This work was supported by the EC within the framework of the STIFF-FLOP FP7-ICT-2011.2.1 European Project (#287728).

Nota: Questo protocollo descrive le fasi di fabbricazione di un singolo modulo, che comprende le camere fluidici, irrigidimento canale, tubazioni di azionamento e guaina esterna. La seguente procedura deve essere eseguita sotto una cappa aspirante e indossando camice e guanti per motivi di sicurezza. Come precedentemente accennato, il processo di fabbricazione del dispositivo elastomerico è basato sull&#39;uso sequenziale di stampi progettati con software CAD. Sono composte da 13 pezzi mostrati nella Figura 2 e elencati nella tabella 1. 1. Preparazione del silicone <ol><li> Pesare 12 g di parte A e 12 g di parte B nello stesso bicchiere di plastica o una scatola Petri e mescolare insieme, mescolando. Nota: proporzioni dei materiali possono variare a seconda del silicone specifico utilizzato, in questo caso è costituito da due parti: la parte A (base) e parte B (catalizzatore). Essi sono utilizzati in proporzione 1A: 1B in peso. </li><li> Posizionare il vetro contenente la miscelamateriali siliconici Ed in una macchina di degasaggio a 1 bar di pressione di vuoto. Mantenere il vetro sotto vuoto fino a quando tutte le bolle vengono rimossi dal materiale siliconico. Per il silicone impiegato il processo di degasaggio dura circa 10 min. Una volta che i materiali sono completamente libero dalla presenza di bolle, ripristinare la pressione atmosferica nella macchina e utilizzare il silicone. </li></ol> 2. Realizzazione del modulo siliconico <ol><li> Assemblaggio dello stampo. <ol><li> Inserire il cilindro irrigidimento e la parte superiore delle camere in cap_A (Figura 3a). </li><li> Chiudere i gusci intorno al secondo strato di cap_A. </li></ol></li><li> Primo casting silicone. <ol><li> Versare il silicone nello stampo assemblato fino al bordo dei gusci (Figura 3b). </li><li> Collocare lo stampo in un forno a 60 ° C per circa 30 min. </li></ol></li><li> Riassetto dello stampo. <ol><li> Rimuovere i gusci esterni e cap_A (Figura 3c). </li><li> Inserire i cilindri dalle basi delle camere e il cilindro irrigidimento interno cap_B (Figura 3d). </li><li> Chiudere i gusci ancora attorno al modulo, facendoli scorrere di 10 mm verso l&#39;alto in modo da avere uno spazio di 10 mm tra la superficie superiore del modulo ed i bordi dei gusci (figura 3e). </li></ol></li><li> Seconda colata di silicone. <ol><li> Versare il silicone nello stampo riarrangiato fino al bordo delle conchiglie sul lato superiore (cioè anche fino al cilindro irrigidimento) (Figura 3f). </li><li> Mettere lo stampo in un forno a 60 ° C per circa 30 min. </li><li> Rimuovere i gusci esterni, cap_B e le camere (ad eccezione del cilindro di irrigidimento) (Figura 3g). </li></ol></li></ol> 3. Inserimento dei tubi <ol><li> Tagliare 3 tubi alla stessa lunghezza desiderata (300 mm per esempio). </li><li> Mettere colla siliconica intorno una estremità di ciascun tubo di 10 mm, senza obstructing i tubi. </li><li> Inserire i tubi all&#39;interno dei 2 mm canali nell&#39;unità siliconico (Figura 3 ore) dedicati. </li><li> Dopo un tempo di maturazione di 12 