ABSTRACT
In this study, the Autokit Total Ketone Bodies kit (Wako Pure Chemical, Japan), a total ketone measurement assay using an enzymatic method, was evaluated using a Roche Cobas e702 instrument (Roche Diagnostics, Germany). Precision, linearity, carryover, and reference range verification were evaluated with reference to Clinical Laboratory Standards Institute guidelines. Standard materials provided by the manufacturer and patient samples were used for the evaluation. The precision and carryover of the evaluation result satisfied the acceptance criteria. Linearity was also acceptable at more than 0.99. The quantitative Autokit Total Ketone Bodies kit is precise, and can be widely used in clinical laboratories.
Subject(s)
Humans , 3-Hydroxybutyric Acid , Evaluation Studies as Topic , Ketone Bodies , Methods , Reference ValuesABSTRACT
BACKGROUND:The poly(hydroxybutyrate-co-hydroxyoctanoate) osteochondral scaffold which has been constructed in previous experiments has good biocompatibility and biodegradability and generates non-toxic degradation products. OBJECTIVE:To observe the vascularization of rabbit renal microvascular endothelial cels co-cultured with poly(hydroxybutyrate-co-hydroxyoctanoate) osteochondral scaffold. METHODS:The poly(hydroxybutyrate-co-hydroxyoctanoate) osteochondral scaffold having a three-layer structure (layer of bone/bone and cartilage interface layer/layer of cartilage) was prepared by solvent casting/particle leaching method. The renal microvascular endothelial cels at passage 3 were seeded onto the scaffold of bone layer. The proliferation of the renal microvascular endothelial cels growing on the scaffolds was examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method, the growth of cels in the scaffold was observed by hematoxylin-eosin staining under electron microscope after 10 days. RESULTS AND CONCLUSION:The integrated osteochondral scaffold had a clear appearance of three-layer structure, which had closed connections between the three layers. Porous bone layer was visible as wel as uniform and interlinked pores, and the porosity was 78%. The renal microvascular endothelial cels seeded onto the scaffold proliferated wel and presented a three-dimensional growth after 10 days of co-culture, but there were no cels on the interface layer. Cels which adhered and grew between the pores of the bone layer were observed through hematoxylin-eosin staining. Cels showed a luminal-like structure growing on the scaffold with the porous structure, but they did not grow into the interface layer of bone and cartilage.
ABSTRACT
BACKGROUND:Many experiments have demonstrated that tissue engineering scaffolds prepared by polymer materials alone or biomaterials cannot meet the requirement of tissue engineering research. OBJECTIVE:To evaluate biological characteristics and cel affinity of poly(hydroxybutyrate-co-hydroxyoctanoate)/col agen composite scaffold. METHODS:Tissue engineering scaffolds were prepared by combination of poly(hydroxybutyrate-co-hydroxyoctanoate) and col agen at different proportions (2%, 4%, 6%, 8%and 10%) using solvent casting/particulate leaching method. Inner structure and apertures were observed by scanning electron microscope, and the porosity was determined by liquid displacement method. Rabbit chondrocytes were co-cultured with poly(hydroxybutyrate-co-hydroxyoctanoate)/col agen composite scaffold and poly(hydroxybutyrate-co-hydroxyoctanoate) scaffold. Growth curve of cel s was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and cel adhesion on the scaffolds was observed by scanning electron microscope. RESULTS AND CONCLUSION:The pore size and porosity of the composite scaffold were about 200μm and (85±2)%, respectively. The cel affinity dynamical y increased with the increasing of proportion of col agen. Compared with the poly(hydroxybutyrate-co-hydroxyoctanoate) scaffold, the poly(hydroxybutyrate-co-hydroxyoctanoate)/col agen composite scaffolds are better to improve cel adhesion and proliferation, with favorable cel ular affinity.
ABSTRACT
BACKGROUND:Polyhydroxybutyrate-co-volerate (PHBV) is a noticeable tissue engineering material of polyhydroxyalkanoates family. It has the properties of low immune rejection response and good biocompatibility, and its degradation products are non-toxic. OBJECTIVE:To investigate the biocompatibility of PHBV membrane material and human bone marrow mesenchymal stem cel s in vitro. METHODS:Human bone marrow mesenchymal stem cel s at passage 3 were seeded upon PHBV membrane as experimental group and upon conventional culture plates as control group. Then we calculated the adherent cel number of two groups at 1, 2 and 4 hours and got the cel adherent rate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay was used at days 2, 4, 6, 8 to observe the cel proliferation of two groups. Fluorimetric method with the fluorescent dye Hoechst 33258 was used to detect the DNA content of cel s at days 3, 6, 9 and 12 in both groups. After cel s were seeded upon PHBV membrane for 5 days, the cel growth upon the material was examined under a scanning electron microscope. RESULTS AND CONCLUSION:When the cel s were cultured for 1 hour, the adherent rate in the experimental group was lower than that in the control group;but there were no significant differences between two groups at the other two periods. No difference was found in the cel proliferation and the DNA content between the two groups. Human bone marrow mesenchymal stem cel s seeded upon PHBV membrane for 5 days grew wel with spindle morphology and the intercel ular connections were tight and more extracel ular matrices were observed by scanning electron microscopy. Taken together, PHBV membrane material shows a good biocompatibility with human bone marrow mesenchymal stem cel s.
ABSTRACT
BACKGROUND: Diabetes mellitus and alcohol consumption are the most common causes of ketoacidosis in adults. Recently, beta-hydroxybutyric acid (betaHBA) was reported to be a potential serum biomarker in the diagnosis and monitoring of ketoacidosis. We evaluated the performance of T-KB-H and 3-HB kits for the measurement of ketone bodies [acetoacetate (AcAc)+betaHBA] and betaHBA, respectively. METHODS: Quantitative enzymatic assays were performed using the T-KB-H and 3-HB kits (Nittobo Medical Co., Japan) and the Architect ci16200 Integrated System (Abbott Laboratories, USA). Simultaneously, the ketone body levels in these serum samples were determined by gas chromatography-mas spectrometry (GC-MS). We evaluated precision and linearity of these kits and correlation with GC-MS, and established reference intervals in children and adults. RESULTS: The coefficients of variation for the T-KB-H and 3-HB kits were less than 4.0% at analyte levels of 50, 100, and 400 micromol/L. Linearity was observed for AcAc and betaHBA over a 0-1,000 micromol/L range (R2<0.99). Results from the T-KB-H and 3-HB kits were in good agreement with those from the GC-MS analysis, with correlation coefficients of 0.94 for AcAc and 0.96 for betaHBA. Reference intervals determined for the T-KB-H kit were 9.8-270.1 micromol/L and 18.5-531.8 micromol/L in children and adults, respectively. For the 3-HB kit, the reference intervals were 6.4-234.0 micromol/L and 16.0-437.2 micromol/L in children and adults, respectively. CONCLUSIONS: The T-KB-H and 3-HB kits displayed good precision, clinically acceptable linearity, and reliable correlation with an established assay. This indicates that the kits can be used clinically for measuring serum ketone bodies.