Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (9): 853-862.doi: 10.3736/jcim20100909

• Original Experimental Research • Previous Articles     Next Articles

Polymeric nanoparticle encapsulation of a naturally occurring plant scopoletin and its effects on human melanoma cell A375

Khuda-Bukhsh ARa,Bhattacharyya SSa,Paul Sa,Boujedaini Nb   

  1. a Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India
    b Boiron Laboratory, 20, rue de la Liberation, Sainte-Les-Foy-Lyon, France
  • Received:2010-04-28 Accepted:2010-07-05 Online:2010-09-20 Published:2010-09-15


We formulated nano-encapsulation of a naturally occurring coumarin — scopoletin (7-hydroxy-6-methoxy coumarin, HMC, C10H8O4), isolated from plant Gelsemium sempervirens having anticancer potentials, with a bio-adhesive agent — polylactic-co-glycolic acid (PLGA) and tested if its cellular uptake, bioavailability and apoptotic (anticancer) potentials could thus be increased vis-vis unencapsulated HMC.

A375 melanoma cancer cells were used for testing cellular entry and anticancer potentials of HMC and nano-7-hydroxy-6-methoxy coumarin (NHMC) through several standard protocols. Characterization of NHMC was done by dynamic light scattering for determination of particle size, polydispersity index (PDI), and zeta potential. Surface morphology of nanoparticles was determined by scanning electron microscopy and atomic force microscopy.

HMC was encapsulated with more than 85% entrapment efficiency, the average particle size of NHMC being less than 110 nm and a PDI 0.237, which resulted in enhanced cellular entry and greater bioavailability. NHMC showed a faster cellular uptake (15 min) than its unencapsulated counterpart (30 min). Study of signal molecules through mRNA expressions revealed that NHMC caused down-regulation of cyclin-D1, proliferating cell nuclear antigen (PCNA), survivin and Stat-3, and up-regulation of p53 and caspase-3, that in turn induced a greater number of apoptosis vis-vis unencapsulated HMC.

The formulation yielded small-sized NHMC by biodegradable PLGA that took less time for cellular entry, and caused more apoptosis to cancer cells, but apparently had negligible cytotoxicity against normal skin cells. Nano-encapsulation of bioactive plant ingredients can be a strategy worth trying for designing effective chemopreventive drug products.

Key words: Polylactic-co-glycolic acid, Nanoparticles, Scopoletin, Melanoma, Apoptosis, Antineoplastic agents, Phytogenic, In vitro

Figure 1

Chemical structure of 7-hydroxy-6-methoxy coumarin (HMC)"


Figure 2

Surface images of NHMC and interaction of NHMC with A375 cells (A) SEM photograph of nanoparticles of HMC prepared from PLGA. (B) Surface topography of NHMC obtained by AFM. (C) Corresponding 3-D image of NHMC obtained by tapping mode AFM and visualized in amplitude mode."


Figure 3

(A) Average particle size obtained from DLS data for PLGA-encapsulated NHMC. (B) Zeta potential of NHMC"

Figure 4

In vitro release kinetics of NHMC"



Figure 5

Cell viability of blank nanoparticles, HMC and NHMC with different doses against A375 cell line tested by MTT assay. **P<0.01, vs HMC group. All results were expressed with vehicle control as 100%."

Figure 6

Cellular uptakes of HMC and NHMC in A375 cells"



Figure 7

Effects of HMC and NHMC on A375 cells Ln1 contains a standard DNA ladder; Ln2 contains DNA from untreated cells; Ln3 contains DNA from HMC (30 μmol/L); Ln4 contains DNA from HMC (60 μmol/L); Ln5 from NHMC (30 μmol/L) and Ln6 from NHMC (60 μmol/L) -treated cells."

Figure 8

Expressions of β-actin, cyclin-D1, survivin, Stat-3, PCNA, caspase-3, and p53 analyzed by RT-PCR Ln1: Control; Ln2: HMC (30 μmol/L); Ln3: HMC (60 μmol/L); Ln4: NHMC (30 μmol/L); Ln5: NHMC (60 μmol/L)."


