Bicomponent Scaffolds Research Articles

Gelatin/EGDE Ultrafine Composite Fibers Reinforced with 3D Spacer Fabric as Bicomponent Scaffolds for Tissue Engineering.

Protein-based ultrafine fibrous scaffolds can mimic the native extracellular matrices (ECMs) with regard to the morphology and chemical composition but suffer from poor mechanical and wet stability. As a result, cells cannot get a true three-dimensional (3D) environment as they find in native ECMs. In this study, an epoxide, ethylene glycol diglycidylether (EGDE), with high reactivity to active hydrogen is introduced to gelatin solution, serving as an effective cross-linker. The gelatin/EGDE 3D-ultrafine (∼500 nm in diameter) fibrous composite scaffolds are made by an ultralow-concentration phase separation technique (ULCPS). The effects of the polymer content and modification conditions on the morphology and wet stability of the constructs are investigated. It is revealed that ultrafine fibers with 3D random orientation could be formed at low concentrations (0.01, 0.05, and 0.1 wt %, respectively). The wet stability of the constructs could be effectively improved by introducing EGDE into the gelatin system. The shrinkage is reduced to merely 2.14% after the modification at 120 °C for 2 h and could be maintained for up to 3 days. In order to improve the compression properties, the same technique is utilized with the presence of a poly(lactic acid) (PLA) spacer fabric to produce a bicomponent scaffold. The mechanical property and cell viability of the bicomponent scaffolds are investigated, and it is found that cells could enter deep inside and orient themselves randomly at the central area of the bicomponent scaffold. The modification and design approach presented in this study has the potential to provide various protein-based ultrafine fibrous biomaterials for a variety of biomedical applications.

First Advanced Bilayer Scaffolds for Tailored Skin Tissue Engineering Produced via Electrospinning and Melt Electrowriting

AbstractIn vitro skin models are validated methods for screening cosmetics and pharmaceuticals, but still have limitations. The bilayer poly(ε‐caprolactone) scaffold/membrane model described here overcomes some of these deficits by integrating a solution electrospun (SES) membrane at the dermoepidermal interface and a melt electrowritten (MEW) scaffold that provides an optimal open‐pore environment for the dermis. To the knowledge, this scaffold/membrane model is the only one capable of creating a properly differentiated, full thickness skin model with neosynthesized extracellular matrix (ECM) in only 18 days. Both the wavy and straight fiber scaffold designs create a well‐organized dermis, but dermal collagen organization differs between designs. Adding cells and vitamin C to the scaffolds improves the mechanical properties to more closely mimic native human skin. These findings establish bicomponent scaffolds as a promising advancement for rapidly creating different skin models with varied properties. The versatility and adaptability of the described model can be used for studying how the biological and physical microenvironment impact skin, and testing dermo‐cosmetics and pharmaceutical treatments on different ages of skin. Furthermore, it can be an excellent new tool for studying wound healing and development into its use as a graft or wound dressing is ongoing.

Electrospun/3D-Printed Bicomponent Scaffold Co-Loaded with a Prodrug and a Drug with Antibacterial and Immunomodulatory Properties.

This work reports the construction of a bicomponent scaffold co-loaded with both a prodrug and a drug (BiFp@Ht) as an efficient platform for wound dressing, by combining the electrospinning and 3D-printing technologies. The outer component consisted of a chitosan/polyethylene oxide-electrospun membrane loaded with the indomethacin-polyethylene glycol-indomethacin prodrug (Fp) and served as a support for printing the inner component, a gelatin methacryloyl/sodium alginate hydrogel loaded with tetracycline hydrochloride (Ht). The different architectural characteristics of the electrospun and 3D-printed layers were very well highlighted in a morphological analysis performed by Scanning Electron Microscopy (SEM). In vitro release profile studies demonstrated that both Fp and Ht layers were capable to release the loaded therapeutics in a controlled and sustained manner. According to a quantitative in vitro biological assessment, the bicomponent BiFp@Ht scaffold showed a good biocompatibility and no cytotoxic effect on HeLa cell cultures, while the highest proliferation level was noted in the case of HeLa cells seeded onto an Fp nanofibrous membrane. Furthermore, the BiFp@Ht scaffold presented an excellent antimicrobial activity against the E. coli and S. aureus bacterial strains, along with promising anti-inflammatory and proangiogenic activities, proving its potential to be used for wound dressing.

