Biology subjects

Rolandi, M.

Publications and source records attributed to Rolandi, M..

15 recordsLinked to original sources

bioRxiv2025

Self-Organizing Neural Networks in Organoids Reveal Principles of Forebrain Circuit Assembly

The mouse cortex is a canonical model for studying how functional neural networks emerge, yet it remains unclear which topological features arise from intrinsic cellular organization versus external regional cues. Mouse forebrain organoids provide a powerful system to investigate these intrinsic mechanisms. We generated dorsal (DF) and ventral (VF) forebrain organoids from mouse pluripotent stem cells and tracked their development using longitudinal electrophysiology. DF organoids showed progressively stronger network-wide correlations, while VF organoids developed more refined activity patterns, enhanced small-world topology, and increased modular organization. These differences emerged without extrinsic inputs and may be driven by the increased generation of Pvalb+ interneurons in VF organoids. Our findings demonstrate how variations in cellular composition influence the self-organization of neural circuits, establishing mouse forebrain organoids as a tractable platform to study how neuronal populations shape cortical network architecture.

neuroscience↗

bioRxiv2025

Fluoxetine Delivery for Wound Treatment Through an Integrated Bioelectronic Device - Pharmacokinetic Parameters and Safety Profile in Swine

Wound infections are a significant medical challenge, often leading to chronicity or systemic infection. Selective serotonin reuptake inhibitors (SSRIs) have emerged as potential non-antibiotic candidates with demonstrated ability to limit growth and biofilm formation in Gram-negative bacteria, in addition to their pro-healing activity. Here, we compared direct delivery of the SSRI fluoxetine by topical bolus dosing to delivery from an iontophoresis bandage device with an actuator for temporally controlled drug delivery, in a porcine excisional wound model. Device delivery of fluoxetine resulted in a maximum concentration of 12.25 ng fluoxetine per mg tissue, compared to 2.926 ng/mg following bolus dosing, and tissue fluoxetine levels were higher after application using the device than after bolus dosing across the range of doses tested (p=0.0041). The half-life of fluoxetine in the wound tissue was 0.988 {+/-} 0.256 days. Fluoxetine was not detected in the pig plasma, and plasma serotonin levels were not affected by the topical application. Fluoxetine delivery using the device, but not bolus delivery, produced tissue concentrations above the minimum inhibitory concentration (MIC) for some clinically important species of bacteria. The experimental device can effectively deliver topical fluoxetine to the wound, producing higher tissue concentrations of fluoxetine at lower cumulative doses compared to bolus dosing, and with minimal risk of off-target effects. The device may simplify wound treatment by reducing the burden for daily drug application, possibly increasing adherence to a prescribed treatment regimen.

bioengineering↗

bioRxiv2024

Design and Validation of a Wearable Imaging System for Automated Wound Monitoring in Porcine Model

Effective wound monitoring has the potential to guide treatment regiments and improve healing outcomes, yet current clinical assessment methods remain largely subjective and labor-intensive. To address this challenge, we present a high-resolution wearable imaging system designed for continuous wound monitoring. The system integrates a 64 MP camera with a plano-convex lens housed in an enclosure measuring 73 mm in diameter and 36.1 mm in height, and features a custom printed circuit board (PCB) for programmable LED illumination. The 3D-printed device enclosure is designed to accommodate a silicone bioelectronic device and can be securely attached using a commercially available ostomy skin barrier. In porcine model validation studies, the system successfully captured daily wound progression over periods up to 7 days. The captured images were wirelessly transmitted to a processing unit where DeepMapper, a machine learning algorithm, processed z-stacked images and performed multi-level feature extraction to predict wound healing stages and indicate potential complications such as infection. This imaging system enables automated analysis of wound progression and supports the development of smart wound care platforms for personalized treatment strategies. The integrated design approach demonstrates the feasibility of creating compact, high-resolution imaging systems suitable for clinical wound monitoring applications.

bioengineering↗

bioRxiv2024

Accelerating Wound Healing Through Deep Reinforcement Learning: A Data-Driven Approach to Optimal Treatment

