Body Fluid Test Research Articles

Polydopamine-assisted chlorhexidine immobilization on medical grade stainless steel 316L: Apatite formation and in vitro osteoblastic evaluation

Immobilization of chlorhexidine (CHX) on stainless steel 316L (SS316L), assisted by a polydopamine film as an intermediate layer is projected as an approach in combating infection while aiding bone regeneration for coating development on orthopedic and dental implants. This study aimed to investigate the ability of CHX coating to promote apatite layer, osteoblast cells viability, adhesion, osteogenic differentiation and mineralization. Stainless steel 316L disks were pre-treated, grafted with a polydopamine film and immobilized with different concentrations of CHX (10–30mM). The apatite layer formation was determined through an in vitro simulated body fluid (SBF) test by ATR-FTIR and SEM-EDX analyses. The osteoblastic evaluations including cells viability, cells adhesion, osteogenic differentiation and mineralization were assessed with human fetal osteoblast cells through MTT assay, morphology evaluation under FESEM, ALP enzyme activity and Alizarin Red S assay. The apatite layer was successfully formed on the CHX coated disks, demonstrating potential excellent bioactivity property. The CHX coatings were biocompatible with the osteoblast cells at low CHX concentration (<20mM) with good adhesion on the metal surfaces. The increment of ALP activity and calcium deposition testified that the CHX coated disks able to support osteoblastic maturation and mineralization. These capabilities give a promising value to the CHX coating to be implied in bone regeneration area.

Study of In Vitro Bioactivity of Nano Hydroxyapatite Composites Doped by Various Cations

Nano hydroxyapatite (nHA) samples were synthesized and doped with chitosan (CS), magnesium (Mg), manganese (Mn), strontium (Sr) and cerium (Ce), using the sol–gel method. The addition of cations dopant to synthetic HA is considered in relation to improving bioactivity. According to the XRD results, it was indicated that the type of dopant has a perceptible effect on the lattice parameters and cell volume. In vitro bioactivity of the samples was studied employing a simulated body fluid (SBF) test. Hence, FESEM images showed the samples with grained particles, different shapes and dimensions after SBF test. On the basis of EDX results, there was no detectable impurity in all the synthesized samples. The results of the cytotoxicity assay revealed an acceptable limit for the cell growth and proliferation.

An approach to analytical validation and testing of body fluid assays for the automated clinical laboratory

BackgroundBiochemical analysis of body fluids may lend insight into the pathogenesis of disease. Most commercially-available clinical laboratory methods are intended for blood-derived specimens and/or urine and laboratories must characterize the analytical performance of these methods for other specimen types such as body fluids. The aim of this work is to demonstrate one approach for characterizing analytical performance of assays for clinical analysis of body fluids. MethodsResidual waste samples were obtained from clinically ordered testing. Validation studies were performed for 8 chemistry analytes on Roche Cobas 6000 c501 (Roche Diagnostics, Inc.) analyzers. Accuracy, precision, analytical measurable range, reportable range, analytic sensitivity, interferences, analyte stability were assessed. Laboratory workflow for body fluid handling was designed based on results obtained. ResultsSample matrix interferences were not observed for the body fluids tested and assay reportable ranges used for serum were validated for body fluids. Dilution of body fluid specimens with saline demonstrated non-linear recovery of some enzyme activities. Assay imprecision was comparable to the manufacturer's claims for serum. The serum index thresholds for interference from hemoglobin and lipemia were lower for body fluids compared to manufacturer's stated limits for serum. Pretreatment of body fluid samples with hyaluronidase caused a 28% false increase in lipase activity, and 13% increase in total protein concentration. Ambient temperature analyte stability in body fluids was ≤24 h for most analytes compared to manufacturer's stated stability of 7 days for serum. In contrast to serum, lactate dehydrogenase (LDH) was labile in frozen body fluid specimens. ConclusionsValidation of analytical methods for body fluid testing is a necessary exercise to exclude potential matrix effects and set pre-analytic specimen quality criteria.

