胶原蛋白神经酰胺脂质体的制备及质量评价

doi:10.3976/j.issn.1002-4026.20230144

摘要/Abstract

摘要：

以蛋黄卵磷脂、胆固醇和神经酰胺为成膜材料,采用薄膜分散法制备胶原蛋白神经酰胺脂质体(CO-CS)。以包封率为响应值,以药脂比、膜材比、超声时间和水合时间为影响因素对工艺进行优化。结果表明,当药脂比为1:16、超声时间为8.5 min、膜材比为4.3:1时,包封率可达90.73%。在此条件下,样品的粒径为(206.63±2.06) nm,聚合物分散性指数(polymer dispersity index,PDI)为0.187±0.010,Zeta电位为(-34.90±0.82) mV。避光保存60 d后,CO-CS的包封率为87.2%,粒径为(223.70±1.85) nm、PDI为0.174±0.013、Zeta电位为(-33.51±2.10) mV。结果表明,本研究制备的CO-CS包封率高,稳定性良好,制备工艺合理可行。

关键词: 胶原蛋白, 神经酰胺脂质体, 响应面, 包封率

Abstract:

In this study, egg yolk lecithin, cholesterol,and ceramide were used as film-forming materials to prepare collagen ceramide liposomes (CO-CS) by the thin film dispersion method. To optimize the process, encapsulation efficiency was used as the response value, and it was evaluated by influencing factors such as drug-lipid ratio, film-material ratio, ultrasonic time, and hydration time. Results showed that encapsulation efficiency reached 90.73% when the drug-lipid ratio, film-material ratio, and ultrasonic time were 1:16, 4.3:1, and 8.5 min, respectively. Under these conditions, the particle size, polymer dispersity index (PDI), and zeta potential of CO-CS were(206.63±2.06) nm, 0.187±0.010, and (34.90±0.82) mV, respectively. After 60 days of storage at room temperature in the dark, the encapsulation efficiency was 87.2 %. Furthermore, the particle size, PDI, and zeta potential of CO-CS changed to (223.70±1.85) nm, 0.174±0.013, and (33.51±2.10) mV, respectively. In summary, CO-CS produced in this study revealed high encapsulation efficiency and good stability, and the preparation process is reasonable and feasible.

Key words: collagen, ceramide liposome, response surface, encapsulation efficiency

中图分类号:

TQ658.2

唐春慧, 张振博, 吴冬琴, 环飞, 张静姝, 王玉邦. 胶原蛋白神经酰胺脂质体的制备及质量评价[J]. 山东科学, 2024, 37(3): 18-26.

TANG Chunhui, ZHANG Zhenbo, WU Dongqin, HUAN Fei, ZHANG Jingshu, WANG Yubang. Preparation and quality evaluation of collagen ceramide liposomes[J]. Shandong Science, 2024, 37(3): 18-26.

图/表 11

表1

表2

图1

表3

表4

回归模型的方差分析及其对包封率的影响"

差异来源	平方和	自由度	均方	F值	P值	显著性
模型	780.18	9	0.314 65	91.60	< 0.000 1	**
A	42.64	1	0.281 25	45.06	0.000 3	**
B	11.14	1	0.061 25	11.77	0.011 0	*
C	142.38	1	0.005	150.45	< 0.000 1	**
AB	0.10	1	0.16	0.11	0.751 8	不显著
AC	1.29	1	0.062 5	1.36	0.281 5	不显著
BC	16.48	1	0.002 5	17.42	0.004 2	**
A²	197.81	1	0.725 158	209.03	< 0.000 1	**
B²	163.25	1	0.114 632	172.50	< 0.000 1	**
C²	145.79	1	1.227 789	154.05	< 0.000 1	**
残差	6.62	7	0.030 071
失拟项	4.31	3	0.067 5	2.49	0.199 5	不显著
纯误差	2.31	4	0.002
总和	786.80	16
R²					0.991 6
$R a d j 2$					0.980 8
$R p r e d 2$					0.907 7

