Imperfect osteogenesis is a hereditary disorder that affects the skeleton, characterized by fragile bones, a high frequency of fractures, bone deformities, and growth deficiencies. Because the production of type I collagen is compromised, people with this disorder may also have other clinical symptoms, such as brittle teeth, blue sclera, hearing loss, reduced respiratory function, and heart problems, such as valve regurgitation.
This disease is caused by the presence of mutations in the genes involved in the production of type I collagen. Its severity can range from mild presentation to severe form. The first is sometimes diagnosed in adulthood. While the second is fatal during the perinatal period.
Diagnosis of imperfect osteogenesis
The severe disorder can be diagnosed by prenatal ultrasonography and can be confirmed by various imaging techniques, such as computed tomography and magnetic resonance imaging. These imaging studies, in addition to detecting imperfect osteogenesis accurately, allow prediction of lethality before birth.
Similarly, during the prenatal period there is the possibility of using genetic tests, which can be invasive or non-invasive, to confirm the diagnosis. After birth, it is possible to do:
- X-rays: They show changes, such as weak or deformed bones and fractures.
- Laboratory tests: include genetic testing.
- Bone density test: This test can show how weak the bones are.
- Bone biopsy: A sample of bone from the hip is examined. This test requires general anesthesia.
General procedure for bone tissue processing and use of the flotation bath
To study changes in bone tissue, proper tissue preparation is needed, this means the chemical preservation or fixation of the material, its support in a solid medium, to be able to cut it into very thin sections and stain it. In general, the steps to perform this study are as follows.
The tissue sample must be carefully obtained and handled, then fixed, usually in formalin and taking care that this substance penetrates the whole tissue; then, the water must be removed from the sample, by means of increasing concentrations of ethanol, this in order to facilitate its infiltration with paraffin. Then, the tissue must be rinsed with xylene because paraffin and ethanol are immiscible, so we must remove the latter. Subsequently, it is infiltrated with paraffin at 60°C, and then included in a paraffin block, which will allow the sample to be cut into a microtome.
Importance of the flotation bath
Once the sample has been cut, sections are included in the flotation bath. This is a laboratory equipment that facilitates the extension of the tissue sections that are obtained from the microtome.
This device is necessary because immediately after cutting, the samples are wrinkled, and thus cannot be placed on the slide. Before introducing the cut sections, the bath must have been prepared with hot distilled water and kept in a temperature range between 40 to 50C°.
The inclusion of the tissue sections in the flotation bath, facilitates their placement on the glass slide, prevents the appearance of folds, wrinkles or deformations in the sample and favors the adhesion of the paraffin on the glass slide.
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