Large Tissue Volumes Research Articles

Treatment of a patient with COVID-19 and adverse premorbid background: A case report

BACKGROUND: In the presence of severe comorbid pathology, children get sick with coronavirus disease 2019 (COVID-19) as severe as adults.
 CASE REPORT: This paper describes the treatment of a 17-year-old patient with severe bilateral pneumonia caused by severe acute respiratory syndrome coronavirus 2 that damage a large volume of the lung tissue (69% on the right, 87.1% on the left, and grade IV lung lesions according to computed tomography) with an unfavorable premorbid background (grade IV obesity with a body mass index of 54.5 kg/m2, Down syndrome, moderate mental retardation, and primary hypothyroidism). The child was admitted to the hospital on day 10 of illness in an extremely serious condition. On admission, she received artificial lung ventilation and then high-flow mask ventilation with an oxygen flow of 12 L/min. In addition to respiratory support, she received etiotropic (hydroxychloroquine), anticoagulant (enoxaparin), antibacterial, (ceftriaxone), and antihypertensive treatments. Levothyroxine sodium was administered for hypothyroidism. Permanent monitoring of the acidbase balance, general and biochemical blood tests, and coagulography were performed. Gradual positive dynamics of the respiratory status was observed, and oxygen flow decreased. The case was constantly consulted to a pulmonologist, endocrinologist, and cardiologist, who corrected antihypertensive therapy depending on blood pressure indicators. Oxygen support was given for 13 days. After receiving a double-negative PCR test for COVID-19, the patient was transferred to the pulmonology department, from which she was discharged in satisfactory condition for outpatient observation.
 CONCLUSION: Our patient with COVID-19, severe lung damage with a combination of comorbid pathologies, extremely unfavorable prognosis (grade IV obesity, Down syndrome, and hypothyroidism), who received active complex treatment in accordance with modern guidelines, recovered despite late admission to the hospital.

Electron microscopy for imaging organelles in plants and algae.

Recent developments in both instrumentation and image analysis algorithms have allowed three-dimensional electron microscopy (3D-EM) to increase automated image collections through large tissue volumes using serial block-face scanning EM (SEM) and to achieve near-atomic resolution of macromolecular complexes using cryo-electron tomography (cryo-ET) and sub-tomogram averaging. In this review, we discuss applications of cryo-ET to cell biology research on plant and algal systems and the special opportunities they offer for understanding the organization of eukaryotic organelles with unprecedently resolution. However, one of the most challenging aspects for cryo-ET is sample preparation, especially for multicellular organisms. We also discuss correlative light and electron microscopy (CLEM) approaches that have been developed for ET at both room and cryogenic temperatures.

Vegfr3-tdTomato, a reporter mouse for microscopic visualization of lymphatic vessel by multiple modalities.

Lymphatic vessels are indispensable for tissue fluid homeostasis, transport of solutes and dietary lipids and immune cell trafficking. In contrast to blood vessels, which are easily visible by their erythrocyte cargo, lymphatic vessels are not readily detected in the tissue context. Their invisibility interferes with the analysis of the three-dimensional lymph vessel structure in large tissue volumes and hampers dynamic intravital studies on lymphatic function and pathofunction. An approach to overcome these limitations are mouse models, which express transgenic fluorescent proteins under the control of tissue-specific promotor elements. We introduce here the BAC-transgenic mouse reporter strain Vegfr3-tdTomato that expresses a membrane-tagged version of tdTomato under control of Flt4 regulatory elements. Vegfr3-tdTomato mice inherited the reporter in a mendelian fashion and showed selective and stable fluorescence in the lymphatic vessels of multiple organs tested, including lung, kidney, heart, diaphragm, intestine, mesentery, liver and dermis. In this model, tdTomato expression was sufficient for direct visualisation of lymphatic vessels by epifluorescence microscopy. Furthermore, lymph vessels were readily visualized using a number of microscopic modalities including confocal laser scanning, light sheet fluorescence and two-photon microscopy. Due to the early onset of VEGFR-3 expression in venous embryonic vessels and the short maturation time of tdTomato, this reporter offers an interesting alternative to Prox1-promoter driven lymphatic reporter mice for instance to study the developmental differentiation of venous to lymphatic endothelial cells.

