A silicone-based support material eliminates interfacial instabilities in 3D silicone printing.

Among the diverse areas of 3D printing, high-quality silicone printing is one of the least available and most restrictive. However, silicone-based components are integral to numerous advanced technologies and everyday consumer products. We developed a silicone 3D printing technique that produces precise, accurate, strong, and functional structures made from several commercially available silicone formulations. To achieve this level of performance, we developed a support material made from a silicone oil emulsion. This material exhibits negligible interfacial tension against silicone-based inks, eliminating the disruptive forces that often drive printed silicone features to deform and break apart. The versatility of this approach enables the use of established silicone formulations in fabricating complex structures and features as small as 8 micrometers in diameter.

Modeling bystander effects that cause growth delay of breast cancer xenografts in bone marrow of mice treated with radium-223.

RATIONALE: The role of radiation-induced bystander effects in cancer therapy with alpha-particle emitting radiopharmaceuticals remains unclear. With renewed interest in using alpha-particle emitters to sterilize disseminated tumor cells, micrometastases, and tumors, a better understanding of the direct effects of alpha particles and the contribution of the bystander responses they induce is needed to refine dosimetric models that help predict clinical benefit. Accordingly, this work models and quantifies the relative importance of direct effects (DE) and bystander effects (BE) in the growth delay of human breast cancer xenografts observed previously in the tibiae of mice treated with 223RaCl2. METHODS: A computational model of MDA-MB-231 and MCF-7 human breast cancer xenografts in the tibial bone marrow of mice administered 223RaCl2 was created. A Monte Carlo radiation transport simulation was performed to assess individual cell absorbed doses. The responses of the breast cancer cells to direct alpha particle irradiation and gamma irradiation were needed as input data for the model and were determined experimentally using a colony-forming assay and compared to the responses of preosteoblast MC3T3-E1 and osteocyte-like MLO-Y4 bone cells. Using these data, a scheme was devised to simulate the dynamic proliferation of the tumors in vivo, including DE and BE propagated from the irradiated cells. The parameters of the scheme were estimated semi-empirically to fit experimental tumor growth. RESULTS: A robust BE component, in addition to a much smaller DE component, was required to simulate the in vivo tumor proliferation. We also found that the relative biological effectiveness (RBE) for cell killing by alpha particle radiation was greater for the bone cells than the tumor cells. CONCLUSION: This modeling study demonstrates that DE of radiation alone cannot explain experimental observations of 223RaCl2-induced growth delay of human breast cancer xenografts. Furthermore, while the mechanisms underlying BE remain unclear, the addition of a BE component to the model is necessary to provide an accurate prediction of the growth delay. More complex models are needed to further comprehend the extent and complexity of 223RaCl2-induced BE.

Radium-223-Induced Bystander Effects Cause DNA Damage and Apoptosis in Disseminated Tumor Cells in Bone Marrow.

Radiation-induced bystander effects have been implicated in contributing to the growth delay of disseminated tumor cells (DTC) caused by 223RaCl2, an alpha particle-emitting radiopharmaceutical. To understand how 223RaCl2 affects the growth, we have quantified biological changes caused by direct effects of radiation and bystander effects caused by the emitted radiations on DTC and osteocytes. Characterizing these effects contribute to understanding the efficacy of alpha particle-emitting radiopharmaceuticals and guide expansion of their use clinically. MDA-MB-231 or MCF-7 human breast cancer cells were inoculated intratibially into nude mice that were previously injected intravenously with 50 or 600 kBq/kg 223RaCl2. At 1-day and 3-days postinoculation, tibiae were harvested and examined for DNA damage (γ-H2AX foci) and apoptosis in osteocytes and cancer cells located within and beyond the range (70 µm) of alpha particles emitted from the bone surface. Irradiated and bystander MDA-MB-231 and MCF-7 cells harbored DNA damage. Bystander MDA-MB-231 cells expressed DNA damage at both treatment levels while bystander MCF-7 cells required the higher administered activity. Osteocytes also had DNA damage regardless of inoculated cancer cell line. The extent of DNA damage was quantified by increases in low (1-2 foci), medium (3-5 foci), and high (5+ foci) damage. MDA-MB-231 but not MCF-7 bystander cells showed increases in apoptosis in 223RaCl2-treated animals, as did irradiated osteocytes. In summary, radiation-induced bystander effects contribute to DTC cytotoxicity caused by 223RaCl2. IMPLICATIONS: This observation supports clinical investigation of the efficacy of 223RaCl2 to prevent breast cancer DTC from progressing to oligometastases.