Alpha-fetoprotein in pericardial, peritoneal, and pleural fluids: A body fluid matrix evaluation

ObjectivesAlpha-fetoprotein (AFP) measurement in pericardial, peritoneal (ascites), and pleural fluids is sometimes requested by clinicians as supportive evidence in the evaluation of suspected malignancy. As commercially available, Food and Drug Administration (FDA)-cleared AFP assays are not validated for these fluid types, laboratories must complete additional validation studies to comply with regulatory requirements for body fluid testing. The objective of this study was therefore to conduct a matrix evaluation for these body fluid types using the Beckman Access AFP assay on the UniCel DxI 800 immunoassay system. Design and methodsUsing an Institutional Review Board (IRB) approved protocol, previously collected pericardial fluid, peritoneal fluid, pleural fluid, and serum specimens were de-identified and frozen at −20 °C prior to matrix evaluation experiments. Spiked recovery, mixed recovery/linearity, and precision studies were conducted. ResultsIn spiked and mixed recovery studies, the average percent (%) recovery was within predefined acceptable limits (±15%) for all three body fluids. Linearity was observed over the analytical measurement range (AMR) for all three body fluids (slope, intercept, systematic error): pericardial 0.988, −0.1, 6.1%; peritoneal 0.986, 0.0, 4.1%; and pleural 1.016, 0.0, 1.6%. Imprecision was ≤6.0% CV for all three body fluids at both high and low AFP concentrations. ConclusionsMatrix interference with AFP testing was not observed for pericardial, peritoneal, or pleural fluids on the Beckman UniCel DxI 800 system.

Solution Precursor Plasma Spraying (SPPS): A novel and simple process to obtain bioactive glass coatings

This research addresses the use of an emerging plasma spray technique known as Solution Precursor Plasma Spraying (SPPS) to obtain bioactive glass coatings. In SPPS, the feedstock is a sol precursor of the bioactive glass instead of glass particles. Precursor feedstock composition (presence or not of acid catalyst), plasma torch enthalpy and stand-off distance of the spraying process have been varied to produce bioactive glass coatings of nominally 45S5 composition. A good quality coating was obtained when catalyst (0.2 M acid nitric) was used in the precursor and the plasma spraying took place with shorter stand-off distance and higher plasma enthalpy. Bioactivity in Simulated Body Fluid test was confirmed and an effect of the coating interconnected porosity to enhance bioactivity was observed.

Synthesis, characterization and solubility analysis of amorphous SiO2-CaO-Na2O-P2O5 scaffolds for hard tissue repair

The substitution of TiO2 with network modifiers (sodium, Na+ and calcium, Ca2+) within a bioactive glass series was investigated to synthesize amorphous scaffolds from the SiO2-CaO-Na2O-P2O5 glass system. Three glasses were formulated for this study, a control (BG) and two experimental formulations with 9 wt% TiO2 substituted for CaO (SC-1) and Na2O (SC-2) respectively. X-ray diffraction presented an amorphous pattern for BG and SC-1, with extensive crystallinity present in SC-2. Differential thermal analysis presented a higher Tg for SC-1 (559 °C) and SC-2 (627 °C) compared to BG (531 °C), however a greater distribution of Q-species was evident in SC-1 and SC-2, particularly in the Q1–Q2 regions as evidenced by MAS-NMR. Simulated Body Fluid (SBF) testing presented calcium phosphate (CaP) deposition on BG however no visible CaP layer was observed on the surface of SC-1 and SC-2. Ion release profiles exhibited a much slower Si release rate with respect to time for SC-1 and SC-2, which may be the limiting factor when tested in SBF. The addition of TiO2 to the glass stabilized the pH during SBF testing and significantly increased the hardness compared to the control BG. Scaffold fabrication was conducted with SC-1, and a porous amorphous structure was synthesized at 635 °C.

Body Fluid Processing Workspace Quality Improvement Initiative in a High-Volume Reference Laboratory.

