Tuning Myogenesis by Controlling Gelatin Hydrogel Properties through Hydrogen Peroxide-Mediated Cross-Linking and Degradation

Abstract

Engineering skeletal muscle tissue in vitro is important to study the mechanism of myogenesis, which is crucial for regenerating muscle cells. The physicochemical properties of the cellular microenvironment are known to govern various cell behaviours. Yet, most studies utilised synthetic materials to model the extracellular matrix that suffers from cytotoxicity to the cells. We have previously reported that the physicochemical property of hydrogels obtained from horseradish peroxidase (HRP)-catalysed cross-linking could be controlled by a simple adjustment to the exposure time to air containing H2O2. In this study, we evaluated the influence of physicochemical properties dynamics in the gelatin possessing phenol groups (Gelatin-Ph) hydrogel to regulate the myogenesis in vitro. We controlled the Young’s modulus of the Gelatin-Ph hydrogel by tuning the air containing 16 ppm H2O2 exposure time for 15–60 min. Additionally, prolonged exposure to air containing H2O2 also induced Gelatin-Ph degradation. Myoblasts showed higher adhesion and myotube formation on stiff hydrogel (3.53 kPa) fabricated through 30 min of exposure to air containing H2O2 compared to those on softer hydrogel (0.77–2.79 kPa) fabricated through 15, 45, and 60 min of the exposure. These results demonstrate that the myogenesis can be tuned by changes in the physicochemical properties of Gelatin-Ph hydrogel mediated by H2O2.

Sterilization of Polymeric Implants: Challenges and Opportunities

“…Benefits of H 2 O 2 sterilization are similar to those of other chemical sterilants, including relatively low operating temperatures and better compatibility with heat sensitive polymers. , Compared to EtO gas, however, vaporized H 2 O 2 has less effective penetration of porous scaffolds ,,,, Furthermore, although deliberate H 2 O 2 incorporation the in polymeric scaffold may be leveraged for certain drug delivery or tissue engineering applications, the hydroxyl free radical generated during H 2 O 2 sterilization has the potential to oxidatively cross-link polymers, resulting in deleterious changes. ,, Studies characterizing the morphological and biochemical changes induced by H 2 O 2 sterilization in UHMWPE revealed changes similar to those induced by gamma irradiation, including embrittlement . H 2 O 2 sterilization has also been observed to induce mechanical and degradative property changes to degradable polymers such as PLA. ,, Lastly, it is of note that H 2 O 2 sterilization was found to be ineffective when paired with cellulose-based products due to the high tendency of cellulose to absorb H 2 O 2 , thereby reducing its effective concentration .…”

“…2,35,65 Benefits of H 2 O 2 sterilization are similar to those of other chemical sterilants, including relatively low operating temperatures and better compatibility with heat sensitive polymers. 2,34 Compared to EtO gas, however, vaporized H 2 O 2 has less effective penetration of porous scaffolds 2,34,35,65,66 Furthermore, although deliberate H 2 O 2 incorporation the in polymeric scaffold may be leveraged for certain drug delivery 67 or tissue engineering applications, 68 the hydroxyl free radical generated during H 2 O 2 sterilization has the potential to oxidatively cross-link polymers, resulting in deleterious changes. 35,68,69 Studies characterizing the morphological and biochemical changes induced by H 2 O 2 sterilization in UHMWPE revealed changes similar to those induced by gamma irradiation, including embrittlement.…”

“…2,34 Compared to EtO gas, however, vaporized H 2 O 2 has less effective penetration of porous scaffolds 2,34,35,65,66 Furthermore, although deliberate H 2 O 2 incorporation the in polymeric scaffold may be leveraged for certain drug delivery 67 or tissue engineering applications, 68 the hydroxyl free radical generated during H 2 O 2 sterilization has the potential to oxidatively cross-link polymers, resulting in deleterious changes. 35,68,69 Studies characterizing the morphological and biochemical changes induced by H 2 O 2 sterilization in UHMWPE revealed changes similar to those induced by gamma irradiation, including embrittlement. 70 H 2 O 2 sterilization has also been observed to induce mechanical and degradative property changes to degradable polymers such as PLA.…”

Tissue engineering modalities in skeletal muscles: focus on angiogenesis and immunomodulation properties

“…High-rate functionality of enzymes in physiological conditions and specific biocatalyst activity on certain substrates can reduce the side effects of applied substrates [ 184 ]. In general, the selection of enzymes for chemical cross-linking is based on the recognition of a specific substrate involved in the progression of a polymeric network [ 185 , 186 ]. Among different enzymes, tyrosinase is a copper-based enzyme that generates covalent linkages via the oxidation of phenolic groups of tyrosine following the production of quinones [ 187 , 188 ].…”

Editorial on the Special Issue “Modification of Hydrogels and Their Applications in Biomedical Engineering”

“…This Special Issue focuses on the modification of hydrogels and their applications in biomedical engineering, aiming to provide a reference for scholars in related fields. Up until now, ten research articles and one review article have been collected, which mainly involve the application of hydrogels in biomedical fields such as phantoms [ 1 ], wound healing [ 2 , 3 ], skin [ 4 ], cartilage [ 5 , 6 ], muscle [ 7 ], and drug loading [ 8 , 9 , 10 ].…”