Release behavior of icariin-chitosan/hydroxyapatite scaffolds in vitro

doi:10.3969/j.issn.2095-4344.2014.52.007

Abstract

Abstract:

BACKGROUND: Icariin has stable nature, which can promote the proliferation of osteoblasts. As a result, it can be used as a new kind of bone induction activity factor for bone tissue engineering.

OBJECTIVE: To study the icariin release behavior of icariin-chitosan/hydroxyapatite scaffolds in vitro.

METHODS: Icariin-chitosan/hydroxyapatite scaffolds (10^-⁷, 10^-⁶, 10^-⁵ mol) were soaked in PBS at 37 °C and shaken gently. The samples collected at 1, 2, 3, 5, 10, 15, 20, 30, 60 and 90 days were analyzed by ultra performance liquid chromatography. The precision and sensitivity of ultra performance liquid chromatography were evaluated by precision test, repeatability test, icariin recovery test and stability test. Icariin released from scaffolds was calculated according to the standard curve and the percentage of icariin released was accumulated to draw a release curve which was fitted based on the Logarithmic model.

RESULTS AND CONCLUSION: The precision, accuracy and repeatability of the experimental method were

good, and icariin exhibited a good stability that could be saved darkly at room temperature at least for 90 days. There was a good linear relationship between icariin dose (1-2 000 ng) and chromatographic peak area. At day 1 to day 3, approximately 25% icariin was released; then the release speed was decreased and 40%-60% icariin was released by the end of 20 days. After 90 days, there was still a certain amount of icariin remained in the scaffolds. The release behavior of icariin-chitosan/hydroxyapatite followed the first-order equation, and there were significant difference among all the models (P = 0). These findings indicate that icariin-chitosan/hydroxyapatite scaffolds have a good control effects on icariin release, and the icariin release behavior can be fitted with the first-order equation.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

全文链接：

Key words: drugs, Chinese herbal, chitosan, hydroxyapatites

CLC Number:

R318

Wu Tao, Wu Jin-hui, Zheng Guo-dong, Lu Zhi-qin, Zeng Hai-yan, Lv Jun, Xing Jian-zhou. Release behavior of icariin-chitosan/hydroxyapatite scaffolds in vitro[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(52): 8399-8404.

Figures/Tables 3

2.1 线性范围与标准曲线在1-2 000 ng质量范围内，淫羊藿苷的色谱峰面积与进样量之间呈良好的线性关系，回归方程为：Y = 7 877.254X+ 202 422.5，R2 = 0.998 (图1)。 2.2 精密度实验和重复性实验结果经取同一标准品溶液重复进样测定后淫羊藿苷相对标准偏差为0.636%，表明本法精密度良好；重复性实验的相对标准偏差为3.245%，表明本法重复性良好(表1)。 2.3 淫羊藿苷加样回收率实验结果淫羊藿苷加样回收率试验结果见表2，平均回收率达96.667%，相对标准偏差为2.139%，说明该结果符合实验要求。 2.4 淫羊藿苷稳定性实验结果对保存1-90 d的淫羊藿苷进行了稳定性检测(图2)，结果表明，虽然1 000 ng淫羊藿苷在1-90 d内的实测值随时间有部分下降，但是并无统计学意义(F=2.209， P=0.079), ，这说明淫羊藿苷性质稳定，可在室温避光的环境下保存至少90 d。 2.5 淫羊藿苷-壳聚糖/羟基磷灰石复合材料的体外释药行为和分析对3种载药量的淫羊藿苷-壳聚糖/羟基磷灰石复合材料进行了体外释药行为研究。从UPLC谱图来看(图3)，供试品与对照品溶液均在相同保留时间内出现吸收峰，淫羊藿苷峰保留时间为1.30 min，壳聚糖/羟基磷灰石材料峰保留时间为0.56 min，在淫羊藿苷峰相应位置处无吸收峰(理论塔板数大于11 000)。通过UPLC图计算淫羊藿苷的峰面积积分值，根据标准曲线换算出每次释药量，再将结果按释药百分率累积得(图4)：在释药初期(0-3 d)，药物从支架材料中爆发性地释放出来，约达载药量的24%；而后释药速度迅速下降，至第20天有40%-65%的药物释出，之后以低速持续释放，90 d后仍有20%-35%的药物存留于支架材料中，这说明淫羊藿苷-壳聚糖/羟基磷灰石复合材料具有良好的控释效果。释药行为拟合曲线结果表明(图5)：3种剂量的淫羊藿苷-壳聚糖/羟基磷灰石复合材料的释药行为遵循一级方程模型，所有模型均有统计学意义(P=0)。"

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