Immune Procedures and Imaging Techniques in Biomaterials Research

<p>Gram's Stain</p> Signup and view all the answers

<p>10% buffered formalin</p> Signup and view all the answers

<p>6 to 8 microns</p> Signup and view all the answers

<p>Embedding</p> Signup and view all the answers

<p>HEMATOXYLIN &amp; EOSIN</p> Signup and view all the answers

<p>The ability of a material to perform with an appropriate host response in a specific application</p> Signup and view all the answers

<p>To regulate the safety of medical devices</p> Signup and view all the answers

<p><a href="https://www.fda.gov/">https://www.fda.gov/</a></p> Signup and view all the answers

<p>Bedpans</p> Signup and view all the answers

<p>Class III</p> Signup and view all the answers

<p>To evaluate the risk level of a medical device</p> Signup and view all the answers

<p>A product comprised of two or more regulated components</p> Signup and view all the answers

<p>Immunofluorescence</p> Signup and view all the answers

<p>Immunohistochemistry</p> Signup and view all the answers

<p>Fluorescence activated cell sorter (FACS)</p> Signup and view all the answers

<p>Immunoelectron microscopy</p> Signup and view all the answers

<p>Hemolysis</p> Signup and view all the answers

<p>Turnover is fast</p> Signup and view all the answers

<p>Immunotoxicity</p> Signup and view all the answers

<p>Ability to analyze multiple parameters simultaneously</p> Signup and view all the answers

<p>Agar Diffusion Test</p> Signup and view all the answers

<p>Direct Contact Test</p> Signup and view all the answers

<p>Hemolysis Testing</p> Signup and view all the answers

<p>Cell Proliferation Assay</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>False</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>True</p> Signup and view all the answers

<p>The ability of a material to perform with an appropriate host response in a specific application.</p> Signup and view all the answers

<p>To ensure the premarket testing, manufacture, and postmarket experience of medical devices.</p> Signup and view all the answers

<ol> <li> <a href="http://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/Overview/ClassifyYourDevice/ucm051530.htm">http://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/Overview/ClassifyYourDevice/ucm051530.htm</a>, 2. <a href="https://www.fda.gov/media/82395/download">https://www.fda.gov/media/82395/download</a> </li> </ol> Signup and view all the answers

<p>Tissue processing is done to convert fixed tissue into a form that can be made into thin microscopic sections for further examination.</p> Signup and view all the answers

<p>Tissue embedding is important because it ensures that the tissues are properly aligned and oriented in the block of paraffin for sectioning.</p> Signup and view all the answers

<p>The tissue sections are cut using a microtome, which is a knife with a mechanism for advancing a paraffin block across it.</p> Signup and view all the answers

<p>Staining is done to reverse the embedding process, remove the paraffin wax, and allow water soluble dyes to penetrate the tissue sections for better visualization and analysis.</p> Signup and view all the answers

<p>To measure the toxic effect of test materials in direct contact with tissue culture cells without an overlayer of agar or agarose gel.</p> Signup and view all the answers

<p>To measure the compatibility of a material with animal blood by evaluating the amount of hemolysis in the supernatant after exposure.</p> Signup and view all the answers

<p>To evaluate the cytotoxicity of an extract of material by preparing it in various dilutions and determining cell responses over time.</p> Signup and view all the answers

<p>Healthy vs. diseased animals (if applicable), animal species, implant site, surgical technique, implant form (size, conditioning, dose, texture), and evaluation of tissue response (histology, immunohistology, microscopy, biochemistry, mechanical testing).</p> Signup and view all the answers

<p>p53, PARP, BAX, caspases</p> Signup and view all the answers

<p>Angiogenesis is vital for both bone formation and inflammation and has numerous applications in biomaterials and tissue engineering.</p> Signup and view all the answers

<p>Immunohistochemistry allows for the visualization and detection of specific antigens in cells exposed to biomaterials.</p> Signup and view all the answers

<p>Altered morphology of endothelial cells with heavy particulate load can indicate cellular stress and potential adverse effects of biomaterials.</p> Signup and view all the answers

<p>To make a device for implantation into a human, such as a new type of vascular graft, the raw chemical needs to undergo several steps. This includes material synthesis, processing, sterilization, and fabrication into the final device. To get FDA approval, you need to submit a Premarket Approval application (501K) to the FDA, which includes safety, efficacy, and clinical utility of the device.</p> Signup and view all the answers

<p>The estimated cost of getting a device like a new type of vascular graft to the bench, through clinical trials, and then to the FDA for approval can vary significantly. It depends on factors such as research and development costs, manufacturing costs, clinical trial expenses, regulatory fees, and other associated costs. It is difficult to provide an exact estimate without specific details about the device and the development process.</p> Signup and view all the answers

