We define Regenerative Engineering as a Convergence of Advanced Materials Science, Stem Cell Science, Physics, Developmental Biology, and Clinical Translation. We believe that an “un-siloed’ technology approach will be important in the future to realize grand challenges such as limb and organ regeneration. We also believe that biomaterials will play a key role in achieving overall translational goals. Through convergence of a number of technologies, with advanced materials science playing an important role, we believe the prospect of engaging future grand challenges is possible. Regenerative Engineering as a field is particularly suited for solving clinical problems that are relevant today. The paradigms utilized can be applied to the regeneration of tissue in the shoulder where tendon and muscle currently have low levels of regenerative capability, and the consequences, especially in alternative surgical solutions for massive tendon and muscle loss at the shoulder have demonstrated significant morbidity. Polymer, polymer-cell, and polymer biological factor, and polymer-physical systems can be utilized to propose a range of solutions to shoulder tissue regeneration. The approaches, possibilities, limitations and future strategies, allow for a variety of clinical solutions in musculoskeletal disease treatment.

Tissue engineering and regenerative medicine are increasingly taking advantage of active materials, allowing to provide specific clues to the cells. In particular, the use of electroactive polymers that deliver an electrical signal to the cells upon mechanical solicitation, open new scientific and technological opportunities, as they in fact mimic signals and effects that occur in living tissues, allowing the development of suitable microenvironments for tissue regeneration. Thus, a novel overall strategy for bone and muscle tissue engineering was developed based on the fact that these cells type are subjected to mechano-electrical stimuli in their in vivo microenvironment and that piezo- and magnetoelectric polymers, used as scaffolds, are suitable for delivering those cues. The processing and functional characterizations of piezoelectric and magnetoelectric polymers based on poly(vinylindene fluoride) and poly-L-lactic acid in a variety of shapes, from microspheres to electrospun mats and three dimensional scaffolds, are shown as well as their performance in the development of novel bone and muscle tissue engineering.

A novel freeze-cast porous chitosan conduit for peripheral nerve repair with highly-aligned, double layered porosity, which provides the ideal mechanical and chemical properties was designed, manufactured, and assessed in vivo. Efficacies of the conduit and the control inverted nerve autograft were evaluated in bridging 10-mm Lewis rat sciatic nerve gap at 12 weeks post-implantation. Biocompatibility and regenerative efficacy of the porous chitosan conduit were evaluated through the histomorphometric analysis of longitudinal and transverse sections. The porous chitosan conduit was found to have promising regenerative characteristics, promoting the desired neovascularization, and axonal ingrowth and alignment through a combination of structural, mechanical and chemical cues.

Current FDA-approved permanent female sterilization procedures are invasive and/or require the implantation of non-biodegradable materials. These techniques pose risks and complications, such as device migration, fracture, and tubal perforation. We propose a safe, non-invasive biodegradable tissue scaffold to effectively occlude the Fallopian tubes within 30 days of implantation. Specifically, the Fallopian tubes are mechanically de-epithelialized, and a tissue scaffold is placed into each tube. It is anticipated that this procedure can be performed in less than 30 minutes by an experienced obstetrics and gynaecology practitioner. Advantages of this method include the use of a fully bio-resorbable polymer, low costs, lower risks, and the lack of general anaesthesia. The scaffold devices are freeze-cast allowing for the custom-design of structural, mechanical, and chemical cues through material composition, processing parameters, and functionalization. The performance of the biomaterial and de-epithelialization procedure was tested in an in vivo rat uterine horn model. The scaffold response and tissue-biomaterial interactions were characterized microscopically post-implantation. Overall, the study resulted in the successful fabrication of resilient, easy-to-handle devices with an anisotropic scaffold architecture that encouraged rapid bio-integration through notable angiogenesis, cell infiltration, and native collagen deposition. Successful tubal occlusion was demonstrated at 30 days, revealing the great promise of a sterilization biomaterial.

In this study, the spontaneous formation of hydrogels from a water-soluble and amphiphilic ABA-type block copolymer in an aqueous system was examined. The block copolymers were composed of poly(2-methacryloyloxyethyl phosphorylcholine) (MPC) as the B segment and poly(n-butyl methacrylate) (BMA) as the A segment. A two-step living radical polymerization of the corresponding monomers yielded a well-defined ABA-type block copolymer. By regulating the polymerization degree of each segment, the water solubility and amphiphilic natures of the resulting block copolymer could be controlled. The block copolymers spontaneously formed hydrogels when the composition of water in the medium increased to 80 vol% in the water/ethanol mixture. This was due to molecular aggregation of the block copolymers driven by the hydrophobic interactions between the poly(BMA) segments in the polymer. This hydrogel formation depended on the composition and polymerization degree of the MPC units. The poly(MPC) also showed good biocompatibility, thereby rendering the block copolymers applicable for the immobilization of biological components such as antibodies and enzymes and the encapsulation of cells under mild conditions.

