Cellular microenvironments are important in coaxing cells to behave collectively as functional, structured tissues. Important cues in this microenvironment are the chemical, mechanical and spatial arrangement of the supporting matrix in the extracellular space. In engineered tissues, synthetic scaffolding provides many of these microenvironmental cues. Key requirements are that synthetic scaffolds should recapitulate the native three-dimensional (3D) hierarchical fibrillar structure, possess biomimetic surface properties and demonstrate mechanical integrity, and in some tissues, anisotropy. Electrospinning is a popular technique used to fabricate anisotropic nanofiber scaffolds. However, it suffers from relatively low production rates and poor control of fiber alignment without substantial modifications to the fiber collector mechanism. Additionally, many biomaterials are not amenable for fabrication via high-voltage electrospinning methods. Hence, we reasoned that we could utilize rotary jet spinning (RJS) to fabricate highly aligned hybrid protein-polymer with tunable chemical and physical properties. In this study, we engineered highly aligned nanofiber constructs with robust fiber alignment from blends of the proteins collagen and gelatin, and the polymer poly-ε-caprolactone via RJS and electrospinning. RJS-spun fibers retain greater protein content on the surface and are also fabricated at a higher production rate compared to those fabricated via electrospinning. We measured increased fiber diameter and viscosity, and decreasing fiber alignment as protein content increased in RJS hybrid fibers. RJS nanofiber constructs also demonstrate highly anisotropic mechanical properties mimicking several biological tissue types. We demonstrate the bio-functionality of RJS scaffold fibers by testing their ability to support cell growth and maturation with a variety of cell types. Our highly anisotropic RJS fibers are therefore able to support cellular alignment, maturation and self-organization. The hybrid nanofiber constructs fabricated by RJS therefore have the potential to be used as scaffold material for a wide variety of biological tissues and organs, as an alternative to electrospinning.

The bulk production of polymeric nanofibers is important for fabricating high performance, nano-scale materials. Rotary Jet-Spinning (RJS) enables the mass production of nanostructured fibers by centrifugal forces but may result in inconsistent surface morphologies. Because nanofiber performance is dependent on its surface features, we asked which parameters must be optimized during production to control fiber morphology. We developed and tested a mathematical model that describes how the competition between fluid instability and solvent removal in RJS regulates the degree of beading in fibers. Our data suggests that solvent evaporation during the spinning process causes an increase in jet viscosity and that these changes inhibit both bead formation and jet thinning. The RJS was used to vary experimental parameters showing that fiber beading can be reduced by increasing solvent volatility, solution viscosity, and spinning velocity. Collectively, our results demonstrate that nanofiber morphology and diameter can be precisely controlled during RJS manufacturing.

Wounds in the fetus can heal without scarring. Consequently, biomaterials that attempt to recapitulate the biophysical and biochemical properties of fetal skin have emerged as promising pro-regenerative strategies. The extracellular matrix (ECM) protein fibronectin (Fn) in particular is believed to play a crucial role in directing this regenerative phenotype. Accordingly, Fn has been implicated in numerous wound healing studies, yet remains untested in its fibrillar conformation as found in fetal skin. Here, we show that high extensional (∼1.2 ×10 s) and shear (∼3 ×10 s) strain rates in rotary jet spinning (RJS) can drive high throughput Fn fibrillogenesis (∼10 mL/min), thus producing nanofiber scaffolds that are used to effectively enhance wound healing. When tested on a full-thickness wound mouse model, Fn nanofiber dressings not only accelerated wound closure, but also significantly improved tissue restoration, recovering dermal and epidermal structures as well as skin appendages and adipose tissue. Together, these results suggest that bioprotein nanofiber fabrication via RJS could set a new paradigm for enhancing wound healing and may thus find use in a variety of regenerative medicine applications.

Nanofiber production platforms commonly rely on volatile carrier solvents or high voltages. Production of nanofibers comprised of charged polymers or polymers requiring nonvolatile solvents thus typically requires customization of spinning setup and polymer dope. In severe cases, these challenges can hinder fiber formation entirely. Here, a versatile system is presented which addresses these challenges by employing centrifugal force to extrude polymer dope jet through an air gap, into a flowing precipitation bath. This voltage-free approach ensures that nanofiber solidification occurs in liquid, minimizing surface tension instability that results in jet breakup and fiber defects. In addition, nanofibers of controlled size and morphology can be fabricated by tuning spinning parameters including air gap length, spinning speed, poly mer concentration, and bath composition. To demonstrate the versatility of our platform, para-aramid (e.g., Kevlar) and biopolymer (e.g., DNA, alginate) nanofibers are produced that cannot be readily produced using standard nanofiber production methods.

One of the goals of soft robotics is the ability to interface with the human body. Traditionally, silicone materials have dominated the field of soft robotics. In order to shift to materials that are more compatible with the body, developments will have to be made into biodegradable and biocompatible soft robots. This investigation focused on developing gummy actuators which are biodegradable, edible, and tasty. Creating biodegradable and edible actuators can be both sold as an interactive candy product and also inform the design of implantable soft robotic devices. First, commercially available gelatin-based candies were recast into pneumatic actuators utilizing molds. Edible robotic devices were pneumatically actuated repeatedly (up to n=8 actuations) using a 150 psi power inflator. To improve upon the properties of actuators formed from commercially available candy, a novel gelatin-based formulation, termed the “Fordmula” was also developed and used to create functional actuators. To investigate the mechanics and functionality of the recast gummy material and the Fordmula, compression testing and biodegradation studies were performed. Mechanical compression tests showed that recast gummy materials had similar properties to commercially available candies and at low strain had similar behavior to traditional silicone materials. Degradation studies showed that actuation was possible within 15 minutes in a biologically relevant solution followed by complete dissolution of the actuator afterwards. A taste test with elementary aged children demonstrated the fun, edible, and educational appeal of the candy actuators. Edible actuator development was an entry and winning submission in the High School Division of the Soft Robotics Toolkit Design Competition hosted by Harvard University. Demonstration of edible soft robotic actuators created by middle and high school aged students shows the applicability of the Soft Robotics Toolkit for K12 STEM education.