Dose-Dependent Growth Delay of Breast Cancer Xenografts in the Bone Marrow of Mice Treated with 223Ra: The Role of Bystander Effects and Their Potential for Therapy.

The role of radiation-induced bystander effects in radiation therapy remains unclear. With renewed interest in therapy with α-particle emitters, and their potential for sterilizing disseminated tumor cells (DTCs), it is critical to determine the contribution of bystander effects to the overall response so they can be leveraged for maximum clinical benefit. Methods: Female Foxn1nu athymic nude mice were administered 0, 50, or 600 kBq/kg 223RaCl2 to create bystander conditions. At 24 hours after administration, MDA-MB-231 or MCF-7 human breast cancer cells expressing luciferase were injected into the tibial marrow compartment. Tumor burden was tracked weekly via bioluminescence. Results: The MDA-MB-231 xenografts were observed to have a 10-day growth delay in the 600 kBq/kg treatment group only. In contrast, MCF-7 cells had 7- and 65-day growth delays in the 50 and 600 kBq/kg groups, respectively. Histologic imaging of the tibial marrow compartment, α-camera imaging, and Monte Carlo dosimetry modeling revealed DTCs both within and beyond the range of the α-particles emitted from 223Ra in bone for both MCF-7 and MDA-MB-231 cells. Conclusion: Taken together, these results support the participation of 223Ra-induced antiproliferative/cytotoxic bystander effects in delayed growth of DTC xenografts. They indicate that the delay depends on the injected activity and therefore is dose-dependent. They suggest using 223RaCl2 as an adjuvant treatment for select patients at early stages of breast cancer.

CXCR1- or CXCR2-modified CAR T cells co-opt IL-8 for maximal antitumor efficacy in solid tumors.

Chimeric antigen receptor (CAR) T-cell therapy targeting solid tumors has stagnated as a result of tumor heterogeneity, immunosuppressive microenvironments, and inadequate intratumoral T cell trafficking and persistence. Early (≤3 days) intratumoral presentation of CAR T cells post-treatment is a superior predictor of survival than peripheral persistence. Therefore, we have co-opted IL-8 release from tumors to enhance intratumoral T-cell trafficking through a CAR design for maximal antitumor activity in solid tumors. Here, we demonstrate that IL-8 receptor, CXCR1 or CXCR2, modified CARs markedly enhance migration and persistence of T cells in the tumor, which induce complete tumor regression and long-lasting immunologic memory in pre-clinical models of aggressive tumors such as glioblastoma, ovarian and pancreatic cancer.

Three-Dimensional Printing for Construction of Tissue-Equivalent Anthropomorphic Phantoms and Determination of Conceptus Dose.