Sustained Biochemical Signaling and Contact Guidance by Electrospun Bicomponents as Promising Scaffolds for Nerve Tissue Regeneration.

Electrospun fibers are excellent delivery vehicles enabling a sustained release of growth factors to elicit favorable cell responses and are increasingly used in tissue engineering. Scaffolds with specific physical/topographical features can also guide cell migration and maturation. Therefore, growth factor-loaded electrospun scaffolds with a designed topography are promising for tissue regeneration. In this investigation, aligned-fiber scaffolds composed of poly(lactic-co-glycolic acid) nanofibers incorporating a glial cell line-derived growth factor and poly (d,l-lactic acid) nanofibers incorporating a nerve growth factor were produced by electrospinning. The scaffolds provided an aligned fibrous topography and a dual release of growth factors. The rat pheochromocytoma cell (PC12 cell) response to produced non-woven and aligned-fiber scaffolds with/without growth factors was studied. The dual release of growth factors and topographical cues provided by aligned-fiber bicomponent scaffolds induced significant neurite extension, neuronal differentiation, and neurite alignment in a synergistic manner. The scaffolds with predesigned biochemical/topographical cues demonstrated in this study might be promising for nerve tissue repair.

Nanofibrous bicomponent scaffolds for the dual delivery of NGF and GDNF: controlled release of growth factors and their biological effects

Electrospun fibrous scaffolds capable of providing dual growth factor delivery in a controlled manner have distinctive advantages for tissue engineering. In this study, we have investigated the formation, structure, and characteristics/properties of fibrous bicomponent scaffolds for the dual delivery of glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) for peripheral nerve tissue regeneration. GDNF and NGF were incorporated into core-shell structured poly(lactic-co-glycolic acid) (PLGA) and poly (d,l-lactic acid) (PDLLA) nanofibers, respectively, through emulsion electrospinning. Using dual-source dual-power electrospinning, bicomponent scaffolds composed of GDNF/PLGA fibers and NGF/PDLLA fibers with different fiber component ratios were produced. The structure, properties, and in vitro release behavior of mono- and bicomponent scaffolds were systematically investigated. Concurrent and sustained release of GDNF and NGF from bicomponent scaffolds was achieved and their release profiles could be tuned. In vitro biological investigations were conducted. Rat pheochromocytoma cells were found to attach, spread, and proliferate on all scaffolds. The release of growth factors from scaffolds could induce much improved neurite outgrowth and neural differentiation. GDNF and NGF released from GDNF/PLGA scaffolds and NGF/PDLLA scaffolds, respectively, could induce dose-dependent neural differentiation separately. GDNF and NGF released from bicomponent scaffolds exerted a synergistic effect on promoting neural differentiation.

Biomimetic mineralization of novel hydroxyethyl cellulose/soy protein isolate scaffolds promote bone regeneration in vitro and in vivo

