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1.
ACS Biomater Sci Eng ; 8(12): 5101-5109, 2022 Dec 12.
Article in English | MEDLINE | ID: mdl-36374748

ABSTRACT

The addition of poly(ethylene glycol) (PEG) to biomolecules and biomaterials is a well-established approach to modify their properties for therapeutic applications. For biomaterials, the approach is typically to blend or electrospray the synthetic polymer with the biomaterial. Effective surface modification approaches such as surface-initiated polymer brushes are challenging since the harsh solvents required for brush synthesis may destroy the biomaterial. Herein, we describe the PEGylation of collagen fibers by surface-initiated PEG brushes using a living anionic grafting-from mechanism. This brush synthesis is done in the absence of solvents to minimize the degradation of the native collagen structure. We quantify the effect the brush synthesis has on the native structure of the collagen fiber using differential scanning calorimetry (DSC) and find that even at long reaction times a significant fraction of the native structure remains. Dynamic mechanical analysis indicates the collagen undergoes only modest structural degradation, while adhesion studies find a significant improvement of antifouling properties. Further, our approach opens the way for further chemistry, as the growing polymer chain is a potassium alkoxy group that can be functionalized by termination or by subsequent reaction by a wide variety of molecules.


Subject(s)
Polyethylene Glycols , Polymers , Solvents , Polyethylene Glycols/chemistry , Biocompatible Materials , Collagen
2.
J Pediatr Urol ; 18(2): 160-167, 2022 04.
Article in English | MEDLINE | ID: mdl-35120811

ABSTRACT

INTRODUCTION: Despite the prevalence of hypospadias surgery and the common use of postoperative urethral stents, there has been no evaluation of the material properties of common stents. Our study sets out to close this gap with a literature review of recent publications comparing outcomes after hypospadias surgery for different urethral stent types and an evaluation of the material properties of four common urethral stents. STUDY DESIGN: A review of the English language literature from 2011 to 2021 was performed. Thermal analysis and mechanical analysis of the Zaontz Urethral Stent, the Firlit-Kluge Urethral Stent, the Koyle Diaper Stent, and the Bard Premature Infant Feeding Tube was also undertaken. RESULTS: Out of 165 papers, four met inclusion criteria. There was limited research on this topic, and no significant evidence that different stent materials impacted surgical complication rates. One study found improved comfort with the Zaontz stent, and another found a reduction emergency room visits with the Koyle stent. Using a foley balloon was associated with increased fistula rates, though this was likely due to the balloon design and not the material. Analysis of stents shows that all four are rubbery polymers at body temperature (Summary Table). The Zaontz and Koyle stents are thermoplastic elastomers with strong melting transitions above body temperature, but the Firlit-Kluge stent is amorphous at 37 °C and is likely covalently cross-linked to generate the network. The Bard feeding tube was the stiffest, with a Young's Modulus of 14.0 ± 0.78 (compared to 4.12 ± 0.56 for Zaontz, 4.92 ± 0.63 for Firlit-Kluge, and 4.09 ± 0.49 for Koyle). The Bard Feeding Tube is also the least extensible, fracturing at just over 300% strain compared to the other stents that can be stretched to greater than 2000% strain before fracture. Cyclic deformation studies demonstrate that the Zaontz, Firlit-Kluge, and Koyle stents are able to stretch and recover their shape more completely, a finding determined by the lower amount of plastic deformation those stents display compared to the Bard Feeding Tube. DISCUSSION: While there is little information associating urethral stent type with outcomes after hypospadias surgery, material properties may account for findings of prior studies. Stiffer stents may contribute to decreased postoperative comfort, while a stent that is too soft and extensible may have issues with dislodgement, kinking and breaking. CONCLUSION: This study provides the foundation for future work optimizing urethral stents, designing support for regenerative medicine applications, and improving hypospadias outcomes.