OBJECTIVE: The purpose of this study was to develop a road map for rapid construction of anthropomorphic phantoms from computational human phantoms for use in diagnostic imaging dosimetry studies. These phantoms are ideal for performing pregnant-patient dosimetry because the phantoms imitate the size and attenuation properties of an average-sized pregnant woman for multiple gestational periods. MATERIALS AND METHODS: The method was derived from methods and materials previously described but adapted for 3D printing technology. A 3D printer was used to transform computational models into a physical duplicate with small losses in spatial accuracy and to generate tissue-equivalent materials characterized for diagnostic energy x-rays. A series of pregnant abdomens were selected as prototypes because of their large size and complex modeling. The process involved the following steps: segmentation of anatomy used for modeling; transformation of the computational model into a printing file format; preparation, characterization, and introduction of phantom materials; and model removal and phantom assembly. RESULTS: The density of the homogenized soft tissue-equivalent substitute was optimized by combining 9.0% by weight of urethane filler powder and 91.0% urethane polymer, which resulted in a mean density of 1.041 g/cm3 measured over 20 samples. Density varied among all of the samples by 0.0026 g/cm3. The total variation in density was 0.00261 g/cm3. The half-value layer of the bone material was measured to be 1.7 mm of bone material at 120 kVp and when simulated by use of the density of the bone tissue-equivalent substitute (1.60 g/cm3) was determined to be 1.61 mm of bone tissue. For dosimetry purposes the phantom provided excellent results for evaluating a site's protocol based on scan range. CONCLUSION: The 3D printing technology is applicable to the fabrication of phantoms used for performing dosimetry. The tissue-equivalent materials used to substitute for the soft tissue were developed to be highly adaptable for optimization based on the dosimetry application. Use of this method resulted in more automated phantom construction with decreased construction time and increased out-of-slice spatial resolution of the phantoms.

An image-based skeletal dosimetry model for the ICRP reference adult female-internal electron sources.

An image-based skeletal dosimetry model for internal electron sources was created for the ICRP-defined reference adult female. Many previous skeletal dosimetry models, which are still employed in commonly used internal dosimetry software, do not properly account for electron escape from trabecular spongiosa, electron cross-fire from cortical bone, and the impact of marrow cellularity on active marrow self-irradiation. Furthermore, these existing models do not employ the current ICRP definition of a 50 µm bone endosteum (or shallow marrow). Each of these limitations was addressed in the present study. Electron transport was completed to determine specific absorbed fractions to both active and shallow marrow of the skeletal regions of the University of Florida reference adult female. The skeletal macrostructure and microstructure were modeled separately. The bone macrostructure was based on the whole-body hybrid computational phantom of the UF series of reference models, while the bone microstructure was derived from microCT images of skeletal region samples taken from a 45 years-old female cadaver. The active and shallow marrow are typically adopted as surrogate tissue regions for the hematopoietic stem cells and osteoprogenitor cells, respectively. Source tissues included active marrow, inactive marrow, trabecular bone volume, trabecular bone surfaces, cortical bone volume, and cortical bone surfaces. Marrow cellularity was varied from 10 to 100 percent for active marrow self-irradiation. All other sources were run at the defined ICRP Publication 70 cellularity for each bone site. A total of 33 discrete electron energies, ranging from 1 keV to 10 MeV, were either simulated or analytically modeled. The method of combining skeletal macrostructure and microstructure absorbed fractions assessed using MCNPX electron transport was found to yield results similar to those determined with the PIRT model applied to the UF adult male skeletal dosimetry model. Calculated skeletal averaged absorbed fractions for each source-target combination were found to follow similar trends of more recent dosimetry models (image-based models) but did not follow results from skeletal models based upon assumptions of an infinite expanse of trabecular spongiosa.

Do tumor and ventricular volume predict the need for postresection shunting in colloid cyst patients?

OBJECTIVE Many colloid cyst patients present with obstructive hydrocephalus that resolves after resection of the cyst. However, a proportion of patients with these cysts will require cerebrospinal fluid shunting after tumor resection, despite resolution of the obstruction at the foramina of Monro. The goal of this study was to determine if colloid cyst size or preoperative ventricular volume predicted the need for postresection shunting. METHODS In a retrospective study design, ICD-9 codes 742.2 (colloid cyst) and 348.0 (brain cyst) were used to identify patients who had undergone resection of a colloid cyst at the University of Florida over the last 20 years. Preoperative imaging (CT or MRI) with a stereotactic software program developed at the University of Florida was used to measure volumes of the colloid cyst and the lateral ventricles. The relationships among ventricular volume, colloid cyst volume, and postoperative shunting were analyzed. RESULTS The number of patients included in the study was 67, and their mean age was 37.7 years. Forty percent of the patients were female. Overall, 49.2% of the patients had a transcallosal approach, 35.8% a transcortical approach, and 14.9% an endoscope-assisted surgery. Mean preoperative ventricular volume was 76.5 cc in patients who never received a ventriculoperitoneal shunt (VPS) and 98.1 cc in those who were eventually treated with a VPS (p = 0.305). Patients with a postoperative VPS had an initial mean colloid cyst volume of 1.8 cc compared with 0.9 cc in patients without a VPS postoperatively (p = 0.019). Patients with colloid cysts larger than 0.6 cc (1-cm diameter) had a 12.8 increased odds of needing a VPS postoperatively (95% CI 1.81-275). CONCLUSIONS Larger colloid cysts are associated with an increased need for postresection shunting independent of preoperative ventricular size. Prospective studies of patients with colloid cysts are necessary to further identify risks of permanent hydrocephalus.