Although various strategies have been utilized to accelerate bone regeneration in bone tissue engineering (BTE), the treatment and repair of large bone defects remains a clinical challenge worldwide. Inspired by the natural extracellular matrix of bone tissue, organic-inorganic composite scaffolds with three-dimensional (3D) porous structures, sufficient mechanical properties, excellent cytocompatibility, osteoconductivity, and osteogenic potential have received considerable attention within the field of bone engineering. In this work, a novel epichlorohydrin (ECH)-crosslinked hydroxyethyl cellulose (HEC)/soy protein isolate (SPI) porous bi-component scaffold (EHSS) with hydroxyapatite (HAp) functionalization (EHSS/HAp) was constructed for bone defect repair via the combination of lyophilization and in situ biomimetic mineralization. Systematic characterization experiments were performed to assess the morphology, HAp-forming properties, mechanical properties and degradation rate of the scaffold. The results indicated that the prepared scaffolds exhibited an interconnected porous structure, a biomimetic HAp coating on their surfaces, improved mechanical properties in compression and a controllable degradation rate. In particular, semiquantitative analysis showed that the calcium/phosphorus (Ca/P) ratio of EHSS/HAp with 70% SPI content (1.65) was similar to that of natural bone tissue (1.67) according to energy dispersive X-ray spectroscopy analysis. In vitro cell culture experiments indicated that the EHSS/HAp with 70% SPI content showed improved cytocompatibility and was suitable for MC3T3-E1 cell attachment, proliferation and growth. Consistently, in vitro osteogenic differentiation studies showed that EHSS/HAp with 70% SPI content can significantly accelerate the expression of osteogenesis-related genes (Col-1, Runx2, OPN, and OCN) during osteogenic differentiation of MC3T3-E1 cells. Furthermore, when applied to the repair of critical-sized cranial defects in a rat model, EHSS/HAp with 70% SPI was capable of significantly promoting tissue regeneration and integration with native bone tissue. Microscopic computed tomography (micro-CT) results demonstrated that the bone defect site was nearly occupied with newly formed bone at 12 weeks after implantation of EHSS/HAp with 70% SPI content into the defect. Hematoxylin and eosin (H&E) staining and Masson's trichrome staining of histological sections further confirmed that EHSS/HAp with 70% SPI markedly promoted new bone formation and maturation. Collectively, our results demonstrate the potential of EHSS/HAp scaffolds with 70% SPI for successful bone defect repair and regeneration.

Viscoelastic Behavior of Embroidered Scaffolds for ACL Tissue Engineering Made of PLA and P(LA-CL) After In Vitro Degradation.

A rupture of the anterior cruciate ligament (ACL) is the most common knee ligament injury. Current applied reconstruction methods have limitations in terms of graft availability and mechanical properties. A new approach could be the use of a tissue engineering construct that temporarily reflects the mechanical properties of native ligament tissues and acts as a carrier structure for cell seeding. In this study, embroidered scaffolds composed of polylactic acid (PLA) and poly(lactic-co-ε-caprolactone) (P(LA-CL)) threads were tested mechanically for their viscoelastic behavior under in vitro degradation. The relaxation behavior of both scaffold types (moco: mono-component scaffold made of PLA threads, bico: bi-component scaffold made of PLA and P(LA-CL) threads) was comparable to native lapine ACL. Most of the lapine ACL cells survived 32 days of cell culture and grew along the fibers. Cell vitality was comparable for moco and bico scaffolds. Lapine ACL cells were able to adhere to the polymer surfaces and spread along the threads throughout the scaffold. The mechanical behavior of degrading matrices with and without cells showed no significant differences. These results demonstrate the potential of embroidered scaffolds as an ACL tissue engineering approach.

Bicomponent nanofibrous scaffolds with dual release of anticancer drugs and biomacromolecules

Abstract

Spatio-temporal release of NGF and GDNF from multi-layered nanofibrous bicomponent electrospun scaffolds

Scaffolds capable of providing dual neurotrophic factor (NTF) delivery with different release kinetics, spatial delivery of NTFs at different loci and topographical guidance are promising for enhanced peripheral nerve regeneration. In this study, we have designed and fabricated multi-layered aligned-fiber scaffolds through combining emulsion electrospinning, sequential electrospinning and high-speed electrospinning (HS-ES) to modulate the release behavior of glial cell line-derived growth factor(GDNF) and nerve growth factor (NGF). GDNF and NGF were incorporated into poly(lactic-co-glycolic acid) (PLGA) fibers and poly(D,L-lactic acid) (PDLLA) fibers, respectively. Aligned fibers were obtained in each layer of multi-layered scaffolds and relatively thick tri-layered and tetra-layered scaffolds with controlled layer thickness were obtained. Their morphology, structure, properties, and the in vitro release of growth factors were examined. Dual and spatio-temporal release of GDNF and NGF with different release kinetics from multi-layered scaffolds was successfully demonstrated. High separation efficiency by PDLLA fibrous barrier layer for spatial neurotrophic factor delivery from both tri-layered scaffolds and tetra-layered scaffolds was achieved.

