Effect of CO2 Preservation Treatments on the Sensory Quality of Pomegranate Juice.

Due to the interest in identifying cost-effective techniques that can guarantee the microbiological, nutritional, and sensorial aspects of food products, this study investigates the effect of CO2 preservation treatment on the sensory quality of pomegranate juice at t0 and after a conservation period of four weeks at 4 °C (t28). The same initial batch of freshly squeezed non-treated (NT) juice was subjected to non-thermal preservation treatments with supercritical carbon dioxide (CO2), and with a combination of supercritical carbon dioxide and ultrasound (CO2-US). As control samples, two other juices were produced from the same NT batch: A juice stabilized with high pressure treatment (HPP) and a juice pasteurized at high temperature (HT), which represent an already established non-thermal preservation technique and the conventional thermal treatment. Projective mapping and check-all-that-apply methodologies were performed to determine the sensory qualitative differences between the juices. The volatile profile of the juices was characterized by gas chromatography-mass spectrometry. The results showed that juices treated with supercritical CO2 could be differentiated from NT, mainly by the perceived odor and volatile compound concentration, with a depletion of alcohols, esters, ketones, and terpenes and an increase in aldehydes. For example, in relation to the NT juice, limonene decreased by 95% and 90%, 1-hexanol decreased by 9% and 17%, and camphene decreased by 94% and 85% in the CO2 and CO2-US treated juices, respectively. Regarding perceived flavor, the CO2-treated juice was not clearly differentiated from NT. Changes in the volatile profile induced by storage at 4 °C led to perceivable differences in the odor quality of all juices, especially the juice treated with CO2-US, which underwent a significant depletion of all major volatile compounds during storage. The results suggest that the supercritical CO2 process conditions need to be optimized to minimize impacts on sensory quality and the volatile profile.

High-efficiency cellular reprogramming with microfluidics.

We report that the efficiency of reprogramming human somatic cells to induced pluripotent stem cells (hiPSCs) can be dramatically improved in a microfluidic environment. Microliter-volume confinement resulted in a 50-fold increase in efficiency over traditional reprogramming by delivery of synthetic mRNAs encoding transcription factors. In these small volumes, extracellular components of the TGF-ß and other signaling pathways exhibited temporal regulation that appears critical to acquisition of pluripotency. The high quality and purity of the resulting hiPSCs (µ-hiPSCs) allowed direct differentiation into functional hepatocyte- and cardiomyocyte-like cells in the same platform without additional expansion.

Long-term microfluidic tracking of coccoid cyanobacterial cells reveals robust control of division timing.

BACKGROUND: Cyanobacteria are important agents in global carbon and nitrogen cycling and hold great promise for biotechnological applications. Model organisms such as Synechocystis sp. and Synechococcus sp. have advanced our understanding of photosynthetic capacity and circadian behavior, mostly using population-level measurements in which the behavior of individuals cannot be monitored. Synechocystis sp. cells are small and divide slowly, requiring long-term experiments to track single cells. Thus, the cumulative effects of drift over long periods can cause difficulties in monitoring and quantifying cell growth and division dynamics. RESULTS: To overcome this challenge, we enhanced a microfluidic cell-culture device and developed an image analysis pipeline for robust lineage reconstruction. This allowed simultaneous tracking of many cells over multiple generations, and revealed that cells expand exponentially throughout their cell cycle. Generation times were highly correlated for sister cells, but not between mother and daughter cells. Relationships between birth size, division size, and generation time indicated that cell-size control was inconsistent with the "sizer" rule, where division timing is based on cell size, or the "timer" rule, where division occurs after a fixed time interval. Instead, single cell growth statistics were most consistent with the "adder" rule, in which division occurs after a constant increment in cell volume. Cells exposed to light-dark cycles exhibited growth and division only during the light period; dark phases pause but do not disrupt cell-cycle control. CONCLUSIONS: Our analyses revealed that the "adder" model can explain both the growth-related statistics of single Synechocystis cells and the correlation between sister cell generation times. We also observed rapid phenotypic response to light-dark transitions at the single cell level, highlighting the critical role of light in cyanobacterial cell-cycle control. Our findings suggest that by monitoring the growth kinetics of individual cells we can build testable models of circadian control of the cell cycle in cyanobacteria.

High Temporal Resolution Detection of Patient-Specific Glucose Uptake from Human ex Vivo Adipose Tissue On-Chip.