Mixed reality ventriculostomy simulation: experience in neurosurgical residency.

BACKGROUND: Medicine and surgery are turning toward simulation to improve on limited patient interaction during residency training. Many simulators today use virtual reality with augmented haptic feedback with little to no physical elements. In a collaborative effort, the University of Florida Department of Neurosurgery and the Center for Safety, Simulation & Advanced Learning Technologies created a novel "mixed" physical and virtual simulator to mimic the ventriculostomy procedure. The simulator contains all the physical components encountered for the procedure with superimposed 3-D virtual elements for the neuroanatomical structures. OBJECTIVE: To introduce the ventriculostomy simulator and its validation as a necessary training tool in neurosurgical residency. METHODS: We tested the simulator in more than 260 residents. An algorithm combining time and accuracy was used to grade performance. Voluntary postperformance surveys were used to evaluate the experience. RESULTS: Results demonstrate that more experienced residents have statistically significant better scores and completed the procedure in less time than inexperienced residents. Survey results revealed that most residents agreed that practice on the simulator would help with future ventriculostomies. CONCLUSION: This mixed reality simulator provides a real-life experience, and will be an instrumental tool in training the next generation of neurosurgeons. We have now implemented a standard where incoming residents must prove efficiency and skill on the simulator before their first interaction with a patient.

Mixed-reality simulation for neurosurgical procedures.

BACKGROUND: Surgical education is moving rapidly to the use of simulation for technical training of residents and maintenance or upgrading of surgical skills in clinical practice. To optimize the learning exercise, it is essential that both visual and haptic cues are presented to best present a real-world experience. Many systems attempt to achieve this goal through a total virtual interface. OBJECTIVE: To demonstrate that the most critical aspect in optimizing a simulation experience is to provide the visual and haptic cues, allowing the training to fully mimic the real-world environment. METHODS: Our approach has been to create a mixed-reality system consisting of a physical and a virtual component. A physical model of the head or spine is created with a 3-dimensional printer using deidentified patient data. The model is linked to a virtual radiographic system or an image guidance platform. A variety of surgical challenges can be presented in which the trainee must use the same anatomic and radiographic references required during actual surgical procedures. RESULTS: Using the aforementioned techniques, we have created simulators for ventriculostomy, percutaneous stereotactic lesion procedure for trigeminal neuralgia, and spinal instrumentation. The design and implementation of these platforms are presented. CONCLUSION: The system has provided the residents an opportunity to understand and appreciate the complex 3-dimensional anatomy of the 3 neurosurgical procedures simulated. The systems have also provided an opportunity to break procedures down into critical segments, allowing the user to concentrate on specific areas of deficiency.

Survival of tumor and normal cells upon targeting with electron-emitting radionuclides.

