Ovarian Cell Encapsulation in an Enzymatically Crosslinked Silk-Based Hydrogel with Tunable Mechanical Properties

Abstract

An artificial ovary is a promising approach for preserving fertility in prepubertal girls and women who cannot undergo current cryopreservation strategies. However, this approach is in its infancy, due to the possible challenges of creating a suitable 3D matrix for encapsulating ovarian follicles and stromal cells. To maintain the ovarian stromal cell viability and proliferation, as a first step towards developing an artificial ovary, in this study, a double network hydrogel with a high water swelling capacity (swelling index 15–19) was developed, based on phenol conjugated chitosan (Cs-Ph) and silk fibroin (SF) through an enzymatic crosslinking method using horseradish peroxidase. The addition of SF (1%) to Cs (1%) decreased the storage modulus (G’) from 3500 Pa (Cs1) to 1600 Pa (Cs-SF1), and the hydrogels with a rapid gelation kinetic produced a spatially homogeneous distribution of ovarian cells that demonstrated 167% proliferation after 7 days. This new Cs-SF hydrogel benefits from the toughness and flexibility of SF, and phenolic chemistry could provide the potential microstructure for encapsulating human ovarian stromal cells.

Synergistic complexation of phenol functionalized polymer induced in situ microfiber formation for 3D printing of marine-based hydrogels

“…In our previous study, the addition of silk fibroin with lower G′ compared to that of chitosan significantly lowered the G′ of the hydrogel (3 times). 36 However, in this study, the in situ microfibers could maintain the hydrogel elasticity in addition to increasing the G″ of the DCCA.…”

Anionic exopolysaccharide from Cryptococcus laurentii 70766 as an alternative for alginate for biomedical hydrogels

“…EPS and Na-Alg were dissolved in MES buffer (1% w/v), and EDC (0.5% w/v) and NHS (0.3% w/v) were added to each solution to activate the carboxylic acid groups of EPS and Na-Alg for 30 minutes under stirring [38,39]. Tyramine HCl (0.35 g/10 ml of MES buffer) was added dropwise to the solutions.…”

“…Tyramine HCl (0.35 g/10 ml of MES buffer) was added dropwise to the solutions. The pH of the solutions was then maintained at 6 by dropwise adding 1 M HCl and allowing it to react with tyramine HCl at room temperature for 24 hours with stirring [38]. The resultant solutions were dialyzed for three days against deionized water in dialysis bags (cutoff: 14 kDa), and the water was changed every eight hours.…”

“…The network microstructure of hydrogels is one of the essential features controlling protein adsorption, cell proliferation, mass transfer, and nutrition transfer [38,68]. Furthermore, hydrogels containing interconnected porose networks can influence cell activity by promoting cell adhesion and cell-cell interaction [69].…”

Modeling Tunable Fracture in Hydrogel Shell Structures for Biomedical Applications

“…Their hydrogel shows a capability of free-floating on the water surface, indicating its potential use in floating pH detection devices. In the same vein, Jafari et al [ 74 ] developed hydrogels of controllable gelation time and adjustable degradation rate. Their new hydrogel exhibited remarkable prospects for the reconstruction of artificial ovaries.…”