Tissue engineering is a very promising field of regenerative medicine. Life expectancy has been increasing, and tissue replacement is increasingly needed in patients suffering from various degenerative disorders of the organs. The use of adult mesenchymal stem cells (e.g. from adipose tissue or from bone marrow) in tissue engineering seems to be a promising approach for tissue replacements. Clinical applications can make direct use of the large secretome of these cells, which can have a positive influence on other cells around. Another advantage of adult mesenchymal stem cells is the possibility to differentiate them into various mature cells via appropriate culture conditions (i.e. medium composition, biomaterial properties, and dynamic conditions). This review is focused on current and future ways to carry out tissue replacement of damaged bones and blood vessels, especially with the use of suitable adult mesenchymal stem cells as a potential source of differentiated mature cells that can later be used for tissue replacement. The advantages and disadvantages of different stem cell sources are discussed, with a main focus on adipose-derived stem cells. Patient factors that can influence later clinical applications are taken into account.
Advanced interdisciplinary scientific field of tissue engineering has been developed to meet increasing demand for safe, functional and easy available substitutes of irreversibly damaged tissues and organs. First biomaterials were constructed as “two-dimensional” (allowing cell adhesion only on their surface), and durable (non-biodegradable). In contrast, biomaterials of new generation are characterized by so-called three dimensional porous or scaffold-like architecture promoting attachment, growth and differentiation of cells inside the material, accompanied by its gradual removal and replacement with regenerated fully functional tissue. In order to control these processes, these materials are endowed with a defined spectrum of bioactive molecules, such as ligands for adhesion receptors on cells, functional parts of natural growth factors, hormones and enzymes or synthetic regulators of cell behavior, incorporated in defined concentrations and spatial distribution against a bioinert background resistant to uncontrolled protein adsorption and cell adhesion.
Migration and proliferation of smooth muscle cells (SMC) were studied in cultures prepared from the aorta of Wistar male rats (170—200 g b.w., 8 weeks old) raised under conventional (CC) or specific pathogen-free (SPF) conditions. In primary cultures, higher movement of cells from explants was found in CC raised donors, namely in samples cultured in serum incomplete medium. In the following subcultures (passage 3—16), the growth curves were steeper and the doubling time shorter in CC type of cultures. The faster growth of SMC population from conventional donors was found to be due to a shorter cell cycle and a higher proportion of dividing cells. As a consequence, the maximum population densities were also higher in the latter type of cultures. The differences in growth, that were dependent on raising conditions, were evident for 16 passages, i.e. 7 months after explantation of cells into culture. The data suggest that breeding conditions may affect the activation of growth of SMC in blood vessels in situ.
Sodium borocaptate (BSH, Na2Bi2HnSH), a slow neutron-capture compound, was injected into the left forebrain ventricle of 1-week-old rats (150 fig BSH/3 p\ phosphate buffered saline). After 90 min, the animals were irradiated by epithermal neutrons (LVR-15 nuclear reactor in Řež near Prague, flux density 8.8 x 107 neutrons cm-2 s'1, 8 MW reactor power, 8.2 cGy/min) for 5,10 or 20 min. The brains were examined histologically 8 h after irradiation. In animals irradiated for 5 to 10 min (41 and 82 cGy-Eq, respectively) lethal damage of cells was found in the external granular layer of the cerebellum and the subependymal layer of the forebrain. Irradiation for 20 min (164 cGy-Eq) caused more extensive destruction of cell populations in these regions and, in addition, dead cells appeared also in the more differentiated postmitotic compartments, namely the deeper layers of the cerebellum, layers II/III of the cerebral cortex and corpus callosum. In the forebrain periventricular layer, the extent of cell damage was declining towards the olfactory bulbs. In intact animals, as well as in those injected only with the 150 p\ phosphate buffered saline, the radiation damage was low and limited only to the most sensitive dividing populations of the cerebellum and the forebrain. The study demonstrates a differentiation-dependent damage of the rat brain cells by alpha particles and presents a simple model for evaluation of the biological effectiveness of slow neutron beams constructed for neutron-capture therapy of tumors.
The morphology and proliferation of vascular smooth muscle cells (VSMC) were studied in cultures prepared from the aorta of newborn male and female Wistar rats. The doubling times (DT) of the male-derived population were 16.4 ±0.7 h and 30.0 ±2.2 h in the exponential and post-exponential growth phases, respectively. In the female donor cells, the corresponding DT values were significantly longer, i.e. 21.9 ± 1.8 h and 38.0 ±2.2 h. In addition, the period of growth was shorter in the female-derived cultures. The percentage of 3H-thymidine labelled cells in male cultures was 61.0±3.1, 92.8± 1.9 and 98.7±0.6 % at 2, 27 and 52 h, respectively. In the female-derived populations, only 24.6 ±4.4, 66.1 ±3.8 and 82.8 ±2.0 % of cells were labelled at the corresponding incubation intervals. As a consequence, the final population density in male cultures was 5.6 times higher. In addition, the male-derived VSMC were mainly spindle-shaped and bulgy in appearance while those from female donors were flat and polygonal which means that the cells were adhering to the growth support to a different extent. The study revealed early determination and long-term persistence of lower adhesiveness as well as higher growth potential of male VSMC, i.e. properties which may be of importance for explaining the higher incidence of vascular wall disorders in males.
The growth capacity of cultured vascular smooth muscle cells (VSMC) obtained from the thoracic aorta of 8-week-old male and female spontaneously hypertensive rats (SHR) was compared. Explants from the intima- media complex were cultured in Dulbecco minimum essential medium supplemented with fetal calf serum (10 %). The migration of VSMC out of the explants started on day 2 in both sexes but on day 18 the number of explants with VSMC migration was 100 ±32 explants/flask in male VSMC and only 24 ±5 explants/flask in female ones. The doubling time at the early exponential phase of growth was shorter (13.5 ±0.5 h) and the p H]-thymidine Labelling Index was higher (34.0±2.3 %) in male VSMC than in those from females (19.9±0.6 h and 23.9±1.9 %, p<0.01, respectively). The difference in the doubling time became even more apparent in the late exponential phase of growth (male VSMC: 51.8±2.0 h, female VSMC: 91.5±5.8 h, p<0.001). Moreover, at the end of the exponential growth phase, the male VSMC reached significantly higher (pcO.OOl) maximum population density than VSMC from females. Our data provide evidence of different growth characteristics of cultured VSMC isolated from male and female SHR aortas.
The growth response to angiotensin II (Ang II) was studied using cultured vascular smooth muscle cells (VSMC) isolated from the aortae of male and female spontaneously hypertensive rats (SHR). Systolic and mean arterial blood pressure of 10-week-old males was significantly higher when compared to age-matched females. The specific growth rate of male VSMC was significantly higher on the third and sixth day after synchronisation. Angiotensin II in concentration 10~7 M stimulated the specific growth rate only in male VSMC during the exponential phase of growth. Moreover, doubling time was 3 hours shorter in male VSMC in comparison with the females. Our results suggest that both the increased specific growth rate and augmented growth-response of male VSMC to Ang II may explain the higher sensitivity of males to hypertensive stimuli.