表4

图2

表5

图3

图4

图5

表6

参考文献 18

[1]	GU L S, SHAN T T, MA Y X, et al. Novel biomedical applications of crosslinked collagen[J]. Trends in Biotechnology, 2019, 37(5): 464-491. DOI: 10.1016/j.tibtech.2018.10.007. pmid: 30447877
[2]	AL-NIMRY S, DAYAH A A, HASAN I, et al. Cosmetic, biomedical and pharmaceutical applications of fish gelatin/hydrolysates[J]. Marine Drugs, 2021, 19(3): 145. DOI: 10.3390/md19030145.
[3]	张燕. 鹿皮胶原蛋白多肽脂质体水凝胶的制备及其在小鼠光老化中的保护作用[D]. 长春: 吉林大学, 2022.
[4]	KOVÁČIK A, KOPEČNÁ M, VÁVROVÁ K. Permeation enhancers in transdermal drug delivery: benefits and limitations[J]. Expert Opinion on Drug Delivery, 2020, 17(2): 145-155. DOI: 10.1080/17425247.2020.1713087.
[5]	孙娅楠, 赵静, 李超华, 等. 二次谐波结合双光子荧光成像方法观察人源胶原蛋白透皮吸收情况[J]. 激光生物学报, 2017, 26(1): 24.
[6]	SUN Y N, LI L S, MA S H, et al. In vivo visualization of collagen transdermal absorption by second-harmonic generation and two-photon excited fluorescence microscopy[J]. Frontiersin Chemistry, 2022, 10: 925931. DOI: 10.3389/fchem.2022.925931.
[7]	ALBASH R, YOUSRY C, AL-MAHALLAWI A M, et al. Utilization of PEGylated cerosomes for effective topical delivery of fenticonazole nitrate: in-vitro characterization, statistical optimization, and in-vivo assessment[J]. Drug Delivery, 2021, 28(1): 1-9. DOI: 10.1080/10717544.2020.1859000.
[8]	王玉真, 高爽, 孙越, 等. 菊苣酸脂质体制备工艺研究[J]. 山东科学, 2020, 33(1): 8-15. DOI: 10.3976/j.issn.1002-4026.2020.01.002.
[9]	马永强, 王姿淇, 黎晨晨, 等. 番茄红素肝靶向脂质体制备工艺研究及肝靶向性评价[J]. 中国食品添加剂, 2023, 34(4): 106-115. DOI: 10.19804/j.issn1006-2513.2023.04.013.
[10]	苏林豫, 朱效素, 张蓬, 等. 新型蛋白降解靶向嵌合体LC-2脂质体的制备和表征[J]. 中国医药工业杂志, 2022, 53(5): 679-687. DOI: 10.16522/j.cnki.cjph.2022.05.013.
[11]	龚频, 方文静, 赵文婧, 等. 灵芝多糖脂质体制备工艺[J]. 食用菌学报, 2022, 29(5): 101-108. DOI: 10.16488/j.cnki.1005-9873.2022.05.011.
[12]	CHEN W T, WU H T, CHANG I C, et al. Preparation of curcumin-loaded liposome with high bioavailability by a novel method of high pressure processing[J]. Chemistry and Physics of Lipids, 2022, 244: 105191. DOI: 10.1016/j.chemphyslip.2022.105191.
[13]	张婷, 温鹤迪, 宋敬一, 等. 基于乙醇注入-高压均质的蛋清肽脂质体制备及体内外缓释效果[J]. 食品科学, 2021, 42(13): 79-86. DOI: 10.7506/spkx1002-6630-20200624-331.
[14]	RAMLAN N, ZUBAIRI S I, MASKAT M Y. Response surface optimisation of polydimethylsiloxane (PDMS) on borosilicate glass and stainless steel (SS316) to increase hydrophobicity[J]. Molecules, 2022, 27(11): 3388. DOI: 10.3390/molecules27113388.
[15]	马永强, 隋佳琦, 黎晨晨, 等. 响应面法优化高顺式虾青素的制备工艺研究[J]. 食品与发酵工业, 2023: 1-15.
[16]	国家药典委员会. 中华人民共和国药典四部2020年版[M]. 北京: 中国医药科技出版社, 2020.
[17]	GULDIKEN B, GIBIS M, BOYACIOGLU D, et al. Physical and chemical stability of anthocyanin-rich black carrot extract-loaded liposomes during storage[J]. Food Research International, 2018, 108: 491-497. DOI: 10.1016/j.foodres.2018.03.071. pmid: 29735084
[18]	ZHAO L S, TEMELLI F. Preparation of anthocyanin-loaded liposomes using an improved supercritical carbon dioxide method[J]. Innovative Food Science & Emerging Technologies, 2017, 39: 119-128. DOI: 10.1016/j.ifset.2016.11.013.

处方组成	用量	处方组成	用量
胶原蛋白原液	1.3 mL	神经酰胺	0.02 g
蛋黄卵磷脂	0.1 g	无水乙醇	5 mL
胆固醇	0.02 g	1%吐温-80	0.5 mL

因素	水平
因素	-1	0	1
药脂比 A	1:20	1:16	1:12
超声时间B/min	5	10	15
膜材比 C	3:1	5:1	7:1

实验号	药脂比A	超声时间B/min	膜材比C	包封率Y/ %	实验号	药脂比A	超声时间B/min	膜材比C	包封率Y/ %
1	1:12	5	5:1	75.55	10	1:16	15	7:1	73.95
2	1:20	5	5:1	80.39	11	1:16	5	3:1	84.46
3	1:12	15	5:1	72.34	12	1:16	15	3:1	79.21
4	1:20	15	5:1	76.54	13	1:16	10	5:1	90.15
5	1:12	10	7:1	70.98	14	1:16	10	5:1	89.45
6	1:20	10	7:1	74.56	15	1:16	10	5:1	89.66
7	1:12	10	3:1	77.40	16	1:16	10	5:1	89.04
8	1:20	10	3:1	83.25	17	1:16	10	5:1	88.13
9	1:16	5	7:1	71.08

实验次数	粒径/nm	PDI	电位/mV	包封率/%
1	206.63	0.185	-34.1	88.54
2	207.33	0.172	-35.5	92.82
3	203.87	0.185	-33.9	91.17
4	205.80	0.195	-34.7	88.57
5	209.50	0.196	-35.7	92.53
均数±标准差	206.63±2.06	0.187±0.010	-34.9±0.8	90.73±2.08

时间/d	外观性状	包封率/%	粒径/nm	PDI	Zeta 电位/mV
7	白色混悬液,流动性良好	90.83±0.40	210.80±0.51	0.172±0.006	-34.50±0.38
14	白色混悬液,流动性良好	90.32±2.71	213.02±3.63	0.180±0.028	-35.44±0.64
21	白色混悬液,流动性良好	89.64±1.01	218.61±1.69	0.169±0.009	-34.60±1.14
28	白色混悬液,流动性良好	89.08±0.99	220.14±1.52	0.171±0.012	-34.72±1.59
60	白色混悬液,流动性良好	87.20±1.09	223.70±1.85	0.174±0.013	-33.51±2.10