Gelatin nanospheres incorporating siRNA for controlled intracellular release

The objective of this study is to develop a sustained release system of small interfering RNA (siRNA) inside cells aiming at a prolonged time period of gene suppression. Gelatin aqueous solution containing luciferase siRNA was coacelvated by acetone addition, followed by the glutaraldehyde (GA) crosslinking of gelatin to prepare gelatin nanospheres incorporating siRNA. The nanospheres were degraded with time in phosphate-buffered saline solution containing collagenase to release siRNA incorporated. The nanospheres were degraded more slowly as the GA concentration become higher, and consequently the rate of siRNA become lower. siRNA was released from the nanospheres as a result of nanospheres degradation. The nanospheres were internalized into colon 26 cells luciferase stably expressed, irrespective of the GA concentration. The gene expression was suppressed by the nanospheres incorporating siRNA capable for the longer-term release, and subsequently the time period of gene suppression was prolonged. The siRNA release inside the cell was observed, while the release period became longer for the slow-degraded nanospheres. It is possible that the intracellular siRNA release for a longer time period contributes to the prolonged time period of gene suppression.