Reinforcing silk scaffolds with silk particles

Silk fibroin is a useful protein polymer for biomaterials and tissue engineering. In this work, porogen leached scaffolds prepared from aqueous and HFIP silk solutions were reinforced through the addition of silk particles. This led to about 40 times increase in the specific compressive modulus and the yield strength of HFIP-based scaffolds. This increase in mechanical properties resulted from the high interfacial cohesion between the silk matrix and the reinforcing silk particles, due to partial solubility of the silk particles in HFIP. The porosity of scaffolds was reduced from ≈90% (control) to ≈75% for the HFIP systems containing 200% particle reinforcement, while maintaining pore interconnectivity. The presence of the particles slowed the enzymatic degradation of silk scaffolds.

Ingrowth of human mesenchymal stem cells into porous silk particle reinforced silk composite scaffolds : an in vitro study

Silk fibroin protein is biodegradable and biocompatible, exhibiting excellent mechanical properties for various biomedical applications. However, porous three-dimensional (3-D) silk fibroin scaffolds, or silk sponges, usually fall short in matching the initial mechanical requirements for bone tissue engineering. In the present study, silk sponge matrices were reinforced with silk microparticles to generate protein-protein composite scaffolds with desirable mechanical properties for in vitro osteogenic tissue formation. It was found that increasing the silk microparticle loading led to a substantial increase in the scaffold compressive modulus from 0.3 MPa (non-reinforced) to 1.9 MPa for 1:2 (matrix:particle) reinforcement loading by dry mass. Biochemical, gene expression, and histological assays were employed to study the possible effects of increasing composite scaffold stiffness, due to microparticle reinforcement, on in vitro osteogenic differentiation of human mesenchymal stem cells (hMSCs). Increasing silk microparticle loading increased the osteogenic capability of hMSCs in the presence of bone morphogenic protein-2 (BMP-2) and other osteogenic factors in static culture for up to 6 weeks. The calcium adsorption increased dramatically with increasing loading, as observed from biochemical assays, histological staining, and microcomputer tomography (μCT) analysis. Specifically, calcium content in the scaffolds increased by 0.57, 0.71, and 1.27 mg (per μg of DNA) from 3 to 6 weeks for matrix to particle dry mass loading ratios of 1:0, 1:1, and 1:2, respectively. In addition, μCT imaging revealed that at 6 weeks, bone volume fraction increased from 0.78% for non-reinforced to 7.1% and 6.7% for 1:1 and 1:2 loading, respectively. Our results support the hypothesis that scaffold stiffness may strongly influence the 3-D in vitro differentiation capabilities of hMSCs, providing a means to improve osteogenic outcomes.

Protein fiber particles for biomedical applications

Ultrafine protein particles have been fabricated from natural fibres, such as silk and wool. Our studies suggested that particles could be used for fabricating tough macro-porous composites scaffolds for tissue engineering. They are also efficient for reversible binding of metal ions. We are currently analyzing the sorption properties, biocompatibility and biodegradability of a range of particles to evaluate possibility for biomedical and healthcare applications.

Three dimensional scaffolds from multilobal fibres for enhanced cell growth

Multilobal fibres are irregularly shaped fibres with several surface channels or grooves. Scaffolds created from these fibres have high surface area which may enhance cell density. This study compared the cell growth of dermal fibroblasts and osteoblast-like SaOS2 cells on multilobal fibre nonwoven scaffolds to the conventional fibre scaffolds. Cells were cultured on round nylon, trilobal nylon, round polyethylene terephthalate (PET) and multilobal PET scaffolds for 14 days. There were more cells growing on trilobal nylon and PET multilobal scaffolds than their round counterparts. The preference to the type of multilobal scaffolds was cell dependent. The density of fibroblast increased 37% on trilobal nylon compared to round nylon scaffolds after 14 days of culture. SaOS2 cells preferred the multilobal PET scaffolds, exhibiting a 66% increase in cell number after 14 days of culture. Scaffolds manufactured from multilobal fibres have the ability to accommodate a high number of cells, demonstrating a great potential in tissue engineering applications.