Human tissue in vitro models on-chip are highly desirable to dissect the complexity of a physio-pathological in vivo response because of their advantages compared to traditional static culture systems in terms of high control of microenvironmental conditions, including accurate perturbations and high temporal resolution analyses of medium outflow. Human adipose tissue (hAT) is a key player in metabolic disorders, such as Type 2 Diabetes Mellitus (T2DM). It is involved in the overall energy homeostasis not only as passive energy storage but also as an important metabolic regulator. Here, we aim at developing a large scale microfluidic platform for generating high temporal resolution of glucose uptake profiles, and consequently insulin sensitivity, under physio-pathological stimulations in ex vivo adipose tissues from nondiabetic and T2DM individuals. A multiscale mathematical model that integrates fluid dynamics and an intracellular insulin signaling pathway description was used for assisting microfluidic design in order to maximize measurement accuracy of tissue metabolic activity in response to perturbations. An automated microfluidic injection system was included on-chip for performing precise dynamic biochemical stimulations. The temporal evolution of culture conditions could be monitored for days, before and after perturbation, measuring glucose concentration in the outflow with high temporal resolution. As a proof of concept for detection of insulin resistance, we measured insulin-dependent glucose uptake by hAT from nondiabetic and T2DM subjects, mimicking the postprandial response. The system presented thus represents an important tool in dissecting the role of single tissues, such as hAT, in the complex interwoven picture of metabolic diseases.

Investigating the Effect of Rosemary Essential Oil, Supercritical CO2 Processing and Their Synergism on the Quality and Microbial Inactivation of Chicken Breast Meat.

Fresh chicken meat is a very perishable good, even at refrigerated storage conditions, due to psychrophilic microbial growth and physicochemical changes. The present study focuses on the use of rosemary (Rosmarinus officinalis L.) essential oil (REO), supercritical CO2 processing and their synergism to increase the microbial inactivation in chicken breast meat. E. coli and L. innocua were inoculated on the chicken breast surface, and the inactivation effects of two different processes, namely SC-CO2 and SC-MAPCO2, were compared with or without the addition of REO. Moreover, the impact of the treatments on the superficial color of the meat was considered. The study demonstrated a synergic effect with 1% REO and supercritical CO2 for the inactivation of E. coli on chicken meat, while for L. innocua, there was no synergism. Regarding SC-CO2 treatment, the E. coli reduction was 1.29 and 3.31 log CFU/g, while for L. innocua, it was 1.42 and 1.11 log CFU/g, respectively, without and with the addition of 1.0% of REO. The same amount of REO allowed us to obtain a reduction of 1.3 log CFU/g of E. coli when coupled with SC-MAPCO2. For L. innocua, no reduction was obtained, either with SC-MAPCO2 or together with REO. The synergism of SC-MAPCO2 with 1% REO was confirmed for the total psychrophilic bacteria, demonstrating a strong dependence on the microorganism. The color modification induced by the SC-MAPCO2 process was lower than the SC-CO2 treatment. Overall, this study demonstrated a possible synergism of the technologies which can support the development of innovative methods to improve the safety and shelf-life of chicken breast meat.

Training in tools to develop quantitative microbial risk assessment along the food chain of Spanish products.

Food safety is a widespread challenge. Every year it is estimated that almost 1 in 10 people in the world fall ill after eating contaminated food resulting in over 400,000 deaths. The risk of outbreaks is higher when consuming ready-to-eat (RTE) products because they are eaten without a further cooking process that could inactivate pathogenic microorganisms. Hence, food processing is essential to increase the safety of RTE products. Microbiological risk assessment (MRA) integrates food science, microbiology and data science to provide a comprehensive understanding of the safety of the food system. MRA provides qualitative and/or quantitative information to decision makers, which might promote the adoption of better food practices. In this contest, this project aims to study and implement tools for quantitative microbial risk assessment (QMRA) of food products along the food chain. A common RTE product (cured ham) from Spain was used as a case study. Following, the exposure assessment model was implemented using mathematical models and statistical software to describe the microbial behaviour along the food chain. The study presents the possibility to identify the risk exposure in different scenarios (e.g. growth during different storage conditions, inactivation induced by traditional or innovative decontamination techniques), showing the flexibility of the predictive tools developed.

Increasing the Safety and Storage of Pre-Packed Fresh-Cut Fruits and Vegetables by Supercritical CO2 Process.