Correlative light and volume electron microscopy (vCLEM): How community participation can advance developing technologies.

Correlative light and electron microscopy is a valuable tool to image samples across resolution scales and link data on structure and function. While studies using this technique have been available since the 1960s, recent developments have enabled applying these workflows to large volumes of cells and tissues. Much of the development in this area has been facilitated through the collaborative efforts of microscopists and commercial companies to bring the methods, hardware and image processing technologies needed into laboratories and core imaging facilities. This is a prime example of how what was once a niche area can be brought into the mainstream of microscopy by the efforts of imaging pioneers who push the boundaries of possibility.

3D imaging of human organs with micrometer resolution - applied to the endocrine pancreas

The possibility to quantitatively study specific molecular/cellular features of complete human organs with preserved spatial 3D context would have widespread implications for pre-clinical and clinical medicine. Whereas optical 3D imaging approaches have experienced a formidable revolution, they have remained limited due to current incapacities in obtaining specific labelling within large tissue volumes. We present a simple approach enabling reconstruction of antibody labeled cells within entire human organs with preserved organ context. We demonstrate the utility of the approach by providing volumetric data and 3D distribution of hundreds of thousands of islets of Langerhans within the human pancreas. By assessments of pancreata from non-diabetic and type 2 diabetic individuals, we display previously unrecognized features of the human islet mass distribution and pathology. As such, this method may contribute not only in unraveling new information of the pancreatic anatomy/pathophysiology, but it may be translated to essentially any antibody marker or organ system.

Expansion tomography for large volume tissue imaging with nanoscale resolution.

Expansion microscopy enables conventional diffraction limit microscopy to achieve super-resolution imaging. However, the enlarged tissue lacks an objective lens with sufficient working distance that can image tissues with whole-brain-scale coverage. Here, we present expansion tomography (ExT) to solve this problem. We have established a modified super-absorbent hydrogel (ExT gel) that possesses high mechanical strength and enables serial sectioning. ExT gel enables tissue and cell imaging and is compatible with various fluorescent labeling strategies. Combining with the high-throughput light-sheet tomography (HLTP) system, we have shown the capability of large volume imaging with nanoscale resolution of mouse brain intact neuronal circuits. The ExT method would allow image samples to support super-resolution imaging of intact tissues with virtually unlimited axial extensions.

Technical note: On the impact of the kV imaging configuration on doses from planar images during motion-synchronized treatments on Radixact®.

Kilovoltage radiographs are acquired during motion‐synchronized treatments on Radixact to localize the tumor during the treatment. Several previous publications have provided estimates of patient dose from these planar radiographs. However, a recent hardware update changed several aspects of the kV imaging system, including a new X‐ray tube, an extended source‐to‐axis distance (SAD), and a larger field size. This is denoted the extended configuration. The purpose of this work was to assess the impact of the configuration change on patient dose from these procedures. Point doses in water were measured using the TG‐61 protocol for tube potentials between 100 and 140 kVp for both the standard and extended configurations under the same water tank setup. Comparisons were made for equal mAs since the same protocols (kVp, mAs) will be used for both configurations. In comparison to the standard configuration, doses per mAs from the extended configuration were found to be ~66% less and falloff less steep due to the increased SAD. However, a larger volume of tissue is irradiated due to the larger field size. Beam quality for a given tube potential was the same as determined by half‐value layer measurements. Both kV configurations are available from the vendor, therefore, the values in this work can be used to compare values previously published in the literature for the standard configuration or to intercompare doses from these two system configurations.