min a temperatura ambiente o mettere il modulo in un forno ad una temperatura superiore (50 ° - 60 °) per accelerare il processo di essiccazione. </li></ol> 4. Realizzazione della pressare intrecciato guaina <ol><li> Tagliare 700 mm di una guaina intrecciata espandibile (circa 15 volte l&#39;altezza del modulo). </li><li> Inserire un cilindro metallico di 30 mm di diametro e 250 mm di lunghezza all&#39;interno della guaina. </li><li> Abbassare e forzare la guaina facendo scorrere sopra il cilindro, al fine di creare piegature. </li><li> Fissare meccanicamente la guaina in posizione con un morsetto e calore con una pistola di riscaldamento a 350 ° C per 2-3 minuti fino ad ottenere una deformazione permanente. </li><li> Lasciare la guaina raffreddare e togliere il cilindro interno. </li></ol> 5. Integrazione della guaina esterna <ol><li> Passa iltubi attraverso i fori di cap_C. </li><li> Versate 3 g di silicone in cap_C. </li><li> Bloccare cap_C ad un supporto che è superiore al piano di lavoro. </li><li> Inserire il lato inferiore del modulo precedentemente fabbricato in cap_C. </li><li> Far scorrere la guaina crimpato intorno al modulo. </li><li> Spingere le prime tacche della guaina interna cap_C e immergerli nel silicone appena versato (Figura 3i). </li><li> Mettere lo stampo in un forno a 60 ° C per circa 20 min. </li><li> Ripetere la stessa procedura dal punto 5,1-5,6 per fissare la guaina sul lato superiore, utilizzando cap_D (Figura 3 undecies). </li><li> Rimuovere cap_C e cap_D. </li><li> Rimuovere il cilindro centrale (Figura 3k). </li></ol> 6. Realizzazione della Granular Jamming membrana <ol><li> Versare 5 g di lattice liquido in un bicchiere di plastica. </li><li> Immergere il cilindro per la membrana (ultimo pezzo illustrato in figura 2) nel lattice liquido finché la superficieè completamente coperto. </li><li> Lasciare asciugare sotto cappa per 20 min. </li><li> Ripetere i punti 6.2 e 6.3. </li><li> Rimuovere la membrana dallo stampo. </li></ol> 7. Inserimento del Granular Jamming membrana <ol><li> Tagliare un tubo (2 mm di diametro) alla lunghezza desiderata (300 mm per esempio). </li><li> Tagliare un pezzo quadrato di circa 100 mm 2 di tessuto di nylon e chiudere una estremità del tubo con questo tessuto con una pellicola di plastica o colla di paraffina. </li><li> Pesare 4 g di polvere di caffè e riempire la membrana. </li><li> Inserire il tubo (l&#39;estremità con il filtro) all&#39;interno della membrana riempito e fissarlo attorno al tubo con una pellicola di plastica di paraffina. </li><li> Applicare il vuoto sul lato opposto del tubo (la membrana diventa più rigido). </li><li> Inserire la membrana all&#39;interno del canale centrale vuota del modulo siliconico (figura 3L). </li><li> Incollare le estremità della membrana irrigidimento al modulo silicone. </li><li> Chiudere gli anelli intorno alla parte superiorelato del modulo (Figura 3m). </li><li> Versare 2 g di silicone negli anelli per livellare la superficie. </li><li> Lasciare il silicone secca sotto cappa o in forno a 60 °. </li><li> Rimuovere gli anelli. </li><li> Ripetere da punti 7.8 a 7.11 per il lato inferiore (Figura 3n). </li></ol>