[1] Newman JDS, Blanchard GJ . Formation and encapsulation of gold nanoparticles using a polymeric amine reducing agent[J]. J Nanopart Res, 2007,9(5):861-868
doi: 10.1007/s11051-006-9145-y
[2] Bisht S, Feldmann G, Soni S, Ravi R, Karikar C, Maitra A, Maitra A .Polymeric nanoparticle-encapsulated curcumin(“nanocurcumin”): a novel strategy for human cancer therapy[J].J Nanobiotechnology, 2007, 5: 3
doi: 10.1186/1477-3155-5-3
[3] Anand P, Nair HB, Sung B, Kunnumakkara AB, Yadav VR, Tekmal RR, Aggarwal BB . Design of curcumin-loaded PLGA nanoparticles formulation with enhanced cellular uptake, and increased bioactivity in vitro and superior bioavailability in vivo[J]. Biochem Pharmacol, 2010,79(3):330-338
doi: 10.1016/j.bcp.2009.09.003
[4] Allemann E, Gurny R, Doelker E . Drug loaded nano-particles preparation methods and drug targeting tissues[J]. Eur J Pharm Biopharm, 1993,39(5):173-191
[5] Bhattacharyya SS, Mandal SK, Biswas R, Paul S, Pathak S, Boujedaini N, Belon P, Khuda-Bukhsh AR . In vitro studies demonstrate anticancer activity of an alkaloid of the plant Gelsemium sempervirens[J]. Exp Biol Med, 2008,233(12):1591-1601
doi: 10.3181/0805-RM-181
[6] Bhattacharyya SS, Paul S, Dutta S, Boujaedaini N, Khuda-Bukhsh AR . Anti-oncogenic potentials of a plant coumarin(7-hydroxy-6-methoxy coumarin) against 7,12-dimethylbenz[a]anthracene-induced skin papilloma in mice: the possible role of several key signal proteins[J]. J Chin Integr Med, 2010,8(7):645-654
Bhattacharyya SS, Paul S, Dutta S, Boujaedaini N, Khuda-Bukhsh AR. 东莨菪亭(7-羟基-6-甲氧基香豆素)对7,12-二甲基苯并蒽诱导的皮肤乳头状瘤小鼠相关关键信号蛋白的影响[J]. 中西医结合学报, 2010,8(7):645-654
[7] Bhattacharyya SS, Paul S, Khuda-Bukhsh AR . Encapsulated plant extract(Gelsemium sempervirens) poly(lactide-co-glycolide) nanoparticles enhance cellular uptake and increase bioactivity in vitro[J]. Exp Biol Med, 2010,235(6):678-688
doi: 10.1258/ebm.2010.009338
[8] Panyam J, Dali MM, Sahoo SK, Ma W, Chakravarthi SS, Amidon GL, Levy RJ, Labhasetwar V.Labhasetwar V . Polymer degradation and in vitro release of a model protein from poly(D,L-lactide-co-glycolide) nano- and microparticles[J]. J Control Release, 2003,92(1-2):173-187
doi: 10.1016/S0168-3659(03)00328-6
[9] Fessi H, Puisieux F, Devissaguet JP, Ammoury N, Benita S . Nanocapsule formation by interfacial polymer deposition following solvent displacement[J]. Int J Pharm, 1989,55(1):1-4
doi: 10.1016/0378-5173(89)90270-6
[10] Mosmann T . Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays[J]. J Immunol Methods, 1983,65(1-2):55-63
doi: 10.1016/0022-1759(83)90303-4
[11] Darzynkiewicz Z, Galkowski D, Zhao H . Analysis of apoptosis by cytometry using TUNEL assay[J]. Methods, 2008,44(3):250-254
doi: 10.1016/j.ymeth.2007.11.008 pmid: 2295206
[12] Kawaii S, Tomono Y, Ogawa K, Sugiura M, Yano M, Yoshizawa Y . The antiproliferative effect of coumarins on several cancer cell lines[J]. Anticancer Res, 2001,21(2A):917-923
[13] Ankola DD, Viswanad B, Bhardwaj V, Ramarao P, Kumar MN . Development of potent oral nanoparticulate formulation of coenzyme Q10 for treatment of hypertension: can the simple nutritional supplements be used as first line therapeutic agents for prophylaxis/therapy?[J]. Eur J Pharm Biopharm, 2007,67(2):361-369
doi: 10.1016/j.ejpb.2007.03.010
[14] Kim TY, Kim DW, Chung JY, Shin SG, Kim SC, Heo DS, Kim NK, Bang YJ . Phase I and pharmacokinetic study of Genexol-PM, a cremophor-free, polymeric micelle-formulated paclitaxel, in patients with advanced malignancies[J]. Clin Cancer Res, 2004,10(11):3708-3716
doi: 10.1158/1078-0432.CCR-03-0655
[15] Vasey PA, Kaye SB, Morrison R, Twelves C, Wilson P, Duncan R, Thomson AH, Murray LS, Hilditch TE, Murray T, Burtles S, Fraier D, Frigerio E, Cassidy J . Phase I clinical and pharmacokinetic study of PK1 [N-(2-hydroxypropyl)methacrylamide copolymer doxorubicin]: first member of a new class of chemotherapeutic agents-drug-polymer conjugates[J]. Clin Cancer Res, 1999,5(1):83-94
[1] Cai-lian Fana, Wan-jun Caib, Meng-nan Ye, Miao Chen, Yi Dai. Qili Qiangxin, a compound herbal medicine formula, alleviates hypoxia-reoxygenation-induced apoptotic and autophagic cell death via suppression of ROS/AMPK/mTOR pathway in vitro. Journal of Integrative Medicine, 2022, 20(4): 365-375.
