Microvascular and tumor cell alterations during continuous hyperfractionated irradiation: an electron microscopic investigation on the rat R1H rhabdomyosarcoma

Abstract

Purpose: Conventionally fractionated γ-irradiation results in severe damage of tumor capillaries associated with decreasing oxygen partial pressure within the tumor. The present study was undertaken to assess whether vasculo-connective changes are less pronounced after continuous hyperfractionated irradiation, implying better tumor oxygenation and improved radiosensitivity. Materials and Methods: Twenty rats with an isotransplanted R1H rhabdomyosarcoma were irradiated for 12 days with 2 daily fractions of 2.5 Gy (Δt = 6 h). After 0, 15, 30, 45, and 60 Gy, tumor tissue of 4 rats each was analyzed histologically and electron-microscopically. Results: Untreated rhabdomyosarcomas were composed of spindle-shaped tumor cells with numerous mitoses. There were many apoptotic nuclei and a large central necrosis. Tumor capillaries showed a continuous lining of flattened endothelial cells with broad overlapping cell contacts overlying a delicate continuous basal lamina. During irradiation, mean tumor volume declined from 1.9 cm 3 to 1.2 cm 3. The number of atypical mitoses and apoptoses increased and numerous giant tumor cells appeared. The proportion occupied by necrotic tumor tissue rose from 30% to 60%. After 15 Gy (3 days), a marked vasodilatation was apparent accompanied by an interstitial edema. Occasionally, endothelial cells were rounded up and showed indented nuclei, with the underlying basal lamina disintegrated. These changes progressed with increasing radiation doses. After 30 Gy (6 days), leukocytes started to adhere to the endothelial wall. Electron-dense fine fibrillar and basal lamina-like deposits appeared in the perivascular space. Endothelial cell edema was only observed after 60 Gy (12 days). Cell contact areas were shortened, however, the endothelial lining was not interrupted. No signs of radiation fibrosis were observed. Conclusion: Continuous hyperfractionated irradiation induces relatively discrete alterations of the vasculo-connective tumor tissue as compared to conventional irradiation. This may be an advantage with respect to tumor blood flow, oxygenation, and thus, radiosensitivity.