Deep learning-based autofocus method enhances image quality in light-sheet fluorescence microscopy.

Light-sheet fluorescence microscopy (LSFM) is a minimally invasive and high throughput imaging technique ideal for capturing large volumes of tissue with sub-cellular resolution. A fundamental requirement for LSFM is a seamless overlap of the light-sheet that excites a selective plane in the specimen, with the focal plane of the objective lens. However, spatial heterogeneity in the refractive index of the specimen often results in violation of this requirement when imaging deep in the tissue. To address this issue, autofocus methods are commonly used to refocus the focal plane of the objective-lens on the light-sheet. Yet, autofocus techniques are slow since they require capturing a stack of images and tend to fail in the presence of spherical aberrations that dominate volume imaging. To address these issues, we present a deep learning-based autofocus framework that can estimate the position of the objective-lens focal plane relative to the light-sheet, based on two defocused images. This approach outperforms or provides comparable results with the best traditional autofocus method on small and large image patches respectively. When the trained network is integrated with a custom-built LSFM, a certainty measure is used to further refine the network's prediction. The network performance is demonstrated in real-time on cleared genetically labeled mouse forebrain and pig cochleae samples. Our study provides a framework that could improve light-sheet microscopy and its application toward imaging large 3D specimens with high spatial resolution.

Modeling the Effect of Binding Kinetics in Spatial Drug Distribution in the Brain.

A 3-dimensional mathematical model is developed to determine the effect of drug binding kinetics on the spatial distribution of a drug within the brain. The key components, namely, transport across the blood-brain barrier (BBB), drug distribution in the brain extracellular fluid (ECF), and drug binding kinetics are coupled with the bidirectional bulk flow of the brain ECF to enhance the visualization of drug concentration in the brain. The model is developed based on the cubical volume of a brain unit, which is a union of three subdomains: the brain ECF, the BBB, and the blood plasma. The model is a set of partial differential equations and the associated initial and boundary conditions through which the drug distribution process in the mentioned subdomains is described. Effects of drug binding kinetics are investigated by varying the binding parameter values for both nonspecific and specific binding sites. All variations of binding parameter values are discussed, and the results show the improved visualization of the effect of binding kinetics in the drug distribution within the brain. For more realistic visualization, we suggest incorporating more brain components that make up the large volume of the brain tissue.

Dose-dependence of radiotherapy-induced changes in serum levels of choline-containing phospholipids; the importance of lower doses delivered to large volumes of normal tissues

BackgroundConformal radiotherapy is a primary treatment in head and neck cancer, which putative adverse effects depend on relatively low doses of radiation delivered to increased volumes of normal tissues. Systemic effects of such treatment include radiation-induced changes in serum lipid profile, yet dose- and volume-dependence of these changes remain to be established.MethodsHere we analyzed levels of choline-containing phospholipids in serum samples collected consecutively during the radiotherapy used as the only treatment modality. The liquid chromatography–mass spectrometry (LC-MS) approach applied in the study enabled the detection and quantitation of 151 phospholipids, including (lyso)phosphatidylcholines and sphingomyelins.ResultsNo statistically significant differences were found in the pretreatment samples from patients with different locations and stages of cancer. To compensate for potential differences between schemes of radiotherapy, the biologically effective doses were calculated and used in the search of correlations with specific lipid levels. We found that the levels of several phospholipids depended on the maximum dose delivered to the gross tumor volume and total radiation energy absorbed by the patient’s body. Increased doses correlated with increased levels of sphingomyelins and reduced levels of phosphatidylcholines. Furthermore, we observed several phospholipids whose serum levels correlated with the degree of acute radiation toxicity.ConclusionNoteworthy, serum phospholipid levels were associated mainly with volumes of normal tissues irradiated with relatively low doses (i.e., total accumulated dose 20 Gy), which indicated the importance of such effects on the systemic response of the patient’s organism to intensity-modulated radiotherapy (IMRT).