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The authors have nothing to disclose.

The technique described in this protocol enables the fabrication of a pneumatically actuated soft unit usable for modular compliant structures. Thanks to the design of the molds and their simple assembly, it is possible to fabricate one complete module in about 4 hours with 7 main steps. The process of fabrication involves specific materials, which are easily available, and work should be carried out under a fume hood. An external set up including air valves, air compressor and vacuum pump is necessary to activate the mo...

Le recenti tendenze in campo medico premono per una riduzione della invasività delle operazioni chirurgiche. Chirurgia mininvasiva (MIS) è stato migliorato con successo negli ultimi anni per le operazioni addominali. Procedure MIS sono basate sull&#39;uso di strumenti introdotti attraverso quattro o cinque punti di accesso (Trequarti) poste sulla parete addominale. Al fine di ridurre il numero dei trocar, gli strumenti possono essere inseriti da porta singola Laparoscopia (SPL) o la chirurgia Natural Orifice transluminale endoscopica (NOTE) 1. Queste procedure impediscono cicatrici visibili esterni, ma aumentano la difficoltà per i medici nell&#39;esecuzione chirurgia. Questa limitazione è dovuta principalmente ai punti ridotti di accesso e la natura rigida o semi-rigida degli strumenti, che non sono in grado di evitare o passare intorno organi 2, 3. Destrezza e motilità può essere migliorata utilizzando articolato e iper-ridondante robot che possono coprire uno spazio di lavoro più ampio e complesso, thpermettendoci un obiettivo specifico nel corpo per raggiungere più facilmente 4, 5, 6 e per lavorare come sistemi quando necessario 7 retrazione. Un manipolatore flessibile può migliorare il rispetto dei tessuti, rendendo più sicuro di contatto con strumenti tradizionali. Tuttavia, questi manipolatori spesso mancano stabilità quando viene raggiunto l&#39;obiettivo e generalmente non possono controllare il contatto con i tessuti circostanti 8, 9. Studi su strutture biologiche, come il braccio polpo 10 e la proboscide 11, hanno recentemente ispirato la progettazione di manipolatori flessibili, deformabili e compatibili con un numero ridondante di gradi di libertà (DOF) e rigidità controllabile 12. Questi tipi di dispositivi utilizzano molle passivi, materiali intelligenti, elementi pneumatici, o tendini 13, 14, 15. Generalmente, manipolatori fabbricate con materiali morbidi e flessibili non garantiscono la generazione di forze elevate. Tegli Stiff-FLOP (rigidezza controllabile manipolatore flessibile e Programmabile per interventi chirurgici) manipolatore è stato recentemente presentato come un dispositivo chirurgico innovativo per NOTES e SPL ispirati a delle capacità del polpo. Per superare le limitazioni dei precedenti manipolatori morbido, ha un corpo morbido ed elevata destrezza, forza elevata e rigidità controllabile 16. L&#39;architettura del manipolatore è basato su un approccio modulare: unità multiple, con la stessa struttura e funzionalità, sono integrati insieme. La singola unità è mostrato nella Figura 1. Esso si basa su un cilindro elastomerico ottenuto da una fabbricazione multifase. Le fasi di montaggio dei componenti dello stampo e il processo di fusione consentono tre camere vuote (per azionamento fluidico) e un canale centrale cavo 17 (per l&#39;alloggiamento di un meccanismo basato inceppamenti granulare 18) per essere incorporati. Le camere sono poste a 120 °, in modo che ilinflazione combinata ir produce movimento omnidirezionale e l&#39;allungamento. Inoltre una guaina intrecciata esterna è posto esternamente per limitare l&#39;espansione radiale verso l&#39;esterno delle camere fluidici quando pressurizzato, ottimizzando così l&#39;effetto della camera di azionamento nel moto modulo (flessione ed allungamento). Il canale centrale ospita un dispositivo cilindrico costituito da una membrana esterna riempito di materiale granulare. Quando viene applicata una pressione di vuoto, cambia le sue proprietà elastiche causando un irrigidimento che interessa l&#39;intera proprietà del modulo. Prestazioni e rigidezza sono controllati da una configurazione esterna comprendente un compressore d&#39;aria e tre valvole di pressione per azionare le camere e una pompa a vuoto per attivare il vuoto nel canale di irrigidimento. Un&#39;interfaccia utente intuitiva permette il controllo di azionamento e del vuoto pressione all&#39;interno del modulo. Dettagli Questo documento il fabrication processo del singolo modulo di questo manipolatore e riporta i risultati più significativi sulla capacità di base di movimento. Considerando la natura modulare del dispositivo, la valutazione della fabbricazione e le prestazioni di un solo modulo singolo permette anche i risultati vengono estese e per prevedere il comportamento di base di un manipolatore multi-modulo integrando due o più moduli.

<table border="0">
	<tbody>
		<tr>
			<td>Ecoflex 00-50 Trial Kit</td>
			<td>SmoothOn</td>
			<td></td>
			<td>Used for the fabrication of the soft unit, combining equal amounts of liquid parts A (the base) and B (the catalyst)</td>
		</tr>
		<tr>
			<td>Latex</td>
			<td>Antichit&#224; Belsito</td>
			<td></td>
			<td>Used for the fabrication of the granular jamming membrane</td>
		</tr>
		<tr>
			<td>Peroxide-Cured Silicone Tubing</td>
			<td>Cole Parmer</td>
			<td>T-06411-59</td>
			<td>Used for actuating the chambers and applying vacuum</td>
		</tr>
		<tr>
			<td>PET expandable braided sleeving</td>
			<td>RS</td>
			<td>408-249</td>
			<td>Used for the fabrication of the external braided sheath</td>
		</tr>
		<tr>
			<td>Silicone Rubber</td>
			<td>Momentive</td>
			<td>127374</td>
			<td>Used to fix the actuation tubes to the module</td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>Cole Parmer</td>
			<td>EW-06720-40</td>
			<td>Used to fix the latex membrane to the vacuum tube</td>
		</tr>
		<tr>
			<td>Fume hood Secuflow</td>
			<td>Groupe Waldner</td>
			<td></td>
			<td>Working space</td>
		</tr>
		<tr>
			<td>Precision scale</td>
			<td>KERN EW</td>
			<td></td>
			<td>Used to weight silicone, latex and coffee powder</td>
		</tr>
		<tr>
			<td>Oven/degasser</td>
			<td>Heraeus</td>
			<td></td>
			<td>Used to degass the silicone and reduce its cure time</td>
		</tr>
		<tr>
			<td>Vacuum pump</td>
			<td>DVP Vacuum Technology</td>
			<td></td>
			<td>Used to apply vacuum to the latex membrane</td>
		</tr>
	</tbody>
</table>

