Kyle J. Lynch

Novel Technique for Repair of Severed Peripheral Nerves in Rats Using Polyurea Crosslinked Silica Aerogel Scaffold

ABSTRACT Purpose/Aim: To design, synthesize, and test in vivo an aerogel-based top-open peripheral nerve scaffold to simultaneously support and guide multiple completely severed peripheral nerves in a rat model. Also, to explore options for immobilizing severed nerves on the aerogel material without the use of sutures resulting in reduced surgical time. Materials and Method: A novel material and approach was developed for the reattachment of severed peripheral nerves. Nerve confinement and alignment in this case relies on the surface properties of a lightweight, highly porous, polyurea crosslinked silica aerogel scaffold. The distal and proximal ends of completely transected nerve terminals were positioned inside prefabricated “top-open” corrugated channels that cradled approximately two thirds of the circumference of the nerve trunk and connectivity of the severed nerves was evaluated using sciatic function index (SFI) technique for five months post-surgery on 10 female Sprague–Dawley rats then compared with the gold standard for peripheral nerve repair. The interaction of nerves with the surface of the scaffold was investigated also. Results and Conclusion: Multichannel aerogel-based nerve support scaffold showed similar SFI recovery trend as the case suture repair technique. Usage of an adhesion-promoting coating reduced the friction between the nerve and the scaffold leading to slippage and lack of attachment between nerve and surface. The aerogel scaffold used in this study did not collapse under pressure during the incubation period and allowed for a rapid and non-invasive peripheral nerve repair approach without the demands of microsurgery on both time and surgical expertise. This technique may allow for simultaneous repair and reconnection of multiple severed nerves particularly relevant to nerve branching sites.

Polymer-Encapsulated Phosphor Particles for In Vivo Phosphor Luminescence Applications

Phosphor thermometry is a highly sensitive, rapid, and portable thermal sensing technique that offers advantages over traditional contact-based thermometry techniques. Phosphor particles would however require an encapsulation medium that is biocompatible and yet optically transparent to permit optical access to the embedded phosphor particles. Here, phosphor-doped silicone implants with varying concentrations were prepared and tested in a rat model. Results indicate that such phosphor-doped polymeric implants are stable, produce a detectable signal, and demonstrate the feasibility of phosphor thermometry as a noninvasive remote thermal sensing technique for in vivo applications. Also, encapsulation in silicone did not lead to significant attenuation of the incoming signal. GRAPHICAL ABSTRACT

In Vivo X-Ray Imaging of Phosphor-Doped PDMS and Phosphor-Doped Aerogel Biomaterials

Noninvasive rapid in vivo imaging and detection of biomedical implants is a critical part of the design and implementation of smart implants. Thermographic phosphors offer a precise and remotely accessible sensing method that has been utilized here. We present the first in vivo X-ray images of La2O2S:Eu-doped crosslinked silica aerogels and polydimethylsiloxane (PDMS) with increasing dopant concentrations. Results show that native PDMS and crosslinked silica aerogel do not show noticeable attenuation of X-rays while image analysis yields values of the absorption coefficient of 0.014 for the doped aerogel and a range of 0.015–0.017 for the doped PDMS. GRAPHICAL ABSTRACT

Optimal digestion theory does not predict the effect of pathogens on intestinal plasticity

One prediction of optimal digestion theory is that organisms will increase the relative length of their digestive tracts when food resources become limited. We used theory of optimal digestion to test whether tadpoles can adjust the relative length of their intestines when challenged with the fungal pathogen Batrachochytrium dendrobatidis (Bd). The degree of tadpole mouthpart damage, a symptom of Bd infections that reduces food consumption, was associated positively with the length of tadpole intestines relative to their body size, consistent with optimal digestion theory. After controlling for mouthpart damage, tadpoles exposed to Bd had shorter intestines relative to their body size, opposite to the predictions of optimal digestion theory. One explanation of why tadpoles with higher Bd loads have shorter relative intestinal lengths is that they divert energy from maintaining intestinal and overall growth towards anti-parasite defences.