Conditioning the microenvironment for soft tissue regeneration in a cell free scaffold

The use of cell-free scaffolds for the regeneration of clinically relevant volumes of soft tissue has been challenged, particularly in the case of synthetic biomaterials, by the difficulty of reconciling the manufacturing and biological performance requirements. Here, we investigated in vivo the importance of biomechanical and biochemical cues for conditioning the 3D regenerative microenvironment towards soft tissue formation. In particular, we evaluated the adipogenesis changes related to 3D mechanical properties by creating a gradient of 3D microenvironments with different stiffnesses using 3D Poly(Urethane-Ester-ether) PUEt scaffolds. Our results showed a significant increase in adipose tissue proportions while decreasing the stiffness of the 3D mechanical microenvironment. This mechanical conditioning effect was also compared with biochemical manipulation by loading extracellular matrices (ECMs) with a PPAR-γ activating molecule. Notably, results showed mechanical and biochemical conditioning equivalency in promoting adipose tissue formation in the conditions tested, suggesting that adequate mechanical signaling could be sufficient to boost adipogenesis by influencing tissue remodeling. Overall, this work could open a new avenue in the design of synthetic 3D scaffolds for microenvironment conditioning towards the regeneration of large volumes of soft and adipose tissue, with practical and direct implications in reconstructive and cosmetic surgery.

Penumbra Consumption Rates Based on Time-to-Maximum Delay and Reperfusion Status: A Post Hoc Analysis of the DEFUSE 3 Trial

In patients with acute large vessel occlusion, the natural history of penumbral tissue based on perfusion time-to-maximum (Tmax) delay is not well established in relation to late-window endovascular thrombectomy. In this study, we sought to evaluate penumbra consumption rates for Tmax delays in patients with large vessel occlusion evaluated between 6 and 16 hours from last known normal. This is a post hoc analysis of the DEFUSE 3 trial (The Endovascular Therapy Following Imaging Evaluation for Ischemic Stroke), which included patients with an acute ischemic stroke due to anterior circulation occlusion within 6 to 16 hours of last known normal. The primary outcome is percentage penumbra consumption, defined as (24-hour magnetic resonance imaging infarct volume-baseline core infarct volume)/(Tmax 6 or 10 s volume-baseline core volume). We stratified the cohort into 4 categories based on treatment modality and Thrombolysis in Cerebral Infarction (TICI score; untreated, TICI 0-2a, TICI 2b, and TICI3) and calculated penumbral consumption rates in each category. We included 141 patients, among whom 68 were untreated. In the untreated versus TICI 3 patients, a median (interquartile range) of 53.7% (21.2%-87.7%) versus 5.3% (1.1%-14.6%) of penumbral tissue was consumed based on Tmax >6 s (P<0.001). In the same comparison for Tmax>10 s, we saw a difference of 165.4% (interquartile range, 56.1%-479.8%) versus 25.7% (interquartile range, 3.2%-72.1%; P<0.001). Significant differences were not demonstrated between untreated and TICI 0-2a patients for penumbral consumption based on Tmax >6 s (P=0.52) or Tmax >10 s (P=0.92). Among extended window endovascular thrombectomy patients, Tmax >10-s mismatch volume may comprise large volumes of salvageable tissue, whereas nearly half the Tmax >6-s mismatch volume may remain viable in untreated patients at 24 hours.

High Resolution Optical Coherence Tomography

Imaging cellular and extracellular structures and processes in situ and in vivo is highly desired for the understanding and diagnosis of human diseases. High resolution optical coherence tomography (HR-OCT) is particularly suitable for this task because it can provide real-time, 3-D images of a large tissue volume at subcellular resolution. Over the past two decades, tremendous technical advances have been made to tackle the fundamental and practical limitations of HR-OCT for applications in various clinical fields. Meanwhile, novel scientific and clinical applications of HR-OCT have also been proposed and validated. This review aims to provide an update on the progress of technology development, with a focus on axial focus extension, aberration correction, and fiber-optic probe development. This review also aims to summarize the newly established capabilities of HR-OCT in fundamental and clinical research, in an effort to promote multidisciplinary research using this powerful imaging tool.