PURPOSE: Previous studies have shown that the mean absorbed dose to a tissue element may not be a suitable quantity for correlating with the biological response of cells in that tissue element. Cell survival can depend strongly on the distribution of radioactivity at the cellular and multicellular levels. Furthermore, when cellular absorbed doses are examined, the cross-dose from neighbor cells can be less radiotoxic than the self-dose component. To better understand how the nonuniformity of activity among cells can affect the dose response, a computer model of a 3D tissue culture was previously constructed and showed that activity distribution among cells is significantly more relevant than the mean absorbed dose for low-energy-electron emitters. The present work greatly expands upon those findings. METHODS: In the present study, we used this same computer model but restricted the number of labeled cells to a fraction of the whole cell population (50%, 10%, and 1%, respectively). The labeled cells were randomly distributed among the whole cell population. RESULTS: While the activity distribution is an important factor in determining the tissue response for low-energy-electron emitters, the fraction of labeled cells has an even more pronounced effect on survival response. For all electron energies studied, reducing the percentage of cells labeled significantly increases the surviving fraction of the whole population. CONCLUSIONS: This study provides abundant information on killing tumor and normal cells under some conditions relevant to targeted radionuclide therapy of isolated tumor cells and micrometastases. The percentage of cells labeled, activity distribution among the labeled cells, and electron energy play key roles in determining their response. Most importantly, and not previously demonstrated, lognormal activity distributions can have a profound impact on the response of the tumor cells even when the radionuclide emits high-energy electrons.

Monte Carlo simulation of irradiation and killing in three-dimensional cell populations with lognormal cellular uptake of radioactivity.

PURPOSE: The biological response of tissue exposed to radiations emitted by internal radioactivity is often correlated with the mean absorbed dose to a tissue element. However, experimental studies show that even when the mean absorbed dose to the tissue element is constant, the response of the cell population within the tissue element can vary significantly depending on the distribution of radioactivity at the cellular and multicellular levels. The present work develops theoretical models to simulate these observations. MATERIALS AND METHODS: Two theoretical models were created to simulate experimental three-dimensional cell culture models with homogeneous and inhomogeneous tissue environments. The cells were assigned activities according to lognormal distributions of an alpha particle emitter or a monoenergetic electron emitter. Absorbed doses to the cell nuclei were assessed with point-kernel geometric-factor and Electron Gamma Shower version nrc (EGSnrc) Monte Carlo radiation transport simulations, respectively. The self- and cross-dose to individual cell nuclei were calculated and a Monte Carlo method was used to determine their fate. Survival curves were produced after tallying the live and dead cells. RESULTS: Both percent cells labeled and breadth of lognormal distribution affected the dose distribution at the cellular level, which in turn, influenced the shape of the cell survival curves. CONCLUSIONS: Multicellular Monte Carlo dosimetry-models offer improved capacity to predict response to radiopharmaceuticals compared to approaches based on mean absorbed dose to the tissue.

Lognormal distribution of cellular uptake of radioactivity: Monte Carlo simulation of irradiation and cell killing in 3-dimensional populations in carbon scaffolds.

UNLABELLED: The biologic response of tissue exposed to radiation emitted by internal radioactivity is often correlated with the mean absorbed dose to a tissue element. However, studies show that even when the macroscopic absorbed dose to the tissue element is constant, the response of the cell population within the tissue element can vary significantly, depending on the distribution of radioactivity at the cellular and multicellular levels. These variations are believed to be the consequence of nonuniform distributions of activity among the cells or subcellular compartments that comprise the tissue element. Furthermore, the self-dose received by a cell containing radioactivity can be more radiotoxic than the cross-dose from neighboring cells. To study how the nonuniformity of activity among cells can affect the dose response, a 3-dimensional model of cells in a heterogeneous carbon scaffold was used to assess response. METHODS: A theoretic model of a 3-dimensional cell culture was constructed, and Monte Carlo radiation transport was performed to assess self- and cross-doses for each cell nucleus in a population of 10(6) cells. On the basis of these individual doses and on empiric models of radiation-induced cell death (i.e., reproductive failure), survival curves were simulated with different electron energies and activity distributions among the cells. RESULTS: Nonuniformity of cell activities are responsible for nonuniformity of the dose at the cellular level, which in turn causes a change in the surviving fraction of the cell population from that expected on the basis of uniform activity and dose. CONCLUSION: The macroscopic dose received by a tissue cannot be used to anticipate its biologic response. The dose distribution among individual cells, because of both their nonuniform activity and geometric environment, is an important factor in determining biologic response of the tissue at the macroscopic level.

An image-based skeletal dosimetry model for the ICRP reference adult male--internal electron sources.

