Thermo-responsive methylcellulose hydrogels as temporary substrate for cell sheet biofabrication

Abstract

Methylcellulose (MC), a water-soluble polymer derived from cellulose, was investigated as a possible temporary substrate having thermo-responsive properties favorable for cell culturing. MC-based hydrogels were prepared by a dispersion technique, mixing MC powder (2, 4, 6, 8, 10, 12 % w/v) with selected salts (sodium sulphate, Na2SO4), sodium phosphate, calcium chloride, or phosphate buffered saline, to evaluate the influence of different compositions on the thermo-responsive behavior. The inversion test was used to determine the gelation temperatures of the different hydrogel compositions; thermo-mechanical properties and thermo-reversibility of the MC hydrogels were investigated by rheological analysis. Gelation temperatures and rheological behavior depended on the MC concentration and type and concentration of salt used in hydrogel preparation. In vitro cytotoxicity tests, performed using L929 mouse fibroblasts, showed no toxic release from all the tested hydrogels. Among the investigated compositions, the hydrogel composed of 8 % w/v MC with 0.05 M Na2SO4 had a thermo-reversibility temperature at 37 °C. For that reason, this formulation was thus considered to verify the possibility of inducing in vitro spontaneous detachment of cells previously seeded on the hydrogel surface. A continuous cell layer (cell sheet) was allowed to grow and then detached from the hydrogel surface without the use of enzymes, thanks to the thermo-responsive behavior of the MC hydrogel. Immunofluorescence observation confirmed that the detached cell sheet was composed of closely interacting cells.

Bioreactor mechanically guided 3D mesenchymal stem cell chondrogenesis using a biocompatible novel thermo-reversible methylcellulose-based hydrogel

“…These data are in accordance to previous literature showing the lack of MC toxicity20212223242526 and thus its suitability as dedicated material for tissue engineering purposes. Moreover, according to in vitro 28 and here presented in vivo results, it can be concluded that the use of Na 2 SO 4 salt in combination with MC does not lead to any inflammatory reaction.…”

“…2B and C). The physical gelation of MC hydrogels was visually observed using the inversion method already described in the literature2628. The measurements showed that the gelation temperature of the selected MC hydrogel occurs between 34 and 37 °C, allowing us to handle cell constructs for some minutes without losing stability as previously shown28.…”

“…The MC hydrogel in vitro cytocompatibility was previously reported28; therefore, in order to propose it as medical tool for cartilage regeneration, its in vivo biocompatibility was tested after 3 and 6 weeks implantation (as schematized in Fig. 3A).…”

Synthesis and investigation of a self-assembled hydrogel based on hydroxyethyl cellulose and itsin vitroibuprofen drug release characteristics

“…Herein, HEC, which is a kind of non-ionic cellulose ether with good water solubility and can be easily manufactured using natural cellulose as a raw material, is used as the hydrogel matrix, instead of synthetic polymers, taking advantage of cellulose resources, including its renewable nature, biocompatibility and biodegradability. [33][34][35] To our knowledge, no studies have been performed on the formation of self-assembled supramolecular hydrogels between hydrophobically modied HEC and b-CDP.…”

“…[41][42][43][44] First, 10 g of b-CD was dissolved in 15 mL of 15 wt% NaOH by stirring overnight at room temperature. Then the mixture was heated to 35 C and 7 mL of EPI was added in a 1 : 10 b-CD : EPI molar ratio. Aer 200 min, the mixture was poured into a dialysis bag (MWCO 8-12 kDa, Biosharp, USA) and dialyzed for 7 days.…”