Recent innovations in silk biomaterials

Silk contains a fibre forming protein, fibroin, which is biocompatible, particularly after removing the potentially immunogenic non-fibroin proteins. Silk can be engineered into a wide range of materials with diverse morphologies. Moreover, it is possible to regenerate fibroin with a desired amount of crystallinity, so that the biodegradation of silk materials can be controlled. These advantages have sparked new interest in the use of silk fibroin for biomedical applications, including tissue engineering scaffolds and carriers for sustained release of biologically active molecules. This article summarizes the current research related to the formation of silk materials with different morphologies, their biocompatibility, and examples of their biomedical applications. Recent work on the preparation of silk particles by mechanical milling and their applications in silk composite scaffolds is also discussed.

Protein fibre powder and new applications

Protein fibers such as silk and wool have been used as textile fibers for centuries. It is only in recent years that these fibers have been converted into fine powder forms for non-textile applications. This presentation will cover our recent research in protein fiber powders. Ultra-fine powders from different protein fibers have been produced using a combination of media and non media milling techniques. New application examples of these fine powders are discussed. These applications include hybrid fibers combining the advantages of natural and synthetic polymer fibers, tissue engineering composite scaffolds with enhanced biomechanical properties, and metal ion absorption.

3D fibrous structures as cardiovascular implants

Medical textiles are a highly specialised stream of technical textiles industry with a growing range of applications. A significant advancement has been achieved in surgical products or biomedical textiles (implantable/non-implantable) with the advent of 3D textile manufacturing techniques. Cardiovascular soft tissue implants (vascular grafts) have been a field of interest over decades for use of innovative 3D tubular structures in treatment of cardiovascular diseases. In the field of soft tissue implants, knitted and woven tubular structures are being used for large diameter blood vessel replacements. Advent of electrospinning and tissue engineering techniques has been able to provide promising answers to small diameter vascular grafts. The aim of this review is to outline the approaches in vascular graft development utilising different 3D tubular structure forming techniques. The emphasis is on vascular graft development techniques that can help improve treatment efficacy in future.

Electromagnetic field and other physical methods influencing cell growth in mammal cell culture systems

The growth rate of cultured mammalian cells can be influenced by chemical and physical methods such as electromagnetic fields (EMF), light, temperature and plasma. These physical methods have a number of well documented effects on mammalian cells including modification of gene expression, cell cycle, invasion, motility, cell viability, proliferation, apoptosis and mammosphere numbers. A study of the existing literature confirms that the impact of physical method on mammalian cells depends on the cell type, culture environment, exposure time, frequency, wave shape, and amount of dose. The modification of cell proliferation and apoptosis is necessary for cells products, tissue engineering, and therapy. In this article, we reviewed the impact of four physical methods on the growth rate and viability of cells. Plasma is the best method among fours because we can get desired result ranging from increasing cell proliferation to inducing apoptosis depending on the dose.

Enhanced cell growth using non-woven scaffolds of multilobal fibres

Multilobal fibres contain several grooves and have higher surface area than round fibres. Cell density can be enhanced when cultured on scaffolds manufactured with multilobal fibres. This study compared the cell growth of dermal fibroblasts and osteoblast-like SaOS2 cells on polymeric scaffolds produced from multilobal fibres to the conventional round-fibred scaffolds. Cells were cultured on round nylon 6,6, trilobal nylon 6,6, round polyethylene terephthalate (PET) and multilobal PET scaffolds for 14 days. There were more cells cultured on trilobal nylon 6,6 and PET multilobal scaffolds than their round counterparts. Preference to the type of multilobal scaffolds was cell dependent. Fibroblasts increased by 21.8 ± 1.9 fold to 6.3 × 105 cells (p < 0.001) when cultured on trilobal nylon 6,6 scaffolds while SaOS2 cells exhibited a 16.7 ± 2.8 fold increase (2.9 × 105 cells, p < 0.001) on the multilobal PET scaffolds after 14 days of culture. The ability of multilobal fibres to accommodate large quantities of cells presents an excellent alternative to round fibres as scaffolds for tissue engineering.