design and fabrication of an elastomeric unit for soft modular robots in minimally invasive surgery

Negli ultimi anni, le tecnologie robotica morbidi hanno suscitato crescente interesse in campo medico a causa della loro interazione sicurezza intrinseca in ambienti non strutturati. Allo stesso tempo, nuove procedure e tecniche sono stati sviluppati per ridurre l&#39;invasività delle operazioni chirurgiche. Chirurgia mininvasiva (MIS) è stato impiegato con successo per gli interventi addominali, procedure MIS però standard sono basati principalmente su strumenti rigidi o semirigidi che limitano la destrezza del clinico. Questo articolo presenta un manipolatore abile morbido e alto per MIS. Il manipolatore è ispirato le capacità biologiche del braccio di polpo, ed è stato progettato con un approccio modulare. Ogni modulo presenta le stesse caratteristiche funzionali, ottenendo così elevata destrezza e versatilità quando più moduli sono integrati. Dettagli La carta progettazione, processo di fabbricazione ed i materiali necessari per lo sviluppo di una singola unità, che è fabbricato da casting silicone all&#39;interno di stampi specifici. Il risultato consiste in un cilindro elastomerico compresi tre attuatori pneumatici flessibili che consentono l&#39;allungamento e flessione omnidirezionale dell&#39;unità. Una guaina intrecciata esterna migliora il movimento del modulo. Nel centro di ogni modulo un meccanismo basato inceppamenti granulare varia la rigidezza della struttura durante l&#39;attività. Le prove dimostrano che il modulo è in grado di piegare fino a 120 ° e di allungare fino al 66% della lunghezza iniziale. Il modulo genera una forza massima di 47 N, e la sua rigidità può aumentare fino al 36%.

Le varie fasi di fabbricazione, descritto nel protocollo, sono illustrati nella figura 3. Per valutare l&#39;efficacia della tecnica ei risultati del prototipo finale, il modulo è stato testato in diverse condizioni di lavoro. Un&#39;installazione esterna permette il controllo sia l&#39;azionamento e la rigidità del modulo. Esso comprende un compressore d&#39;aria che attiva tre valvole. Essi sono collegati ai tubi siliconici integrati nelle camere e consentire loro pressu...

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Design and Fabrication of an Elastomeric Unit for Soft Modular Robots in Minimally Invasive Surgery

This paper describes the design and fabrication of a soft unit for surgical manipulators. The base module includes three flexible fluidic actuators to achieve omnidirectional bending and elongation, and a granular jamming-based mechanism to enable stiffness control. A complete mechanical characterization is also reported.

Progettazione e Realizzazione di un&#39;unità elastomerico per soft robot modulari in Chirurgia Mini-Invasiva

In recent years, soft robotics technologies have aroused increasing interest in the medical field due to their intrinsically safe interaction in ...

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8.9K Views.  Scuola Superiore Sant'Anna. This paper describes the design and fabrication of a soft unit for surgical manipulators. The base module includes three flexible fluidic actuators to achieve omnidirectional bending and elongation, and a granular jamming-based mechanism to enable stiffness control. A complete mechanical characterization is also reported. 