In this study, a comprehensive electron dosimetry model of the adult male skeletal tissues is presented. The model is constructed using the University of Florida adult male hybrid phantom of Lee et al (2010 Phys. Med. Biol. 55 339-63) and the EGSnrc-based Paired Image Radiation Transport code of Shah et al (2005 J. Nucl. Med. 46 344-53). Target tissues include the active bone marrow, associated with radiogenic leukemia, and total shallow marrow, associated with radiogenic bone cancer. Monoenergetic electron emissions are considered over the energy range 1 keV to 10 MeV for the following sources: bone marrow (active and inactive), trabecular bone (surfaces and volumes), and cortical bone (surfaces and volumes). Specific absorbed fractions are computed according to the MIRD schema, and are given as skeletal-averaged values in the paper with site-specific values reported in both tabular and graphical format in an electronic annex available from http://stacks.iop.org/0031-9155/56/2309/mmedia. The distribution of cortical bone and spongiosa at the macroscopic dimensions of the phantom, as well as the distribution of trabecular bone and marrow tissues at the microscopic dimensions of the phantom, is imposed through detailed analyses of whole-body ex vivo CT images (1 mm resolution) and spongiosa-specific ex vivo microCT images (30 µm resolution), respectively, taken from a 40 year male cadaver. The method utilized in this work includes: (1) explicit accounting for changes in marrow self-dose with variations in marrow cellularity, (2) explicit accounting for electron escape from spongiosa, (3) explicit consideration of spongiosa cross-fire from cortical bone, and (4) explicit consideration of the ICRP's change in the surrogate tissue region defining the location of the osteoprogenitor cells (from a 10 µm endosteal layer covering the trabecular and cortical surfaces to a 50 µm shallow marrow layer covering trabecular and medullary cavity surfaces). Skeletal-averaged values of absorbed fraction in the present model are noted to be very compatible with those weighted by the skeletal tissue distributions found in the ICRP Publication 110 adult male and female voxel phantoms, but are in many cases incompatible with values used in current and widely implemented internal dosimetry software.

An image-based skeletal dosimetry model for the ICRP reference newborn--internal electron sources.

In this study, a comprehensive electron dosimetry model of newborn skeletal tissues is presented. The model is constructed using the University of Florida newborn hybrid phantom of Lee et al (2007 Phys. Med. Biol. 52 3309-33), the newborn skeletal tissue model of Pafundi et al (2009 Phys. Med. Biol. 54 4497-531) and the EGSnrc-based Paired Image Radiation Transport code of Shah et al (2005 J. Nucl. Med. 46 344-53). Target tissues include the active bone marrow (surrogate tissue for hematopoietic stem cells), shallow marrow (surrogate tissue for osteoprogenitor cells) and unossified cartilage (surrogate tissue for chondrocytes). Monoenergetic electron emissions are considered over the energy range 1 keV to 10 MeV for the following source tissues: active marrow, trabecular bone (surfaces and volumes), cortical bone (surfaces and volumes) and cartilage. Transport results are reported as specific absorbed fractions according to the MIRD schema and are given as skeletal-averaged values in the paper with bone-specific values reported in both tabular and graphic format as electronic annexes (supplementary data). The method utilized in this work uniquely includes (1) explicit accounting for the finite size and shape of newborn ossification centers (spongiosa regions), (2) explicit accounting for active and shallow marrow dose from electron emissions in cortical bone as well as sites of unossified cartilage, (3) proper accounting of the distribution of trabecular and cortical volumes and surfaces in the newborn skeleton when considering mineral bone sources and (4) explicit consideration of the marrow cellularity changes for active marrow self-irradiation as applicable to radionuclide therapy of diseased marrow in the newborn child.

Rapid fabrication of custom patient biopsy guides.