Incorporation and release of dual growth factors for nerve tissue engineering using nanofibrous bicomponent scaffolds

Electrospun fibrous scaffolds have been extensively used as cell-supporting matrices or delivery vehicles for various biomolecules in tissue engineering. Biodegradable scaffolds with tunable degradation behaviors are favorable for various resorbable tissue replacements. In nerve tissue engineering, delivery of growth factors (GFs) such as nerve growth factor (NGF) and glial cell line-derived neurotrophic factor (GDNF) from scaffolds can be used to promote peripheral nerve repair. In this study, using the established dual-source dual-power electrospinning technique, bicomponent scaffolds incorporated with NGF and GDNF were designed and demonstrated as a strategy to develop scaffolds providing dual GF delivery. NGF and GDNF were encapsulated in poly(D, L-lactic acid) (PDLLA) and poly(lactic-co-glycolic acid) (PLGA) nanofibers, respectively, via emulsion electrospinning. Bicomponent scaffolds with various mass ratios of GDNF/PLGA fibers to NGF/PDLLA fibers were fabricated. Their morphology, structure, properties, and the in vitro degradation were examined. Both types of core–shell structured fibers were evenly distributed in bicomponent scaffolds. Robust scaffolds with varying component ratios were fabricated with average fiber diameter ranging from 307 ± 100 nm to 688 ± 129 nm. The ultimate tensile stress and elastic modulus could be tuned ranging from 0.23 ± 0.07 MPa to 1.41 ± 0.23 MPa, 11.1 ± 3.0 MPa to 75.9 ± 3.3 MPa, respectively. Adjustable degradation was achieved and the weight loss of scaffolds ranged from 9.2% to 44.0% after 42 day degradation test. GDNF and NGF were incorporated with satisfactory encapsulation efficiency and their bioactivity were well preserved. Sustained release of both types of GFs was also achieved.

Polylactic Acid Nanofiber Scaffold Decorated with Chitosan Islandlike Topography for Bone Tissue Engineering.

In this work, a bicomponent scaffold with a core-shell and islandlike structure that combines the respective advantages of polylactic acid (PLA) and chitosan (CS) was prepared via electrospinning accompanied by automatic phase separation and crystallization. The objective of this research was to design nanosized topography with highly bioactive CS onto PLA electrospun fiber surface to improve the cell biocompatibility of the PLA fibrous membrane. The morphology, inner structure, surface composition, crystallinity, and thermodynamic analyses of nanofibers with various PLA/CS ratios were carried out, and the turning mechanism of a core-shell or islandlike topography structure was also speculated. The mineralization of hydroxyapatite and culture results of preosteoblast (MC3T3-E1) cells on the modified scaffolds indicate that the outer CS component and rough nanoscale topography on the surface of the nanofibers balanced the hydrophilicity and hydrophobicity of the fibers, enhanced their mineralization ability, and made them more beneficial for the attachment and growth of cells. Moreover, CS and "islandlike" protrusions on the fiber surface increased the alkaline phosphatase activity of the MC3T3-E1 cells seeded on the fibrous membrane and provided a more appropriate interface for cell adhesion and proliferation. These results illustrate that this kind of PLA/CS membrane has the potential in tissue engineering. More importantly, our study provides a new approach to designing PLA scaffolds, with combined topographic and bioactive modification effects at the interface between cells and materials, for biomedicine.

Bicomponent fibrous scaffolds made through dual-source dual-power electrospinning: Dual delivery of rhBMP-2 and Ca-P nanoparticles and enhanced biological performances.