Video: Design and Fabrication of an Elastomeric Unit for Soft Modular Robots in Minimally Invasive Surgery

Elastomeric PGS Scaffolds in Arterial Tissue Engineering

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and increasing obesity1. Currently, bypass surgery using autologous vessels, allografts, and synthetic grafts are known as a commonly used for arterial substitutes2. However, these grafts have limited applications when an inner diameter of arteries is less than 6 mm due to low availability, thrombotic complications, compliance mismatch, and late intimal hyperplasia3,4. To overcome these limitations, tissue engineering has been successfully applied as a promising alternative to develop small-diameter arterial constructs that are nonthrombogenic, robust, and compliant. Several previous studies have developed small-diameter arterial constructs with tri-lamellar structure, excellent mechanical properties and burst pressure comparable to native arteries5,6. While high tensile strength and burst pressure by increasing collagen production from a rigid material or cell sheet scaffold, these constructs still had low elastin production and compliance, which is a major problem to cause graft failure after implantation. Considering these issues, we hypothesized that an elastometric biomaterial combined with mechanical conditioning would provide elasticity and conduct mechanical signals more efficiently to vascular cells, which increase extracellular matrix production and support cellular orientation.
The objective of this report is to introduce a fabrication technique of porous tubular scaffolds and a dynamic mechanical conditioning for applying them to arterial tissue engineering. We used a biodegradable elastomer, poly (glycerol sebacate) (PGS)7 for fabricating porous tubular scaffolds from the salt fusion method. Adult primary baboon smooth muscle cells (SMCs) were seeded on the lumen of scaffolds, which cultured in our designed pulsatile flow bioreactor for 3 weeks. PGS scaffolds had consistent thickness and randomly distributed macro- and micro-pores. Mechanical conditioning from pulsatile flow bioreactor supported SMC orientation and enhanced ECM production in scaffolds. These results suggest that elastomeric scaffolds and mechanical conditioning of bioreactor culture may be a promising method for arterial tissue engineering.

Elastomeric PGS scaffolds with vascular smooth muscle cells cultured in a pulsatile flow bioreactor may lead to promising small-diameter arterial constructs with native ECM production in a relatively short culture period.

PGS elastomerico Ponteggi in Ingegneria tessuto arterioso

Cardiovascular disease is one of the leading cause of mortality in the US and especially, coronary artery disease increases with an aging population and ...

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Polymeric Microneedle Array Fabrication by Photolithography

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a photomask consisting of embedded micro-lenses. Embedded micro-lenses were found to influence MN geometry (sharpness). Robust MN arrays with tip diameters ranging between 41.5 &#181;m &#177; 8.4 &#181;m and 71.6 &#181;m &#177; 13.7 &#181;m, with two different lengths (1,336 &#181;m &#177; 193 &#181;m and 957 &#181;m &#177; 171 &#181;m) were fabricated. These MN arrays may provide potential applications in delivery of low molecular and macromolecular therapeutic agents through skin.

Here, we present a protocol describing a mold-free fabrication process of the polymeric microneedles by photolithography.

Polimerici Microneedle Array Fabrication per Fotolitografia

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a ...

Rod-based Fabrication of Customizable Soft Robotic Pneumatic Gripper Devices for Delicate Tissue Manipulation

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations at the point of contact. It can be achieved by complementing traditional rigid grippers with soft robotic pneumatic gripper devices. This manuscript describes a rod-based approach that combined both 3D-printing and a modified soft lithography technique to fabricate the soft pneumatic gripper. In brief, the pneumatic featureless mold with chamber component is 3D-printed and the rods were used to create the pneumatic channels that connect to the chamber. This protocol eliminates the risk of channels occluding during the sealing process and the need for external air source or related control circuit. The soft gripper consists of a chamber filled with air, and one or more gripper arms with a pneumatic channel in each arm connected to the chamber. The pneumatic channel is positioned close to the outer wall to create different stiffness in the gripper arm. Upon compression of the chamber which generates pressure on the pneumatic channel, the gripper arm will bend inward to form a close grip posture because the outer wall area is more compliant. The soft gripper can be inserted into a 3D-printed handling tool with two different control modes for chamber compression: manual gripper mode with a movable piston, and robotic gripper mode with a linear actuator. The double-arm gripper with two actuatable arms was able to pick up objects of sizes up to 2 mm and yet generate lower compressive forces as compared to elastomer-coated and non-coated rigid grippers. The feasibility of having other designs, such as single-arm or hook gripper, was also demonstrated, which further highlighted the customizability of the soft gripper device, and it's potential to be used in delicate surgical manipulation to reduce the risk of tissue grip damage.