Advancements in bioelectronic sensors and actuators have paved the way for real-time monitoring and control of wound healing progression. Real-time monitoring allows for precise adjustment in treatment strategies that align with an individuals unique biological response. However, due to the complexities of human-drug interactions and a lack of predictive models it is challenging to determine just how one should adjust drug dosage to achieve the desired biological response. This work proposes an adaptive closed-loop control framework that integrates deep learning, optimal control, and reinforcement learning to update treatment strategies in real-time with the goal of accelerating wound closure. The proposed approach eliminates the need for mathematical modeling of complex nonlinear wound healing dynamics. We demonstrate the convergence of the controller via an in silico experimental setup, where the proposed approach successfully accelerates the wound healing process by 17.71%. Finally, we share the experimental setup and results of an in vivo implementation to highlight the translational potential of our work. Our data-driven model estimates a 40% acceleration in wound closure.

systems biology↗

bioRxiv2024

Remote-Controlled Wireless Bioelectronics for Fluoxetine Therapy to Promote Wound Healing in a Porcine Model

Wound healing presents a significant challenge in biomedical science, requiring precise therapeutic delivery and real-time monitoring. Bioelectronic systems offer a promising solution but remain largely unexplored for wound care, particularly in large animal models that reflect human healing dynamics. This study introduces a remote controlled wireless bioelectronic platform equipped with an iontophoretic pump to deliver fluoxetine, a selective serotonin reuptake inhibitor that promotes wound repair. In vitro and ex-vivo testing validated efficient on demand fluoxetine delivery. In vivo experiments in a porcine wound model demonstrated clear therapeutic efficacy over 3-day and 7-day periods. The system enhanced healing outcomes, increasing re-epithelialization by 37% (H&E staining), reducing the M1/M2 macrophage ratio by 33%, and stimulating neuronal growth at the wound site. This bioelectronic platform delivers fluoxetine in a controlled, remotely-controlled manner while allowing for wound direct wound imaging that can be used to monitor wound healing progress. Additionally, it allows precise dose and temporal delivery of treatment to enhance the outcome of future large animal wound healing studies.

bioengineering↗

bioRxiv2024

DeepMapper: Attention-Based AutoEncoder for System Identification in Wound Healing and Stage Prediction

systems biology↗

bioRxiv2024

Wireless bioelectronic device for wound healing

Wireless bioelectronic actuators have been developed to deliver targeted treatments over multiple days while continuously monitoring delivery, thereby improving wound healing. Specifically, these devices can deliver charged biomolecules such as fluoxetine cations (Flx+) and electric field (EF) in freely moving pigs. Treatments can be controlled and monitored in real time via WiFi, with options for both user-specified delivery rates and durations, as well as automated closed-loop (CL) control. The devices are engineered to handle various failure scenarios that may arise in dynamic, real-world experiments--such as communication or power interruptions--ensuring that valuable experimental data is collected with minimal disruption. The ion pump features eight drug reservoirs and channels arranged around a 20 mm diameter-wound, with a central ground electrode (0 V). When voltages above 0 V are applied to the outer channels, currents flow from the reservoirs and channels into the wound, delivering Flx+ and/or EF depending on the reservoir solution. The device records applied voltages and currents locally to a microSD card at a high sampling rate, while simultaneously transmitting real-time measurements via a local WiFi network to a wound healing algorithm running on a nearby laptop. CL control of current/delivery rate is performed by an onboard microcontroller unit (MCU) and current-source microchips, based on instructions received from the wound healing algorithm. A graphical user interface (GUI) provides intuitive user control and real-time data visualization, with support for multiple devices. In vivo studies over seven days showed that Flx+-treated wounds had a 20% lower M1/M2 macrophage ratio and 41.67% greater re-epithelialization compared to controls (standard-of-care), demonstrating the actuators potential to enhance wound healing.

biochemistry↗

bioRxiv2023

Programmable delivery of fluoxetine via wearable bioelectronics for wound healing in vivo

The ability to deliver drugs with precise dosages at specific time points can significantly improve disease treatment while reducing side effects. Drug encapsulation for gradual delivery has opened up the doors for superior treatment regimen. To expand on this ability, programming bioelectronic devices to deliver small molecules enables ad-hoc personalized therapeutic profiles that are more complex than simple gradual release. Here, we introduce a wearable bioelectronic bandage with an integrated electrophoretic ion pump that affords on-demand drug delivery with precise dose control. Delivery of fluoxetine to wounds in mice resulted in a 27.2% decrease in the macrophage ratio (M1/M2) and a 39.9% increase in re-epithelialization, indicating a shorter inflammatory phase and faster overall healing. Programmable drug delivery using wearable bioelectronics in wounds introduces a broadly applicable strategy for the long-term delivery of a prescribed treatment regimen with minimal external intervention.