Tissue Engineering has been studied to develop tissues as an alternative approach to the organ regeneration. Successful artificial tissue growth in regenerative medicine depends on the precise scaffold fabrication as well as the cell-cell and cell-scaffold interaction. Scaffolds are extracellular matrices that guide cells to grow in 3D to regenerate the tissues. Cell-seeded scaffolds must be implanted to the damaged tissues to do the tissue regeneration. Scaffolds’ mechanical properties and porosities are the two main scaffold fabrication parameters as the scaffolds must be able to hold the pressure due to the surrounding tissues after the implantation process. In this research, scaffolds were fabricated by photolithography and Poly(ethylene glycol) Diacrylate (PEGDA) which is a biocompatible and biodegradable material was used as a fabrication material. In order to compare the compressive properties of PEGDA only with the compressive properties of drug delivered PEGDA, firstly, PEGDA only solutions were prepared. Then, PEGDA was mixed with Meloxicam 15 mg, Hydrochlorothiazide 12.5 mg, Cyclobenzaprine 10 mg and Spironolactone-hctz 25-25 mg respectively and they were placed under the UV light for about 15 minutes to solidify the cylindrical shaped hydrogels. 5 samples from each group were fabricated under the same conditions. Laboratory temperature, photoinitiator concentration and UV light intensity was kept constant during the fabrication process. After the fabrication was completed, Instron 3369 universal mechanical testing machine with the 5 mm/min compression rate was used to do the compression tests to compare the drug effects on PEGDA hydrogels. Our results indicate that average ultimate strength of PEGDA only samples was 3.820 MPa. Also, due to the fact that Meloxicam 15 mg and PEGDA mixture did not solidify under the UV light at all, compression test could not be performed for PEGDA- Meloxicam 15 mg mixture. However, Hydrochlorothiazide 12.5 mg, Cyclobenzaprine 10 mg and Spironolactone-hctz 25-25 mg dissolved within the PEGDA completely and our compression results show that average ultimate strengths were 3.372 MPa, 1.602 MPa, 1.999 MPa respectively. This preliminary research showcases that compressive properties of the PEGDA-based photopolymerized scaffolds can be altered with the control of the drug type and drug concentration.

Hydroxyapatite (HA)/nanotubular titanium dioxide (TiO2) composite coatings loaded with antibiotics were developed to combine biocompatibility and antibacterial property. TiO2 nanotubes were first fabricated on Ti plates using anodization techniques. Then HA nanocrystals were synthesized on the TiO2 nanotubes by electrochemical deposition, followed by loading of a model drug compound, streptomycin. The streptomycin release profile of the composite coating was investigated. Bacterial tests demonstrate that the streptomycin-loaded composite coatings were highly effective in inhibiting bacterial growth. Simulated body fluid (SBF) experiments indicated that the composite coatings possessed good osseointegration capability.

Biomimetic Method has been widely used to prepare calcium phosphate coatings on Ti and its alloys. This modification is based on a Synthetic/simulated Body Fluid (BSF) which facilitates the mimicking of the biological process in order to provide hard tissue repairs. The formation of HA and other calcium phosphates under biological medium and SBF occurs in the presence of Ca2+ and PO43- ions, as well as essential ions such as: Mg2+, HCO3-, K+ and Na+. Ti-15Mo alloy samples were irradiated by pulsed Yb: YAG pulsed laser beam under air and atmospheric pressure. Sequentially, calcium phosphate coatings were deposited on the irradiated surfaces by the biomimetic method. The biomimetic calcium phosphates-based surfaces were submitted to heat treatment conditions at 350°C and 600°C. The present study correlates two conditions of fluency (1,91 and 5,54 J.cm-2) as established have a sufficient energy to promote ablation on the laser beam irradiated surfaces. Likewise, it has been demonstrated the processes of fusion and fast solidification from the laser beam irradiation, under ambient atmosphere, inducing the formation of stoichiometric TiO2 and non-stoichiometric titanium oxides, including Ti3O5, TiO, Ti3O and Ti6O with different oxide percentages depending on the fluency applied. Besides that, laser modification has allowed a clean and reproducible process, providing no traces of contamination, an important feature for clinical applications. The physico-chemical and morphological analysis indicated the formation of a multiphase coatings depending on the heat treatment temperature performed to 350 °C (ACP1 and 2, HAD, HA phases) and 600 °C (HAD, HA and β-TCP phases). It is worth noting that multiphasic bioceramic systems has been gaining attention for biomedical applications. Thus, the laser beam irradiation associated to bioactive coatings of calcium phosphates of biological interest have shown to be promising and economically feasible for use in dental and orthopedic implants.

Poloxamer 407 (P407) is a biocompatible thermo-setting polymer, while agarose is a biocompatible thermo-softening material. It is interesting to mix them to examine any possible synergy in thermomechanical properties. In this study, rotational rheometer was adopted to study rheological properties of the mixtures of agarose/P407 gels with different concentrations at various frequencies, strain rates and temperatures. It has revealed that the addition of P407 decreased the gel stiffness by an order of magnitude. For the given combinations in this study, the increase in agarose concentration would increase both the storage modulus and loss modulus of the gel mixtures. The variation in P407 concentration (2.5%-10%) minimally changes the composite moduli. These agarose/P407 gel mixtures also exhibited shear thinning behavior. However, the addition of P407 (2.5%-10%) to agarose gel only has very small effect on thermomechanical properties of agarose gels. The overall transition temperature for these gel mixtures is governed by P407 melting point where the phase change starts around 55°C and the gels completely collapse at the melting temperature of agarose.

Dental pulp stem cells (DPSCs) can differentiate into bone cells when provided the correct environment, potentially generating cells to repair non-union fractures. Polylactic Acid (PLA) is a biocompatible polymer for 3-D printing of scaffolds, but DPSCs do not proliferate well on PLA. With the goal of making PLA more conducive for DPSC growth, Graphene Oxide (GO); partially reduced Graphene Oxide (pRGO); GO with iron nanoparticles (FeGO) or Fe-pRGO were incorporated into PLA and spun cast as thin films onto silicon wafers for DPSC plating. DPSCs on Fe-pRGO displayed the fastest doubling time and the highest cell modulus; Fe-pRGO with exterior magnets produced high cell density. SEM demonstrated DPSC mineralization, whereas PLA-only DPSC cultures showed none. Results suggest that PLA/Fe-pRGO and PLA/pRGO enhance DPSC proliferation and possibly differentiation with the potential for use as a 3-D printed scaffold for tissue engineering.