Implantation of Neuropixels probes for chronic recording of neuronal activity in freely behaving mice and rats.

How dynamic activity in neural circuits gives rise to behavior is a major area of interest in neuroscience. A key experimental approach for addressing this question involves measuring extracellular neuronal activity in awake, behaving animals. Recently developed Neuropixels probes have provided a step change in recording neural activity in large tissue volumes with high spatiotemporal resolution. This protocol describes the chronic implantation of Neuropixels probes in mice and rats using compact and reusable 3D-printed fixtures. The fixtures facilitate stable chronic in vivo recordings in freely behaving rats and mice. They consist of two parts: a covered main body and a skull connector. Single-, dual- and movable-probe fixture variants are available. After completing an experiment, probes are safely recovered for reimplantation by a dedicated retrieval mechanism. Fixture assembly and surgical implantation typically take 4-5 h, and probe retrieval takes ~30 min, followed by 12 h of incubation in probe cleaning agent. The duration of data acquisition depends on the type of behavioral experiment. Since our protocol enables stable, chronic recordings over weeks, it enables longitudinal large-scale single-unit data to be routinely obtained in a cost-efficient manner, which will facilitate many studies in systems neuroscience.

Blood Flow Within Bioengineered 3D Printed Vascular Constructs Using the Porcine Model.

Recently developed biofabrication technologies are enabling the production of three-dimensional engineered tissues containing vascular networks which can deliver oxygen and nutrients across large tissue volumes. Tissues at this scale show promise for eventual regenerative medicine applications; however, the implantation and integration of these constructs in vivo remains poorly studied. Here, we introduce a surgical model for implantation and direct in-line vascular connection of 3D printed hydrogels in a porcine arteriovenous shunt configuration. Utilizing perfusable poly(ethylene glycol) diacrylate (PEGDA) hydrogels fabricated through projection stereolithography, we first optimized the implantation procedure in deceased piglets. Subsequently, we utilized the arteriovenous shunt model to evaluate blood flow through implanted PEGDA hydrogels in non-survivable studies. Connections between the host femoral artery and vein were robust and the patterned vascular channels withstood arterial pressure, permitting blood flow for 6 h. Our study demonstrates rapid prototyping of a biocompatible and perfusable hydrogel that can be implanted in vivo as a porcine arteriovenous shunt, suggesting a viable surgical approach for in-line implantation of bioprinted tissues, along with design considerations for future in vivo studies. We further envision that this surgical model may be broadly applicable for assessing whether biomaterials optimized for 3D printing and cell function can also withstand vascular cannulation and arterial blood pressure. This provides a crucial step toward generated transplantable engineered organs, demonstrating successful implantation of engineered tissues within host vasculature.

Effectiveness of surface mould brachytherapy in electron era.

