Biodegradable and Bioactive PCL-PGS Core-Shell Fibers for Tissue Engineering.
ABSTRACT: Poly(glycerol sebacate) (PGS) has increasingly become a desirable biomaterial due to its elastic mechanical properties, biodegradability, and biocompatibility. Here, we report microfibrous core-shell mats of polycaprolactone (PCL)-PGS prepared using wet-wet coaxial electrospinning. The anticoagulant heparin was immobilized onto the surface of these electrospun fiber mats, and they were evaluated for their chemical, mechanical, and biological properties. The core-shell structure of PCL-PGS provided tunable degradation and mechanical properties. The slowly degrading PCL provided structural integrity, and the fast degrading PGS component increased fiber elasticity. Young's modulus of PCL-PGS ranged from 5.6 to 15.7 MPa. The ultimate tensile stress ranged from 2.0 to 2.9 MPa, and these fibers showed elongation from 290 to 900%. The addition of PGS and grafting of heparin improved the attachment and proliferation of human umbilical vein endothelial cells. Core-shell PCL-PGS fibers demonstrate improved performance as three-dimensional fibrous mats for potential tissue-engineering applications.
Project description:Electrospinning is a valuable technology for cartilage tissue engineering (CTE) due to its ability to produce fibrous scaffolds mimicking the nanoscale and alignment of collagen fibers present within the superficial zone of articular cartilage. Coaxial electrospinning allows the fabrication of core-shell fibers able to incorporate and release bioactive molecules (e.g., drugs or growth factors) in a controlled manner. Herein, we used coaxial electrospinning to produce coaxial poly(glycerol sebacate) (PGS)/poly(caprolactone) (PCL) aligned nanofibers (core:PGS/shell:PCL). The obtained scaffolds were characterized in terms of their structure, chemical composition, thermal properties, mechanical performance and in vitro degradation kinetics, in comparison to monoaxial PCL aligned fibers and respective non-aligned controls. All the electrospun scaffolds produced presented average fiber diameters within the nanometer-scale and the core-shell structure of the composite fibers was clearly confirmed by TEM. Additionally, fiber alignment significantly increased (>2-fold) the elastic modulus of both coaxial and monoxial scaffolds. Kartogenin (KGN), a small molecule known to promote mesenchymal stem/stromal cells (MSC) chondrogenesis, was loaded into the core PGS solution to generate coaxial PGS-KGN/PCL nanofibers. The KGN release kinetics and scaffold biological performance were evaluated in comparison to KGN-loaded monoaxial fibers and respective non-loaded controls. Coaxial PGS-KGN/PCL nanofibers showed a more controlled and sustained KGN release over 21 days than monoaxial PCL-KGN nanofibers. When cultured with human bone marrow MSC in incomplete chondrogenic medium (without TGF-?3), KGN-loaded scaffolds enhanced significantly cell proliferation and chondrogenic differentiation, as suggested by the increased sGAG amounts and chondrogenic markers gene expression levels. Overall, these findings highlight the potential of using coaxial PGS-KGN/PCL aligned nanofibers as a bioactive scaffold for CTE applications.
Project description:The development of living heart valves that grow with the patient is a promising strategy for heart valve replacements in pediatric patients. Despite active research in the field of tissue engineered heart valves there have been limited efforts to optimize the balance between biodegradation of the scaffolds and de novo extracellular matrix (ECM) synthesis by cells and study their consequences on the mechanical properties of the cell-seeded construct. This study investigates the effect of in vitro degradation and ECM secretion on the mechanical properties of hybrid polyester scaffolds. The scaffolds were synthesized from blends of fast degrading polyglycerol sebacate (PGS) and slowly degrading polycaprolactone (PCL). PGS-PCL scaffolds were electrospun using a 2:1 ratio of PGS to PCL. Accelerated hydrolytic degradation in 0.1 mM sodium hydroxide revealed 2-fold faster degradation of PGS-PCL scaffolds compared with PCL scaffolds. Thermal analysis and scanning electron microscopy demonstrated marginal change in PCL scaffold properties, while PGS-PCL scaffolds showed preferential mass loss of PGS and thinning of the individual fibers during degradation. Consequently, the mechanical properties of PGS-PCL scaffolds decreased gradually with no significant change for PCL scaffolds during accelerated degradation. Valvular interstitial cells (VICs) seeded on PGS-PCL scaffolds showed higher ECM protein secretion compared with PCL. Thus the mechanical properties of the cell-seeded PCL scaffolds did not change significantly compared with acellular scaffolds, probably due to slower degradation and ECM deposition by VICs. In contrast, the PGS-PCL scaffolds exhibited a gradual decrease in the mechanical properties of the acellular scaffolds due to degradation, which was compensated for by new matrix secreted by VICs seeded on the scaffolds. Our study demonstrated that the faster degrading PGS component of PGS-PCL accelerated the degradation rate of the scaffolds. VICs, on the other hand, were able to remodel the synthetic scaffold, depositing new matrix proteins and maintaining the mechanical properties of the scaffolds.