bioengineering↗

bioRxiv2023

Delivering biochemicals with precision using bioelectronic devices enhanced with feedback control

Precision medicine tailors treatment in a way that accounts for variations in patient response. Treatment strategies can be determined based on factors such as genetic mutations, age, and diet. Another way of implementing precision medicine in a dynamic fashion is through bioelectronics equipped with real-time sensing and intelligent actuation. Bioelectronic devices such as ion pumps can be utilized to deliver therapeutic drugs. To be able to perform precision medicine, medical devices need to be able to deliver drugs with high precision. For this, closed-loop control is required to be able to change the treatment strategy as new information about the response and progression of the biological system is received. To this end, a sliding mode controller is utilized given its ability to perform satisfactory control actions when there is model uncertainty. The controller is used in an experiment with the goal of delivering a pre-determined dosage of fluoxetine throughout a period of time.

systems biology↗

bioRxiv2023

Modulation of neuronal activity in cortical organoids with bioelectronic delivery of ions and neurotransmitters

Precise modulation of brain activity is fundamental for the proper establishment and maturation of the cerebral cortex. To this end, cortical organoids are promising tools to study circuit formation and the underpinnings of neurodevelopmental disease. However, the ability to manipulate neuronal activity with high temporal resolution in brain organoids remains limited. To overcome this challenge, we introduce a bioelectronic approach to control cortical organoid activity with the selective delivery of ions and neurotransmitters. Using this approach, we sequentially increased and decreased neuronal activity in brain organoids with the bioelectronic delivery of potassium ions (K+) and {gamma}-aminobutyric acid (GABA), respectively, while simultaneously monitoring network activity. This works highlights bioelectronic ion pumps as tools for high-resolution temporal control of brain organoid activity toward precise pharmacological studies that can improve our understanding of neuronal function.

bioengineering↗

bioRxiv2023

DNA nanopores as artificial membrane channels for origami based bioelectronics

Biological membrane channels mediate information exchange between cells and facilitate molecular recognition1-4. While tuning the shape and function of membrane channels for precision molecular sensing via de-novo routes is complex, an even more significant challenge is interfacing membrane channels with electronic devices for signal readout5-8. This challenge at the biotic-abiotic interface results in low efficiency of information transfer - one of the major barriers to the continued development of high-performance bioelectronic devices9. To this end, we integrate membrane spanning DNA nanopores with bioprotonic contacts to create programmable, modular, and efficient artificial ion-channel interfaces that resolve the iono-electronic disparity between the biotic environment and electronics. Through simulations and experiments, we show that cholesterol modified DNA nanopores spontaneously and with remarkable affinity span the lipid bilayer formed over the planar bio-protonic electrode surface and mediate proton transport across the bilayer. Using the ability to easily modify DNA nanostructures, we illustrate that this bioelectronic device can be programmed for electronic recognition of biomolecular signals such as presence of Streptavidin, without disrupting the native environment of the biomolecule. We anticipate this robust biotic-abiotic interface will allow facile electronic measurement of inter-cellular ionic communication and also open the door for active control of cell behavior through externally controlled selective gating of the channels.

bioengineering↗

bioRxiv2022

Modular automated microfluidic cell culture platform reduces glycolytic stress in cerebral cortex organoids

Organ-on-a-chip systems combine microfluidics, cell biology, and tissue engineering to culture 3D organ-specific in vitro models that recapitulate the biology and physiology of their in vivo counterparts. Here, we have developed a multiplex platform that automates the culture of individual organoids in isolated microenvironments at user-defined media flow rates. Programmable workflows allow the use of multiple reagent reservoirs that may be applied to direct differentiation, study temporal variables, and grow cultures long term. Novel techniques in polydimethylsiloxane (PDMS) chip fabrication are described here that enable features on the upper and lower planes of a single PDMS substrate. RNA sequencing (RNA-seq) analysis of automated cerebral cortex organoid cultures shows benefits in reducing glycolytic and endoplasmic reticulum stress compared to conventional in vitro cell cultures.