This protocol describes a rod-based approach, combining 3D-printing and soft lithography techniques for fabricating the soft gripper devices. This approach eliminates the need for an external air source by incorporating a chamber component and reduces the chance of occlusion during the sealing process, particularly for miniaturized pneumatic channels.

Fabrication Rod a base di personalizzabili morbide robotizzati pneumatici Gripper dispositivi per la manipolazione dei tessuti delicati

Soft compliant gripping is essential in delicate surgical manipulation for minimizing the risk of tissue grip damage caused by high stress concentrations ...

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; represented by &#34;devices&#34; created as combinations of individual genetic components. However, manual assembly of such large numbers of devices is time-intensive, error-prone, and costly. The increasing sophistication and scale of synthetic biology research necessitates an efficient, reproducible way to accommodate large-scale, complex, and high throughput device construction.
Here, a DNA assembly protocol using the Type-IIS restriction endonuclease based Modular Cloning (MoClo) technique is automated on two liquid-handling robotic platforms. Automated liquid-handling robots require careful, often times tedious optimization of pipetting parameters for liquids of different viscosities (e.g. enzymes, DNA, water, buffers), as well as explicit programming to ensure correct aspiration and dispensing of DNA parts and reagents. This makes manual script writing for complex assemblies just as problematic as manual DNA assembly, and necessitates a software tool that can automate script generation. To this end, we have developed a web-based software tool, http://mocloassembly.com, for generating combinatorial DNA device libraries from basic DNA parts uploaded as Genbank files. We provide access to the tool, and an export file from our liquid handler software which includes optimized liquid classes, labware parameters, and deck layout. All DNA parts used are available through Addgene, and their digital maps can be accessed via the Boston University BDC ICE Registry. Together, these elements provide a foundation for other organizations to automate modular cloning experiments and similar protocols.
The automated DNA assembly workflow presented here enables the repeatable, automated, high-throughput production of DNA devices, and reduces the risk of human error arising from repetitive manual pipetting. Sequencing data show the automated DNA assembly reactions generated from this workflow are ~95% correct and require as little as 4% as much hands-on time, compared to manual reaction preparation.

Here, an automated workflow to perform modular DNA &#34;device&#34; assembly using a modular cloning DNA assembly method on liquid-handling robots is presented. The protocol uses an intuitive software tool for generating liquid handler picklists for combinatorial DNA device library generation, which we demonstrate using two liquid handling platforms.

Automatizzato robot Liquid Handling Assembly di dispositivi modulari del DNA

Recent advances in modular DNA assembly techniques have enabled synthetic biologists to test significantly more of the available &#34;design space&#34; ...

Biomechanical Characterization of Human Soft Tissues Using Indentation and Tensile Testing

Regenerative medicine aims to engineer materials to replace or restore damaged or diseased organs. The mechanical properties of such materials should mimic the human tissues they are aiming to replace; to provide the required anatomical shape, the materials must be able to sustain the mechanical forces they will experience when implanted at the defect site. Although the mechanical properties of tissue-engineered scaffolds are of great importance, many human tissues that undergo restoration with engineered materials have not been fully biomechanically characterized. Several compressive and tensile protocols are reported for evaluating materials, but with large variability it is difficult to compare results between studies. Further complicating the studies is the often destructive nature of mechanical testing. Whilst an understanding of tissue failure is important, it is also important to have knowledge of the elastic and viscoelastic properties under more physiological loading conditions.
This report aims to provide a minimally destructive protocol to evaluate the compressive and tensile properties of human soft tissues. As examples of this technique, the tensile testing of skin and the compressive testing of cartilage are described. These protocols can also be directly applied to synthetic materials to ensure that the mechanical properties are similar to the native tissue. Protocols to assess the mechanical properties of human native tissue will allow a benchmark by which to create suitable tissue-engineered substitutes.

Tissue biomechanics is important for maintaining cell shape and function and for determining phenotype. This report demonstrates non-destructive mechanical protocols for characterizing elastic and viscoelastic properties of human soft tissues, which can be directly applied to tissue-engineered substrates to allow a close matching of engineered materials to native tissue.

Caratterizzazione biomeccanica di morbidi tessuti umani usando una indentazione e prove di trazione

Regenerative medicine aims to engineer materials to replace or restore damaged or diseased organs. The mechanical properties of such materials should ...