Subject(s)
Hypospadias , Plastic Surgery Procedures , Humans , Hypospadias/surgery , Infant , Male , Postoperative Period , Stents , Urethra/surgery
3.
J Mater Chem B ; 8(45): 10392-10406, 2020 12 07.
Article in English | MEDLINE | ID: mdl-33112356

ABSTRACT

Bombyx mori silk fibroin is a fibrous protein whose tunable properties and biocompatibility have resulted in its utility in a wide-variety of applications, including as drug delivery vehicles, wound dressings, and tissue engineering scaffolds. Control of protein and cell attachment is vital to the performance of biomaterials, but silk fibroin is mostly hydrophobic and interacts nonspecifically with cells and proteins. Silk functionalised with hydrophilic polymers reduces attachment, but the low number of reactive sites makes achieving a uniform conjugation a persistent challenge. This work presents a new approach to grow brush-like polymers from the surface of degradable silk films, where the films were enriched with hydroxyl groups, functionalised with an initiator, and finally reacted with acrylate monomers using atom transfer radical polymerisation. Two different routes to hydroxyl enrichment were investigated, one involving reaction with ethylene oxide (EO) and the other using a two-step photo-catalysed oxidation reaction. Both routes increased surface hydrophilicity, and hydrophilic monomers containing either uncharged (poly(ethylene glycol), PEG) pendant groups or zwitterionic pendant groups were polymerised from the surfaces. The initial processing of the films to induce beta sheet structures was found to impact the success of the polymerizations. Compared to the EO modified or unmodified silk surfaces, the oxidation reaction resulted in more polymer conjugation and the surfaces appear more uniform. Mesenchymal stem cell and protein attachment were the lowest on polymers grown from oxidised surfaces. PEG-containing brush-like polymers displayed lower protein attachment than surfaces conjugated with PEG using a previously reported "grafting to" method, but polymers containing zwitterionic side chains displayed both the lowest contact angles and the lowest cell and protein attachment. This finding may arise from the interactions of the zwitterionic pendant groups through their permanent dipoles and is an important finding because PEG is susceptible to oxidative damage that can reduce efficacy over time. These modified silk materials with lower cell and protein attachments are envisioned to find utility when enhanced diffusion around surfaces is required, such as in drug delivery implants.


Subject(s)
Bombyx/chemistry , Fibroins/chemistry , Polyethylene Glycols/chemistry , Tissue Scaffolds/chemistry , Animals , Cell Adhesion , Cell Line , Humans , Hydrophobic and Hydrophilic Interactions , Polymerization , Serum Albumin, Bovine/isolation & purification
4.
Biomed Mater ; 15(5): 055037, 2020 09 12.
Article in English | MEDLINE | ID: mdl-32924975

ABSTRACT

A reactive silk fibroin ink formulation designed for extrusion three-dimensional (3D) printing of protein-based hydrogels at room temperature is reported. This work is motivated by the need to produce protein hydrogels that can be printed into complex shapes with long-term stability using extrusion 3D printing at ambient temperature without the need for the addition of nanocomposites, synthetic polymers, or sacrifical templates. Silk fibroin from the Bombyx mori silkworm was purified and synthesized into reactive inks by enzyme-catalyzed dityrosine bond formation. Rheological and printing studies showed that tailoring the peroxide concentration in the reactive ink enables the silk to be extruded as a filament and printed into hydrogel constructs, supporting successive printed layers without flow of the construct or loss of desired geometry. To enable success of longer-term in vitro studies, 3D printed silk hydrogels were found to display excellent shape retention over time, as evidenced by no change in construct dimensions or topography when maintained for nine weeks in culture medium. Caco-2 (an intestinal epithelial cell line) attachment, proliferation, and tight junction formation on the printed constructs was not found to be affected by the geometry of the constructs tested. Intestinal myofibroblasts encapsulated within reactive silk inks were found to survive shearing during printing and proliferate within the hydrogel constructs. The work here thus provides a suitable route for extrusion 3D printing of protein hydrogel constructs that maintain their shape during printing and culture, and is expected to enable longer-term cellular studies of hydrogel constructs that require complex geometries and/or varying spatial distributions of cells on demand via digital printing.