Ex vivo expansion of haematopoietic stem/progenitor cells from human umbilical cord blood on acellular scaffolds prepared from MS-5 stromal cell line

Lineage-specific expansion of haematopoietic stem/progenitor cells (HSPCs) from human umbilical cord blood (UCB) is desirable because of their several applications in translational medicine, e.g. treatment of cancer, bonemarrowfailure and immunodeficiencies. The currentmethods forHSPC expansion use either cellular feeder layers and/or soluble growth factors and selected matrix components coated on different surfaces. The use of cell-free extracellular matrices from bone marrow cells for this purpose has not previously been reported. We have prepared insoluble, cell- free matrices from a murine bone marrow stromal cell line (MS-5) grown under four different conditions, i.e. in presence or absence of osteogenic medium, each incubated under 5% and 20% O2 tensions. These acellularmatrices were used as biological scaffolds for the lineage-specific expansion of magnetically sorted CD34+ cells and the results were evaluated by flow cytometry and colony-forming assays. We could get up to 80-fold expansion of some HSPCs on one of the matrices and our results indicated that oxygen tension played a significant role in determining the expansion capacity of the matrices. A comparative proteomic analysis of the matrices indicated differential expression of proteins, such as aldehyde dehydrogenase and gelsolin, which have previously been identified as playing a role in HSPC maintenance and expansion. Our approach may be of value in identifying factors relevant to tissue engineering-based ex vivo HSPC expansion, and itmay also provide insights into the constitution of the niche in which these cells reside in the bone marrow.

Crosslinking neat ultrathin films and nanofibres of pH-responsive poly(acrylic acid) by UV radiation

Electrospun polyelectrolyte hydrogel nanofibres are being developed for many applications including artificial muscles, scaffolds for tissue engineering, wound dressings and controlled drug release. For electrospun polyelectrolytes, a post-spinning crosslinking process is necessary for producing a hydrogel. Typically, radiation or thermal crosslinking routines are employed that require multifunctional crosslinking molecules and crosslink reaction initiators (free radical producers). Here, ultraviolet subtype-C (UVC) radiation was employed to crosslink neat poly(acrylic acid) (PAA) nanofibres and films to different crosslink densities. Specific crosslink initiators or crosslinking molecules are not necessary in this fast and simple process providing an advantage for biological applications. Scanning probe microscopy was used for the first time to measure the dry and wet dimensions of hydrogel nanofibres. The diameters of the swollen fibres decrease monotonically with increasing UVC radiation time. The fibres could be reversibly swollen/contracted by treatment with solutions of varying pH, demonstrating their potential as artificial muscles. The surprising success of UVC radiation exposure to achieve chemical crosslinks without a specific initiator molecule exploits the ultrathin dimensions of the PAA samples and will not work with relatively thick samples.

Self-assembled peptides : characterisation and in vivo response

The fabrication of tissue engineering scaffolds is a well-established field that has gained recent prominence for the in vivo repair of a variety of tissue types. Recently, increasing levels of sophistication have been engineered into adjuvant scaffolds facilitating the concomitant presentation of a variety of stimuli (both physical and biochemical) to create a range of favourable cellular microenvironments. It is here that self-assembling peptide scaffolds have shown considerable promise as functional biomaterials, as they are not only formed from peptides that are physiologically relevant, but through molecular recognition can offer synergy between the presentation of biochemical and physio-chemical cues. This is achieved through the utilisation of a unique, highly ordered, nano- to microscale 3-D morphology to deliver mechanical and topographical properties to improve, augment or replace physiological function. Here, we will review the structures and forces underpinning the formation of self-assembling scaffolds, and their application in vivo for a variety of tissue types.