Extracellular matrix regulation of cell spheroid invasion in a 3D bioprinted solid tumor-on-a-chip.

Tumor organoids and tumors-on-chips can be built by placing patient-derived cells within an engineered extracellular matrix (ECM) for personalized medicine. The engineered ECM influences the tumor response, and understanding the ECM-tumor relationship accelerates translating tumors-on-chips into drug discovery and development. In this work, we tuned the physical and structural characteristics of ECM in a 3D bioprinted soft-tissue sarcoma microtissue. We formed cell spheroids at a controlled size and encapsulated them into our gelatin methacryloyl (GelMA)-based bioink to make perfusable hydrogel-based microfluidic chips. We then demonstrated the scalability and customization flexibility of our hydrogel-based chip via engineering tools. A multiscale physical and structural data analysis suggested a relationship between cell invasion response and bioink characteristics. Tumor cell invasive behavior and focal adhesion properties were observed in response to varying polymer network densities of the GelMA-based bioink. Immunostaining assays and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) helped assess the bioactivity of the microtissue and measure the cell invasion. The RT-qPCR data showed higher expressions of HIF-1α, CD44, and MMP2 genes in a lower polymer density, highlighting the correlation between bioink structural porosity, ECM stiffness, and tumor spheroid response. This work is the first step in modeling STS tumor invasiveness in hydrogel-based microfluidic chips. STATEMENT OF SIGNIFICANCE: We optimized an engineering protocol for making tumor spheroids at a controlled size, embedding spheroids into a gelatin-based matrix, and constructing a perfusable microfluidic device. A higher tumor invasion was observed in a low-stiffness matrix than a high-stiffness matrix. The physical characterizations revealed how the stiffness is controlled by the density of polymer chain networks and porosity. The biological assays revealed how the structural properties of the gelatin matrix and hypoxia in tumor progression impact cell invasion. This work can contribute to personalized medicine by making more effective, tailored cancer models.

Cell encapsulation in gelatin methacryloyl bioinks impairs microscale diffusion properties.

Light-assisted bioprinted gelatin methacryloyl (GelMA) constructs have been used for cell-laden microtissues and organoids. GelMA can be loaded by desired cells, which can regulate the biophysical properties of bioprinted constructs. We study how the degree of methacrylation (MA degree), GelMA mass concentration, and cell density change mass transport properties. We introduce a fluorescent-microscopy-based method of biotransport testing with improved sensitivity compared to the traditional particle tracking methods. The diffusion capacity of GelMA with a higher MA significantly decreased compared to a lower MA. Opposed to a steady range of linear elastic moduli, the diffusion coefficient in GelMA varied when cell densities ranged from 0 to 10 × 106 cells/ml. A comparative study of different cell sizes showed a higher diffusivity coefficient for the case of larger cells. The results of this study can help bioengineers and scientists to better control the biotransport characteristics in light-assisted bioprinted microtissues and organoids.

MR Imaging of hypoxic ischemic encephalopathy - Distribution Patterns and ADC value correlations.

BACKGROUND AND PURPOSE: Neonatal hypoxic-ischemic encephalopathy causes hypoxic brain injury. Due to differences in brain maturity at time of insult, severity of hypotension and duration of insult, there are four distinct patterns of brain injury. Magnetic resonance imaging is the most sensitive modality for evaluating these patterns of brain injury. Additional role of Diffusion weighted imaging and ADC values can be useful in the evaluation of such cases. We conducted this study to analyse the usefulness of ADC values in the brain tissue affected by hypoxic-ischemic injury. MATERIALS AND METHODS: We conducted a prospective study of all the patients referred to our department for magnetic resonance scanning of brain with history of hypoxic ischemic encephalopathy and clinical features cerebral palsy. 23 Cases with imaging manifestations of hypoxic ischemic encephalopathy were included in the study. We studied distribution patterns of HIE in our cases and calculated the ADC values of involved as well as normal grey and white matter. Further, sensitivity, specificity, predictive values, and likelihood ratios for each dichotomized diffusion and ADC values were obtained Wilson Score method. RESULTS: The most common distribution pattern in our study was involvement of peri-rolandic area (15 cases, 65%). ADC values were significantly (p < 0.005) increased in abnormal white matter. No significant changes (p = 0.8) were seen in ADC values of normal and abnormal grey matter. CONCLUSIONS: Due to significant increase in ADC values of affected white matter, ADC value can be used as a marker to detect chronic sequel of hypoxic ischaemic brain injury. Another observation was the perirolandic brain tissue being most common area of involvement in the cases with cerebral palsy.

Superior semicircular canal dehiscence: A new perspective.

OBJECTIVE: To determine the use of multi-detector computed tomography (MDCT) in the diagnostic interpretation of superior semicircular canal dehiscence (SSCD) or thinning and its association with ear pathologies and to find whether it is an acquired condition and its association with increase in age. MATERIALS AND METHODS: study was performed in a tertiary care institute present in a village, following approval of the institutional ethical committee. Retrospective review of temporal bone CT examinations performed between September 2016 and March 2017 was done. 1 mm interval axial images with sagittal and coronal reformatted images were reviewed for the presence of canal dehiscence and thinning by investigators. We characterised the Superior semicircular canal status as normal, frank dehiscence or thinning. Frank dehiscence was further classified anatomically as anterior limb, apex and posterior limb dehiscence.The patient list was then subcategorized into 5 age groups, and the prevalence of SSCD was calculated for each group. RESULTS: Retrospective review yielded 80 positive cases which included SSC dehiscence (N = 39) and thinning (N = 41). 80 normal scans were selected as control group retrospectively. Statistical analysis was performed to assess for differences between the groups studied. Pearson chi-square test applied. there was a significant association of SSC pathologies prevalence with increasing age (p = < 0.001). No significant relationship was found between SSCD and presence of either CSOM or Cholesteatoma (p = 0.285). Vertigo rather than Tullio phenomenon was the statistically significant complaint (p = <0.001). which brought the patient to the hospital. CONCLUSIONS: The SSCD and thinning belong to the same spectrum and are acquired conditions. Increasing prevalence in old age suggests it to be an acquired condition rather than a congenital one. No significant association of these condition was seen with CSOM and cholesteatoma. Vertigo is the predominat symptom bringing the patient to hospital along with Tullio phenomenon.