[2] Hong-xiao Li, Ling Shi, Shang-jie Liang Chen-chen Fang, Qian-qian Xu, Ge Lu, Qian Wang, Jie Cheng, Jie Shen, Mei-hong Shen. Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway. Journal of Integrative Medicine, 2022, 20(2): 163-172.
[3] Mohamed S. Othman, Sofian T. Obeidat, Amal H. Al-Bagawi, Mohamed A. Fareid, Alaa Fehaid, Ahmed E. Abdel Moneim. Green-synthetized selenium nanoparticles using berberine as a promising anticancer agent#br#
. Journal of Integrative Medicine, 2022, 20(1): 65-72.
[4] Hyongjun Jeon, Hee-Young Kim, Chang-Hwan Bae, Yukyung Lee, Sungtae Koo, Seungtae Kim. Korean red ginseng decreases 1-methyl-4-phenylpyridinium-induced mitophagy in SH-SY5Y cells. Journal of Integrative Medicine, 2021, 19(6): 537-544.
[5] Porwornwisit Tritripmongkol, Tullayakorn Plengsuriyakarn, Mayuri Tarasuk, Kesara Na-Bangchang. In vitro cytotoxic and toxicological activities of ethanolic extract of Kaempferia galanga Linn. and its active component, ethyl-p-methoxycinnamate, against cholangiocarcinoma. Journal of Integrative Medicine, 2020, 18(4): 326-333.
[6] Varuni Colamba Pathiranage, Ira Thabrew, Sameera R. Samarakoon, Kamani H. Tennekoon, Umapriyatharshini Rajagopalan, Meran K. Ediriweera. Evaluation of anticancer effects of a pharmaceutically viable extract of a traditional polyherbal mixture against non-small-cell lung cancer cells. Journal of Integrative Medicine, 2020, 18(3): 242-252.
[7] Maged Mohamed Maher Abou-Hashem, Dina Mohamed Abo-elmatty, Noha Mostafa Mesbah, Ahmed Mohamed Abd EL-Mawgoud. Induction of sub-G0 arrest and apoptosis by seed extract of Moringa peregrina (Forssk.) Fiori in cervical and prostate cancer cell lines. Journal of Integrative Medicine, 2019, 17(6): 410-422.
[8] Supakanya Kumkarnjana, Rutt Suttisri, Ubonthip Nimmannit, Apirada Sucontphunt, Mattaka Khongkow, Thongchai Koobkokkruad, Nontima Vardhanabhuti. Flavonoids kaempferide and 4,2′-dihydroxy-4′,5′,6′-trimethoxychalcone inhibit mitotic clonal expansion and induce apoptosis during the early phase of adipogenesis in 3T3-L1 cells. Journal of Integrative Medicine, 2019, 17(4): 288-295.
[9] Shriniwas S. Basaiyye, Sanjay Kashyap, Kannan Krishnamurthi, Saravanadevi Sivanesan. Induction of apoptosis in leukemic cells by the alkaloid extract of garden cress (Lepidium sativum L.). Journal of Integrative Medicine, 2019, 17(3): 221-228.
[10] Carlos R. Oliveira, Daniel G. Spindola, Daniel M. Garcia, Adolfo Erustes, Alexandre Bechara, Caroline Palmeira-dos-Santos, Soraya S. Smaili, Gustavo J.S. Pereira, André Hinsberger, Ezequiel P. Viriato, Maria Cristina Marcucci, Alexandra C.H.F. Sawaya, Samantha L. Tomaz, Elaine G. Rodrigues, Claudia Bincoletto. Medicinal properties of Angelica archangelica root extract: Cytotoxicity in breast cancer cells and its protective effects against in vivo tumor development. Journal of Integrative Medicine, 2019, 17(2): 132-140.
[11] Bruno José Martins Da Silva, Sandro Wilson Gomes Pereira, Ana Paula Drummond Rodrigues, José Luiz Martins Do Nascimento, Edilene Oliveira Silva. In vitro antileishmanial effects of Physalis angulata root extract on Leishmania infantum. Journal of Integrative Medicine, 2018, 16(6): 404-410.
[12] Eric Wei Chiang Chan, Siu Kuin Wong, Hung Tuck Chan . Casticin from Vitex species: A short review on its anticancer and anti-inflammatory properties. Journal of Integrative Medicine, 2018, 16(3): 147-152.
[13] Jesmin Mondal, Jayeeta Das, Rajesh Shah, Anisur Rahman Khuda-Bukhsh. A homeopathic nosode, Hepatitis C 30 demonstrates anticancer effect against liver cancer cells in vitro by modulating telomerase and topoisomerase II activities as also by promoting apoptosis via intrinsic mitochondrial pathway. Journal of Integrative Medicine, 2016, 14(3): 209-218.
[14] Jesmin Mondal, Asmita Samadder, Anisur Rahman Khuda-Bukhsh. Psorinum 6× triggers apoptosis signals in human lung cancer cells. Journal of Integrative Medicine, 2016, 14(2): 143-153.
[15] Qing-qing Zang, Lu Zhang, Ning Gao, Cheng Huang. Ophiopogonin D inhibits cell proliferation, causes cell cycle arrest at G2/M, and induces apoptosis in human breast carcinoma MCF-7 cells. Journal of Integrative Medicine, 2016, 14(1): 51-59.
Full text