This work presents a feasibility lab-scale study for a new preservation method to inactivate microorganisms and increase the shelf life of pre-packed fresh-cut products. Experiments were conducted on coriander leaves and fresh-cut carrots and coconut. The technology used the combination of hydrostatic pressure (<15 MPa), low temperature (≤45 °C), and CO2 modified atmosphere packaging (MAP). The inactivation was achieved for the naturally present microorganisms (total mesophilic bacteria, yeasts and molds, total coliforms) and inoculated E. coli. Yeasts and molds and coliform were under the detection limit in all the treated samples, while mesophiles were strongly reduced, but below the detection limit only in carrots. Inoculated E. coli strains were completely inactivated (>6.0 log CFU/g) on coconut, while a reduction >4.0 log CFU/g was achieved for carrots and coriander. For all the treated products, the texture was similar to the fresh ones, while a small alteration of color was detected. Microbiological stability was achieved for up to 14 days for both fresh-cut carrots and coconut. Overall, the results are promising for the development of a new mild and innovative food preservation technique for fresh food.

Promoting the preservation of strawberry by supercritical CO2 drying.

This work aimed to investigate the supercritical CO2 (ScCO2) drying of strawberries and its effect on enzymatic, chemical and microbial stability. Process conditions influenced the final weight loss (WL), water activity (aw) and the inactivation of polyphenol oxidase (PPO) and peroxidase (POD). At 40 °C, an efficient drying (WL > 92 %, aw < 0.34) and a complete enzymatic (POD and PPO activity) inactivation can be achieved using several combinations of pressure, time and flow rate. ScCO2 dried strawberry at 40 °C, 13.3 MPa, 7 h and 19 kg/h flow rate maintain the total content of Vitamin C (358.5 mg/100 g), 95 % of total anthocyanin (61.68 mg/100 g) and 76 % of total flavonoids (25.85 mg/100 g) in comparison with fresh samples. Foodborne pathogens (E.coli O157:H7, Salmonella enterica and Listeria monocytogenes) inoculated at high concentration (≥6 log CFU/g) were undetected after the process. Overall results are promising for the development of a novel low temperature drying process for the production of healthy and safe snack.

Optimization of the Appearance Quality in CO2 Processed Ready-to-Eat Carrots through Image Analysis.

A high-pressure CO2 process applied to ready-to-eat food products guarantees an increase of both their microbial safety and shelf-life. However, the treatment often produces unwanted changes in the visual appearance of products depending on the adopted process conditions. Accordingly, the alteration of the visual appearance influences consumers' perception and acceptability. This study aims at identifying the optimal treatment conditions in terms of visual appearance by using an artificial vision system. The developed methodology was applied to fresh-cut carrots (Daucus carota) as the test product. The results showed that carrots packaged in 100% CO2 and subsequently treated at 6 MPa and 40 °C for 15 min maintained an appearance similar to the fresh product for up to 7 days of storage at 4 °C. Mild appearance changes were identified at 7 and 14 days of storage in the processed products. Microbiological analysis performed on the optimal treatment condition showed the microbiological stability of the samples up to 14 days of storage at 4 °C. The artificial vision system, successfully applied to the CO2 pasteurization process, can easily be applied to any food process involving changes in the appearance of any food product.

Research Note: Microbial inactivation of raw chicken meat by supercritical carbon dioxide treatment alone and in combination with fresh culinary herbs.

The objective of the present study was to assess the potential synergistic effect between supercritical carbon dioxide (SC-CO2) and fresh culinary herbs (Coriandrum sativum and Rosmarinus officinalis) on the microbial inactivation of raw chicken meat. The microbiological inactivation was performed on Escherichia coli and natural flora (total mesophilic bacteria, yeasts, and molds). High pressure treatments were carried out at 40°C, 80 or 140 bar from 15 to 45 min. Microbial inactivation had a strong dependence on treatment time, achieving 1.4 log CFU/g reduction of E. coli after 15 min, and up to 5 log after 45 min, while a pressure increase from 80 up to 140 bar was not significant on the microbial inactivation. Mesophilic microorganisms were strongly reduced (>2.6 log CFU/g) after 45 min, and yeasts and molds were below the detection limits of the technique (<100 CFU/g) in most cases. The combination of fresh herbs together with SC-CO2 treatment did not significantly increase the inactivation of either E. coli or natural flora, which was similar to the SC-CO2 alone. The synergistic effect was obtained on the inactivation of E. coli using a proper concentration of coriander essential oil (EO) (0.5% v/w), while rosemary EO did not show a significant effect. Color analysis after the treatment showed an increment of lightness (L*), and a decrease of redness (a*) on the surface of the sample, making the product visually similar to cooked meat. Texture analysis demonstrated the modification of the texture parameters as a function of the process pressure making the meat more similar to the cooked one.