Batrachochytrium dendrobatidis exposure effects on foraging efficiencies and body size in anuran tadpoles.

Chytridiomycosis, the amphibian disease caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd), is fatal to adults of many species. Bd is largely sublethal to amphibian larvae; however, it is known to reduce larval (i.e. tadpole) growth rates, with possible long-term effects on population dynamics and fitness. We conducted an experiment to test how Bd altered southern leopard frog Lithobates sphenocephalus tadpole mouthpart damage, percentage of food ingested, and subsequent body size. We examined our results using path analyses. We hypothesized that Bd would increase mouthpart damage, causing less food to be ingested, and ultimately reduce body size. In our model, both Bd exposure and increased mouthpart damage significantly reduced food ingested and subsequent body size. However, our study provides evidence against the long-standing hypothesis of mouthpart damage as a pathway for Bd-induced reductions in larval group. Here we provide evidence for reduced foraging efficiency (percentage of food ingested) as a mechanism for Bd-induced reductions in body size. This work highlights the importance of studying the sublethal effects of Bd on larval amphibians.

Investigation of surface topography and stiffness on adhesion and neurites extension of PC12 cells on crosslinked silica aerogel substrates

Fundamental understanding and characterization of neural response to substrate topography is essential in the development of next generation biomaterials for nerve repair. Aerogels are a new class of materials with great potential as a biomaterial. In this work, we examine the extension of neurites by PC12 cells plated on matrigel-coated and collagen-coated mesoporous aerogel surfaces. We have successfully established the methodology for adhesion and growth of PC12 cells on polyurea crosslinked silica aerogels. Additionally, we have quantified neurite behaviors and compared their response on aerogel substrates with their behavior on tissue culture (TC) plastic, and polydimethylsiloxane (PDMS). We found that, on average, PC12 cells extend longer neurites on crosslinked silica aerogels than on tissue culture plastic, and, that the average number of neurites per cluster is lower on aerogels than on tissue culture plastic. Aerogels are an attractive candidate for future development of smart neural implants and the work presented here creates a platform for future work with this class of materials as a substrate for bioelectronic interfacing.

Mouthparts of Southern Leopard Frog, Lithobates sphenocephalus , Tadpoles not Affected by Exposure to a Formulation of Glyphosate

This study evaluated the impacts of Roundup® on tadpole mouthpart damage as a mechanism for reduced growth and developmental rates in Lithobates sphenocephalus (Southern leopard frog) tadpoles. We did not find evidence that Roundup® damages larval mouthparts, nor was there a significant relationship between mouthpart damage and either body condition or developmental rate. However, the highest concentration of Roundup® significantly stunted development compared to all other treatments. Although we observed a significant effect of Roundup® on developmental rate, we conclude that mouthpart damage is likely not a mechanism for this life history response.

Growing Neural PC-12 Cell on Crosslinked Silica Aerogels Increases Neurite Extension in the Presence of an Electric Field

Externally applied electrical stimulation (ES) has been shown to enhance the nerve regeneration process and to influence the directionality of neurite outgrowth. In addition, the physical and chemical properties of the substrate used for nerve-cell regeneration is critical in fostering regeneration. Previously, we have shown that polyurea-crosslinked silica aerogels (PCSA) exert a positive influence on the extension of neurites by PC-12 cells, a cell-line model widely used to study neurite extension and electrical excitability. In this work, we have examined how an externally applied electric field (EF) influences the extension of neurites in PC-12 cells grown on two substrates: collagen-coated dishes versus collagen-coated crosslinked silica aerogels. The externally applied direct current (DC) bias was applied in vitro using a custom-designed chamber containing polydimethysiloxane (PDMS) embedded copper electrodes to create an electric field across the substrate for the cultured PC-12 cells. Results suggest orientation preference towards the anode, and, on average, longer neurites in the presence of the applied DC bias than with 0 V DC bias. In addition, neurite length was increased in cells grown on silica-crosslinked aerogel when compared to cells grown on regular petri-dishes. These results further support the notion that PCSA is a promising material for nerve regeneration.