Subject(s)
Bioprinting/methods , Fibroins/chemistry , Printing, Three-Dimensional , Silk/chemistry , Alginates/chemistry , Animals , Bombyx , Caco-2 Cells , Cell Adhesion , Cell Proliferation , Coculture Techniques , Humans , Hydrogels/chemistry , In Vitro Techniques , Microscopy, Confocal , Microscopy, Fluorescence , Myofibroblasts/metabolism , Nanocomposites , Oscillometry , Rheology , Robotics , Spectrometry, Fluorescence , Spectroscopy, Fourier Transform Infrared , Temperature , Tissue Engineering/methods , Tissue Scaffolds/chemistry , Tyrosine/analogs & derivatives , Tyrosine/chemistry , Water
5.
Bioconjug Chem ; 31(5): 1307-1312, 2020 05 20.
Article in English | MEDLINE | ID: mdl-32378886

ABSTRACT

Silk fibroin is a semicrystalline protein used as a renewable polymer source and as a biomaterial platform, but existing methods to synthetically modify fibroin suffer from low efficiencies that can limit the protein's utility. This work reports on a mild synthesis that results in a 2-fold increase in carboxylation through the disruption of noncovalent interactions during the reaction. Importantly, silk fibroin maintains its ability to form ß-sheets that are critical for tailoring mechanical and degradation properties, as well as for rendering solid constructs (e.g., films and scaffolds) insoluble in water. Increasing carboxyl functionalization affords control over protein charge, which permits tailoring the loading and release of small molecules using electrostatic interactions. Disruption of noncovalent interactions during aqueous carbodiimide coupling also significantly enhances conjugation efficiency of molecules containing primary amine groups, thus enabling high degrees of functionalization with biological molecules, such as proteins and peptides, for biomaterial applications.


Subject(s)
Carboxylic Acids/chemistry , Fibroins/chemistry , Amines/chemistry , Mechanical Phenomena , Models, Molecular , Protein Conformation, beta-Strand , Protein Stability , Solubility , Water/chemistry
6.
Biomacromolecules ; 21(6): 2365-2375, 2020 06 08.
Article in English | MEDLINE | ID: mdl-32374159

ABSTRACT

Hydrogels are frequently utilized as three-dimensional matrices for the culture and regeneration of soft tissues, but one challenge with the existing hydrogels is that, though the natural extracellular matrix of tissues may be ordered, there are few biocompatible ways to incorporate anisotropy within hydrogels. Liquid crystalline (LC) polymers are well suited for this because of their combination of molecular ordering and polymer elasticity; however, the hydrophobic nature of LC monomers has hindered their polymerization into hydrogels under cytocompatible conditions. This work reports on the generation of main-chain LC hydrogels in aqueous media and the ability of LC phases to affect mesenchymal stem cell behavior. The synthesis results in high gel fraction materials, and calorimetry, thermomechanical analysis, and X-ray scattering show that the networks organize into LC phases in the dry and hydrogel states. Human mesenchymal stem cells (hMSCs) cultured within the hydrogels show excellent viability, and hMSC proliferation proceeds at a faster rate in LC hydrogels compared to non-LC hydrogels. TThe result is a new synthetic approach for calamitic liquid crystalline hydrogels, which support the encapsulation and culture of human stem cells and are expected to enable applications as anisotropic and responsive substrates for tissue engineering and regenerative medicine.


Subject(s)
Hydrogels , Mesenchymal Stem Cells , Cell Culture Techniques , Extracellular Matrix , Humans , Tissue Engineering
7.
ACS Biomater Sci Eng ; 5(7): 3246-3259, 2019 Jul 08.
Article in English | MEDLINE | ID: mdl-33405568

