Characterization of engineered tissue construct mechanical function by magnetic resonance imaging.

Non-invasive magnetic resonance imaging (MRI) is a technology that enables the characterization of multiple physical phenomena in living and engineered tissues. The mechanical function of engineered tissues is a primary endpoint for the successful regeneration of many biological tissues, such as articular cartilage, spine and heart. Here we demonstrate the application of MRI to characterize the mechanical function of engineered tissue. Phase contrast-based methods were demonstrated to characterize detailed deformation fields throughout the interior of native and engineered tissue, using an articular cartilage defect model as a study system. MRI techniques revealed that strain fields varied non-uniformly, depending on spatial position. Strains were highest in the tissue constructs compared to surrounding native cartilage. Tissue surface geometry corresponded to strain fields observed within the tissue interior near the surface. Strain fields were further evaluated with respect to the spatial variation in the concentration of glycosaminoglycans ([GAG]), critical proteoglycans in the extracellular matrix of cartilage, as determined by gadolinium-enhanced imaging. [GAG] also varied non-uniformly, depending on spatial position and was lowest in the tissue constructs compared to the surrounding cartilage. The use of multiple MRI techniques to assess tissue mechanical function provides complementary data and suggests that deformation is related to tissue geometry, underlying extracellular matrix constituents and the lack of tissue integration in the model system studied. Specialized and advanced MRI phase contrast-based methods are valuable for the detailed characterization and evaluation of mechanical function of tissue-engineered constructs.

Micronuclei in lymphocytes of prostate cancer patients undergoing radiation therapy.

To further verify the applicability of the micronucleus (MN) assay in biodosimetry, we measured the MN yield in cytokinesis-blocked (CB) peripheral blood lymphocytes (PBL) of eight prostate cancer (PC) patients. These patients had no previous chemotherapy or radiotherapy (xRT). They were treated with standardized schemes of fractionated pelvic xRT. Before xRT, and at one random time-point during the course of xRT, blood samples were collected from each patient for the following purposes: (1) to verify the relationship between the MN yield in PBL and the estimated equivalent (EQ) total-body absorbed dose; and (2) to evaluate the individual differences of ex vivo radiation dose-response (1-4 Gy) relationship of MN yield in PBL before xRT. The number of xRT fractions, cumulative tumor dose, and EQ total-body absorbed doses of these patients represented a wide range. We found in PBL of these patients that (1) MN yield (Y) increased linearly with the estimated EQ total-body absorbed dose as Y=14.6+9.2D (R(2)=0.7, p=0.007); the distributions of MN yield were overdispersed; the ratio of relative increment of MN yield per 1000 binucleated (BN) PBL ranged from 0.9 to 8.2 (median: 4.1) folds above that of the respective baseline levels; and (2) before xRT, the MN yields also increased linearly with the ex vivo radiation dose; at each radiation dose level, the distributions of MN yield were overdispersed in most patients. In two of the three patients with xRT-induced early side effects (cystitis, diarrhea), the MN yield in PBL induced by ex vivo irradiation before xRT was significantly higher than in the other patients without xRT-induced side effects. These findings suggest that MN yields in CB PBL can be used as an in vivo biodosimeter. Since the differences in individual ex vivo radiation dose-response relationship of MN yield in PBL before xRT appeared to be significant, our preliminary results also suggest that it may be possible to identify individual intrinsic radiosensitivity before the start of xRT.

Phase III study of interstitial thermoradiotherapy compared with interstitial radiotherapy alone in the treatment of recurrent or persistent human tumors. A prospectively controlled randomized study by the Radiation Therapy Group.

