Assessing effectiveness of Komagataeibacter strains for producing surface-microstructured cellulose via guided assembly-based biolithography.

In this study, a medical device made of surface microstructured bacterial cellulose was produced using cellulose-producing acetic acid bacteria wild-type strains in combination with guided assembly-based biolithography. The medical device aims at interfering with the cell's focal adhesion establishment and maturation around implantable devices placed in soft tissues by the symmetrical array on its surface. A total of 25 Komagataeibacter strains was evaluated over a three-step selection. In the first step, the ability of strains to produce a suitable bacterial cellulose layer with high production yield was examined, then nine strains, with a uniform and smooth layer of bacterial cellulose, were cultured in a custom-made silicone bioreactor and finally the characteristics of the symmetrical array of topographic features on the surface were analysed. Selected strains showed high inter and intra species variability in bacterial cellulose production. The devices obtained by K2G30, K1G4, DSM 46590 (Komagataeibacter xylinus), K2A8 (Komagataeibacter sp.) and DSM 15973T (Komagataeibacter sucrofermentas) strains were pouched-formed with hexagonal surface pattern required for reducing the formation of fibrotic tissue around devices, once they are implanted in soft tissues. Our findings revealed the effectiveness of the selected Komagataeibacter wild-type strains in producing surface microstructured bacterial cellulose pouches for making biomedical devices.

On the function of biosynthesized cellulose as barrier against bacterial colonization of VAD drivelines.

Bacterial colonization of drivelines represents a major adverse event in the implantation of left ventricular assist devices (L-VADs) for the treatment of congestive heart failure. From the external driveline interface and through the skin breach, pathogens can ascend to the pump pocket, endangering the device function and the patient's life. Surface Micro-Engineered Biosynthesized cellulose (BC) is an implantable biomaterial, which minimizes fibrotic tissue deposition and promotes healthy tissue regeneration. The topographic arrangement of cellulose fibers and the typical material porosity support its potential protective function against bacterial permeation; however, this application has not been tested in clinically relevant animal models. Here, a goat model was adopted to evaluate the barrier function of BC membranes. The external silicone mantle of commercial L-VAD drivelines was implanted percutaneously with an intervening layer of BC to separate them from the surrounding soft tissue. End-point evaluation at 6 and 12 weeks of two separate animal groups revealed the local bacterial colonization at the different interfaces in comparison with unprotected driveline mantle controls. The results demonstrate that the BC membranes established an effective barrier against the bacterial colonization of the outer driveline interface. The containment of pathogen infiltration, in combination with the known anti-fibrotic effect of BC, may promote a more efficient immune clearance upon driveline implantation and support the efficacy of local antibiotic treatments, therefore mitigating the risk connected to their percutaneous deployment.

Microengineered biosynthesized cellulose as anti-fibrotic in vivo protection for cardiac implantable electronic devices.

Upon cardiac implantable electronic device (CIED) exchange, upgrade, or revision surgery patients are exposed to a considerable risk of adverse events. The presence of firm fibrotic tissue endangers these procedures. Leads can be damaged in the attempt of freeing them from fibrotic tissue. Hematoma can form as result of capsulectomy, pocket debridement and leads dissection. Due to the increasing number of CIED exchange, upgrade and revision surgeries, the incidence of related complications is expected to rise in the near future.The aim of the study was to evaluate the feasibility, safety, and performance of a rationally micro-engineered non-resorbable biosynthesized cellulose (BC) membrane as conformal wrapping protection around CIED implants. Protective membranes were generated by means of a recently established method to transfer on-demand microscale geometries onto the surface of BC. A chronic minipig animal model was selected to investigate the performance of the BC anti-fibrotic protection, directly measured as reduction of fibrotic tissue formation. Sixteen (n = 16) animals received each one BC coated pacemaker (PMC) and one native pacemaker (BI) at equivalent anatomical sites. BC protective layers were juxtaposed around pacemakers through a fast and well-repeatable procedure. Explants were performed at 3 and 12 months after implantation. Endpoint analysis showed that the BC protective layers were 100% integer, with no sign of chemical or mechanical degradation and appeared as a thin layer of white-tan material, adherent to the surrounding thin fibrous capsule, from which it could be peeled off by gently pulling with forceps. The protective effect of micro-engineered BC yielded an average thickness reduction of 66% of the fibrotic tissue thickness generated around PMC, as compared to that measured around the naked counterpart (i.e. the BI). When protected by in BC, both the generator and the proximal parts of the leads were completely free from fibrotic tissue. The insertion of an anti-adhesive, non-resorbable and well-tolerated BC interface between the implant and the surrounding tissue in the surgical pocket significantly reduced the formation of fibrotic tissue, ensuring an easy access to the device pocket, and thus creating the conditions for simplified CIED revision surgeries.