ABSTRACT

Compared to conventional wound closure methods like sutures and staples, polymer-based tissue adhesives afford some distinct advantages, such as greater ease of deployment in spatially constrained surgical sites. One way to achieve aqueous adhesion is by introducing catechol functional groups that form coordinate and covalent bonds with a variety of substrates. This approach, inspired by marine organisms, has been applied to biopolymers and synthetic polymers, but one key challenge is that compositions that are soluble in water are often susceptible to high swelling ratios that can result in undesired compression of neighboring tissues. This work sought to synthesize aqueous adhesive gels that are capable of two modes of association: (1) adhesion and covalent cross-linking reactions arising from catechol oxidation and (2) noncovalent cross-linking arising from self-assembly of polymer backbones within the gelled adhesive. The network's self-assembly after gelation was envisioned to afford control over swelling and reinforce its strength. Bombyx mori silk fibroin was selected as the backbone of the adhesive network because it can be processed into an aqueous solution yet later be rendered insoluble in water through the assembly of its hydrophobic protein core. Distinct from a previous approach to functionalize silk directly with catechol groups, this work investigated in situ generation of catechol on silk fibroin by enzymatically modifying phenolic side chains, where it was found that this enzymatic approach led to conjugates with higher degrees of catechol functionalization and aqueous solubility. Silk fibroin was functionalized with tyramine to enrich the protein's phenolic side chains, which were subsequently oxidized into catechol groups using mushroom tyrosinase (MT). The gelation of the silk conjugates with MT was monitored by rheology, and the gels exhibited low water uptake. Phenolic enrichment increased the rate of chemical cross-linking leading to gelation but did not interrupt assembly of silk's secondary structures. Adhesion of the tyramine-silk conjugates to porcine intestine was found to be superior to fibrin sealant, and induction of ß sheet secondary structures was found to further enhance adhesive strength through a second mode of cross-linking. Neither the chemical functionalization nor phenol oxidation affected the ability of intestinal epithelial cells (Caco-2) to attach and proliferate. Phenolic functionalization and oxidative cross-linking of silk fibroin was found to afford a new route to water-soluble, catechol-functionalized polymers, which were found to display excellent adhesion to mucosal tissue and whose secondary structure provides an additional mode to control strength and swelling of adhesive gels.

8.
Soft Matter ; 14(48): 9885-9900, 2018 Dec 12.
Article in English | MEDLINE | ID: mdl-30511082

ABSTRACT

Liquid crystalline polymer networks (LCNs) couple polymer chain organization to molecular ordering, the switching of which has been shown to impart stimuli-responsive properties, including actuation and one-way shape memory, to the networks. While LCNs have long been proposed as artificial muscles, recent reports have also suggested potential as dynamic biomaterial substrates. In contrast to many existing LCNs synthesized using hydrophobic spacers, this work investigates networks synthesized using more hydrophilic spacers to promote interaction with water. A challenge with such materials is liquid crystalline phases could be disrupted in hydrated networks. This work thus investigates the impact of polyether spacers and mesogen composition on the phase behavior of LCNs. Main-chain LCNs were synthesized using alkyne-azide cycloaddition ("click" chemistry), where two different mesogens (5yH and 5yMe) and a non-LC monomer (5yTe) were coupled with one of two different polyether spacers, poly(ethylene glycol) and poly(propylene glycol), and a crosslinker. The chemistry led to high gel fraction materials, the workup of which resulted in networks that displayed no difference in cellular toxicity due to leachable components compared to tissue culture plastic control. Calorimetric analysis, dynamic mechanical analysis, and X-ray scattering revealed the LC microstructure and temperature-responsive properties of the networks. The use of low molecular weight polyether spacers was found to prevent their crystallization within the LC network, and adjusting mesogen composition to enhance its LC phase stability allowed the use of spacers with larger molecular weights and pendant groups. Hydrated networks were found to rearrange their structure compared to dry networks, while maintaining their LC phases. Like other crosslinked LC materials, the networks display shape changes (actuation) that are tied to changes in LC ordering. The result is a new synthetic approach for polydomain networks that form stable LC phases that are tailorable using polyether spacers and may enable future application as hydrated, stimuli-responsive materials.

9.
Med Confl Surviv ; 34(2): 95-120, 2018 Jun.
Article in English | MEDLINE | ID: mdl-30016122

ABSTRACT

This study explores how one social-structural variable, control over Sunni-Arab communities during the Iraq civil war, affected the types of violence used by insurgent/non-government actors that killed and injured civilians in these communities from January 2004 to December 2009. The study classifies three levels of control: (1) incumbent (government-supported) control, (2) insurgent control and (3) actively contested control. It uses Iraq Body Count (IBC) fatality data to characterize two general types of violence (selective and indiscriminate) evident during the Iraq conflict. It demonstrates that the type of violence committed by non-government actors was significantly (P > .01) different as related to the level of control insurgents had over territory. Primarily, insurgents/non-governmental actors used more selective forms of violence when insurgents controlled territory and more indiscriminate violence when incumbent (government-supported) forces controlled territory. Also, acts of indiscriminate violence cause considerably more injuries and death per act as compared to selective violence. Importantly, if control over territory has broadly generalizable effects on the types of violence that civilian's experience during civil war, than understanding this relationship could be useful when determining the types of medical assistance, medical supplies and training most needed in combat zones.