In vitro metabolic zonation through oxygen gradient on a chip.

Among the multiple metabolic signals involved in the establishment of the hepatic zonation, oxygen could play a key role. Indeed, depending on hepatocyte position in the hepatic lobule, gene expression and metabolism are differently affected by the oxygen gradient present across the lobule. The aim of this study is to understand whether an oxygen gradient, generated in vitro in our developed device, is sufficient to instruct a functional metabolic zonation during the differentiation of human embryonic stem cells (hESCs) from endoderm toward terminally differentiated hepatocytes, thus mimicking the in vivo situation. For this purpose, a microfluidic device was designed for the generation of a stable oxygen gradient. The oxygen gradient was applied to differentiating hESCs at the pre-hepatoblast stage. The definitive endoderm and hepatic endoderm cells were characterized by the expression of the transcription factor SOX-17 and alpha-fetoprotein (AFP). Immature and mature hepatocytes were characterized by hepatocyte nuclear factor 4-alpha (HNF-4α) and albumin (ALB) expression and also analyzed for cytochrome P450 (CYP3A4) zonation and glycogen accumulation through PAS staining. Metabolic zonated genes expression was assessed through quantitative real time PCR. Application of the oxygen gradient during differentiation induced zonated glycogen storage, which was higher in the hepatocytes grown in high pO2 compared to those grown in low pO2. The mRNA levels of glutamine synthetase (GLUL), beta-catenin (CTNNB) and its direct target cyclin D1 (CCND1) showed significantly higher expression in the cells grown in low pO2 compared to those grown in high pO2. On the contrary, carbamoyl-phosphate synthetase 1 (CPS1), ALB, the proliferative marker ki67 (MKI67) and cyclin A (CCNA) resulted to be significantly higher expressed in cells cultured in high pO2 compared to those cultured in low pO2. These results indicate that the oxygen gradient generated in our device can instruct the establishment of a functional metabolic zonation in differentiating hESCs. The possibility to obtain differentiated hepatocytes in vitro may allow in the future to deepen our knowledge about the physiology/pathology of hepatocytes in relation to the oxygen content.

Cross-talk of healthy and impaired human tissues for dissection of disease pathogenesis.

Systemic diseases affect multiple tissues that interact with each other within a network difficult to explore at the body level. However, understanding the interdependences between tissues could be of high relevance for drug target identification, especially at the first stages of disease development. In vitro systems have the advantages of accessibility to measurements and precise controllability of culture conditions, but currently have limitations in mimicking human in vivo systemic tissue response. In this work, we present an in vitro model of cross-talk between an ex vivo culture of adipose tissue from an obese donor and a skeletal muscle in vitro model from a healthy donor. This is relevant to understand type 2 diabetes mellitus pathogenesis, as obesity is one of its main risk factors. The human adipose tissue biopsy was maintained as a three-dimensional culture for 48 h. Its conditioned culture medium was used to stimulate a human skeletal muscle-on-chip, developed by differentiating primary cells of a patient's biopsy under topological cues and molecular self-regulation. This system has been characterized to demonstrate its ability to mimic important features of the normal skeletal muscle response in vivo. We then found that the conditioned medium from a diseased adipose tissue is able to perturb the normal insulin sensitivity of a healthy skeletal muscle, as reported in the early stages of diabetes onset. In perspective, this work represents an important step toward the development of technological platforms that allow to study and dissect the systemic interaction between unhealthy and healthy tissues in vitro. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2766, 2019.

Simulating Inflammation in a Wound Microenvironment Using a Dermal Wound-on-a-Chip Model.