<p>Biomaterials can fail due to various reasons. The pathogenesis of biomaterial failure involves a sequence of events that lead to structural or functional abnormalities. This can include factors such as improper material selection, inadequate biocompatibility, mechanical stress, infection, inflammation, immune response, and other complications. Understanding the pathogenic mechanisms is crucial for improving the design and performance of biomaterials.</p> Signup and view all the answers

<p>The steps involved in the biocompatibility evaluation of medical devices include: 1) Performing in vitro tests to assess cytotoxicity, genotoxicity, hemocompatibility, and other relevant parameters. 2) Conducting in vivo tests using animal models to evaluate systemic toxicity, implantation response, and other biological effects. 3) Analyzing the results of these tests to assess the compatibility of the device with the intended use and the host response. 4) Documenting and submitting the biocompatibility data as part of the FDA approval process.</p> Signup and view all the answers

<p>Some advantages of using immune procedures in biomaterials research include the ability to specifically identify and quantify proteins and other molecules on surfaces, as well as the potential for multiplexing to analyze multiple targets simultaneously. However, there are also disadvantages such as the need for specific antibodies and reagents, the possibility of non-specific binding, and the potential for interference from other components in complex biological samples. In clinical research and development, immune procedures can be used to study biomaterials in relevant biological systems and assess their interactions with cells and tissues, providing valuable information for evaluating biocompatibility and efficacy.</p> Signup and view all the answers

<p>SEM stands for scanning electron microscopy. It is a powerful imaging technique that uses a focused beam of electrons to scan the surface of a sample. This produces high-resolution images that can reveal the topography, morphology, and composition of biomaterials. In biomaterials research, SEM can be used to study the surface characteristics of materials, such as roughness, porosity, and coating integrity. It can also be used to visualize the interactions between biomaterials and cells, such as cell adhesion, spreading, and morphology. Additionally, SEM can provide information about the structural integrity and degradation of biomaterials over time.</p> Signup and view all the answers

<p>Flow cytometry is a technique that allows for the analysis of individual cells or particles in a fluid stream. In biomaterials applications, flow cytometry can be used to quantify and characterize cell populations interacting with biomaterials. It can provide information about cell viability, proliferation, apoptosis, and differentiation, as well as surface marker expression and intracellular signaling. This can help researchers evaluate the biocompatibility and immunogenicity of biomaterials, as well as study their effects on cell behavior and function. In clinical research and development, flow cytometry can be used to analyze patient samples and assess the response to biomaterials in a personalized medicine approach. It can also be used to monitor immune responses, evaluate drug efficacy, and identify biomarkers for diagnosis and prognosis.</p> Signup and view all the answers

<p>The general principles of biological evaluation of medical devices include considering the chemical composition of the materials, including the conditions of exposure; the nature, degree, frequency, and duration of exposure of the device or its constituents to the body; and the properties and characteristics of the final product. It is important to identify and quantify any extractable chemical entities of the final product before conducting biological evaluation. Testing should take into account the potential short-term and long-term effects, such as acute toxicity, irritation, hemolysis, sensitization, genotoxicity, carcinogenicity, and effects on reproduction. The choice of tests and interpretation of results should also consider the relevance to the overall biological evaluation and be based on end-use applications. In vitro tests should be followed by in vivo testing whenever possible, and careful observations for unexpected adverse reactions or events in humans during clinical use should be conducted.</p> Signup and view all the answers

<p>Advantages of immunofluorescence in biomaterial research include specific localization of antigens in situ, the ability to co-localize several antigens on the same cell, and the visualization of cells grown on light impenetrable materials. However, immunofluorescence has disadvantages such as fluorescence fading over time and the requirement of a relatively expensive fluorescent microscope.</p> Signup and view all the answers

<p>The applications of flow cytometry in biomaterial research include the detection and quantification of cell-bound antigens (surface or intracellular), the detection and quantification of bead-bound antigens, the identification and enumeration of specific cell types within a complex mixture of cells, and the separation of a specific cell type within a complex mixture of cells.</p> Signup and view all the answers

<p>The purpose of fluorescence activated cell sorting (FACS) is to quantitatively analyze and sort cells based on the expression of certain antigens. FACS allows the quantification of the proportion of cells expressing a certain antigen or antigens.</p> Signup and view all the answers

<p>Advantages of immunoelectron microscopy in biomaterial research include high magnification allowing subcellular location of antigens and the ability to be performed on light impenetrable materials. However, immunoelectron microscopy is expensive, time-consuming, and requires electron microscopy.</p> Signup and view all the answers

<p>Biocompatibility = The ability of a material to perform with an appropriate host response in a specific application Hemostasis/Thrombosis = Biomaterials-Tissue Interactions In Vitro, In Vivo, Ex Vivo Testing = Tissue Procurement and Material Evaluation Regulatory Issues = Safety</p> Signup and view all the answers