Subject(s)
Armed Conflicts/statistics & numerical data , Violence/classification , War-Related Injuries/mortality , Arabs , Humans , Iraq/epidemiology , Islam
10.
Materials (Basel) ; 10(7)2017 Jul 08.
Article in English | MEDLINE | ID: mdl-28773132

ABSTRACT

Recent advancements in metal fibers have introduced a promising new type of stainless steel fiber with high stiffness, high failure strain, and a thickness < 100 µm (<0.00394 in.) that can be utilized in a steel fiber-reinforced polymer. However, stainless steel is known to be susceptible to pitting corrosion. The main goal of this study is to compare the impact of corrosion on the mechanical properties of steel fiber-reinforced composites with those of conventional types of stainless steel. By providing experimental evidences, this study may promote the application of steel fiber-reinforced composite as a viable alternative to conventional metals. Samples of steel fiber-reinforced polymer and four different types of stainless steel were subjected to 144 and 288 h of corrosion in ferric chloride solution to simulate accelerated corrosion conditions. The weight losses due to corrosion were recorded. The corroded and control samples were tested under monotonic tensile loading to measure the ultimate stresses and strains. The effect of corrosion on the mechanical properties of the different materials was evaluated. The digital image correlation (DIC) technique was used to investigate the failure mechanism of the corrosion-damaged specimens. Overall, steel fiber-reinforced composites had the greatest corrosion resistance.

11.
Polymers (Basel) ; 9(4)2017 Apr 23.
Article in English | MEDLINE | ID: mdl-30970830

ABSTRACT

While conventional fiber-reinforced polymer composites offer high strength and stiffness, they lack ductility and the ability to absorb energy before failure. This work investigates hybrid fiber composites for structural applications comprised of polymer, steel fiber, and glass fibers to address this shortcoming. Varying volume fractions of thin, ductile steel fibers were introduced into glass fiber reinforced epoxy composites. Non-hybrid and hybrid composite specimens were prepared and subjected to monolithic and half-cyclic tensile testing to obtain stress-strain relationships, hysteresis behavior, and insight into failure mechanisms. Open-hole testing was used to assess the vulnerability of the composites to stress concentration. Incorporating steel fibers into glass/epoxy composites offered a significant improvement in energy absorption prior to failure and material re-centering capabilities. It was found that a lower percentage of steel fibers (8.2%) in the hybrid composite outperformed those with higher percentages (15.7% and 22.8%) in terms of energy absorption and re-centering, as the glass reinforcement distributed the plasticity over a larger area. A bilinear hysteresis model was developed to predict cyclic behavior of the hybrid composite.

12.
ACS Appl Mater Interfaces ; 8(25): 16218-26, 2016 Jun 29.
Article in English | MEDLINE | ID: mdl-27322909

ABSTRACT

Regenerated silk fibroin is a high molecular weight protein obtained by purifying the cocoons of the domesticated silkworm, Bombyx mori. This report exploits the aqueous processing and tunable ß sheet secondary structure of regenerated silk to produce nanostructures (i.e., inverse opals) that can be used as colorimetric immunosensors. Such sensors would enable direct detection of antigens by changes in reflectance spectra induced by binding events within the nanostructure. Silk inverse opals were prepared by solution casting and annealing in a humidified atmosphere to render the silk insoluble. Next, antigen sensing capabilities were imparted to silk through a three step synthesis: coupling of avidin to silk surfaces, coupling of biotin to antibodies, and lastly antibody attachment to silk through avidin-biotin interactions. Varying the antibody enables detection of different antigens, as demonstrated using different protein antigens: antibodies, red fluorescent protein, and the beta subunit of cholera toxin. Antigen binding to sensors induces a red shift in the opal reflectance spectra, while sensors not exposed to antigen showed either no shift or a slight blue shift. This work constitutes a first step for the design of biopolymer-based optical systems that could directly detect antigens using commercially available reagents and environmentally friendly chemistries.