Electrospun scaffolds incorporated with both calcium phosphates (Ca-P) and bone morphogenetic protein-2 (BMP-2) have been used for bone tissue regeneration. However, in most cases BMP-2 and Ca-P were simply mixed and loaded in a monolithic structure, risking low BMP-2 loading level, reduced BMP-2 biological activity, uncontrolled BMP-2 release and inhomogeneous Ca-P distribution. In this investigation, novel bicomponent scaffolds having evenly distributed rhBMP-2-containing fibers and Ca-P nanoparticle-containing fibers were made using an established dual-source dual-power electrospinning technique with the assistance of emulsion electrospinning and blend electrospinning. The release behavior of rhBMP-2 and Ca2+ ions could be separately tuned and the released rhBMP-2 retained a 68% level for biological activity. MC3T3-E1 cells showed high viability and normal morphology on scaffolds. Compared to monocomponent scaffolds, enhanced cell proliferation, alkaline phosphatase activity, cell mineralization, and gene expression of osteogenic markers were achieved for bicomponent scaffolds due to the synergistic effect of rhBMP-2 and Ca-P nanoparticles. Bicomponent scaffolds with a double mass elicited further enhanced cell adhesion, spreading, proliferation, and osteogenic differentiation. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2199-2209, 2017.

Chondroitin Sulfate- and Decorin-Based Self-Assembling Scaffolds for Cartilage Tissue Engineering

Cartilage injury and degenerative tissue progression remain poorly understood by the medical community. Therefore, various tissue engineering strategies aim to recover areas of damaged cartilage by using non-traditional approaches. To this end, the use of biomimetic scaffolds for recreating the complex in vivo cartilage microenvironment has become of increasing interest in the field. In the present study, we report the development of two novel biomaterials for cartilage tissue engineering (CTE) with bioactive motifs, aiming to emulate the native cartilage extracellular matrix (ECM). We employed a simple mixture of the self-assembling peptide RAD16-I with either Chondroitin Sulfate (CS) or Decorin molecules, taking advantage of the versatility of RAD16-I. After evaluating the structural stability of the bi-component scaffolds at a physiological pH, we characterized these materials using two different in vitro assessments: re-differentiation of human articular chondrocytes (AC) and induction of human adipose derived stem cells (ADSC) to a chondrogenic commitment. Interestingly, differences in cellular morphology and viability were observed between cell types and culture conditions (control and chondrogenic). In addition, both cell types underwent a chondrogenic commitment under inductive media conditions, and this did not occur under control conditions. Remarkably, the synthesis of important ECM constituents of mature cartilage, such as type II collagen and proteoglycans, was confirmed by gene and protein expression analyses and toluidine blue staining. Furthermore, the viscoelastic behavior of ADSC constructs after 4 weeks of culture was more similar to that of native articular cartilage than to that of AC constructs. Altogether, this comparative study between two cell types demonstrates the versatility of our novel biomaterials and suggests a potential 3D culture system suitable for promoting chondrogenic differentiation.

Morphology and surface chemistry of bicomponent scaffolds in terms of mesenchymal stromal cell viability

Biological interaction between cells and scaffolds is mediated through events at surfaces. Proteins present in the culture medium adsorb on substrates, generating a protein adlayer that triggers further downstream events governing cell adhesion. Polymer blends often combine the properties of the individual components, for example, can provide mechanical as well as surface properties in one fibre. Therefore, mixtures of synthetic polycaprolactone and gelatin as a denatured form of collagen were electrospun at selected conditions and polymer weight ratios. Fibre morphologies and chemical properties of the surfaces were analysed. These scaffolds were seeded with human mesenchymal stromal cells and their viability was studied. Gelatin addition to polycaprolactone leads to a reduction in fibre diameter. A linear increase in gelatin at the fibre surface was observed in function of the weighed polymers, except for polycaprolactone/gelatin fibres incorporating equal weight ratios. Thereby, a depletion of gelatin at the fibre surface is stated for equally mixed polymers. The depletion of gelatin at the fibre surface is most probably due to hydrophobic interactions between hydrophobic segments of polycaprolactone and gelatin, affecting the spinning mechanism and thus fibre structure. Furthermore, polycaprolactone/gelatin blends show enhanced wettability properties compared to pure gelatin, at least partly due to molecular segregation. Results of in vitro studies reveal an increase in cellular viability and proliferation for cells cultivated on nanofibres containing gelatin, caused by the cell-attractive surface composition as well as the hydrophilic nature of the scaffolds. Contact guidance of cells seeded on parallelised fibres is observed, and DNA tests show evidently enhanced cell numbers on nanofibres containing 20 wt% of gelatin.