In a clinical laboratory, the design of the workspace directs the workflow and significantly affects the productivity of clinical laboratory scientists (CLS). With the chronic shortage of CLS, a well-designed workspace is essential to take full advantage of available staff, especially in high-volume laboratories. Through the use of quality improvement tools, a manual body fluid testing workspace was redesigned to address weaknesses in the layout that led to excessive physical steps taken by staff. System engineering tools such as a fishbone diagram, spaghetti diagrams, Plan-Do-Study-Act cycles, and a counterbalance measure were all used in a CLS-led quality improvement initiative to redesign a workspace in the manual body fluid processing area of a clinical laboratory at Mayo Clinic. After the redesign, physical steps taken and time to process body fluids were reduced by an average of 40% and 32%, respectively, demonstrating the utility of quality improvement tools in clinical laboratory settings.

Detection of Malignancy in Body Fluids: A Comparison of the Hematology and Cytology Laboratories

An increasing burden is loaded on clinical laboratories for daily testing of body fluids, including pleural fluid, peritoneal fluid (ascites), pericardial fluid, and cerebrospinal fluid. The request for detection of cellular components in the body fluid samples is usually delivered simultaneously to the hospital’s hematology laboratory and cytology laboratory. The medical technologists of hematology laboratory provide the screening result for suspicious malignant cells; meanwhile, the pathologists of cytology laboratory provide the presence of cancer cells at a diagnostic level. However, unfortunately, neither of these results reaches 100% sensitivity or specificity. To assess the clinical practice values, the present study performed a comparison of the hematology and cytology laboratories on the detection of malignancy in body fluids.

A Retrospective Study on Sensitivity, Specificity, Negative Predictive Value, Positive Predictive Value of TB PCR Versus TB Culture in Diagnosing Tuberculosis in Filipino Children Aged 3 Months to 18 Years at a Tertiary Care Center

Objectives: This study aimed to establish the accuracy of TB PCR versus TB culture and rifampicin resistance detection by PCR versus conventional susceptibility testing of body fluids in diagnosing tuberculosis in pediatric patients 3 months to 18 years with suspected tuberculous disease at a tertiary care center. Methods: This is a retrospective analytical study of patients seen between January 1, 2012 to May 31, 2017, with clinical and radiographic features suggestive of tuberculosis, who had diagnostic testing of body fluids for TB PCR and TB culture. Results: Among 159 patients suspected of TB, 46 (28%) tested positive by PCR, of which one was rifampicin-resistant. The sensitivity, specificity, positive predictive value and negative predictive values of TB PCR, using TB culture as the gold standard were 90%, 91.6%, 78.3%, and 96.5% respectively. The sensitivity, specificity, positive predictive value, and negative predictive values of TB PCR for detecting rifampicin resistance, using TB culture and sensitivity as the gold standard, were 33%, 100%, 100%, and 95%, respectively. Overall, the accuracy of TB PCR in detecting TB disease is 91.2% and the accuracy of TB PCR in detecting rifampicin resistance is 95%. Conclusion: Findings in our study suggest that TB PCR play an important role in TB disease diagnosis, but clinical and radiological assessment continue to be essential in the diagnosis of childhood tuberculosis. The accuracy of TB PCR in detecting TB disease in children is 91.2% and the accuracy of TB PCR in detecting Rifampicin resistance is 95%.

Effects of platelet-rich plasma on biological activity and bone regeneration of brushite-based calcium phosphate cement.

In this study, we focused on identifying the effects of platelet-rich plasma (PRP) on bioactivity and bone ingrowths incorporated into brushite cement. We introduced PRP as a series of substitutions with an aqueous citrate-ion solution, and the optimized cement with PRP showed no disintegration of the paste consistency. Incorporating PRP showed that the setting time decreased with the increasing of PRP ratios, although the compressive strength was not significantly changed. We evaluated in vitro degradation and bioactivity with the simulated body fluid test, and the result showed that adding PRP accelerated the carbonated apatite nucleation and markedly improved the surface reactivity of the cement. The in vitro studies demonstrated that incorporating PRP into the brushite cement improved cell adhesion and proliferation. The in vivo effects of PRP were faster degradation, improved tissue response in the early stage, and bone ingrowths. We demonstrated based on our results of this study, incorporation of PRP into brushite cement could be helpful for improving biological activity of the cement as well as bone regeneration. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2316-2326, 2018.