Project description:Small-diameter vascular grafts perform poorly as arterial bypasses. We developed a cell-free, resorbable graft intended to remodel in situ into a living vessel. The graft consisted of a soft electrospun poly(glycerol sebacate) (PGS) core, a PGS prepolymer (pPGS) coating, and a reinforcing electrospun poly(ε-caprolactone) (PCL) sheath. The core contained 4.37 ± 1.95 μm fibers and had a porosity of 66.4 ± 3.2%, giving it large pores to encourage cellular infiltration and pro-healing macrophages. The sheath contained 6.63 ± 0.89 μm fibers and had a porosity of 80.5 ± 2.1%. in vitro testing suggested that the stress achieved at arterial pressure would be 13-fold lower than the yield stress of the graft. Grafts were implanted as 7 cm carotid interpositions in two sheep. Sheep were maintained on dual antiplatelet therapy and followed with duplex ultrasound. One graft ruptured at 13 days. The second animal was euthanized with a dilated graft at 15 days. Histology showed near-total degradation of the core and a robust inflammatory response within the sheath. Little neotissue had formed within the graft wall or lumen, but the graft had become surrounded by fibroblast-rich and vascularized connective tissue. Because PCL is commonly used in resorbable grafts, this mechanical destabilization was unexpected. We speculate that the inflammatory response instigated by the rapidly degrading PGS intensified degradation of the PCL and that the large pores enabled a prolonged acute host-graft reaction which attacked the entire bulk of the material, speeding weakening. Future work will focus on how to moderate inflammation and improve remodeling of grafts in large animals.
Project description:Novel biodegradable multiblock copolymers of [PCL-<i>b</i>-P(THF-<i>co</i>-CL)]<i><sub>m</sub></i> with PCL fractions of 53.3 and 88.4 wt % were prepared by Janus polymerization of ε-caprolactone (CL) and tetrahydrofuran (THF). Their electrospun mats were obtained with optimized parameters containing bead-free nanofibers whose diameters were between 290 and 520 nm. The mechanical properties of the nanofiber scaffolds were measured showing the tensile strength and strain at break of 8⁻10 MPa and 123⁻161%, respectively. Annealing improved their mechanical properties and their tensile strength and strain at break of the samples increased to 10⁻13 MPa and 267⁻338%, respectively. Due to the porous structure and crystallization in nanoscale confinement, the mechanical properties of the nanofiber scaffolds appeared as plastics, rather than as the elastomers observed in bulk thermal-molded film.
Project description:Matching material degradation with host remodeling, including endothelialization and muscular remodeling, is important to vascular regeneration. We fabricated 3D PGS-PCL vascular grafts, which presented tunable polymer components, porosity, mechanical strength, and degrading rate. Furthermore, highly porous structures enabled 3D patterning of conjugated heparin-binding peptide, dimeric thymosin β4 (DTβ4), which played key roles in antiplatelets, fibrinogenesis inhibition, and recruiting circulating progenitor cells, thereafter contributed to high patency rate, and unprecedentedly acquired carotid arterial regeneration in rabbit model. Through single-cell RNA sequencing analysis and cell tracing studies, a subset of endothelial progenitor cells, myeloid-derived CD93<sup>+</sup>/CD34<sup>+</sup> cells, was identified as the main contributor to final endothelium regeneration. To conclude, DTβ4-inspired porous 3DVGs present adjustable physical properties, superior anticoagulating, and re-endothelializing potentials, which leads to the regeneration of small-caliber artery, thus offering a promising tool for vessel replacement in clinical applications.
Project description:Polymeric nanofibers are of great interest in biomedical applications, such as tissue engineering, drug delivery and wound healing, due to their ability to mimic and restore the function of natural extracellular matrix (ECM) found in tissues. Electrospinning has been heavily used to fabricate nanofibers because of its reliability and effectiveness. In our research, we fabricated poly(ε-caprolactone)-(PCL), magnesium oxide-(MgO) and keratin (K)-based composite nanofibers by electrospinning a blend solution of PCL, MgO and/or K. The electrospun nanofibers were analyzed by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), mechanical tensile testing and inductively-coupled plasma optical emission spectroscopy (ICP-OES). Nanofibers with diameters in the range of 0.2-2.2 µm were produced by using different ratios of PCL/MgO and PCL-K/MgO. These fibers showed a uniform morphology with suitable mechanical properties; ultimate tensile strength up to 3 MPa and Young's modulus 10 MPa. The structural integrity of nanofiber mats was retained in aqueous and phosphate buffer saline (PBS) medium. This study provides a new composite material with structural and material properties suitable for potential application in tissue engineering.
