kidney is characterized by the accumulation of extracellular matrix and usually results in a loss of function when normal parenchyma is replaced by fibrotic tissue. Abnormal kidney function will accumulate waste, toxins and useless moisture in the kidney tissue, may cause renal tubular dilation, glomerular sclerosis, proteinuria, or renal interstitial fibrosis, and finally lead to acute kidney injury (AKI).
Stem cells are a kind of undifferentiated cells which may differentiate into other cell types. And they can also divide indefinitely into stem cell line, but may become cancer cells at about the tenth generation. We collect the bone marrow stem cells (BMSCs) for our research.
Platelet-rich fibrin (PRF) is a natural source of growth factors, which contains a large amount of growth factors that can facilitate bone regeneration, Such as: TGFb-1 (Transforming growth factor), PDGF (platelet-derived growth factor) , VEGF (vascular endothelial growth factor) and Matrix proteins (thrombospondin-1, fibronectin, vitronectin). The growth factors secreted from PRF will cause cell transformation by promoting cell proliferation, matrix formation, bone formation and synthesis of collagen.
We co-cultured rat bone marrow stem cells with platelet-rich fibrin releasate (PRFr), to investigate whether the release would improve stem cell proliferation in vitro or not. The results show that proliferation rate of the bone marrow stem cells which co-cultured with releasate were more rapid, and cell viability was much higher. Therefore, we deduced that platelet-rich fiber release solution have significant assistance on the growth and differentiation of bone marrow stem cells.
To evaluate therapeutic efficacy of cell-based therapy in AKI: a bone marrow stem cells (BMSCs) and platelet-rich fibrin releasate (PRFr) were used for the treatment. An AKI was established with a rat model by ischemia reperfusion (IR). Transplantation of BMSCs, PRFr, and BMSCs + PRFr into IR rats for investigating the therapeutic potential for recovery of renal function. IR or sham operations were performed on SD rats at 10-weeks old, which were randomly divided into two parts: SHAM, and IR.6 rats in SHAM group was subjected to sham surgery, and 24 rats in IR group rats will accept four different treatments (1)Control group, injected 0.8ml PBS; (2)PRFr group, injected PRFr 0.8ml; (3)BMSCs group, injected BMSCs 2106 cells combine 0.8 ml PBS; (4)BMSCs + PRFr group, injected BMSCs 2106 cells combine 0.8 ml PRFr once a week for four weeks. At the firth month, urine was collected in one time a week; at the following three months, urine was collected in one time a month. At four months after surgery, the blood was taken for checking the value of blood urea nitrogen (BUN) and creatinine; the kidney was made into a Paraffin section for checking the effectiveness of treatments.
Estimates based on preliminary data, not only the urinalysis (urine test) but also the serum analysis results indicated that BMSCs combined with PRFr group was significant lower than other groups. By three months after injection, the results of BMSCs combined with PRFr group were also measured in normal values, which performed better than those other groups. It was concluded that the promising experimental data are beginning to emerge in support of the use of MSCs combine PRFr for regenerative applications.
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