13.
Biomacromolecules ; 17(1): 237-45, 2016 Jan 11.
Article in English | MEDLINE | ID: mdl-26674175

ABSTRACT

Silk fibroin from the domesticated silkworm Bombyx mori is a naturally occurring biopolymer with charged hydrophilic terminal regions that end-cap a hydrophobic core consisting of repeating sequences of glycine, alanine, and serine residues. Taking inspiration from mussels that produce proteins rich in L-3,4-dihydroxyphenylalanine (DOPA) to adhere to a variety of organic and inorganic surfaces, the silk fibroin was functionalized with catechol groups. Silk fibroin was selected for its high molecular weight, tunable mechanical and degradation properties, aqueous processability, and wide availability. The synthesis of catechol-functionalized silk fibroin polymers containing varying amounts of hydrophilic polyethylene glycol (PEG, 5000 g/mol) side chains was carried out to balance silk hydrophobicity with PEG hydrophilicity. The efficiency of the catechol functionalization reaction did not vary with PEG conjugation over the range studied, although tuning the amount of PEG conjugated was essential for aqueous solubility. Adhesive bonding and cell compatibility of the resulting materials were investigated, where it was found that incorporating as little as 6 wt % PEG prior to catechol functionalization resulted in complete aqueous solubility of the catechol conjugates and increased adhesive strength compared with silk lacking catechol functionalization. Furthermore, PEG-silk fibroin conjugates maintained their ability to form ß-sheet secondary structures, which can be exploited to reduce swelling. Human mesenchymal stem cells (hMSCs) proliferated on the silks, regardless of PEG and catechol conjugation. These materials represent a protein-based approach to catechol-based adhesives, which we envision may find applicability as biodegradable adhesives and sealants.


Subject(s)
Biocompatible Materials/chemical synthesis , Fibroins/chemical synthesis , Mesenchymal Stem Cells/physiology , Tissue Adhesives/chemical synthesis , Animals , Bivalvia , Bombyx , Cells, Cultured , Humans , Hydrophobic and Hydrophilic Interactions , Polyethylene Glycols/chemistry , Protein Structure, Secondary , Proteins/metabolism , Silk/chemistry
14.
ACS Biomater Sci Eng ; 1(4): 260-270, 2015 Apr 13.
Article in English | MEDLINE | ID: mdl-25984573

ABSTRACT

We present a silk biomaterial platform with highly tunable mechanical and degradation properties for engineering and regeneration of soft tissues such as, skin, adipose, and neural tissue, with elasticity properties in the kilopascal range. Lyophilized silk sponges were prepared under different process conditions and the effect of silk molecular weight, concentration and crystallinity on 3D scaffold formation, structural integrity, morphology, mechanical and degradation properties, and cell interactions in vitro and in vivo were studied. Tuning the molecular weight distribution (via degumming time) of silk allowed the formation of stable, highly porous, 3D scaffolds that held form with silk concentrations as low as 0.5% wt/v. Mechanical properties were a function of silk concentration and scaffold degradation was driven by beta-sheet content. Lyophilized silk sponges supported the adhesion of mesenchymal stem cells throughout 3D scaffolds, cell proliferation in vitro, and cell infiltration and scaffold remodeling when implanted subcutaneously in vivo.

