Titanium Phosphate Glass Microspheres as Microcarriers for In Vitro Bone Cell Tissue Engineering - Biointerfaces: Where Material Meets Biology - page 123

considered to contribute to glass degradation at the implant site. 76 Histology
studies have revealed that less soluble titanium glasses are more likely to be
fully surrounded by new bone tissue while more soluble Ti-free glasses show
some empty spaces.
These studies have been carried out using macroscopic glass specimens or
glass granules, but the culturing of cells on microspheres under in vitro
conditions presents unique challenges. We focus on the use of Transwell s
inserts to enable easier handling of microspheres inside the cell culture well
and also to provide improved distribution of medium and nutrients within
the system. These inserts are available in a variety of diameters, membrane
types and pore sizes which can fulfil different research requirements.

d n 3 r 4 n g | 3

5.4.1 Materials and Reagents
MG63 osteosarcoma cell line

CAUTION! All cell culture studies must be carried out with prior ethics
approval from the host institution.

Titanium phosphate glass microspheres
Dulbecco modified Eagle's medium (DMEM; Gibco, catalog no. 31885-049)
Ethanol (50%, 70%, 90% and 100%)
Fetal bovine serum (FBS; Gibco, catalog no. 10106-169)
Glutaraldehyde, 3% E.M. grade (Fisher Scientific, catalog no.

R3281800150)
Hexamethyldisilazane (HMDS; Agar Scientific, catalog no. AGR1228)
Penicillin-streptomycin, 5000 U mL 1 (Gibco, catalog no. 15070-063)
Phosphate-buffered saline (PBS; Lonza, catalog no. BE17-516F)
Trypan Blue solution, 0.4% (Sigma-Aldrich, catalog no. T8154)
Trypsin-EDTA solution, 1 (Gibco, catalog no. R-001-100)
Virkon virucidal surface disinfectant (Dupont, catalog no. 12358667)

.

5.4.2 Equipment
Aluminium foil
Autoclave tape
Carbon adhesive discs (Agar Scientific, UK)
Cell culture well plates (TPP; Helena Biosciences, UK)
Centrifuge tubes (TPP; Helena Biosciences, UK)
Disposable scalpels (VWR, UK)
Forceps
Glass vials, 2 mL (Sigma-Aldrich, UK)
Neubauer haemocytometer
Phase contrast microscope
Pipettes - 10, 200 and 1000 mL (Gilson, UK)
Pipette controller (Easypet; Eppendorf, UK)
Pipette tips - 10, 200 and 1000 mL (Star Labs, UK)