e21562 Background: In India, skin cancers constitute about 1-2% of all diagnosed cancers.Surgery stays main modality of treatment with adequate surgical margin shown excellent local control rates - generally 95%.However, conditions where surgery is not feasible, brachytherapy is alternative approach.Advanced technology improved the ability to deliver safe and effective radiotherapy, resulting in renewed interest in this modality.Brachytherapy has many advantages: they can deposit a significantly higher dose within a tumor, with better sparing of adjacent normal structures over external beam radiotherapy or electron therapy, which sometimes require irradiating a large volume of tissue in order to provide adequately coverage, which in term increase toxicity. The purpose of our paper is to find out the clinical outcomes of skin tumors and superficial tumours of head and neck region treated with Surface mould brachytherapy (SMB), in terms of survival and toxicity.We report on our institution’s experience with using Co-60 based HDR SMB for the treatment of skin and head and neck superficial malignancies. Methods: A retrospective review of all patients treated with surface mould Co-60-based HDR brachytherapy at our center.A total of 23 patients, with 23 lesions, were treated during this period, and included in the analysis. A total of three of these lesions were treated with a palliative intent, and were included. The most common fractionation scheme was 35 Gy in 10 fractions given daily (52%, n = 12), though a range of doses were used, from 8/2 fractions to 36 Gy/12 fractions. Results: Median age at diagnosis was 52 (range = 28-91). The majority were Squamous cell (43%, n = 10) or Basal cell carcinomas (34%, n = 8). Most lesions were located in the head and neck region. The most commonly used RT dose was 3GyX10 fractions; all patients had individualized CT-based planning.The 5-year overall survival (OS) was 86 %( twenty patients). Three patients were died. Most deaths were from unrelated causes. Response was assessed in OPD 2-4 months post-treatment. Our complete response (CR) rate was 73.9% (seventeen patients), with partial response in three patients; two patients could not be assessed for response and one patient died due to other medical condition. We report a 2-year local control (LC) rate of 91.3%, and local recurrence was found in two patients. The procedure was well tolerated, with no grade 3 or more acute or late toxicities. There was one case of grade 3 ulcer (CTCAE). The 100% isodose line median depth was 0.5 cm, and median surface dose = 126.5%. The median V90 = 92.3%. Conclusions: Surface mould brachytherapy is a safe, effective modality for treatment of skin malignancies or tumors. Brachytherapy was overall well tolerated, with no grade 3-5 acute or late toxicities. This treatment is a good alternative option for those patients unwilling or unable to undergo surgery for their skin malignancies

Severe acute respiratory syndrome coronavirus 2 vaccination for patients with solid cancer: Review and point of view of a French oncology intergroup (GCO, TNCD, UNICANCER)

The impacts of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic on cancer care are multiple, entailing a high risk of death from coronavirus disease 2019 (COVID-19) in patients with cancer treated by chemotherapy. SARS-CoV-2 vaccines represent an opportunity to decrease the rate of severe COVID-19 cases in patients with cancer and also to restore normal cancer care. Patients with cancer to be targeted for vaccination are difficult to define owing to the limited contribution of these patients in the phase III trials testing the different vaccines. It seems appropriate to vaccinate not only patients with cancer with ongoing treatment or with a treatment having been completed less than 3 years ago but also household and close contacts. High-risk patients with cancer who are candidates for priority access to vaccination are those treated by chemotherapy. The very high-priority population includes patients with curative treatment and palliative first- or second-line chemotherapy, as well as patients requiring surgery or radiotherapy involving a large volume of lung, lymph node and/or haematopoietic tissue. When possible, vaccination should be carried out before cancer treatment begins. SARS-CoV-2 vaccination can be performed during chemotherapy while avoiding periods of neutropenia and lymphopenia. For organisational reasons, vaccination should be performed in cancer care centres with messenger RNA vaccines (or non-replicating adenoviral vaccines in non-immunocompromised patients). Considering the current state of knowledge, the benefit-risk ratio strongly favours SARS-CoV-2 vaccination of all patients with cancer. To obtain more data concerning the safety and effectiveness of vaccines, it is necessary to implement cohorts of vaccinated patients with cancer.