15.
Methods ; 84: 84-9, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25843606

ABSTRACT

Adipose tissue engineered models are needed to enhance our understanding of disease mechanisms and for soft tissue regenerative strategies. Perfusion systems generate more physiologically relevant and sustainable adipose tissue models, however adipocytes have unique properties that make culturing them in a perfusion environment challenging. In this paper we describe the methods involved in the development of two perfusion culture systems (2D and 3D) to test their applicability for long term in vitro adipogenic cultures. It was hypothesized that a silk protein biomaterial scaffold would provide a 3D framework, in combination with perfusion flow, to generate a more physiologically relevant sustainable adipose tissue engineered model than 2D cell culture. Consistent with other studies evaluating 2D and 3D culture systems for adipogenesis we found that both systems successfully model adipogenesis, however 3D culture systems were more robust, providing the mechanical structure required to contain the large, fragile adipocytes that were lost in 2D perfused culture systems. 3D perfusion also stimulated greater lipogenesis and lipolysis and resulted in decreased secretion of LDH compared to 2D perfusion. Regardless of culture configuration (2D or 3D) greater glycerol was secreted with the increased nutritional supply provided by perfusion of fresh media. These results are promising for adipose tissue engineering applications including long term cultures for studying disease mechanisms and regenerative approaches, where both acute (days to weeks) and chronic (weeks to months) cultivation are critical for useful insight.


Subject(s)
Adipogenesis/physiology , Adipocytes/cytology , Adult Stem Cells/cytology , Animals , Biocompatible Materials/chemistry , Cell Culture Techniques/methods , Humans , Materials Testing , Perfusion/instrumentation , Silk/chemistry , Tissue Engineering/methods , Tissue Scaffolds/chemistry
16.
Nanomedicine (Lond) ; 10(5): 803-14, 2015.
Article in English | MEDLINE | ID: mdl-25816881

ABSTRACT

AIM: Silk-tropoelastin alloys, composed of recombinant human tropoelastin and regenerated Bombyx mori silk fibroin, are an emerging, versatile class of biomaterials endowed with tunable combinations of physical and biological properties. Electrodeposition of these alloys provides a programmable means to assemble functional gels with both spatial and temporal controllability. MATERIALS & METHODS: Tropoelastin-modified silk was prepared by enzymatic coupling between tyrosine residues. Hydrogel coatings were electrodeposited using two wire electrodes. RESULTS & DISCUSSION: Mechanical characterization and in vitro cell culture revealed enhanced adhesive capability and cellular response of these alloy gels as compared with electrogelled silk alone. CONCLUSION: These electro-depositable silk-tropoelastin alloys constitute a suitable coating material for nanoparticle-based drug carriers and offer a novel opportunity for on-demand encapsulation/release of nanomedicine.


Subject(s)
Biocompatible Materials/chemistry , Fibroins/chemistry , Tropoelastin/chemistry , Animals , Biomechanical Phenomena , Cell Adhesion , Cells, Cultured , Coated Materials, Biocompatible/chemistry , Drug Carriers/chemistry , Electrochemical Techniques , Gels , Human Umbilical Vein Endothelial Cells , Humans , Magnetic Resonance Spectroscopy , Materials Testing , Nanocapsules/chemistry , Nanoconjugates/chemistry , Nanomedicine , Oxidation-Reduction , Spectroscopy, Fourier Transform Infrared , Tyrosine/chemistry
17.
Macromol Biosci ; 15(6): 861-74, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25761231

ABSTRACT

The effects of common sterilization techniques on the physical and biological properties of lyophilized silk fibroin sponges are described. Sterile silk fibroin sponges were cast using a pre-sterilized silk fibroin solution under aseptic conditions or post-sterilized via autoclaving, γ radiation, dry heat, exposure to ethylene oxide, or hydrogen peroxide gas plasma. Low average molecular weight and low concentration silk fibroin solutions could be sterilized via autoclaving or filtration without significant loses of protein. However, autoclaving reduced the molecular weight distribution of the silk fibroin protein solution, and silk fibroin sponges cast from autoclaved silk fibroin were significantly stiffer compared to sponges cast from unsterilized or filtered silk fibroin. When silk fibroin sponges were sterilized post-casting, autoclaving increased scaffold stiffness, while decreasing scaffold degradation rate in vitro. In contrast, γ irradiation accelerated scaffold degradation rate. Exposure to ethylene oxide significantly decreased cell proliferation rate on silk fibroin sponges, which was rescued by leaching ethylene oxide into PBS prior to cell seeding.