Developmental validation of the ParaDNA® Body Fluid ID System

LGC has developed a rapid mRNA-based body fluid test that runs on the ParaDNA® System, a direct sampling and instrument platform utilizing HyBeacon® hybridization probes and end-point melt detection. The ParaDNA Body Fluid ID Test can detect and identify six different body fluids; vaginal fluid, seminal fluid, sperm cells, saliva, menstrual blood and peripheral blood. Crime stains or swabs are directly sampled to collect a small percentage of material from the item for delivery into pre-prepared chemistry within each test plate. Here we describe some of the ParaDNA Body Fluid ID Test developmental validation studies. The validation assessed performance of the different body fluid markers in the test, including sensitivity, sample aging/environmental studies, mixtures and exposure to various inhibitors.

Reliable Biomarkers to Identify New and Recurrent Cancer.

Breast cancer is the most frequent cancer detected throughout both the developing and the developed world. Its incidence is on the rise in the developing world. Great strides have been made in developing biomarkers to guide therapy for women diagnosed with breast cancer. Far fewer advances have occurred with biomarker development for the early diagnosis of breast cancer. Standard screening for new and recurrent breast cancer involves clinical breast exam and breast imaging. There are no Food and Drug Administration (FDA) approved noninvasive body fluid tests for the early detection of new or recurrent breast cancer. Promising biomarker approaches include multianalyte testing of tissue for individuals diagnosed with breast cancer and body fluid analysis for both at risk women and to monitor individuals after treatment.

Development of risk reduction behavioral counseling for Ebola virus disease survivors enrolled in the Sierra Leone Ebola Virus Persistence Study, 2015-2016.

BackgroundDuring the 2014–2016 West Africa Ebola Virus Disease (EVD) epidemic, the public health community had concerns that sexual transmission of the Ebola virus (EBOV) from EVD survivors was a risk, due to EBOV persistence in body fluids of EVD survivors, particularly semen. The Sierra Leone Ebola Virus Persistence Study was initiated to investigate this risk by assessing EBOV persistence in numerous body fluids of EVD survivors and providing risk reduction counseling based on test results for semen, vaginal fluid, menstrual blood, urine, rectal fluid, sweat, tears, saliva, and breast milk. This publication describes implementation of the counseling protocol and the key lessons learned.Methodology/Principal findingsThe Ebola Virus Persistence Risk Reduction Behavioral Counseling Protocol was developed from a framework used to prevent transmission of HIV and other sexually transmitted infections. The framework helped to identify barriers to risk reduction and facilitated the development of a personalized risk-reduction plan, particularly around condom use and abstinence. Pre-test and post-test counseling sessions included risk reduction guidance, and post-test counseling was based on the participants’ individual test results. The behavioral counseling protocol enabled study staff to translate the study’s body fluid test results into individualized information for study participants.Conclusions/SignificanceThe Ebola Virus Persistence Risk Reduction Behavioral Counseling Protocol provided guidance to mitigate the risk of EBOV transmission from EVD survivors. It has since been shared with and adapted by other EVD survivor body fluid testing programs and studies in Ebola-affected countries.