Adjusting the mechanical behavior of embroidered scaffolds to lapin anterior cruciate ligaments by varying the thread materials

Traumatic rupture of the anterior cruciate ligament (ACL) can cause local destabilization and loss of mobility. Reconstruction using engineered ACL grafts is rarely successful due to sub-optimal material choice and mechanical performance. Thus, the presented work demonstrates the fabrication of various embroidered single- and bi-component scaffolds made of two commercially available monofilament threads (polydioxanone, poly(lactic acid-co-ɛ- caprolactone)) as well as a novel melt spun poly(L-lactic acid) multifilament and their mechanical analysis by tensile tests and under cyclic loading. Selected scaffolds, adjusted by material composition and textile parameters, revealed a load–strain behavior comparable to native lapin ACL tissue exhibiting a sufficient amount of elastic deformation within the toe-region of 1.7%, scaffold stiffness of 123 N/mm and adequate maximum tensile load (300 N) and strain (20%). Therefore, the design of resorbable embroidered bi-component scaffolds represents a promising approach to replace artificial non-resorbable ligament grafts and allows for innovative tissue engineering strategies.

Preparation of chitosan/silk fibroin/hydroxyapatite porous scaffold and its characteristics in comparison to bi‐component scaffolds

Composite porous scaffolds have attracted extensive attention in the biomedical material field. The aim of this research was to prepare a novel tri-component composite porous scaffold and to evaluate its relevant properties. The porous scaffold was composed of chitosan (CS), silk fibroin (SF), and nanohydroxyapatite particles (nHA), which we named CS/SF/nHA scaffold and prepared via salt fractionation method combined with lyophilization. The porous structure was achieved using a porogen (salt), and the pore size was controlled by the size of porogen. To evaluate the characteristics of the tri-component scaffold, three bi-component scaffolds, CS/SF, CS/nHA, and SF/nHA, were simultaneously prepared for comparison. The scaffolds were subjected to morphological, micro-structural, and biodegradation analyses. Results demonstrated that all of the scaffolds had pore sizes of 100-300 μm and a porosity of 90.5-96.1%. The biodegradation characteristics of all scaffolds meet the requirements of good biomedical materials. The investigation of the mechanical properties showed that the tri-component scaffold has better properties than the bi-component scaffolds. The in vitro biocompatibility with osteoblast-like MG-63 cells showed that all the scaffolds are suitable for cell attachment and proliferation; however, the CS/SF/nHA composite porous scaffold is much more effective than the others.

Preparation of chitosan/silk fibroin/hydroxyapatite porous scaffold and its characteristics in comparison to bi-component scaffolds

Preparation of chitosan/silk fibroin/hydroxyapatite porous scaffold and its characteristics in comparison to bi-component scaffolds

Dual-source dual-power electrospinning and characteristics of multifunctional scaffolds for bone tissue engineering.

Electrospun tissue engineering scaffolds are attractive due to their distinctive advantages over other types of scaffolds. As both osteoinductivity and osteoconductivity play crucial roles in bone tissue engineering, scaffolds possessing both properties are desirable. In this investigation, novel bicomponent scaffolds were constructed via dual-source dual-power electrospinning (DSDPES). One scaffold component was emulsion electrospun poly(d,l-lactic acid) (PDLLA) nanofibers containing recombinant human bone morphogenetic protein (rhBMP-2), and the other scaffold component was electrospun calcium phosphate (Ca–P) particle/poly(lactic-co-glycolic acid) (PLGA) nanocomposite fibers. The mass ratio of rhBMP-2/PDLLA fibers to Ca–P/PLGA fibers in bicomponent scaffolds could be controlled in the DSDPES process by adjusting the number of syringes used to supply solutions for electrospinning. Through process optimization, both types of fibers could be evenly distributed in bicomponent scaffolds. The structure and properties of each type of fibers in the scaffolds were studied. The morphological and structural properties and wettability of scaffolds were assessed. The effects of emulsion composition for rhBMP-2/PDLLA fibers and mass ratio of fibrous components in bicomponent scaffolds on in vitro release of rhBMP-2 from scaffolds were investigated. In vitro degradation of scaffolds was also studied by monitoring their morphological changes, weight losses and decreases in average molecular weight of fiber matrix polymers.