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices. Whole organ culture of the FSU preserves the native extracellular matrix, cell differentiation phenotypes, and cellular-matrix interactions. Thus, organ culture techniques are particularly useful for investigating the complex biological mechanisms of the IVD. Here, we describe a high-throughput approach for culturing whole lumbar mouse FSUs that provides an ideal platform for studying disease mechanisms and therapies for the IVD. Furthermore, we describe several applications that utilize this organ culture method to conduct further studies including contrast-enhanced microCT imaging and three-dimensional high-resolution finite element modeling of the IVD.

An In Vitro Organ Culture Model of the Murine Intervertebral Disc

Whole organ culture of the intervertebral disc (IVD) preserves the native extracellular matrix, cell phenotypes, and cellular-matrix interactions. Here we describe an IVD culture system using mouse lumbar and caudal IVDs in their functional spinal units and several applications utilizing this system.

Un<em&gt; In Vitro</em&gt; Organ Culture Modello del disco intervertebrale Murine

Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and ...

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver engineering, including in vivo organ decellularization followed by repopulation, has emerged as a promising approach over ex vivo liver engineering. However, postoperative survival was not achieved. The aim of this study is to develop a novel surgical technique of in vivo selective liver lobe perfusion in rats as a prerequisite for in vivo liver engineering. We generate a circuit bypass only through the left lateral lobe. Then, the left lateral lobe is perfused with heparinized saline. The experiment is performed with 4 groups (n = 3 rats per group) based on different perfusion times of 20 min, 2 h, 3 h, and 4 h. Survival, as well as the macroscopically visible change of color and the histologically determined absence of blood cells in the portal triad and the sinusoids, is taken as an indicator for a successful model establishment. After selective perfusion of the left lateral lobe, we observe that the left lateral lobe, indeed, turned from red to faint yellow. In a histological assessment, no blood cells are visible in the branch of the portal vein, the central vein, and the sinusoids. The left lateral lobe turns red after reopening the blocked vessels. 12/12 rats survived the procedure for more than one week. We are the first to report a surgical model for in vivo single liver lobe perfusion with a long survival period of more than one week. In contrast to the previously published report, the most important advantage of the technique presented here is that perfusion of 70% of the liver is maintained throughout the whole procedure. The establishment of this technique provides a foundation for in vivo partial liver engineering in rats, including decellularization and recellularization.

A Novel Surgical Technique As a Foundation for In Vivo Partial Liver Engineering in Rat

We establish a novel surgical technique for an in vivo single liver lobe perfusion model in rat as a prerequisite for further studying in vivo partial liver engineering in the future.

Una nuova tecnica chirurgica come una Fondazione per In Vivo parziale ingegneria del fegato nel ratto

Organ engineering is a novel strategy to generate liver organ substitutes that can potentially be used in transplantation. Recently, in vivo liver ...

Bioinspired Soft Robot with Incorporated Microelectrodes

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics paradigm, especially in biomedical research. Recreating artificial life-like actuation dynamics is crucial for a soft-robotic system. However, the precise control and tuning of actuation behavior still represents one of the main challenges of modern soft robotic systems. This method describes a low-cost, highly scalable, and easy-to-use procedure to fabricate an electrically controllable soft robot with life-like movements that is activated and controlled by the contraction of cardiac muscle tissue on a micropatterned sting ray-like hydrogel scaffold. The use of soft photolithography methods makes it possible to successfully integrate multiple components in the soft robotic system, including micropatterned hydrogel-based scaffolds with carbon nanotubes (CNTs) embedded gelatin methacryloyl (CNT-GelMA), poly(ethylene glycol) diacrylate (PEGDA), flexible gold (Au) microelectrodes, and cardiac muscle tissue. In particular, the hydrogels alignment and micropattern are designed to mimic the muscle and cartilage structure of the sting ray. The electrically conductive CNT-GelMA hydrogel acts as a cell scaffold that improves the maturation and contraction behavior of cardiomyocytes, while the mechanically robust PEGDA hydrogel provides structural cartilage-like support to the whole soft robot. To overcome the hard and brittle nature of metal-based microelectrodes, we designed a serpentine pattern that has high flexibility and can avoid hampering the beating dynamics of cardiomyocytes. The incorporated flexible Au microelectrodes provide electrical stimulation across the soft robot, making it easier to control the contraction behavior of cardiac tissue.