bioengineering↗

bioRxiv2021

Implementing QR Codes in Academia to Improve Sample Tracking, Data Accessibility, and Traceability in Multicampus Interdisciplinary Collaborations

With the growing number of multicampus interdisciplinary projects in academic institutions, there is an increasing need for tracking systems that make device and sample data and associated results instantly accessible to all collaborators involved. This need has become particularly salient with the COVID pandemic and consequent travel restrictions that have hampered in person meetings and laboratory visits. We report a Quick Response (QR) code tracking system that integrates project management tools for seamless communication and tracking of materials and devices between three manufacturing sites (cleanrooms) and six research laboratories across five universities. We use this system to track the design, fabrication, and quality control of bioelectronic devices (in the cleanroom and engineering laboratories), in vitro experimental results (in a biology laboratory), and in vivo testing (in the school of medicine). Setting up a tracking system for multisite interdisciplinary teams improves traceability, efficiency, and the quality of produced results making it easier to accomplish project milestones on a tight timeline. This tracking system is particularly useful to track device issues and ensure engineering device consistency when working with expensive biological samples in vitro and animals in vivo to reduce waste of biological and animal resources associated with device failure.

bioengineering↗

bioRxiv2021

Structural Characteristics and Proton Conductivity of the Gel Within the Electrosensory Organs of Cartilaginous Fishes

The Ampullae of Lorenzini (AoL) of cartilaginous fishes are sensory organs used to detect environmental electric fields. The proximal ends of the organs are externally visible as pores in the skin that lead into gel-filled tubular canals which terminate in rounded chambers filled with highly specialized electrosensory cells. The viscoelastic gel that fills the organs is composed of proteins and polysaccharides that are not yet completely characterized but are thought to play a critical role in the electrosensing mechanism. Although recent studies have identified various components of AoL gel, it has remained unclear how the component molecules are structurally arranged and how their structure influences the overall function of the AoL. Here we present the first microscopic descriptions and x-ray scattering data from AoL gel extracted from spotted ratfish (Hydrolagus colliei). Our results suggest that AoL gel is colloidal in nature and composed of spherical globules that are approximately 10-100 nm in size. We investigated the structural influence of the protein components of the gel specifically by analyzing gel that had been digested in situ via enzymatic proteolysis. By comparing gel before and after digestion using microscopy, x-ray scattering analyses, and proton conductivity measurements, we directly observed the structural and functional influence of proteins in AoL gel. The findings described here represent the first detailed structural analysis of AoL gel and lay the groundwork for more detailed studies into the specific interactions of molecules inside AoL gel at the nanoscale, with particular reference to their mechanistic role in electrosensing.

biophysics↗

bioRxiv2018

Proton Conductivity of Glycosaminoglycans

Proton (H+) conductivity is important in many natural phenomena including oxidative phosphorylation in mitochondria and archea, uncoupling membrane potentials by the antibiotic Gramicidin, and proton actuated bioluminescence in dinoflagellate. In all of these phenomena, the conduction of H+ occurs along chains of hydrogen bonds between water and hydrophilic residues. These chains of hydrogen bonds are also present in many hydrated biopolymers and macromolecule including collagen, keratin, chitosan, and various proteins such as reflectin. All of these materials are also proton conductors. Recently, our group has discovered that the jelly found in the Ampullae of Lorenzini-sharks electrosensing organs- is the highest naturally occurring proton conducting substance. The jelly has a complex composition, but we attributed the conductivity to the glycosaminoglycan keratan sulfate (KS). Here, we have measured the proton conductivity of hydrated keratan sulfate using PdHx contacts to be 0.50 {+/-} 0.11 mS cm -1- consistent to that of Ampullae of Lorenzini jelly, 2 {+/-} 1 mS cm -1. Proton conductivity, albeit with lower values, is also shared by other glycosaminoglycans with similar chemical structures including dermatan sulfate, chondroitin sulfate A, heparan sulfate, and hyaluronic acid. This observation confirms the structure property relationship between proton conductivity and the chemical structure of biopolymers.

bioengineering↗