Porous aerogel scaffold, method of preparing the same and use thereof
A method for prepring a porous aerogel scaffold includes: adding a photoinitiator and polyethylene glycol diacrylate in a buffer solution, dissolving by heating and evenly mixing, adding Pluronic® F127 (Poloxamer 407) into the mixed solution, and standing at a low temperature to obtain an aerogel scaffold material; printing a hydrogel scaffold by using a 3D printing technology, and performing UV irradiation so that a cross-linking of the hydrogel scaffold is caused to form a three-dimensional scaffold with a stable structure, performing low-temperature soaking to remove Pluronic® F127 (Poloxamer 407), and then freeze drying the three-dimensional scaffold to obtain the porous aerogel scaffold. Wherein, Pluronic® F127 (Poloxamer 407) serves as a sacrificial material which is removed after the 3D printing of the hydrogel scaffold is completed, and then a porous structure can be formed in the scaffold in combination with a freeze drying technology, which facilitates the survival, growth and proliferation of cells during the three-dimensional culture.
1 . A method of three-dimensional (3D) cell culture comprising the following steps:
adding a photoinitiator and polyethylene glycol diacrylate in a buffer solution, heating to dissolve and mix evenly, then adding Poloxamer 407 into the mixed solution, and leaving to stand at low temperature to obtain an aerogel scaffold material; and printing a hydrogel scaffold by using a 3D printing technology, and performing an ultraviolet (UV) irradiation on the printed hydrogel scaffold to generate a cross-linkage, then forming a structurally stable 3D scaffold, performing a low-temperature soaking to remove the Poloxamer 407, performing a freeze drying to obtain the porous aerogel scaffold;
wherein, the photoinitiator is a lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) photoinitiator; in the buffer solution, a concentration of the LAP photoinitiator is 0.1-0.25 wt %, a concentration of polyethylene glycol diacrylate is 10-15 wt %, and a concentration of the Poloxamer 407 is 15-25 wt %;
performing the low-temperature soaking of the 3D scaffold by using phosphate buffered saline (PBS), to remove the Poloxamer 407; wherein a temperature of the low-temperature soaking is 0-5° C., a duration time of the low-temperature soaking is 20-30 h; and a temperature of the freeze drying is −60-−50° C., and a duration time of the freeze drying is 20-30 h;
performing a soaking sterilization on the porous aerogel scaffold by using an ethanol solution, soaking the sterilized porous aerogel scaffold in a complete culture medium, incubating at 37° C., 5% CO 2 for 20-30 h, carrying out a functionalization of the porous aerogel scaffold; and, adding a cell suspension solution to the functionalized porous aerogel scaffold and culturing in a cell incubator at 37° C. and 5% CO 2 .
2 . The method according to claim 1 , wherein: the buffer solution is the phosphate buffered saline (PBS).
3 . The method according to claim 1 , wherein: a temperature of the low-temperature standing of the aerogel scaffold material is 0-5° C., and a duration time of the low-temperature standing is 2-5 days.
4 . The method according to claim 1 , wherein: a wavelength of the UV light is 405 nm; an intensity of the UV light is 20-30 mW/cm 2 , and an irradiation time is 5-15 min.
5 . The method according to claim 1 , wherein: the temperature of the low-temperature soaking is 4° C., and the duration time of the low-temperature soaking is 24 h.