A bioinspired scaffold is fabricated by a soft photolithography technique using mechanically robust and electrically conductive hydrogels. The micropatterned hydrogels provide directional cardiomyocyte cell alignment, resulting in a tailored direction of actuation. Flexible microelectrodes are also integrated into the scaffold to bring electrical controllability for a self-actuating cardiac tissue.

Robot morbido bioispirato con microelettrodi incorporati

Bioinspired soft robotic systems that mimic living organisms using engineered muscle tissue and biomaterials are revolutionizing the current biorobotics ...

Simple, Affordable, and Modular Patterning of Cells using DNA

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells of the same or different type, micropatterning techniques have proved useful. DNA Programmed Assembly of Cells (DPAC) is a micropatterning technique that targets the adhesion of cells to a substrate or other cells using DNA hybridization. The most basic operations in DPAC begin with decorating cell membranes with lipid-modified oligonucleotides, then flowing them over a substrate that has been patterned with complementary DNA sequences. Cells adhere selectively to the substrate only where they find a complementary DNA sequence. Non-adherent cells are washed away, revealing a pattern of adherent cells. Additional operations include further rounds of cell-substrate or cell-cell adhesion, as well as transferring the patterns formed by DPAC to an embedding hydrogel for long-term culture. Previously, methods for patterning oligonucleotides on surfaces and decorating cells with DNA sequences required specialized equipment and custom DNA synthesis, respectively. We report an updated version of the protocol, utilizing an inexpensive benchtop photolithography setup and commercially available cholesterol modified oligonucleotides (CMOs) deployed using a modular format. CMO-labeled cells adhere with high efficiency to DNA-patterned substrates. This approach can be used to pattern multiple cell types at once with high precision and to create arrays of microtissues embedded within an extracellular matrix. Advantages of this method include its high resolution, ability to embed cells into a three-dimensional microenvironment without disrupting the micropattern, and flexibility in patterning any cell type.

Here we present a protocol to micropattern cells at single-cell resolution using DNA-programmed adhesion. This protocol uses a benchtop photolithography platform to create patterns of DNA oligonucleotides on a glass slide and then labels cell membranes with commercially available complementary oligonucleotides. Hybridization of the oligos results in programmed cell adhesion.

Modellazione semplice, economica e modulare delle cellule utilizzando il DNA

The relative positioning of cells is a key feature of the microenvironment that organizes cell-cell interactions. To study the interactions between cells ...

Voxel Printing Anatomy: Design and Fabrication of Realistic, Presurgical Planning Models through Bitmap Printing

Most applications of 3-dimensional (3D) printing for presurgical planning have been limited to bony structures and simple morphological descriptions of complex organs due to the fundamental limitations in accuracy, quality, and efficiency of the current modeling paradigm. This has largely ignored the soft tissue critical to most surgical specialties where the interior of an object matters and anatomical boundaries transition gradually. Therefore, the needs of the biomedical industry to replicate human tissue, which displays multiple scales of organization and varying material distributions, necessitate new forms of representation. 
Presented here is a novel technique to create 3D models directly from medical images, which are superior in spatial and contrast resolution to current 3D modeling methods and contain previously unachievable spatial fidelity and soft tissue differentiation. Also presented are empirical measurements of novel, additively manufactured composites that span the gamut of material stiffnesses seen in soft biological tissues from MRI and CT. These unique volumetric design and printing methods allow for deterministic and continuous adjustment of material stiffness and color. This capability enables an entirely new application of additive manufacturing to presurgical planning: mechanical realism. As a natural complement to existing models that provide appearance matching, these new models also allow medical professionals to "feel" the spatially varying material properties of a tissue simulant-a critical addition to a field in which tactile sensation plays a key role.

This method demonstrates a voxel-based 3D printing workflow, which prints directly from medical images with exact spatial fidelity and spatial/contrast resolution. This enables the precise, graduated control of material distributions through morphologically complex, graduated materials correlated to radiodensity without loss or alteration of data.

Anatomia della stampa Voxel: progettazione e fabbricazione di modelli di pianificazione realistici e prechirurgici tramite stampa bitmap

Most applications of 3-dimensional (3D) printing for presurgical planning have been limited to bony structures and simple morphological descriptions of ...