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Thermoresponsive Hyperbranched Copolymer with Multi Acrylate
Functionality for In Situ Cross-linkable Hyaluronic Acid Composite
Semi-IPN Hydrogel
Yixiao Dong1, Waqar Hassan1, Yu Zheng1, Aram Omer Saeed1, Hongliang Cao1, Hongyun Tai2*, Abhay
Pandit1 and Wenxin Wang1*
Network of Excellence for Functional Biomaterials, National University of Ireland, Galway, Ireland.
School of Chemistry, Bangor University, Bangor, LL57 2UW, UK. E-mail:
Cytotoxicity Study by AlamarBlue® Assay
For cellular viability studies, 3T3 mouse fibroblast cells (Passage 12) were utilized to study polymer
cytotoxicity. 15,000 cells in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma, 10% FBS, 1%
P/S,) were previously seeded into each well of a 48-wells tissue culture plate. After 24 h of incubation at
37 oC and 5% CO2, all the cells were attached to the bottom of culture plate and showed stable
morphology under the microscope. Then PEGMEMA-MEO2MA-PEGDA copolymer and QT solutions
(0.5, 1, 5 and 10 mg mL-1, dissolved in culture medium) were added into each well of the culture plate.
After 48 h of incubation, the alamarBlue® reduction method was used to assess cellular metabolic
activity. The absorbance at the lower wavelength filter (550 nm) was measured followed by the higher
wavelength filter (595 nm) via a thermo scientific Varioskan® Flash Plate Reader. The results were
calculated and normalized to the positive control (cells without treatment).
Figure S1. Cellular metabolism viability assessment of 3T3 cells after 48 h treatment with PEGMEMAMEO2MA-PEGDA (entry 6 in Table 2) and QT using alamarBlue® assay. There is no significant difference
between copolymer and QT treatment with control of cell alone (n=3, P < 0.05).