A micron-scale surface topography design reducing cell adhesion to implanted materials.

The micron-scale surface topography of implanted materials represents a complementary pathway, independent of the material biochemical properties, regulating the process of biological recognition by cells which mediate the inflammatory response to foreign bodies. Here we explore a rational design of surface modifications in micron range to optimize a topography comprised of a symmetrical array of hexagonal pits interfering with focal adhesion establishment and maturation. When implemented on silicones and hydrogels in vitro, the anti-adhesive topography significantly reduces the adhesion of macrophages and fibroblasts and their activation toward effectors of fibrosis. In addition, long-term interaction of the cells with anti-adhesive topographies markedly hampers cell proliferation, correlating the physical inhibition of adhesion and complete spreading with the natural progress of the cell cycle. This solution for reduction in cell adhesion can be directly integrated on the outer surface of silicone implants, as well as an additive protective conformal microstructured biocellulose layer for materials that cannot be directly microstructured. Moreover, the original geometry imposed during manufacturing of the microstructured biocellulose membranes are fully retained upon in vivo exposure, suggesting a long lasting performance of these topographical features after implantation.

Surface-structured bacterial cellulose with guided assembly-based biolithography (GAB).

A powerful replica molding methodology to transfer on-demand functional topographies to the surface of bacterial cellulose nanofiber textures is presented. With this method, termed guided assembly-based biolithography (GAB), a surface-structured polydimethylsiloxane (PDMS) mold is introduced at the gas-liquid interface of an Acetobacter xylinum culture. Upon bacterial fermentation, the generated bacterial cellulose nanofibers are assembled in a three-dimensional network reproducing the geometric shape imposed by the mold. Additionally, GAB yields directional alignment of individual nanofibers and memory of the transferred geometrical features upon dehydration and rehydration of the substrates. Scanning electron and atomic force microscopy are used to establish the good fidelity of this facile and affordable method. Interaction of surface-structured bacterial cellulose substrates with human fibroblasts and keratinocytes illustrates the efficient control of cellular activities which are fundamental in skin wound healing and tissue regeneration. The deployment of surface-structured bacterial cellulose substrates in model animals as skin wound dressing or body implant further proves the high durability and low inflammatory response to the material over a period of 21 days, demonstrating beneficial effects of surface structure on skin regeneration.

Morphing surfaces enable acoustophoretic contactless transport of ultrahigh-density matter in air.

The controlled contactless transport of heavy drops and particles in air is of fundamental interest and has significant application potential. Acoustic forces do not rely on special material properties, but their utility in transporting heavy matter in air has been restricted by low power and poor controllability. Here we present a new concept of acoustophoresis, based on the morphing of a deformable reflector, which exploits the low reaction forces and low relaxation time of a liquid with enhanced surface tension through the use of thin overlaid membrane. An acoustically induced, mobile deformation (dimple) on the reflector surface enhances the acoustic field emitted by a line of discretized emitters and enables the countinuos motion of heavy levitated samples. With such interplay of emitters and reflecting soft-structure, a 5 mm steel sphere (0.5 grams) was contactlessly transported in air solely by acoustophoresis.

Assessment of intracranial dynamics in hydrocephalus: effects of viscoelasticity on the outcome of infusion tests.