Considerable progress has been made in the field of microfluidics to develop complex systems for modeling human skin and dermal wound healing processes. While microfluidic models have attempted to integrate multiple cell types and/or 3D culture systems, to date they have lacked some elements needed to fully represent dermal wound healing. This paper describes a cost-effective, multicellular microfluidic system that mimics the paracrine component of early inflammation close to normal wound healing. Collagen and Matrigel are tested as materials for coating and adhesion of dermal fibroblasts and human umbilical vein endothelial cells (HUVECs). The wound-on-chip model consists of three interconnecting channels and is able to simulate wound inflammation by adding tumor necrosis factor alpha (TNF-α) or by triculturing with macrophages. Both the approaches significantly increase IL-1ß, IL-6, IL-8 in the supernatant (p < 0.05), and increases in cytokine levels are attenuated by cotreatment with an anti-inflammatory agent, Dexamethasone. Incorporation of M1 and M2 macrophages cocultured with fibroblasts and HUVECs leads to a stimulation of cytokine production as well as vascular structure formation, particularly with M2 macrophages. In summary, this wound-on-chip system can be used to model the paracrine component of the early inflammatory phase of wound healing and has the potential for the screening of anti-inflammatory compounds.

Enzymatic, physicochemical, nutritional and phytochemical profile changes of apple (Golden Delicious L.) juice under supercritical carbon dioxide and long-term cold storage.

The impact of supercritical carbon dioxide (SCCD) (10-60 MPa/45 °C/30 min) and subsequent 10 weeks storage at 4 °C on polyphenol oxidase (PPO), peroxidase (POD) activities, phenolic profile, vitamin C, sugars, physicochemical properties of cloudy apple juices was investigated. No significant changes in sugars and total polyphenols were observed, whereas significant degradation (≈28%) of vitamin C and individual polyphenols (≈18%) was noted after SCCD treatment. After 4 weeks storage only 34% of vitamin C was retained and no vitamin C was detected after this time. Ten weeks of storage caused hydrolysis of sucrose in 15%, whereas degradation of individual polyphenols ranged from 43 to 50% depending on the pressure applied. The highest pressure was applied the highest retention of polyphenols was observed. The lightness of juice significantly increased by 15% after SCCD and decreased during storage. Moreover, the synergistic effect of both enzymes with chlorogenic acid and catechol was found.

In Situ Raman Analysis of CO2-Assisted Drying of Fruit-Slices.

This work explores the feasibility of applying in situ Raman spectroscopy for the online monitoring of the supercritical carbon dioxide (SC-CO2) drying of fruits. Specifically, we investigate two types of fruits: mango and persimmon. The drying experiments were carried out inside an optical accessible vessel at 10 MPa and 313 K. The Raman spectra reveal: (i) the reduction of the water from the fruit slice and (ii) the change of the fruit matrix structure during the drying process. Two different Raman excitation wavelengths were compared: 532 nm and 785 nm. With respect to the quality of the obtained spectra, the 532 nm excitation wavelength was superior due to a higher signal-to-noise ratio and due to a resonant excitation scheme of the carotenoid molecules. It was found that the absorption of CO2 into the fruit matrix enhances the extraction of water, which was expressed by the obtained drying kinetic curve.

Simvastatin Rapidly and Reversibly Inhibits Insulin Secretion in Intact Single-Islet Cultures.

INTRODUCTION: Epidemiological studies suggest that statins may promote the development or exacerbation of diabetes, but whether this occurs through inhibition of insulin secretion is unclear. This lack of understanding is partly due to the cellular models used to explore this phenomenon (cell lines or pooled islets), which are non-physiologic and have limited clinical transferability. METHODS: Here, we study the effect of simvastatin on insulin secretion using single-islet cultures, an optimal compromise between biological observability and physiologic fidelity. We develop and validate a microfluidic device to study single-islet function ex vivo, which allows for switching between media of different compositions with a resolution of seconds. In parallel, fluorescence imaging provides real-time analysis of the membrane voltage potential, cytosolic Ca2+ dynamics, and insulin release during perfusion under 3 or 11 mM glucose. RESULTS: We found that simvastatin reversibly inhibits insulin secretion, even in high-glucose. This phenomenon is very rapid (<60 s), occurs without affecting Ca2+ concentrations, and is likely unrelated to cholesterol biosynthesis and protein isoprenylation, which occur on a time span of hours. CONCLUSIONS: Our data provide the first real-time live demonstration that a statin inhibits insulin secretion in intact islets and that single islets respond differently from cell lines on a short time scale. FUNDING: University of Padova, EASD Foundation.

Straightforward Micropatterning of Oligonucleotides in Microfluidics by Novel Spin-On ZrO2 Surfaces.