Multi-Electrode Architecture Modeling and Optimization for Homogeneous Electroporation of Large Volumes of Tissue

Electroporation is a phenomenon that consists of increasing the permeability of the cell membrane by means of high-intensity electric field application. Nowadays, its clinical application to cancer treatment is one of the most relevant branches within the many areas of electroporation. In this area, it is essential to apply homogeneous treatments to achieve complete removal of tumors and avoid relapse. It is necessary to apply an optimized transmembrane potential at each point of the tissue by means of a homogenous electric field application and appropriated electric field orientation. Nevertheless, biological tissues are composed of wide variety, heterogeneous and anisotropic structures and, consequently, predicting the applied electric field distribution is complex. Consequently, by applying the parallel-needle electrodes and single-output generators, homogeneous and predictable treatments are difficult to obtain, often requiring several repositioning/application processes that may leave untreated areas. This paper proposes the use of multi-electrode structure to apply a wide range of electric field vectors to enhance the homogeneity of the treatment. To achieve this aim, a new multi-electrode parallel-plate configuration is proposed to improve the treatment in combination with a multiple-output generator. One method for optimizing the electric field pattern application is studied, and simulation and experimental results are presented and discussed, proving the feasibility of the proposed approach.

A Linearized Fit Model for Robust Shape Parameterization of FET-PET TACs.

The kinetic analysis of [Formula: see text]-FET time-activity curves (TAC) can provide valuable diagnostic information in glioma patients. The analysis is most often limited to the average TAC over a large tissue volume and is normally assessed by visual inspection or by evaluating the time-to-peak and linear slope during the late uptake phase. Here, we derived and validated a linearized model for TACs of [Formula: see text]-FET in dynamic PET scans. Emphasis was put on the robustness of the numerical parameters and how reliably automatic voxel-wise analysis of TAC kinetics was possible. The diagnostic performance of the extracted shape parameters for the discrimination between isocitrate dehydrogenase (IDH) wildtype (wt) and IDH-mutant (mut) glioma was assessed by receiver-operating characteristic in a group of 33 adult glioma patients. A high agreement between the adjusted model and measured TACs could be obtained and relative, estimated parameter uncertainties were small. The best differentiation between IDH-wt and IDH-mut gliomas was achieved with the linearized model fitted to the averaged TAC values from dynamic FET PET data in the time interval 4-50 min p.i.. When limiting the acquisition time to 20-40 min p.i., classification accuracy was only slightly lower (-3%) and was comparable to classification based on linear fits in this time interval. Voxel-wise fitting was possible within a computation time ≈ 1 min per image slice. Parameter uncertainties smaller than 80% for all fits with the linearized model were achieved. The agreement of best-fit parameters when comparing voxel-wise fits and fits of averaged TACs was very high (p < 0.001).

Interstitial fluid velocity is decreased around cortical bone vascular pores and depends on osteocyte position in a rat model of disuse osteoporosis.

Muscle paralysis induced with botulinum toxin (Botox) injection increases vascular porosity and reduces osteocyte lacunar density in the tibial cortical bone of skeletally mature rats. These morphological changes potentially affect interstitial fluid flow in the lacunar-canalicular porosity, which is thought to play a role in osteocyte mechanotransduction. The aim of this study was to investigate the effects of disuse-induced morphological changes on interstitial fluid velocity around osteocytes in the bone cortex. Micro-CT images from a previous study that quantified the effects of Botox-induced muscle paralysis on bone microarchitecture in skeletally mature rats were used to create high-resolution, animal-specific finite element models that included the vascular pores and osteocyte lacunae within the tibial metaphysis of Botox-injected (BTX, n = 8) and saline-injected control (CTRL, n = 8) groups. To quantify fluid flow, lacunar and canalicular porosities were modeled as fluid-saturated poroelastic materials, and boundary conditions were applied to simulate physiological loading. This modeling approach allowed a detailed quantification of the fluid flow velocities around osteocytes in a relatively large volume of bone tissue. The analysis demonstrated that interstitial fluid velocity at the vascular pore surfaces was significantly lower in BTX compared to CTRL because of the decreased vascular canal separation. No significant differences in average fluid velocity were observed at the osteocyte lacunae and no correlation was found between the fluid velocity and the lacunar density, which was significantly lower in BTX. Instead, the lacunar fluid velocity was dependent on the osteocyte's specific position in the bone cortex and its proximity to a vascular pore.