PURPOSE: The objectives of this randomized trial were to determine if interstitial thermoradiotherapy (ITRT) improves tumor regression/control in accessible lesions in comparison with interstitial radiotherapy (IRT) alone and to assess the skin and soft tissue complications with either modality. METHODS AND MATERIALS: From January 1986 to June 1992, 184 patients with persistent or recurrent tumors after previous radiotherapy and/or surgery, which were amenable to interstitial radiotherapy, were accessioned to a protocol developed by the Radiation Therapy Oncology Group (RTOG). One hundred seventy-three cases were analyzed (87 patients in the IRT group and 86 in the ITRT arm). The two arms were well balanced regarding stratification criteria. Most tumors were in the head and neck (40% in the IRT group and 46% in the ITRT group), and pelvis (42% and 43%, respectively). Eighty-four percent of patients in both arms had prior radiation therapy (> or = 40 Gy); 50% and 40%, respectively, had prior surgery, and 34% in each arm had prior chemotherapy. The dose of radiation therapy administered was dependent on the previous radiation dose and did not exceed a total cumulative dose of 100 Gy. Hyperthermia was delivered in one or two sessions, either before or before and after interstitial implant. The intended goal of the hyperthermia was to maintain a minimal tumor temperature of 42.5 degrees C for 30 to 60 min. RESULTS: There was no difference in any of the study end points between the two arms. Complete response (CR) was 53% and 55% in both arms. Two-year survival was 34% and 35%, respectively. Complete response rate for persistent lesions was 69% and 63% in the two treatment arms as compared with 40% and 48% for recurrent lesions. A set of minimal adequacy criteria for the delivery of hyperthermia was developed. When these criteria were applied, only one patient had an adequate hyperthermia session. Acute Grade 3 and 4 toxicities were 12% for IRT and 22 % for ITRT. Late Grade 3 and 4 toxicities were 15% for IRT and 20% for ITRT. The difference was not significant. CONCLUSIONS: Interstitial hyperthermia, as applied in this randomized study, did not show any additional beneficial effects over interstitial radiotherapy alone. Delivery of hyperthermia remains a major obstacle (since only one patient met the basic minimum adequacy criteria as defined in this study). The benefit of hyperthermia in addition to radiation therapy still remains to be proven in properly randomized prospective clinical trials after substantial technical improvements in heat delivery and dosimetry are achieved.

Effects of hyperthermia on bone. I. Heating rate patterns induced by microwave irradiation in bone and muscle phantoms.

We describe the initial heating rate patterns generated by microwave irradiation of 915 MHz, with constant power output, in muscle-equivalent phantoms containing a freshly excised bone, and compared with those in phantoms consisting of muscle-equivalent gel only. At 1 cm depth the muscle was cooler in the centre of the field when bone was present underneath. Also, the orientation of the bone in the field had a pronounced effect on the heating rate profiles in the overlying muscle: when the long axis of the bone was parallel to E field, a hot area in the centre of the field was observed; after rotation of the applicator by 90 degrees so that the long axis of the bone was perpendicular to the E field, more homogeneous heating was obtained along most of the field. In contrast, the heating patterns obtained in the cortex of the bone at similar depth (1.3 cm) were not substantially influenced by its orientation in the field. Depending on field location, the heating rate of the cortical bone closest to the applicator was within 50-75% of the SAR in muscle at the same depth. We believe that these data may be useful for the extension of such measurements in vivo, to permit the effective application of hyperthermia, with or without radiation, in the treatment of bone lesions.

Effects of hyperthermia on bone. II. Heating of bone in vivo and stimulation of bone growth.

Previous studies in vitro have shown that it is possible to achieve comparable temperature distribution in bone and the adjacent soft tissues, under appropriate experimental conditions. The objective of the present work was to determine the effects of hyperthermia on bone in vivo. In order to obtain direct temperature measurements in bone, catheters were surgically installed on top of and inside the medullary cavity of the femur of normal rabbits. The thighs were irradiated with 915 MHz microwaves for 45 min, once or twice a week. The temperatures on and inside the bone were maintained between 42.5 and 44.0 degrees C; the resulting temperatures in the muscle were within 1.0 degrees C at depths equidistant from the applicator. After four to six treatments the femora were excised for histopathological examination. New trabecular bone was deposited around the catheters; most bone components including periosteum, osteoid, and fully calcified matrix could be seen. Large numbers of osteoblasts and osteoclasts lined the trabecular surfaces, and numerous cement lines were visible, running in all directions, indicating extensive bone deposition and remodelling. In contrast, control bones (catheters installed--no hyperthermia) showed much less ossification, with many areas of thin incomplete osteoid. Further, bones treated with hyperthermia only (no surgical trauma) showed no such changes. Thus, it appears that following an initial insult, hyperthermia promotes bone deposition.