<p>Hemostasis/Thrombosis = An interaction between biomaterials and tissue Design of Replacement Tissues = Sequence of Events In Vitro, In Vivo, Ex Vivo Testing = Methods of testing tissue procurement and material evaluation FDA Regulatory Program for Devices = Established in 1938 with the goal of premarket testing, manufacture, and postmarket experience of devices</p> Signup and view all the answers

<p>Medical Devices and the FDA = Regulatory program for devices (not materials) Good Manufacturing Practices (GMP) = Standards required for the manufacture and postmarket experience of devices How to Classify Devices = A useful introductory website provided by the FDA 501K and Device Definitions = Information available for download from the FDA website</p> Signup and view all the answers

<p>Immunohistochemistry = Specific localisation of antigens in situ Immunofluorescence = Enables the visualisation of cells grown on light impenetrable materials in situ Western Blotting = Sensitive, specific and semiquantifiable depending on band intensity ELISA = Quantifiable detection of antibody or antigen</p> Signup and view all the answers

<p>Immunohistochemistry = Not suitable for light impenetrable materials Immunofluorescence = Fluorescence fades over time Western Blotting = Cells lysed prior to examination and therefore unable to localise cell specific antigen expression ELISA = Does not allow in situ localisation of antigens</p> Signup and view all the answers

<p>Immunohistochemistry = Commonly used for the examination of pathological samples e.g. disease diagnosis in biopsies Immunofluorescence = Widely used in cellular research both in vivo and in vitro Western Blotting = Commonly used in cell culture research and in vivo digested tissue sections ELISA = Frequently used for the detection of mediators released by cells in cell culture media and for diagnostic detection of antibodies to disease in human serum</p> Signup and view all the answers

<p>Immunoelectron microscopy = High magnification allowing subcellular location of antigens Scanning Electron Microscopy = Produces a largely magnified image by using electrons instead of light to form an image</p> Signup and view all the answers

<p>SEM = A technique used to identify blood cells on materials Biocompatibility Testing = Evaluation of the interaction between a material and a biological system In Vitro = Testing method conducted in a controlled environment outside of a living organism Immunogold Labelling = A method used to identify proteins on surfaces</p> Signup and view all the answers

<p>Cytotoxicity = Testing for cell damage or death Sensitization = Testing for an allergic response Genotoxicity = Testing for damage to genetic information within a cell Hemocompatibility = Testing for compatibility with blood components</p> Signup and view all the answers

<p>Acute toxicity = Adverse effects occurring shortly after exposure Sensitization = Allergic reaction to repeated exposure Carcinogenicity = Tendency to produce cancer Effects on reproduction = Impact on fertility or fetal development</p> Signup and view all the answers

<p>In Vivo = Testing method within a living organism In Vitro = Testing method outside the living organism Ex Vivo = Testing method on organs or tissues from an organism outside the body ISO-10993 = International standard for biological evaluation of medical devices</p> Signup and view all the answers

<p>Hematoxylin = Cells/tissue blue Eosin = Cells/tissues pink H&amp;E = Cytoplasm, Cells, Nucleus Gomori’s silver = Bacteria and Fungi</p> Signup and view all the answers

<p>Pro-inflammatorycytokines (IL-1, IL-6 TNF- ) = Inflammation Integrins and focal contacts = Cell attachment p53, PARP, BAX, caspases = Cell stress Alkaline phosphatase, osteocalcin, Bone morphogenic protein –1 (BMP-1), collagen = Bone formation</p> Signup and view all the answers

<p>Histologic Evaluation = Routine stain used in evaluation of general morphology Immunohistochemistry = Use of substrates and chromogens after antigen binding Immunofluorescence = Use of fluorescent labelled primary antibody Comparative Staining = Use of different stains like H&amp;E, Trichrome, Wright Stain</p> Signup and view all the answers

<p>Inflammatory response = Primary reason for orthopaedic implant failure Cell attachment = Determining substrate biocompatibility in vitro Cell death (apoptosis or necrosis) = Analysis of cell stress exposure to biomaterials Cell activation = Influence on cell phenotypic responses</p> Signup and view all the answers

<p>Gross Examination and Fixation = Involves describing the specimen and placing it into a small plastic cassette for processing to a paraffin block Tissue Processing = The technique of getting fixed tissue into paraffin Tissue Embedding = Process of aligning tissues properly in the block of paraffin Tissue Sectioning = Cutting embedded tissues into sections that can be placed on a slide</p> Signup and view all the answers

<p>Hematoxylin &amp; Eosin = General morphology Gram’s = Bacteria von Kossa = Calcium (CaPO4) Congo red with polarizing light = Amyloid</p> Signup and view all the answers

