FLASH: Fluorescently LAbelled Sensitive Hydrogel to monitor bioscaffolds degradation during neocartilage generation

Onofrillo, Carmine; Duchi, Serena; Francis, Sam L.; O’Connell, Cathal; Aguilar, Lilith M Caballero; Doyle, Stephanie E.; Yue, Zhilian; Wallace, Gordon G.; Choong, Peter; Bella, Claudia Di

doi:10.1016/j.biomaterials.2020.120383

Cited by 38 publications

(37 citation statements)

References 42 publications

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“…Using a previously reported method, we employed fluorescent material to track the degradation of GelMA scaffolds containing transglutaminase with the enzyme collagenase type II added to the PBS for biologically mediated degradation ( 36 ). Fluorescent images of individual scaffolds illustrate that those containing transglutaminase remained stable in the enzymatic solution for longer relative to controls at the same GelMA concentration ( Figure 3A ).…”

Section: Resultsmentioning

confidence: 99%

“…In this study, the addition of microbial transglutaminase did not negatively impact the metabolic activity of hADSCs encapsulated in photocrosslinked GelMA scaffolds in comparison to transglutaminase free controls over a 7 day period. No increase in metabolic activity (indicating proliferation) was observed; however, previous work has shown little growth in metabolic activity of hADSCs in 10% GelMA over a 7 day period (36). It may be that 10% GelMA scaffolds have relatively poor diffusive properties, which may limit nutrient access within the scaffold.…”

Section: Transglutaminase Did Not Negatively Impact Embedded Hadscs Nor Did Cells Prevent Enzymatic Crosslinkingmentioning

confidence: 86%

“…Gelatin type A from porcine skin (~300 Bloom) was purchased from Sigma-Aldrich (St Louis, MO, USA), and gelatin methacryloyl (GelMA) with ~85% degree of functionalization was used, building on previous work using the same GelMA product provided by TRICEP (Wollongong, NSW, Australia) ( 36 ). Fluorescent isothiocyanate isomer I (FITC) was conjugated to GelMA as previously reported ( 36 ). GelMA, GelMA-FITC and gelatin were sterilized using ethylene oxide (EtO) gas based on our previously reported method ( 37 ).…”

Section: Methodsmentioning

confidence: 99%

“…Aliquots were prepared to achieve final GelMA concentrations of 6, 8, and 10% w/v with the remaining components of the gel. These groups were selected based on a previous study where these concentrations showed suitable properties for cartilage regeneration in vitro ( 36 ). Stock LAP solution was added to a final concentration of 0.05%, and for scaffolds containing transglutaminase, the final enzyme concentration was 1 U/ml.…”

Section: Methodsmentioning

confidence: 99%

“…A previously validated method was used to track the enzymatic degradation of the hydrogel using fluorescence (36). In brief, after overnight incubation and swelling, PBS was removed and replaced with 1 ml PBS containing collagenase type II at a concentration of 5 U/ml (Worthington Biochemical Corporation) to commence enzymatic degradation upon incubation at 37 • C. For the first 9 h of the experiment, collections of the PBS containing degraded fluorescent material were made every 90 min, and PBS/collagenase was refreshed.…”

Section: Enzymatic Degradation Of Scaffoldsmentioning

confidence: 99%

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Microbial Transglutaminase Improves ex vivo Adhesion of Gelatin Methacryloyl Hydrogels to Human Cartilage

Trengove

Duchi

Onofrillo

et al. 2021

Front. Med. Technol.

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Current surgical techniques to treat articular cartilage defects fail to produce a satisfactory long-term repair of the tissue. Regenerative approaches show promise in their ability to generate hyaline cartilage using biomaterials in combination with stem cells. However, the difficulty of seamlessly integrating the newly generated cartilage with the surrounding tissue remains a likely cause of long-term failure. To begin to address this integration issue, our strategy exploits a biological enzyme (microbial transglutaminase) to effect bioadhesion of a gelatin methacryloyl implant to host tissue. Mechanical characterization of the bioadhesive material shows that enzymatic crosslinking is compatible with photocrosslinking, allowing for a dual-crosslinked system with improved mechanical properties, and a slower degradation rate. Biocompatibility is illustrated with a 3D study of the metabolic activity of encapsulated human adipose derived stem cells. Furthermore, enzymatic crosslinking induced by transglutaminase is not prevented by the presence of cells, as measured by the bulk modulus of the material. Adhesion to human cartilage is demonstrated ex vivo with a significant increase in adhesive strength (5.82 ± 1.4 kPa as compared to 2.87 ± 0.9 kPa, p < 0.01) due to the addition of transglutaminase. For the first time, we have characterized a bioadhesive material composed of microbial transglutaminase and GelMA that can encapsulate cells, be photo crosslinked, and bond to host cartilage, taking a step toward the integration of regenerative implants.

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Section: Resultsmentioning

confidence: 99%

Section: Transglutaminase Did Not Negatively Impact Embedded Hadscs Nor Did Cells Prevent Enzymatic Crosslinkingmentioning

confidence: 86%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Enzymatic Degradation Of Scaffoldsmentioning

confidence: 99%

See 3 more Smart Citations

Microbial Transglutaminase Improves ex vivo Adhesion of Gelatin Methacryloyl Hydrogels to Human Cartilage

Trengove

Duchi

Onofrillo

et al. 2021

Front. Med. Technol.

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Stimuli‐Responsive Fluorescent Polymeric Hydrogels

Lü

Wei

Chen

2022

Smart Stimuli‐Responsive Polymers, Films, and Gels

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In Situ Forming Hydrogel as a Tracer and Degradable Lacrimal Plug for Dry Eye Treatment

Xiao

Zheng

et al. 2022

Adv Healthcare Materials

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Lacrimal plug is an effective and widely therapeutic strategy to treat dry eye. However, almost all commercialized plugs are fixed in a certain design and associated with many complications, such as spontaneous plug extrusion, epiphora, and granuloma and cannot be traced in the long‐term. Herein, a simple in situ forming hydrogel is developed as a tracer and degradable lacrimal plug to achieve the best match with the irregular lacrimal passages. In this strategy, methacrylate‐modified silk fibroin (SFMA) is served as a network, and a self‐assembled indocyanine green fluorescence tracer nanoparticle (FTN) is embedded as an indicator to develop the hydrogel plug using visible photo‐crosslinking. This SFMA/FTN hydrogel plug has excellent biocompatibility and biodegradability, which can be noninvasively monitored by near‐infrared light. In vivo tests based on dry eye rabbits show that the SFMA/FTN hydrogel plug can completely block the lacrimal passages and greatly improve the various clinical indicators of dry eye. These results demonstrate that the SFMA/FTN hydrogel is suitable as an injectable and degradable lacrimal plug with a long‐term tracking function. The work offers a new approach to the development of absorbable plugs for the treatment of dry eye.

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FLASH: Fluorescently LAbelled Sensitive Hydrogel to monitor bioscaffolds degradation during neocartilage generation

Cited by 38 publications

References 42 publications

Microbial Transglutaminase Improves ex vivo Adhesion of Gelatin Methacryloyl Hydrogels to Human Cartilage

Microbial Transglutaminase Improves ex vivo Adhesion of Gelatin Methacryloyl Hydrogels to Human Cartilage

Stimuli‐Responsive Fluorescent Polymeric Hydrogels

In Situ Forming Hydrogel as a Tracer and Degradable Lacrimal Plug for Dry Eye Treatment

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