Mechanical mapping of the prostate in vivo using Dynamic Instrumented Palpation; towards an in vivo strategy for cancer assessment.

A calibrated palpation sensor has been developed for making instrumented Digital Rectal Examinations (iDREs) with a view to assessing patients for prostate cancer. The instrument measures the dynamic stiffness of the palpable surface of the prostate, and has been trialled on 12 patients in vivo. The patients had been diagnosed with prostate cancer and were scheduled for radical prostatectomy. As far as possible, patients with asymmetric disease were chosen so as to give a variation in gland condition over the palpable surface. The device works by applying an oscillating pressure (force) to a flexible probe whose displacement into the tissue is also measured in order to yield a dynamic stiffness, the static stiffness being incidentally measured at the mean oscillatory force. The device was deployed mounted on the index finger of a urologist and measurements taken at 12-16 positions on each patient using light and firm pressure and palpation frequencies of 1 or 5 Hz. In parallel, conventional DRE assessments were made by a consultant urologist for cancer. After in vivo measurement, the glands were removed and examined histologically with each palpation point being classified as cancerous (C) or not (NC). The work has established the first measurements of static modulus of living prostate tissue to be: 26.8 (13.3) kPa for tissue affected by prostate cancer (C classification), and 24.8 kPa (11.9) for tissue unaffected by cancer (NC classification), values quoted as median (interquartile range). The dynamic properties were characterised by: dynamic modulus, 5.15 kPa (4.86) for the C classification and 4.61 kPa (3.08) for the NC classification and the time lag between force and displacement at 5 Hz palpation frequency, 0.0175 s (0.0078) for the C classification and 0.0186 s (0.0397) for the NC classification, values again quoted as median (interquartile range). With the limited set of features that could be generated, an Artificial Neural Network (ANN) classification yielded a sensitivity of 97%, negative predictive value of 86%, positive predictive value of 67% and accuracy of 70% but with relatively poor specificity (30%). Besides extending the feature set, there are a number of changes in probe design, probing strategy and in mechanics analysis, which are expected to improve the diagnostic capabilities of the method.

Three dimensional in vitro models of cancer: Bioprinting multilineage glioblastoma models.

Three dimensional (3D) bioprinting of multiple cell types within optimised extracellular matrices has the potential to more closely model the 3D environment of human physiology and disease than current alternatives. In this study, we used a multi-nozzle extrusion bioprinter to establish models of glioblastoma made up of cancer and stromal cells printed within matrices comprised of alginate modified with RGDS cell adhesion peptides, hyaluronic acid and collagen-1. Methods were developed using U87MG glioblastoma cells and MM6 monocyte/macrophages, whilst more disease relevant constructs contained glioblastoma stem cells (GSCs), co-printed with glioma associated stromal cells (GASCs) and microglia. Printing parameters were optimised to promote cell-cell interaction, avoiding the 'caging in' of cells due to overly dense cross-linking. Such printing had a negligible effect on cell viability, and cells retained robust metabolic activity and proliferation. Alginate gels allowed the rapid recovery of printed cell protein and RNA, and fluorescent reporters provided analysis of protein kinase activation at the single cell level within printed constructs. GSCs showed more resistance to chemotherapeutic drugs in 3D printed tumour constructs compared to 2D monolayer cultures, reflecting the clinical situation. In summary, a novel 3D bioprinting strategy is developed which allows control over the spatial organisation of tumour constructs for pre-clinical drug sensitivity testing and studies of the tumour microenvironment.

Quantitative mechanical assessment of the whole prostate gland ex vivo using dynamic instrumented palpation.

An instrumented palpation sensor, designed for measuring the dynamic modulus of tissue in vivo, has been developed and trialled on ex vivo whole prostate glands. The sensor consists of a flexible membrane sensor/actuator with an embedded strain gauge and is actuated using a dynamically varying airflow at frequencies of 1 and 5 Hz. The device was calibrated using an indentation stiffness measurement rig and gelatine samples with a range of static modulus similar to that reported in the literature for prostate tissue. The glands were removed from patients with diagnosed prostate cancer scheduled for radical prostatectomy, and the stiffness was measured within 30 min of surgical removal. Each prostate was later examined histologically in a column immediately below each indentation point and graded into one of the four groups; normal, benign prostatic hyperplasia, cancerous and mixed cancer and benign prostatic hyperplasia. In total, 11 prostates were assessed using multiple point probing, and the complex modulus at 1 and 5 Hz was calculated on a point-by-point basis. The device yielded values of quasi-static modulus of 15 ± 0.5 kPa and dynamic modulus of 20 ± 0.5 kPa for whole prostates, and a sensitivity of up to 80% with slightly lower specificity was achieved on diagnosis of prostate cancer using a combination of mechanical measures. This assessment did not take into account some obvious factors such as edge effects, overlap and clinical significance of the cancer, all of which would improve performance. The device, as currently configured, is immediately deployable in vivo. A number of improvements are also identified which could improve the sensitivity and specificity in future embodiments of the probe.