The effects of single doses of X radiation ranging from 200 to 2000 rad were studied by direct morphometry in vivo of the mature, stable microvasculature in rabbit ear chambers. Reproducible observations in vivo of the mature microvasculature were obtained by photomicrography of identical 0.033-mm2 sites in each ear chamber prior to and 1 and 5 days following single doses of X radiation. Measurements were made directly on color photomicrographs at a total magnification of 2000X. The microvessels were divided into two groups according to size: vessels greater than 10 microns in diameter (arterioles and venules), and vessels less than or equal to 10 microns in diameter (capillaries). Vascular length and outer and inner surface areas were measured directly on the projected photomicrographs, and vascular volumes and diameters were calculated from these measured parameters. Measurements of capillary length per unit surface area disclosed a decrease in capillary density with increasing dose, resulting in a calculated intercapillary distance in excess of 300 microns, conceivably associated with a decrease in oxygen delivery by the microvasculature. With this method, radiosensitivity of the capillaries was found to be significantly greater than that of larger vessels. Computerized histogram analysis of vascular length, surface area, and volume as a function of increasing diameter (1-micron bins) confirmed the significant difference in reduction of these measured parameters between the capillaries and the larger vessels. The total microvascular volume profile dominated by the volume of larger vessels did not change much 5 days after irradiation, although capillary volume was markedly reduced. Furthermore, the basic profile of the microvasculature showed a shift to larger diameters following irradiation, thus confirming the calculated dilatation of surviving vessels. Qualitative morphologic observations revealed considerable extravasation from the microvessels and formation of micropetechiae at the site of disrupted capillaries with subsequent inflammatory changes.

The cellular outgrowths from three layers of rabbit and monkey aorta were used as primary cultures. Irradiation of the tissue fragments at the time of explanation resulted in a reduction in outgrowth of 50% with a dose of 200 rad, and in a reduction of over 90% with doses of 300 rad and above. When comparable cultures were irradiated after 2 months in vitro as a mature actively metabolizing but slowly proliferating cell population, radioresistance was increased. Subcultures of medial smooth muscle cells irradiated during their logarithmic growth phase showed a linear dose response in the cell number parameter up to 150 rad. A dose of 250 rad resulted in complete flattening of the growth curve, with a reduction in labeling index, after a 3-hr terminal [3H]TdR pulse. On the other hand, the labeling index indicated some recovery 3 days after irradiation in cultures receiving less than 250 rad. Under the same experimental conditions, cells derived from the intima of the same aorta showed no recovery when increase in cell numbers over time, or the number of labeled cells per area, were used as parameters. Cells derived from adventitia showed a relative increase in the number of labeled cells per area 4 and 7 days after irradiation following an initial decrease on Day 1.

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