Molecular and macro-scale analysis of enzyme-crosslinked silk hydrogels for rational biomaterial design

McGill, Meghan; Coburn, Jeannine M.; Partlow, Benjamin P.; Mu, Xuan; Kaplan, David L.

doi:10.1016/j.actbio.2017.09.020

Cited by 84 publications

(52 citation statements)

References 58 publications

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“…To verify successful gentamicin-conjugation on bactericidal effects against P. aeruginosa , we validated EDC-mediated coupling of gentamicin to silk via liquid chromatography-tandem mass spectrometry (LC-MS/MS), following acid hydrolysis of silk and gentamicin standards and gentamicin-conjugated silk film samples (Figure 3A). We confirmed effective silk hydrolysis based on the presence of tyrosine in the silk standard (precursor ion 182.1 m/z, product ion 136.1 m/z, retention time 3.8 min), which was consistent with previous reports [35]. A standard solution of gentamicin showed a precursor ion at 478.4 m/z, product ion at 322.2 m/z, and a retention time of 5.9 min, consistent with literature reports [36].…”

Section: Resultssupporting

confidence: 92%

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Silk-Based Therapeutics Targeting Pseudomonas aeruginosa

McKay

Parker

Hawker

et al. 2019

JFB

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Pseudomonas aeruginosa (P. aeruginosa) infections may lead to severe damage of the cornea, mucosa, and skin. The highly aggressive nature of P. aeruginosa and the rise in multi-drug resistance, particularly in nosocomial settings, lead to an increased risk for permanent tissue damage and potentially death. Thus, a growing need exists to develop alternative treatments to reduce both the occurrence of bacterial infection and biofilm development, as well as pathological progression post-infection. Silk derived from Bombyx mori silkworms serves as a unique biomaterial that is biocompatible with low immunogenicity and high versatility, and thereby ideal for stabilizing therapeutics. In this study, we assessed the cytotoxicity of P. aeruginosa on human corneal stromal stem cells and two mucosal cell lines (Caco-2 and HT29-MTX). To determine whether antibiotic-immobilized scaffolds can serve as alternative therapeutics to free, diffuse forms, we developed novel gentamicin-conjugated silk films as functional scaffolds and compared antimicrobial effects and free gentamicin. The advantages of generating a surface coating with a covalently-bound antibiotic may reduce potential side-effects associated with free gentamicin, as well as limit the diffusion of the drug. Our results suggest that gentamicin conjugated to native silk and carboxyl-enriched silk inhibits P. aeruginosa growth. Development of stabilized antibiotic treatments with surface toxicity selective against bacteria may serve as an alternative approach to treat active infections, as well as potential prophylactic use as coatings in high-risk cases, such as post-surgical complications or prolonged hospitalization.

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Section: Resultssupporting

confidence: 92%

“…We performed liquid chromatography tandem mass spectrometry (LC-MS/MS) as previously described [35]. Briefly, experiments were performed on an Agilent 1200 series high performance liquid chromatography (HPLC) instrument and Agilent 6410 triple-quadruple mass spectrometer (Agilent Technologies, Santa Clara, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Silk-Based Therapeutics Targeting Pseudomonas aeruginosa

McKay

Parker

Hawker

et al. 2019

JFB

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“…The enzymatically‐induced crosslinking of silk fibroin can be done by use of enzyme such as horseradish peroxidase (HRP) or mushroom tyrosinase. Mushroom tyrosinases is an oxidative enzyme which oxides the tyrosine residue present in silk fibroin protein to reactive o‐quinone‐moieties without disrupting the peptide bond 74 …”

