Frozen human bone: a microscopic investigation.

The taphonomic effects of prolonged extreme cold and freezing on human bone have received little research attention. Questions of specific interest include whether previously frozen bone can be identified and whether freezing alters the structural integrity enough to prevent histological aging. There is no evidence from previous studies that freezing damages the structural integrity, and to date no research investigating the freezing process on bone microstructure has been undertaken. This research attempts to distinguish histologically previously frozen bone from nonfrozen bone by identifying patterned defects. To determine the effects of freezing in bone microstructure using light and scanning electron microscopy (SEM), several human bone sections were subjected to prolonged freezing and allowed to thaw before thin sectioning. Light microscopy failed to demonstrate statistically significant differences between frozen and nonfrozen specimens. SEM analysis revealed fractures, although these lacked pattern and did not occur systematically throughout the section. Evidence of microstructural changes caused by liquid expansion, however, was remarkable but did not alter the structural integrity of the microstructure. The results of this study suggest that freezing does not alter the process of histomorphological analysis.

Effect of selenite treatment on ultrastructural changes in experimental diabetic rat bones.

It is known that streptozotocin (STZ)-induced diabetes causes functional and structural alterations in some types of tissue and organ. A number of methods have been used to characterize the properties of diabetic tissues and their diagnosis. Selenium compounds, playing an antioxidant role, can restore some altered metabolic parameters and diminished functions in experimental diabetes. The first aim of the present study was to investigate the effects of STZ-induced diabetes on structural properties of rat long bones. Electron and light microscopic observations showed deleterious alterations in the structure of the diabetic rat long bones, the most prominent effect being in osteocytic cells. Fine cytoplasmic processes of the osteocytes seemed to be shortened, and diabetes affected the normal cytoplasmic processes in a negative manner. The second aim of the present study was to evaluate the effects of sodium selenite treatment for 4 wk on the long bones of the diabetic rats. Electron and light microscopic observations demonstrated that sodium selenite treatment prevented the STZ-induced structural as well as ultrastructural changes in the long bones of the rats. In conclusion, this study first showed that a period of 5-wk diabetes was enough to cause some important and degenerative changes in the structure of the bone tissues, and, second, it demonstrated that sodium selenite treatment of the diabetic rats could normalize these alterations.

Sustained osteomalacia of long bones despite major improvement in other hypophosphatasia-related mineral deficits in tissue nonspecific alkaline phosphatase/nucleotide pyrophosphatase phosphodiesterase 1 double-deficient mice.

We have shown previously that the hypomineralization defects of the calvarium and vertebrae of tissue nonspecific alkaline phosphatase (TNAP)-deficient (Akp2-/-) hypophosphatasia mice are rescued by simultaneous deletion of the Enpp1 gene, which encodes nucleotide pyrophosphatase phosphodiesterase 1 (NPP1). Conversely, the hyperossification in the vertebral apophyses typical of Enpp1-/- mice is corrected in [Akp2-/-; Enpp1-/-] double-knockout mice. Here we have examined the appendicular skeletons of Akp2-/-, Enpp1-/-, and [Akp2-/-; Enpp1-/-] mice to ascertain the degree of rescue afforded at these skeletal sites. Alizarin red and Alcian blue whole mount analysis of the skeletons from wild-type, Akp2-/-, and [Akp2-/-; Enpp1-/-] mice revealed that although calvarium and vertebrae of double-knockout mice were normalized with respect to mineral deposition, the femur and tibia were not. Using several different methodologies, we found reduced mineralization not only in Akp2-/- but also in Enpp1-/- and [Akp2-/-; Enpp1-/-] femurs and tibias. Analysis of calvarial- and bone marrow-derived osteoblasts for mineralized nodule formation in vitro showed increased mineral deposition by Enpp1-/- calvarial osteoblasts but decreased mineral deposition by Enpp1-/- long bone marrow-derived osteoblasts in comparison to wild-type cells. Thus, the osteomalacia of Akp2-/- mice and the hypomineralized phenotype of the long bones of Enpp1-/- mice are not rescued by simultaneous deletion of TNAP and NPP1 functions.

[La accumulation and microstructure change of leg bones of rats fed with La(NO(3))(3) in low dosage for a long term].

OBJECTIVE: To study La accumulation and microstructure change of leg bones of rats fed with La(NO(3))(3) in low dosage for a long term. METHODS: After the rats were fed by La(NO(3))(3) in dosage of 2 mg x (kg(-1) x d(-1)) for 6 months, the contents of La and Ca,P in the leg bones were determined by ICP MS and spectrophotometry; the microstructure changes of the leg bones were investigated by electron microscopy and X-ray powder diffraction. RESULTS: In the leg bones of tested rats, the contents of La and P increased greatly, and those of Ca did not change obviously, so that Ca/P ratio values decreased in comparison with the control group. CONCLUSION: La was accumulatied in the rat leg bones and the change of bone microstructure induced after the rats were fed with La(NO(3))(3) in low dosage for a long term.

Three dimensional ultrastructure of transverse (Harris) lines in the long bone.

To investigate the occurrence of transverse lines in long bones of adults, we studied 27 cadavers (14 males and 13 females). After confirming the presence of a transverse line, a cross-sectional sample was examined macroscopically, and by soft X-ray and scanning electron microscopy. The following results were obtained: 1. According to roentgenograms, transverse (Harris') lines were observed in 40.7% of the distal half of the femur and in 29.6% of the proximal half of the tibia. 2. Macroscopic examination of the bone cross-sections at the level of the transverse line showed various membranous structures. 3. In scanning electron micrographs, no marked difference in structure was observed between the transverse line trabeculae and the compact bone.