Novel Electrospun Bicomponent Scaffolds for Bone Tissue Engineering: Fabrication, Characterization and Sustained Release of Growth Factor

ABSTRACTElectrospinning is a versatile technique for fabricating three-dimensional (3D) nanofibrous scaffolds and the scaffolds have been found to elicit desirable cellular behavior for tissue regeneration because the nanofibrous structures mimic the nanofibrous extracellular matrix (ECM) of biological tissues. From the material point of view, the ECM of bone is a nanofibrous nanocomposite consisting of an organic matrix (mainly collagen) and inorganic bone apatite nanoparticles. Therefore, for bone tissue engineering scaffolds, it is natural to construct nanofibrous nanocomposites having a biodegradable polymer matrix and nanosized bioactive bioceramics. Our previous studies demonstrated: (1) electrospun nanocomposite fiber loaded with calcium phosphate (Ca-P) were osteoconductive and could promote osteoblastic cell proliferation and differentiation better than pure polymer fibers; (2) The controlled release of recombinant human bone morphogenetic protein (rhBMP-2) from scaffolds provided the scaffolds with desired osteoinductivity. In the current investigation, novel bicomponent scaffolds for bone tissue engineering were produced using our established dual-source dual-power electrospinning technique to achieve both osteoconductivity and osteoinductivity. In the bicomponent scaffolds, one fibrous component was electrospun Ca-P/PLGA nanocomposite fibers and the other component was emulsion electrospun PDLLA nanofibers incorporated with rhBMP-2. Through electrospinning optimization, both fibers were evenly distributed in bicomponent scaffolds. The mass ratio of rhBMP-2/PDLLA fibers to Ca-P/PLGA fibers in bicomponent scaffolds could be controlled using multiple syringes. The structure and morphology of mono- and bicomponent scaffolds were examined. The in vitro release of rhBMP-2 from mono- and bicomponent scaffolds showed different release amount but similar release profile, exhibiting an initial burst release. Blending PDLLA with polyethylene glycol (PEG) could reduce the initial burst release of rhBMP-2.

Mesenchymal Stem Cell Responses to Bone-Mimetic Electrospun Matrices Composed of Polycaprolactone, Collagen I and Nanoparticulate Hydroxyapatite

The performance of biomaterials designed for bone repair depends, in part, on the ability of the material to support the adhesion and survival of mesenchymal stem cells (MSCs). In this study, a nanofibrous bone-mimicking scaffold was electrospun from a mixture of polycaprolactone (PCL), collagen I, and hydroxyapatite (HA) nanoparticles with a dry weight ratio of 50/30/20 respectively (PCL/col/HA). The cytocompatibility of this tri-component scaffold was compared with three other scaffold formulations: 100% PCL (PCL), 100% collagen I (col), and a bi-component scaffold containing 80% PCL/20% HA (PCL/HA). Scanning electron microscopy, fluorescent live cell imaging, and MTS assays showed that MSCs adhered to the PCL, PCL/HA and PCL/col/HA scaffolds, however more rapid cell spreading and significantly greater cell proliferation was observed for MSCs on the tri-component bone-mimetic scaffolds. In contrast, the col scaffolds did not support cell spreading or survival, possibly due to the low tensile modulus of this material. PCL/col/HA scaffolds adsorbed a substantially greater quantity of the adhesive proteins, fibronectin and vitronectin, than PCL or PCL/HA following in vitro exposure to serum, or placement into rat tibiae, which may have contributed to the favorable cell responses to the tri-component substrates. In addition, cells seeded onto PCL/col/HA scaffolds showed markedly increased levels of phosphorylated FAK, a marker of integrin activation and a signaling molecule known to be important for directing cell survival and osteoblastic differentiation. Collectively these results suggest that electrospun bone-mimetic matrices serve as promising degradable substrates for bone regenerative applications.