Analysis of the Basic Characteristics of Osteogenic and Chondrogenic Cell Lines Important for Tissue Engineering Implants.

We isolated and characterized cultures of bone and cartilage tissue cells of laboratory minipigs. The size and morphological features of adherent osteogenic and chondrogenic cells were specified. During long-term culturing under standard conditions, the studied cultures expressed specific markers that were detected by immunohistochemical staining: alkaline phosphatase and calcium deposits in osteoblasts and type II collagen and cartilage extracellular matrix in chondrogenic cells. Proliferative potential (mitotic index) of both cell types was 4.64% of the total cell number. Cell motility, i.e. the mean velocity of cell motion was 49 pixels/h for osteoblasts and 47 pixels/h for chondroblasts; the mean migration distance was 2045 and 2118 pixels for chondroblasts and osteoblasts, respectively. The obtained cell lines are now used as the control for evaluation of optimal biocompatibility of scaffold materials in various models. Characteristics of the motility of the bone and cartilage tissue cells can be used for modeling and estimation of the rate of cells population of 3D scaffolds made of synthetic and biological polymers with different internal structure and physicochemical properties during designing in vitro tissue implants.

The mode of inheritance of Scheuermann's disease.

The mode of Scheuermann's disease inheritance and its phenotypic traits in probands and their relatives were studied in 90 pedigrees (90 probands and 385 relatives). The disorder was identified as a genetically related pathology inherited by autosomal dominant type, controlled by a mutant major gene, as a kyphotic deformity without signs of vertebral bodies' anomaly and torsion. Morphological and biochemical studies showed disturbance in the structure of vertebral growth plate anterior aspects at the level of deformity, defects in proliferation and differentiation of chondrocytes, and change in proteoglycan spectrum in cells and matrix. Twelve candidate genes were studied in chondrocytes isolated from vertebral growth plates of patients with Scheuermann's disease. The study results included disorder in the IHH gene expression and preservation of the expression of PAX1, two aggrecan isoforms, link protein, types I and II collagen, lumican, versican, growth hormone and growth factor receptor genes, and proliferation gene. Preservation of the SOX9 gene (transcription gene) probably indicates posttranscriptional genetic disorders. The study is under way.

[Redistribution of immune defence in male mice exposed by female scent].

Since scent marks of mice are harbored by parasites, their sniffing during olfactory search of the mating partner leads to increase of the infection risk. A hypothesis that sexual signals can induce, along with the reproductive behavior, non-specific immune defense against respiratory infections is tested in the present paper. It was found in the experiments on outbred ICR mice that the scent of soiled bedding from cages with mature females stimulated leukocyte intervention to the upper air-ways. Migration of the white blood cells to lung tissue was accompanied with a more prominent immune and endocrine responeses to intranasal application of the bacterial lipopolysacharide (LPS). In particular, LPS administration to male mice treated by female scent was resulted in much greater amount of leukocyte aggregations in the peribronchial areas than that was found in the males kept isolated from the female signals. The female scent also enhanced adrenocortical response to LPS administration, which was coincided with statistically significant increase of IL-1beta concentration in hypothalamus. So, chemical signals of the mature female induce travel of white blood cells to the upper air-waya in the scent treated male mice. It can increase resistance to respiratory infections, on the one hand, and aggravates stress response to inhalation of the bacterial compounds, on the other hand.

Glycosaminoglycans of the trabecular meshwork of the eye in primary juvenile glaucoma.

Significant changes in the qualitative and quantitative composition of glycosaminoglycans (decreased content of sulfated glycosaminoglycans and increased content of collagen-bound proteoglycanes) in the trabecular meshwork of the eye in primary juvenile glaucoma indicate fibrosis of the juxtacanalicular tissue, which was detected in pathomorphological examination of the operation material.

Aggrecan gene expression disorder as aetiologic factor of idiopathic scoliosis.

A pathogenetic mechanism of the idiopathic scoliosis (IS) has been established on the basis of in-depth morphological and biochemical investigations of structural components of the spine in patients with IS (surgical material). We have shown that IS develops on the basis of disturbance of proteoglycans (PG) synthesis and formation in vertebral growth plates. The found keratan sulphate-related fraction is likely a marker of genetic changes in PGs in IS. Long-term our studies demonstrated a major-gene effect in IS. The study has shown that aggrecan gene expression is significantly decreased in cultivated chondroblasts from patients with IS. The presence of keratan sulphate-related fraction and keratan sulphate increase are associated with lumnican increase.

Glycosaminoglycans of the vertebral body growth plate in patients with idiopathic scoliosis.

The concentrations of keratan sulfates and unmodified keratan sulfates increased in the vertebral body growth plate in patients with idiopathic scoliosis. Sulfation and acetylation of total glycosaminoglycans decreased by 50 and 30%, respectively. These changes reflect the decrease in biological activity of molecules that modulate function of the growth plate.

[Growth plate structure of the vertebral body in children of different age groups].

The purpose of this study was to investigate the regularities of formation and functioning of the structural components of the growth plate (GP) of the vertebral body in children during the postnatal period of ontogenesis: in newborns and in children aged 1, 3-5- 6-10- and 12-14-years. GP samples of were studied using histological, histochemical (demonstration of oxidation-reduction enzyme activity, polysaccharides, glycosaminoglycans) and electron microscopic methods with special reference to the age changes of the cells and matrix in different GP zones of the vertebral body. The process of chondrogenic differentiation is described, which includes the successive stages of cellular modifications starting from an undifferentiated chondroblast through a highly differentiated chondrocyte to a degrading and dying chondrocyte. The changes in synthetic activity of cartilage cells are noted as well as the nature of matrix components that are produced by them in the course of differentiation. Regular age-related changes of spatial distribution of mitotically active cells, forming cellular columns, and of isogenous groups are characterized in relation to the alterations of matrix quantity and chemical content in GP of the vertebral body.