Fatal Meningitis in Swine after Intrathecal Administration of Adeno-associated Virus Expressing Syngeneic Interleukin-10

Interleukin-10 (IL-10) delivered by intrathecal (i.t.) gene vectors is a candidate investigational new drug (IND) for several chronic neurological disorders such as neuropathic pain. We performed a preclinical safety study of IL-10. A syngeneic large animal model was used delivering porcine IL-10 (pIL-10) to the i.t. space in swine by adeno-associated virus serotype 8 (AAV8), a gene vector that was previously found to be nontoxic in the i.t. space. Unexpectedly, animals became ill, developing ataxia, seizures, and an inability to feed and drink, and required euthanasia. Necropsy demonstrated lymphocytic meningitis without evidence of infection in the presence of normal laboratory findings for body fluids and normal histopathology of peripheral organs. Results were replicated in a second animal cohort by a team of independent experimenters. An extensive infectious disease and neuropathology workup consisting of comprehensive testing of tissues and body fluids in a specialized research veterinary pathology environment did not identify a pathogen. These observations raise the concern that i.t. IL-10 therapy may not be benign, that previously used xenogeneic models testing the human homolog of IL-10 may not have been sensitive enough to detect toxicity, and that additional preclinical studies may be needed before clinical testing of IL-10 can be considered.

Systematic comparison of exosomal proteomes from human saliva and serum for the detection of lung cancer

Circulating tumor exosomes harbor plenty of cancer biological information, which have emerged as promising targets for cancer early detection and diagnosis. Human serum and saliva are unique diagnostic body fluids, which contain numerous circulating exosomes. It is necessary to establish standardized isolation method and compare their proteome profiling for translational medicine. High abundant proteins in these body fluids were removed before exosomes isolation and obtained exosomes were further confirmed by morphology analysis and surface biomarker test. Label free quantification was applied to systematically compare the protein profiling in saliva and serum exosomes. 319 and 994 exosomal proteins were identified from saliva and serum by LC-MS/MS, respectively. To explore their utility for cancer proteomics, we systematically compared the proteome of saliva and serum exosomes from healthy subjects and lung cancer patients. In particular, 11 potential candidates were coincidently discovered in both body fluids of lung cancer patients. Our finding enforced the hypothesis that cancer related proteins were presented in saliva and serum exosomes, which promoted the unique features of exosomes in our body fluids. A circulating exosomes based body fluid test could be easily established for monitoring cancer once these candidates were validated.

Microarc oxidation discharge types and bio properties of the coating synthesized on zirconium

This study is an attempt for gaining a better understanding on relationship between microarc oxidation (MAO) coating discharge types and bioactivity of an oxide-based coating synthesized on a Zr substrate. The discharge types and the coating growth mechanism were identified by the examination of the real cross-section image of the coating microstructure. The coating was conducted by using MAO in an electrolyte containing Na2SiO3, Ca(CH3COO)2 and C3H7Na2O6P, for different durations of 2.5, 5, 15, and 30mins. The effect of the process duration on the different discharge model types (Type-A, B, and C) and bioactivity of the coatings were investigated by using X-ray Diffractometry (XRD), Fourier Transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy-Energy-Dispersive X-ray spectroscopy measurements (SEM-EDS) and Optical Surface Profilometry (OSP). It was found that the increasing MAO duration resulted in thicker and rougher coatings. The XRD data revealed that all the samples prepared at different process durations contained the t-ZrO2 (tetragonal zirconia) phase. During the MAO process, non-crystalline hydroxyapatite (HA) formed, which was confirmed from the FTIR data. The surface morphology, the amount and distribution of the features of the coating surface were modified by increasing voltage. The simulated body fluid (SBF) tests showed that the more bioactive surface with more HA crystals formed owing to chemical composition and high surface roughness of the coating. The pore, crack and discharge structures played a key role in apatite nucleation and growth, and provided ingrowth of apatite into discharge channels on the coating surface.

Recent Development of Wearable Microfluidics Applied in Body Fluid Testing and Drug Delivery

Abstract The most recent developments of the wearable microfluidics applied in body fluid testing and drug delivery and their applications in life monitoring was introduced in this review. Wearable microfluidics was widely applied in the medical, sports and military field. With the help of wearable microfluidics, the pH, glucose, lactate, Na + /K + , Ca 2+ and heavy metal in the body fluid could be detected from sweat, tear and saliva without puncture for blood. These information are of great importance for the monitoring of vital signs and disease diagnosis. The up-to-date research development in the drug delivery using wearable microfluidics and the outlook of the emerging trend in this field were also briefly introduced in this review.