Image guided surgery is currently performed using frame-based as well as frameless approaches. In order to reduce the invasive nature of stereotactic guidance as well as to reduce the cost in both equipment and time required within the operating room we investigated the use of rapid prototyping (RP) technology. In our approach we fabricated custom patient specific face-masks and guides that can be applied to the patient during surgery. These guides provide a stereotactic reference for the accurate placement of surgical tools to a pre-planned target along a pre-planned trajectory. While the use of RP machines has previously been shown to be satisfactory for the accuracy standpoint, one of our design criteria, completing the entire built and introduction into the sterile field in less than 120 minutes, was unobtainable. Our primary problems were the fabrication time and the non-resistance of the built material to high-temperature sterilization. In the current study, we have investigated the use of subtractive rapid prototyping (SRP) machines to perform the same quality of surgical guidance while improving the fabrication time and allowing for choosing materials suitable for sterilization. Because SRP technology does not offer the same flexibility as RP in term of prototype shape and complexity, our software program was adapted to provide new guide designs suitable for SRP fabrication. The biopsy guide was subdivided for a more efficient built with the parts being uniquely assembled to form the final guide. The accuracy of the assembly was then assessed using a modified Brown-Roberts-Wells phantom base that allows measuring the position of a biopsy needle introduced into the guide and comparing it with the actual planned target. These tests showed that 1) SRP machines provide an average accuracy of 0.77 mm with a standard deviation of 0.05 mm (plus or minus one image pixel) and 2) SRP allows for fabrication and sterilization within three and a half hours after diagnostic image acquisition and we are confident that that further improvements can reduce this time to less than two hours. Further tests will determine the accuracy of the positioning of the face mask on the patient's head under an IRB-approved trial judged against actual frame-based and frameless systems.

Development of an intramuscular xenograft model of canine osteosarcoma in mice for evaluation of the effects of radiation therapy.

OBJECTIVE: To develop an IM xenograft model of canine osteosarcoma in mice for the purpose of evaluating effects of radiation therapy on tumors. ANIMALS: 27 athymic nude mice. PROCEDURES: Mice were randomly assigned to 1 of 3 groups of 9 mice each: no treatment (control group), radiation at 10 Gy, or radiation at 15 Gy. Each mouse received 5 x 10(5) highly metastasizing parent osteosarcoma cells injected into the left gastrocnemius muscle. Maximum tumor diameter was determined with a metric circles template to generate a tumor growth curve. Conscious mice were restrained in customized plastic jigs allowing local tumor irradiation. The behavior and development of the tumor xenograft were assessed via evaluations of the interval required for tumor-bearing limbs to reach diameters of 8 and 13 mm, extent of tumor vasculature, histomorphology of tumors, degree of tumor necrosis, and existence of pulmonary metastasis and clinical disease in affected mice. RESULTS: Tumor-bearing limbs grew to a diameter of 8 mm (0.2-g tumor mass) in a mean +/- SEM interval of 7.0 +/- 0.2 days in all mice. Interval to grow from 8 to 13 mm was significantly prolonged for both radiation therapy groups, compared with that of the control group. Histologic evaluation revealed the induced tumors were highly vascular and had characteristics consistent with those of osteosarcoma. Pulmonary metastasis was not detected, and there was no significant difference in percentage of tumor necrosis between groups. CONCLUSIONS AND CLINICAL RELEVANCE: A reliable, repeatable, and easily produced IM xenograft model was developed for in vivo assessment of canine osteosarcoma.

Intrinsic radiosensitivity and repair of sublethal radiation-induced damage in canine osteosarcoma cell lines.

