Limapornvanich Araya, Jitpukdeebodintra Suwanna, Hengtrakool Chanothai, Kedjarune-Leggat Ureporn
Dental Unit, Yala Regional Hospital, Yala 95000, Thailand.
J Dent. 2009 Sep;37(9):686-90. doi: 10.1016/j.jdent.2009.05.007. Epub 2009 May 14.
This study aimed to evaluate the effect of adding chitosan (CS) to conventional glass ionomer cement (GIC) on protein release and its cytotoxicity.
Bovine serum albumin (BSA) was used as the released protein from two glass ionomer formulations. One (GIC+BSA) contained fluoro-aluminosilicate glass mixed with BSA, and another (GIC:CS+BSA) used a similar glass and BSA with 20% chitosan. Six disc specimens per group (10mm in diameter, 2mm in height) were prepared and placed in phosphate buffer saline, which was replaced at various times over 2 weeks. The released protein was determined by a BCA assay. Cytotoxicity of the extracts from these materials for 1, 2 and 7 days to dental pulp cells was evaluated using MTT assay.
The GIC:CS+BSA released a burst of BSA in the first 6h, and slowly released at different rates over the 2 weeks. GIC+BSA showed a similar result, but protein could not be detected at the 12h. The protein release rate of GIC:CS+BSA was significantly greater than GIC+BSA (P<0.01); nearly three times higher. The released BSA had the same molecular weight as evaluated by SDS-PAGE. From the MTT assay, the percentages of viable cells were significantly different and can be arranged as: GIC:CS+BSA>GIC:CS>GI+BSA>GI and the cytotoxicity was increased by time of extraction.
Chitosan added in glass ionomer cement can prolong release of BSA as well as not increasing the toxicity to pulp cells. This material may be useful for protein delivery.
本研究旨在评估在传统玻璃离子水门汀(GIC)中添加壳聚糖(CS)对蛋白质释放及其细胞毒性的影响。
使用牛血清白蛋白(BSA)作为两种玻璃离子配方中释放的蛋白质。一种(GIC + BSA)包含与BSA混合的氟铝硅酸盐玻璃,另一种(GIC:CS + BSA)使用类似的玻璃和含20%壳聚糖的BSA。每组制备六个圆盘标本(直径10mm,高2mm),并置于磷酸盐缓冲盐水中,在2周内不同时间更换。通过BCA测定法测定释放的蛋白质。使用MTT测定法评估这些材料的提取物在1、2和7天对牙髓细胞的细胞毒性。
GIC:CS + BSA在最初6小时内爆发性释放BSA,并在2周内以不同速率缓慢释放。GIC + BSA显示出类似的结果,但在12小时时未检测到蛋白质。GIC:CS + BSA的蛋白质释放速率明显大于GIC + BSA(P < 0.01);几乎高出三倍。通过SDS-PAGE评估,释放的BSA具有相同的分子量。从MTT测定法来看,活细胞百分比有显著差异,可排列为:GIC:CS + BSA > GIC:CS > GI + BSA > GI,并且细胞毒性随提取时间增加。
在玻璃离子水门汀中添加壳聚糖可以延长BSA的释放,同时不增加对牙髓细胞的毒性。这种材料可能对蛋白质递送有用。