Temperature Coagulant Tub Variation Effect on the Characteristic of Hollow Fiber Dialyser from Cellulose Acetate Composite D-Glucose Monohydrate

Polysulfone is synthetic polymer widely used as basic material for dialyzer membrane and hydrophobic so it tends to cause fouling. Cellulose acetate is non-synthetic, hydrophilic polymer which has low tendency of fouling and has good thermal stability and permeability so it is considered as alternative material for hollow fiber dialyzer. A proper hollow fiber can be achieved by setting a proper temperature of coagulation bath along the spinning process. This research aims to understand the effect of coagulation bath temperature variations on the physical characteristic such as pore size, tensile strength, swelling rate and creatinine clearance of cellulose acetate – D-glucose monohydrate hollow fibers. Hollow fibers were fabricated using spinneret at temperature variations 5°C, 10°C, 15°C dan 20°C. Physical characteristics were estimated by doing morphology test using SEM, tensile test, swelling test towards Simulated Body Fluid (SBF) and filtration test towards creatinine. Result revealed that the hollow fibers from 5°C coagulation bath temperature gives the best characteristic and performance with tensile strength 27,421 N mm-2, pore size 0,0295–0,0858 nm, swelling rate 4,18%, elongation rate 4,4 %, flux rate 1,6032–1,7956 mL cm-2 min-1 and creatinine clearance rate 40,14–48,30% so it is potential to be applied as dialyzer membrane.

Bioactivity and Cytotoxicity of Oxynitride Glasses

Different kinds of bioactive materials are used as bone substitutes. In particular, bioglasses containing Si, Ca, Na and O, bind to host tissues with a stable chemical bond. The phenomenon of bioactivity is associated with the formation of a crystallized hydroxycarbonated apatite (HCA) layer on the bioglass surface when soaked in a simulated physiological fluid. This layer is similar to the mineral phase of bone. However, they do not have sufficient strength for use in load-bearing situations and therefore improving their mechanical properties would allow their use in more robust applications. Within the previously studies, we have shown that the density, glass transition temperature, hardness and elastic modulus have increased linearly with nitrogen content. The purpose of this work was to study the effects of nitrogen addition on the bioactivity in a simulated body fluid (SBF) and cytotoxicity tests have complemented the study.

Study of in vitro bioactivity and mechanical properties of diopside nano-bioceramic synthesized by a facile method using eggshell as raw material

In this study, diopside bioceramic was synthesized using a mechanical milling process and subsequent heat treatment. The simplicity of experiments and also the high energy available in ball milling lead to rapid synthesis of the products in comparison with other synthesis methods. Magnesium oxide (MgO), silicon dioxide (SiO2) and eggshell (as the calcium source) powders were weighted in stoichiometric conditions and milled to initial activation of the surface of the powder's mixture. Then a sintering process was conducted to complete formation of diopside nanopowder and also evaluates its thermal stability. The mechanisms occurred during the synthesis of this bioceramic were carefully investigated. X-Ray diffraction analysis (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), thermogravimetry (TG), differential thermal analysis (DTA), and inductive coupled plasma atomic emission spectroscopy (ICP-AES) were used for gathering and analyzing data. The ability and rate of apatite formation on the sample surface were evaluated by Simulated Body Fluid (SBF) test, a method that is well recognized to characterize the in vitro bioactivity of ceramic materials. According to the results obtained, the diopside samples had a significant potential to form apatite layer on their surface during soaking in the SBF solution. Besides, the bonding strength of this bioceramic was about 350±7MPa which was almost more than three times of that reported for hydroxyapatite. An excellent fracture toughness of 4±0.3MPam0.5 was also obtained for this ceramic which was higher than that of previously reported works.