OBJECT: The treatment of hydrocephalus requires insight into the intracranial dynamics in the patient. Resistance to CSF outflow (R0) is a clinically obtainable parameter of intracranial fluid dynamics that quantifies the apparent resistance to CSF absorption. It is used as a criterion for the selection of shunt candidates and serves as an indicator of shunt performance. The R0 is obtained clinically by performing 1 of 3 infusion tests: constant flow, constant pressure, or bolus infusion. Among these, the bolus infusion method has the shortest examination times and provides the shortest time of exposure of patients to artificially increased intracranial pressure (ICP) levels. However, for unknown reasons, the bolus infusion method systematically underestimates the R0. Here, the authors have tested and verified the hypothesis that this underestimation is due to lack of accounting for viscoelasticity of the craniospinal space in the calculation of the R0. METHODS: The authors developed a phantom model of the human craniospinal space in order to reproduce in vivo pressure-volume (PV) relationships during infusion testing. The phantom model followed the Marmarou exponential PV equation and also included a viscoelastic response to volume changes. Parameters of intracranial fluid dynamics, such as the R0, could be controlled and set independently. In addition to the phantom model, the authors designed a computational framework for virtual infusion testing in which viscoelasticity can be turned on or off in a controlled manner. Constant flow, constant pressure, and bolus infusion tests were performed on the phantom model, as well as on the virtual computational platform, using standard clinical protocols. Values for the R0 were derived from each infusion test by using both a standard method based on the Marmarou PV equation and a novel method based on a system identification approach that takes into account viscoelastic behavior. RESULTS: Experiments with the phantom model confirmed clinical observations that both the constant flow and constant pressure infusion tests, but not the bolus infusion test, yield correct R0 values when they are determined with the standard method according to Marmarou. Equivalent results were obtained using the computational framework. When the novel system identification approach was used to determine the R0, all of the 3 infusion tests yielded correct values for the R0. CONCLUSIONS" The authors' investigations demonstrate that intracranial dynamics have a substantial viscoelastic component. When this viscoelastic component is taken into account in calculations, the R0, is no longer underestimated in the bolus infusion test.

Craniospinal pressure-volume dynamics in phantom models.

Regulation of intracranial pressure (ICP) is vital to proper brain function. Pathologic conditions such as traumatic brain injury and hydrocephalus can cause lethal changes in ICP through an imbalance of fluid passage into and out of the craniospinal space. The relationship between craniospinal volume and pressure determines to a large extent whether such imbalance can be compensated or if it will lead to neuronal damage. Phantom models are predisposed for the evaluation of medical procedures and devices that alter volume in the spinal or cranial space. However, current phantoms have substantial limitations in the reproduction of craniospinal pressure-volume relationships, which need to be overcome prior to their deployment outside the basic research setting. We present herein a novel feedback controlled phantom for the reproduction of any physiologic or pathologic pressure-volume relation. We compare its performance to those of existing passive methods, showing that it follows reference curves more precisely during both infusion of large volumes and fast oscillatory volume changes.

Phantom model of physiologic intracranial pressure and cerebrospinal fluid dynamics.

We describe herein a novel life-size phantom model of the intracranial cavity and its validation. The cerebrospinal fluid (CSF) domains including ventricular, cysternal, and subarachnoid spaces were derived via magnetic resonance imaging. Brain mechanical properties and cranio-spinal compliance were set based on published data. Both bulk and pulsatile physiologic CSF flow were modeled. Model validation was carried out by comparisons of flow and pressure measurements in the phantom with published in vivo data of healthy subjects. Physiologic intracranial pressure with 10 mmHg mean and 0.4 mmHg peak pulse amplitude was recorded in the ventricles. Peak CSF flow rates of 0.2 and 2 ml/s were measured in the cerebral aqueduct and subarachnoid space, respectively. The phantom constitutes a first-of-its-kind approach to modeling physiologic intracranial dynamics in vitro. Herein, we describe the phantom design and manufacturing, definition and implementation of its operating parameters, as well as the validation of the modeled dynamics.