<p>Low-power magnification (20-40x) = Identification of organ/tissue site and basic disease process High-power magnification (200-400x) = Confirmation of initial impressions and specific diagnosis</p> Signup and view all the answers

<p>Deparaffinized = The process of removing paraffin wax to allow water soluble dyes to penetrate the sections Microtome = Instrument used to cut sections from a paraffin block Coverslipping = Process of covering the stained section on the slide with a thin piece of plastic or glass Fixative = Substance used to preserve tissues from degradation, typically 10% buffered formalin or 2% buffered glutaraldehyde for SEM</p> Signup and view all the answers

<p>Class I = Low risk devices that need not be reviewed by FDA before marketing Class II = Devices that present more risk and require special controls Class III = Devices intended to support or sustain life that require premarket approval Combination Product = Product comprised of two or more regulated components combined or mixed and produced as a single entity</p> Signup and view all the answers

<p>Liver-like, hard, soft, fibrotic, necrotic = Gross Fibrotic, calcific, neoplastic, necrotic, apoptotic, pyknotic = Microscopic CT, MRI, Ultrasound = Imaging K+ level, cholesterol level, BUN, LDH, serum IgG, urine glucose, CSF, Blood count or CBC, differential blood count = Clinical lab findings</p> Signup and view all the answers

<p>Tongue depressors, bedpans, elastic bandages, examination gloves, and some hand-held surgical instruments = Class I X-ray machines, powered wheelchairs, infusion pumps, surgical drapes, surgical needles, suture materials, acupuncture needles, stainless steel bone plate, vascular grafts = Class II Heart valve, artificial heart, assist devices = Class III Device coated or impregnated with a drug or biologic such as drug-eluting stent, pacing lead with steroid-coated tip, catheter with antimicrobial coating; condom with spermicide, skin substitutes with cellular components, prefilled syringes = Combination Product</p> Signup and view all the answers

<p>Explant = Removal of organ or tissue Pathogenic mechanism = Sequence of events that lead to the structural or functional abnormalities Pathologic findings = Examination using eyes, gross &amp; microscopic Morbidity or Mortality = Consequence of implant failure</p> Signup and view all the answers

<p>Agar Diffusion Test = Tissue culture cells with an overlayer of agar or agarose gel. Test materials are added which will/will not diffuse through gel and uptake of a dye is measured. Direct Contact Test = Tissue culture cells without an overlayer of agar or agarose gel. Test materials are added in direct contact with cells. The measurement is done with vital stain over time. Extract Dilution (Elution) Test = An extract of material is prepared and evaluated for cytotoxicity. It is usually done in saline, PBS or culture medium or sometimes solvents like DMSO. Hemolysis Testing = Exposure of material to animal blood (rabbit) for a period of time, ~1 hour. After centrifugation, the supernatant is read for hemolysis (545nm). Less than 5% hemolysis indicates compatibility.</p> Signup and view all the answers

<p>Cell Proliferation Assay = Counting number cells per well by enzymatic removal of cells using trypsin solution. Cell Migration Assay = Cell adhesion to beads and migration of cells from beads onto coated surfaces. Intracellular Function Assay = Assessment of cell cytoskeletal disruption. Cell Co-Culture Assays = Measure smooth muscle cell function in response to NO.</p> Signup and view all the answers

<p>ability</p> Signup and view all the answers

<p>Class I, Class II, and Class III</p> Signup and view all the answers

<p>agar or agarose</p> Signup and view all the answers

<p>paraffin</p> Signup and view all the answers

<p>microtome</p> Signup and view all the answers

<p>hematoxylin</p> Signup and view all the answers

<p>coverslip</p> Signup and view all the answers

<p>agar, agarose</p> Signup and view all the answers

<p>solvents</p> Signup and view all the answers

<p>animal</p> Signup and view all the answers

<p>healthy</p> Signup and view all the answers

<p>in histologic evaluation</p> Signup and view all the answers

<p>nuclei</p> Signup and view all the answers

<p>collagens</p> Signup and view all the answers

<p>glucagon insulin</p> Signup and view all the answers

<p>device</p> Signup and view all the answers

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<p>FACS</p> Signup and view all the answers

<p>fluorescence</p> Signup and view all the answers

<p>Immunocytochemical technique</p> Signup and view all the answers

<p>Scanning Electron Microscopy</p> Signup and view all the answers

<p>application</p> Signup and view all the answers

<p>hematoxylin</p> Signup and view all the answers

<p>paraffin</p> Signup and view all the answers

<p>microscopic</p> Signup and view all the answers

<p>identify</p> Signup and view all the answers

<p>biomaterials</p> Signup and view all the answers

<p>turnover</p> Signup and view all the answers

<p>costly</p> Signup and view all the answers