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Nt validated increased -SMA protein level albeit P-PRP induced more -SMA production than L-PRP. Semi-quantification of the Western blots by ImageJ (e). At least three independent experiments were performed for each analysis. A t test was used to perform PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/14960617 statistical analysis. Significant differences (P 2-[(4S)-4,5-Dihydro-4-isopropyl-2-oxazolyl]pyridine due to differences in the preparation protocols with some resulting in L-PRP containing variable concentrations of leukocytes and some resulting in P-PRP without leukocytes. Therefore, in this study, we investigated whether the presence of leukocytes in PRP affects the proliferation and differentiation of TSCs isolated from young adult rabbits. Our findings demonstrated that both L-PRP and P-PRP preparations induced TSC differentiation into active tenocytes, which were proliferating in culture in a PRP-dose-dependent manner (Figs. 1?). Thus, neither L-PRP nor P-PRP appears to pose safety concerns, in terms of producing non-tendinous tissues in the treated tendons, for their use in clinics to treat tendon journal.pone.0167038 injuries.However, both PRP preparations differed in the PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/9638577 following aspects: L-PRP treatment increased the expression of catabolic genes and proteins (MMP-1, MMP-13, IL-1, IL-6, and TNF-) and the production of PGE2, an inflammatory mediator in tendon cells that, at high concentrations, impairs tendon cell proliferation and induces non-tenocyte differentiation [48]. In contrast, P-PRP specifically induced differentiation of TSCs into active tenocytes, marked by -SMA expression, and stimulated cellular production of collagen types I and III; more importantly, P-PRP affected PGE2 production minimally. These findings indicate that L-PRP and P-PRP exert differential effects on TSCs. Therefore, the type of PRP preparation (L-PRP vs. PPRP) 3-Fluoro-2-(trifluoromethyl)aniline is likely a critical factor in assessing the efficacy of PRP treatment on tendon injuries in clinics because they produce differential effects on tendon cells, as demonstrated by this and other studies. In clinics, PRP is prepared by using commercially available PRP preparation kits. Although most kits yield high platelet concentrations (as expected), the level of leukocytes in the PRP preparations may vary, thus likely contributingZhou et al. Stem Cell Research Therapy (2015)6:Page 9 ofFig. 4 Active tenocytes differentiated from TSCs after L-PRP or P-PRP treatment express collagen types I and III. Immunostaining for collagen types I and III (a-f). Both PRP treatments increased the expression of collagen types I and III (pink/red stain), although cells treated with P-PRP stained more intensely for collagen type I and those treated with L-PRP stained more robustly for collagen type III. Nuclei are stained blue with Hoechst 33342. Western blot analysis (g) on total.