The confined internal space of a liquid marble, as well as its porous and non-adhesive shell, offers an attractive application possibility - accommodating living cells inside liquid marbles. Cancer cells in suspension may aggregate to form three dimensional structures, also known as cancer cell spheroids (CCS). In this study, CCS formation inside liquid marble is investigated. This liquid marble application opens significant and novel avenues for biomedical applications and cancer research.
S1. Materials and Methods S1.1. Materials Carboxymethyl cellulose sodium salt (M w ~ 90 kDa), tyramine hydrochloride (TYR), N-(3dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), Tween 20, Triton-X100, deuterium oxide (D 2 O), dialysis tubing (MWCO 124000), and bovine serum albumin (BSA) were obtained from Sigma-Aldrich Pty. Ltd. (Castle Hill, NSW, Australia). Gelatin (M w = 80-140 kDa, pI = 5) and horseradish peroxidase (HRP; 100 units/mg) were obtained from Wako Pure Chemical Industries (Novachem Pty. Ltd., Collingwood, VIC, Australia). Fetal bovine serum (FBS), LIVE/DEAD Cell Viability Assays, calcein AM, 4',6-diamidino-2-phenylindole (DAPI), Dulbecco's Modified Eagle's Medium (DMEM), penicillin-streptomycin, trypsin-EDTA, and phosphate buffered saline (PBS) were acquired from Invitrogen (Life Technologies Pty. Ltd., Mulgrave, VIC, Australia). Hydrogen peroxide (H 2 O 2) was obtained from Merck Pty. Ltd.
Since rates of tissue growth vary significantly between tissue types, and also between individuals due to differences in age, dietary intake, and lifestyle-related factors, engineering a scaffold system that is appropriate for personalized tissue engineering remains a significant challenge. In this study, a gelatin-hydroxyphenylpropionic acid/carboxylmethylcellulose-tyramine (Gtn-HPA/CMC-Tyr) porous hydrogel system that allows the pore structure of scaffolds to be altered in vivo after implantation is developed. Cross-linking of Gtn-HPA/CMC-Tyr hydrogels via horseradish peroxidase oxidative coupling is examined both in vitro and in vivo. Post-implantation, further alteration of the hydrogel structure is achieved by injecting cellulase enzyme to digest the CMC component of the scaffold; this treatment yields a structure with larger pores and higher porosity than hydrogels without cellulase injection. Using this approach, the pore sizes of scaffolds are altered in vivo from 32-87 μm to 74-181 μm in a user-controled manner. The hydrogel is biocompatible to COS-7 cells and has mechanical properties similar to those of soft tissues. The new hydrogel system developed in this work provides clinicians with the ability to tailor the structure of scaffolds post-implantation depending on the growth rate of a tissue or an individual's recovery rate, and could thus be ideal for personalized tissue engineering.
Brain repair following disease and injury is very limited due to difficulties in recruiting and mobilizing stem cells towards the lesion. More importantly, there is a lack of structural and trophic support to maintain viability of the limited stem/progenitor cells present. This study investigates the effectiveness of an injectable gelatin-based hydrogel in attracting neural progenitor cells (NPCs) from the subventricular zone (SVZ) towards the implant. Glial cell-line-derived neurotrophic factor (GDNF) encapsulated within the hydrogel and porosity within the hydrogel prevents glial scar formation. By directly targeting the hydrogel implant towards the SVZ, neuroblasts can actively migrate towards and along the implant tract. Significantly more doublecortin (DCX)-positive neuroblasts surround implants at 7 d post-implantation (dpi) compared with lesion alone controls, an effect that is enhanced when GDNF is incorporated into the hydrogels. Neuroblasts are not observed at the implant boundary at 21 dpi, indicating that neuroblast migration has halted, and neuroblasts have either matured or have not survived. The development of an injectable gelatin-based hydrogel has significant implications for the treatment of some neurodegenerative diseases and brain injuries. The ability of GDNF and porosity to effectively prevent glial scar formation will allow better integration and interaction between the implant and surrounding neural tissue.
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