[1] Wei-xiong Liang. Problems-solving strategies in clinical treatment guideline for traditional Chinese medicine and integrative medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 1-4
[2] Zhao-guo Li. Discussion on English translation of commonly used sentences in traditional Chinese medicine: part one. Journal of Chinese Integrative Medicine, 2008, 6(1): 107-110
[3] Jun Hu, Jian-ping Liu. Non-invasive physical treatments for chronic/recurrent headache. Journal of Chinese Integrative Medicine, 2008, 6(1): 31
[4] Xue-mei Liu, Qi-fu Huang, Yun-ling Zhang, Jin-li Lou, Hong-sheng Liu, Hong Zheng. Effects of Tribulus terrestris L. saponion on apoptosis of cortical neurons induced by hypoxia-reoxygenation in rats. Journal of Chinese Integrative Medicine, 2008, 6(1): 45-50
[5] . Uniform requirements for manuscripts submitted to biomedical journals: Writing and editing for biomedical publication (Chinese version, part two). Journal of Chinese Integrative Medicine, 2010, 8(11): 1001-1005
[6] Daniel Weber, Janelle M Wheat, Geoffrey M Currie. Inflammation and cancer: Tumor initiation, progression and metastasis,and Chinese botanical medicines. Journal of Chinese Integrative Medicine, 2010, 8(11): 1006-1013
[7] Hong Liu , Guo-liang Zhang, Li Shen , Zhen Zeng, Bao-luo Zhou, Cheng-hai Liu, Guang Nie . Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5Nl avian influenza virus. Journal of Chinese Integrative Medicine, 2010, 8(11): 1036-1040
[8] Zhao-guo Li . A discussion of English translation of 1995 and 1997 Chinese National Standards of Traditional Chinese Medical Terminologies for Clinical Diagnosis and Treatment. Journal of Chinese Integrative Medicine, 2010, 8(11): 1090-1096
[9] Rui Jin, Bing Zhang. A complexity analysis of Chinese herbal property theory: the multiple formations of herbal property (Part 1). Journal of Chinese Integrative Medicine, 2012, 10(11): 1198-1205
[10] Hui-min Liu, Xian-bo Wang, Yu-juan Chang, Li-li Gu. Systematic review and meta-analysis of randomized controlled trials of integrative medicine therapy for treatment of chronic severe hepatitis. Journal of Chinese Integrative Medicine, 2012, 10(11): 1211-1228