Section: Preparation Of Silk Fibroin Based Bioinkmentioning

confidence: 99%

Evaluation of silk‐based bioink during pre and post 3D bioprinting: A review

et al. 2020

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During past few decades, the demand for the replacement of damaged organs is increasing consistently. This is due to the advancement in tissue engineering, which opens the possibility of regeneration of damaged organs or tissues into functional parts with the help of 3D bioprinting. Bioprinting technology presents an excellent potential to develop complex structures with precise control over cell suspension and structure. A brief description of different types of 3D bioprinting techniques, including inkjetbased, laser-based, and extrusion-based bioprinting is presented here. Due to innate advantageous features like tunable biodegradability, biocompatibility, elasticity and mechanical robustness, silk has carved a niche in the realm of tissue engineering. In this review article, the focus is to highlight the possible approach of exploring silk as bioink for fabrication of bioprinted implants using 3D bioprinting. This review discusses different type of degumming, dissolution techniques for extraction of proteins from different sources of silk. Different recently reported 3D bioprinting techniques suitable for silkbased bioink are further elaborated. Postprinting characterization of resultant scaffolds are also describe here. However, there is an astounding progress in 3D bioprinting technology, still there is a need to develop further the current bioprinting technology to make it suitable for generation of heterogeneous tissue construct. The possibility of utilizing the adhesive property of sericin to consider it as bioink is elaborated.

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“…The two primary sources of silk are spider or silkworm (Arntzen and Ritter, 1994 ), and these can also be modified (e.g., by crosslinking tyrosine residues) to provide a more tunable carrier platform (Partlow et al, 2014 ). Silks can also be processed into a number of forms including films (Dutta et al, 2013 ), fibers (Mandal et al, 2012 ; Panda et al, 2015 ), hydrogels (McGill et al, 2017 ), scaffolds (Marolt et al, 2006 ; Zhu et al, 2014 ), and are compatible for fabrication of composites with ceramic (Jin et al, 2015 ) or synthetic polymers. Furthermore, these materials have been used as carriers for rhBMP-2 (Karageorgiou et al, 2006 ; Ma et al, 2016 ) and promoted bridging in a critically-sized rat femur defect model (Kirker-Head et al, 2007 ).…”

Section: Alternative Biomaterials Carriers For Molecules Promote Bone mentioning

confidence: 99%

Biomaterials for the Delivery of Growth Factors and Other Therapeutic Agents in Tissue Engineering Approaches to Bone Regeneration

2018

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Bone fracture followed by delayed or non-union typically requires bone graft intervention. Autologous bone grafts remain the clinical “gold standard”. Recently, synthetic bone grafts such as Medtronic's Infuse Bone Graft have opened the possibility to pharmacological and tissue engineering strategies to bone repair following fracture. This clinically-available strategy uses an absorbable collagen sponge as a carrier material for recombinant human bone morphogenetic protein 2 (rhBMP-2) and a similar strategy has been employed by Stryker with BMP-7, also known as osteogenic protein-1 (OP-1). A key advantage to this approach is its “off-the-shelf” nature, but there are clear drawbacks to these products such as edema, inflammation, and ectopic bone growth. While there are clinical challenges associated with a lack of controlled release of rhBMP-2 and OP-1, these are among the first clinical examples to wed understanding of biological principles with biochemical production of proteins and pharmacological principles to promote tissue regeneration (known as regenerative pharmacology). After considering the clinical challenges with such synthetic bone grafts, this review considers the various biomaterial carriers under investigation to promote bone regeneration. This is followed by a survey of the literature where various pharmacological approaches and molecular targets are considered as future strategies to promote more rapid and mature bone regeneration. From the review, it should be clear that pharmacological understanding is a key aspect to developing these strategies.

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Molecular and macro-scale analysis of enzyme-crosslinked silk hydrogels for rational biomaterial design

Cited by 84 publications

References 58 publications

Silk-Based Therapeutics Targeting Pseudomonas aeruginosa

Silk-Based Therapeutics Targeting Pseudomonas aeruginosa

Evaluation of silk‐based bioink during pre and post 3D bioprinting: A review

Biomaterials for the Delivery of Growth Factors and Other Therapeutic Agents in Tissue Engineering Approaches to Bone Regeneration

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