Subject(s)
Fibroblasts/metabolism , Fibroins/chemistry , Materials Testing , Sterilization/methods , Tissue Scaffolds/chemistry , Fibroblasts/cytology , Humans , Male
18.
Adv Funct Mater ; 24(29): 4615-4624, 2014 Aug 06.
Article in English | MEDLINE | ID: mdl-25395921

ABSTRACT

Elastomeric, fully degradable and biocompatible biomaterials are rare, with current options presenting significant limitations in terms of ease of functionalization and tunable mechanical and degradation properties. We report a new method for covalently crosslinking tyrosine residues in silk proteins, via horseradish peroxidase and hydrogen peroxide, to generate highly elastic hydrogels with tunable properties. The tunable mechanical properties, gelation kinetics and swelling properties of these new protein polymers, in addition to their ability to withstand shear strains on the order of 100%, compressive strains greater than 70% and display stiffness between 200 - 10,000 Pa, covering a significant portion of the properties of native soft tissues. Molecular weight and solvent composition allowed control of material mechanical properties over several orders of magnitude while maintaining high resilience and resistance to fatigue. Encapsulation of human bone marrow derived mesenchymal stem cells (hMSC) showed long term survival and exhibited cell-matrix interactions reflective of both silk concentration and gelation conditions. Further biocompatibility of these materials were demonstrated with in vivo evaluation. These new protein-based elastomeric and degradable hydrogels represent an exciting new biomaterials option, with a unique combination of properties, for tissue engineering and regenerative medicine.

19.
Biomaterials ; 35(1): 83-91, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24099708

ABSTRACT

Over the past 30 years, silk has been proposed for numerous biomedical applications that go beyond its traditional use as a suture material. Silk sutures are well tolerated in humans, but the use of silk for vascular engineering applications still requires extensive biocompatibility testing. Some studies have indicated a need to modify silk to yield a hemocompatible surface. This study examined the potential of low molecular weight heparin as a material for refining silk properties by acting as a carrier for vascular endothelial growth factor (VEGF) and improving silk hemocompatibility. Heparinized silk showed a controlled VEGF release over 6 days; the released VEGF was bioactive and supported the growth of human endothelial cells. Silk samples were then assessed using a humanized hemocompatibility system that employs whole blood and endothelial cells. The overall thrombogenic response for silk was very low and similar to the clinical reference material polytetrafluoroethylene. Despite an initial inflammatory response to silk, apparent as complement and leukocyte activation, the endothelium was maintained in a resting, anticoagulant state. The low thrombogenic response and the ability to control VEGF release support the further development of silk for vascular applications.


Subject(s)
Biocompatible Materials , Blood Vessels/cytology , Heparin , Silk , Tissue Engineering , Humans
20.
Reprod Sci ; 20(8): 929-36, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23271162

ABSTRACT

OBJECTIVE: New therapies to prevent preterm birth are needed. Our objective was to study an injectable biomaterial for human cervical tissue as an alternative to cervical cerclage. STUDY DESIGN: Human cervical tissue specimens were obtained from premenopausal gynecological hysterectomies for benign indications. A 3-part biomaterial was formulated, consisting of silk protein solution blended with a 2-part polyethylene glycol gelation system. The solutions were injected into cervical tissue and the tissue was evaluated for mechanical properties, swelling, cytocompatibility, and histology. RESULTS: The stiffness of cervical tissue more than doubled after injection (P = .02). Swelling properties of injected tissue were no different than native tissue controls. Cervical fibroblasts remained viable for at least 48 hours when cultured on the biomaterial. CONCLUSIONS: We report a silk-based, biocompatible, injectable biomaterial that increased the stiffness of cervical tissue compared to uninjected controls. Animal studies are needed to assess this biomaterial in vivo.


Subject(s)
Biocompatible Materials , Cerclage, Cervical , Cervix Uteri/drug effects , Polyethylene Glycols/administration & dosage , Premature Birth/prevention & control , Silk/administration & dosage , Cell Survival , Cells, Cultured , Cervix Uteri/pathology , Elasticity , Female , Fibroblasts/drug effects , Fibroblasts/pathology , Humans , Injections , Materials Testing , Polyethylene Glycols/chemistry , Polyethylene Glycols/toxicity , Silk/chemistry , Silk/toxicity , Stress, Mechanical , Time Factors
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