Project description:Two-component fibrous materials based on poly(3-hydroxybutyrate) (PHB, Tm = 160 °C) and poly(?-caprolactone) (PCL, Tm = 60 °C) were successfully fabricated by dual-jet electrospinning of their separate spinning solutions. The desired alignment of the fibers that compose PHB/PCL mats was achieved by using three types of rotating collectors-drum (smooth), blade and grid. Additional fiber alignment in the direction of collector rotation was achieved by rotating at 2200 rpm. Moreover, the selected concentration of PCL spinning solution resulted in fibers with spindle-like defects along their length. Thus, "segment" sealing of the PHB (high-melting) fibers by the molten PCL (low-melting) fibers/defects sites was achieved after heating the PHB/PCL mats above the melting temperature (Tm) of PCL. The surface morphology, thermal behavior and mechanical properties of the PHB/PCL mats before and after thermal treatment were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, differential scanning calorimetry (DSC) and mechanical tests. The results indicated that regardless of the cutting direction of the specimens (0° or 90°), thermal treated PHB/PCL mats reveal enhanced mechanical properties. Therefore, this work provides an easily feasible route for the fabrication of electrospun PHB/PCL mats with tunable mechanical properties and improved performance.
Project description:A continuous and scalable method for the wet spinning of cellulose nanofibrils (CNFs) is introduced in a core/shell configuration. Control on the interfacial interactions was possible by the choice of the shell material and coagulant, as demonstrated here with guar gum (GG) and cellulose acetate (CA). Upon coagulation in acetone, ethanol, or water, GG and CA formed supporting polymer shells that interacted to different degrees with the CNF core. Coagulation rate was shown to markedly influence the CNF orientation in the filament and, as a result, its mechanical strength. The fastest coagulation noted for the CNF/GG core/shell system in acetone led to an orientation index of ?0.55 (Herman's orientation parameter of 0.40), Young's modulus of ?2.1 GPa, a tensile strength of ?70 MPa, and a tenacity of ?8 cN/tex. The system that underwent the slowest coagulation rate (CNF/GG in ethanol) displayed a limited CNF orientation but achieved an intermediate level of mechanical resistance, owing to the strong core/shell interfacial affinity. By using CA as the supporting shell, it was possible to spin CNF into filaments with high water absorption capacity (43 g water/g dry filament). This was explained by the fact that water (used as the coagulant for CA) limited the densification of the CNF core structure, yielding filaments with high accessible area and pore density.
Project description:In this study, heparin-loaded poly-ɛ-caprolactone (PCL) fibrous mats were prepared and characterized based on their physical, cytotoxic, thermal, and biological properties. The main objective of the work described here was to test the hypothesis that incorporation of heparin into a PCL carrier could serve as bio-compatible material capable of inhibiting Human Papillomavirus (HPV) infection. The idea of firmly anchoring heparin to capture soluble virus, vs. a slow heparin release to inhibit a virus in solution was tested. Thus, one material was produced via conventional heparin matrix encapsulation and electrohydrodynamic fiber processing in one step. A second type of material was obtained via heparin crosslinking. This was achieved by running a carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling reaction on preformed PCL fibers. In vitro HPV16 L1 protein binding capacity studies were performed. Infectivity assays were done using HPV16 pseudoviruses (PsVs) carrying a GFP plasmid to directly test the ability of the fibrous mats to prevent internalization of HPV PsVs. The crosslinked heparin material presented a dissociation constant (Kd) value comparable to those found in the literature for different heparin-protein L1 peptide interactions. Both materials significantly reduced internalization of HPV PsVs, with a reduction of 94% of PsVs internalization when matrix encapsulated heparin-loaded material was present. Differences in performance between the two proposed structures are discussed.
Project description:Tissue engineered heart valves (TEHV) can be useful in the repair of congenital or acquired valvular diseases due to their potential for growth and remodeling. The development of biomimetic scaffolds is a major challenge in heart valve tissue engineering. One of the most important structural characteristics of mature heart valve leaflets is their intrinsic anisotropy, which is derived from the microstructure of aligned collagen fibers in the extracellular matrix (ECM). In the present study, a directional electrospinning technique is used to fabricate fibrous poly(glycerol sebacate):poly(caprolactone) (PGS:PCL) scaffolds containing aligned fibers, which resemble native heart valve leaflet ECM networks. In addition, the anisotropic mechanical characteristics of fabricated scaffolds are tuned by changing the ratio of PGS:PCL to mimic the native heart valve's mechanical properties. Primary human valvular interstitial cells (VICs) attach and align along the anisotropic axes of all PGS:PCL scaffolds with various mechanical properties. The cells are also biochemically active in producing heart-valve-associated collagen, vimentin, and smooth muscle actin as determined by gene expression. The fibrous PGS:PCL scaffolds seeded with human VICs mimick the structure and mechanical properties of native valve leaflet tissues and would potentially be suitable for the replacement of heart valves in diverse patient populations.