OBJECTIVE: To characterize the radiosensitivity and capacity for sublethal damage repair (SLDR) of radiation-induced injury in 4 canine osteosarcoma cell lines. SAMPLE POPULATION: 4 canine osteosarcoma cell lines (HMPOS, POS, COS 31, and D17). PROCEDURES: A clonogenic colony-forming assay was used to evaluate the cell lines' intrinsic radiosensitivities and SLDR capacities. Dose-response curves for the cell lines were generated by fitting the surviving fractions after radiation doses of 0 (control cells), 1, 2, 3, 6, and 9 Gy to a linear quadratic model. To evaluate SLDR, cell lines were exposed to 2 doses of 3 Gy (split-dose experiments) at an interval of 0 (single 6-Gy dose), 2, 4, 6, or 24 hours, after which the surviving fractions were assessed. RESULTS: Mean surviving fraction did not differ significantly among the 4 cell lines at the radiation doses tested. Mean surviving fraction at 2 Gy was high (0.62), and the alpha/beta ratios (predictor of tissue sensitivity to radiation therapy) for the cell lines were low (mean ratio, 3.47). The split-dose experiments revealed a 2.8- to 3.9-fold increase in cell survival when the radiation doses were applied at an interval of 24 hours, compared with cell survival after radiation doses were applied consecutively (0-hour interval). CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that these canine osteosarcoma cell lines are fairly radioresistant; alpha/beta ratios were similar to those of nonneoplastic, late-responding tissues. Future clinical investigations should involve increasing the fraction size in a manner that maximizes tumor killing without adverse effects on the nonneoplastic surrounding tissues.

Nonspecific immunotherapy with intratumoral lipopolysaccharide and zymosan A but not GM-CSF leads to an effective anti-tumor response in subcutaneous RG-2 gliomas.

PURPOSE: Nonspecific stimulation of cells of the immune system may be useful in generating an anti-tumor response for a variety of cancers and may work synergistically with currently available cytotoxic therapies. In this study we examined the response of syngeneic rat gliomas to treatment with several nonspecific stimulators of dendritic cells and macrophages alone or in combination with radiation therapy. EXPERIMENTAL DESIGN: RG-2 gliomas were implanted subcutaneously and treated with intratumoral (IT) injections of the toll-like receptor (TLR) ligands lipopolysaccharide (LPS) and zymosan A (ZymA) and the cytokine granulocyte-macrophage colony stimulating factor (GM-CSF). Combination treatment with IT LPS and single-fraction external beam radiotherapy (EBRT) was also evaluated. RESULTS: Treatment with IT LPS and ZymA delayed tumor growth compared to saline controls. Multiple doses of both substances were superior to single doses, and led to complete tumor regression in 71% (LPS) and 50% (ZymA) of animals. GM-CSF showed no anti-tumor effects in this study. Combinations of IT LPS and EBRT appeared to have a synergistic effect in delaying tumor growth. Rechallenge studies and IT LPS treatment of RG-2 tumors in nude rats suggested the importance of T cells in this treatment paradigm. CONCLUSIONS: Direct IT treatment with the TLR ligands LPS and ZymA are effective in generating an anti-tumor response. These treatments may synergize with cytotoxic therapies such as EBRT, and appear to require T cells for a successful outcome.

An investigation of the potential of rapid prototyping technology for image guided surgery.

Image-guided surgery can be broken down into two broad categories: frame-based guidance and frameless guidance. In order to reduce both the invasive nature of stereotactic guidance and the cost in equipment and time, we have developed a new guidance technique based on rapid prototyping (RP) technology. This new system first builds a computer model of the patient anatomy and then fabricates a physical reference frame that provides a precise and unique fit to the patient anatomy. This frame incorporates a means of guiding the surgeon along a preplanned surgical trajectory. This process involves (1) obtaining a high-resolution CT or MR scan, (2) building a computer model of the region of interest, (3) developing a surgical plan and physical guide, (4) designing a frame with a unique fit to the patient's anatomy with a physical linkage to the surgical guide, and (5) fabricating the frame using an RP unit. Software was developed to support these processes. To test the accuracy of this process, we first scanned and reproduced a plastic phantom fabricated to validate the system's ability to build an accurate virtual model. A target on the phantom was then identified, a surgical approach planned, a surgical guide designed, and the accuracy and precision of guiding a probe to that target were determined. Steps 1 through 5 were also evaluated using a head phantom. The results show that the RP technology can replicate an object from CT scans with submillimeter resolution. The fabricated reference frames, when positioned on the surface of the phantom and used to guide a surgical probe, can position the probe tip with an accuracy of 1.7 mm at the probe tip. These results demonstrate that the RP technology can be used for the fabrication of customized positioning frames for use in image-guided surgery.