DNA biochip assays often require immobilization of bioactive molecules on solid surfaces. A simple biofunctionalization protocol and precise spatial binding represent the two major challenges in order to obtain localized region specific biopatterns into lab-on-a-chip (LOC) systems. In this work, a simple strategy to anchor oligonucleotides on microstructured areas and integrate the biomolecules patterns within microfluidic channels is reported. A photosensitive ZrO2 system is proposed as an advanced platform and versatile interface for specific positioning and oriented immobilization of phosphorylated DNA. ZrO2 sol-gel structures were easily produced on fused silica by direct UV lithography, allowing a simple and fast patterning process with different geometries. A thermal treatment at 800 °C was performed to crystallize the structures and maximize the affinity of DNA to ZrO2. Fluorescent DNA strands were selectively immobilized on the crystalline patterns inside polydimethylsiloxane (PDMS) microchannels, allowing high specificity and rapid hybridization kinetics. Hybridization tests confirmed the correct probe anchoring and the bioactivity retention, while denaturation experiments demonstrated the possibility of regenerating the surface.

Determination of glucose flux in live myoblasts by microfluidic nanosensing and mathematical modeling.

Glucose is the main energy source for cells in an organism and its blood concentration is tightly regulated in healthy individuals. However, impaired blood glucose control has been found in diseases such as metabolic syndrome and diabetes, and anomalous glucose utilization in cancer tissues. Dissecting the dynamics of the different phenomena involved in glucose handling (extracellular mass transport, membrane diffusion, and intracellular phosphorylation) is very relevant to identify which mechanisms are disrupted under disease conditions. In this work, we developed an effective methodology for quantitatively analyzing these phenomena in living cells. A measurement of steady-state glucose uptake is, by itself, insufficient to determine the dynamics of intracellular glucose. For this purpose, we integrated two types of measurements: cytosolic glucose concentration at the single-cell level, obtained using a cytosolic FRET nanosensor, and cell population glucose uptake, obtained without perturbing culture conditions using a microfluidic perfusion system. Microfluidics enabled accurate temporal stimulation of cells through cyclic pulses of glucose concentration at defined flow rates. We found that both, glucose uptake and phosphorylation, are linearly dependent on glucose concentration in the physiological range. Mathematical modeling enabled precise determination of the kinetic constants of membrane transport (0.27 s(-1)) and intracellular phosphorylation (2.01 s(-1)).

Flow biosensing and sampling in indirect electrochemical detection.

Miniaturization in biological analyses has several advantages, such as sample volume reduction and fast response time. The integration of miniaturized biosensors within lab-on-a-chip setups under flow conditions is highly desirable, not only because it simplifies process handling but also because measurements become more robust and operator-independent. In this work, we study the integration of flow amperometric biosensors within a microfluidic platform when analyte concentration is indirectly measured. As a case study, we used a platinum miniaturized glucose biosensor, where glucose is enzymatically converted to [Formula: see text] that is oxidized at the electrode. The experimental results produced are strongly coupled to a theoretical analysis of fluid dynamic conditions affecting the electrochemical response of the sensor. We verified that the choice of the inlet flow rate is a critical parameter in flow biosensors, because it affects both glucose and [Formula: see text] transport, to and from the electrode. We identify optimal flow rate conditions for accurate sensing at high time resolution. A dimensionless theoretical analysis allows the extension of the results to other sensing systems according to fluid dynamic similarity principles. Furthermore, we developed a microfluidic design that connects a sampling unit to the biosensor, in order to decouple the sampling flow rate from that of the actual measurement.

Microfluidic-driven viral infection on cell cultures: Theoretical and experimental study.

Advanced cell culture systems creating a controlled and predictable microenvironment together with computational modeling may be useful tools to optimize the efficiency of cell infections. In this paper, we will present a phenomenological study of a virus-host infection system, and the development of a multilayered microfluidic platform used to accurately tune the virus delivery from a diffusive-limited regime to a convective-dominated regime. Mathematical models predicted the convective-diffusive regimes developed within the system itself and determined the dominating mass transport phenomena. Adenoviral vectors carrying the enhanced green fluorescent protein (EGFP) transgene were used at different multiplicities of infection (MOI) to infect multiple cell types, both in standard static and in perfused conditions. Our results validate the mathematical models and demonstrate how the infection processes through perfusion via microfluidic platform led to an enhancement of adenoviral infection efficiency even at low MOIs. This was particularly evident at the longer time points, since the establishment of steady-state condition guaranteed a constant viral concentration close to cells, thus strengthening the efficiency of infection. Finally, we introduced the concept of effective MOI, a more appropriate variable for microfluidic infections that considers the number of adenoviruses in solution per cell at a certain time.