Skip to main content

Green synthesis of nanohydroxyapatite trough Elaeagnus angustifolia L. extract and evaluating its anti-tumor properties in MCF7 breast cancer cell line



One of the most common types of cancer in women is breast cancer. There are numerous natural plant-based products, which exert anti-tumoral effects including Elaeagnus Angustifolia (EA). It modulates cell-cycle process, heat-shock proteins expression, anti-proliferative properties, apoptosis induction, blocking of angiogenesis, and cell invasion inhibition. The current study aimed to synthesize and evaluate the anticancer effects of hydroalcoholic EA extract (HEAE), Nanohydroxyapatite (nHAp) and nHAp synthesized trough EA (nHA-EA) in MCF-7 breast cancer cell line.


In the present study, HEAE preparation and green synthesis of nHA-EA was done and phase composition, functional groups, and crystallin phase of nHA-EA and nHAp were determined using Fourier-transform infrared (FTIR) and X-ray diffraction (XRD). The characteristics of synthesized nanoparticles including structural and morphological parameters were investigated using scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) techniques. Then, by using MTT-assay (Dimethylthiazoldiphenyltetrazolium), the in vitro cytotoxic and half maximal inhibitory concentration (IC50) of EA extract, nHAp, and nHA-EA in the MCF-7 breast cancer cell line was evaluated. Next, we assessed the expression of apoptosis-related genes Bax, Bcl2 and p53 using quantitative reverse-transcriptase polymerase-chain-reaction (qRT-PCR) and migration of MCF-7 cells by scratch assay.


The FTIR results demonstrated formation of nHAp and its interaction with HEAE during synthesis process. The XRD results of the synthesized nanoparticles showed similar XRD pattern of nHA-EA and nHAp and purity of synthesized nanomaterials. The average IC50 of HEAE, nHAp, and nHA-EA extract after treatment of cancer cells for 24 h was 400 µg/mL, 200 µg/mL, and 100 µg/mL, respectively. Our results revealed that nHA-EA significantly reduced the migration and invasion of the MCF-7 cells, in comparison to the nHAp and EA extract. Moreover, level of Bax/Bcl2 and p53 was significantly higher in the nHA-EA extract group in comparison to the EA extract and nHAp group.


Taken together, our results demonstrated that bioactive constituents of EA medicinal plant in form of nHA-EA particles, can effectively exerts potential anticancer and chemo preventive effect against breast cancer growth and can be proposed as a promising beneficial candidate for BC therapy. However, further investigations are required to discover what bioactive compounds are responsible for the chemo preventive effect of this extract.

Peer Review reports


One of the most common types of cancer in women is breast cancer (BC) [1], and it is considered a leading cause of death in patients with cancer all around the world [2]. Despite the recent advances in BC diagnostic and therapeutic methods, due to the high rate of mortality and chemoresistance, BC cancer patients' treatment is still a matter of debate [3].

There are numerous natural plant-based products and extracts, which exert anti-tumoral effects and could be useful for the design and development of new anti-cancer drugs [4]. Elaeagnus Angustifolia (EA) (also called oleaster, Russian olive, wild olive, silver berry), is a tree, that belongs to the Elaeagnacea (Araliaceae) family, and its fruits are characterized by small size and reddish-brown color. Different type of EA is commonly growing in Asia, Europe, and some regions of North America [5, 6]. The fruit of EA has been utilized as a medicinal plant, which acts as a potential therapeutic agent in numerous disorders through modulation of the immune system and oxidative stress balance [5]. It also exerts anti-cancer, antibacterial, and antifungal effects and has gastro- and hepatoprotective efficacy [7,8,9,10]. It has been shown that EA fruit is a rich source of different vitamins, carbohydrates, proteins, and minerals [6, 11, 12]. Moreover, EA fruits are a good source of beneficial compounds such as coumarins, tannins, phenolic acids, and flavonoids [5, 13, 14]. Different studies are showing that EA exerts anticancer effects via modulating the cell-cycle process, heat-shock proteins expression, anti-proliferative properties, apoptosis induction, blocking of angiogenesis, and cell invasion inhibition [15,16,17].

Nanomedicine has drawn a lot of interest because of its various and effective utilization in medicine, particularly in drug delivery [18]. Due to the high dispersion stability of nanoparticles (NPs), they have attracted considerable research interest in biomedical applications and drug delivery systems [19, 20]. The biosynthesis of NPs, using natural sources such as plants, without the utilization of any hazardous substances, reduces potential health and environmental threats [21, 22]. Moreover, plant extract allows the preparation of NPs with controlled and defined shape and size [23].

Hydroxyapatite (HAp, Ca10(PO4)6(OH)2), has various biomedical applications in many areas of medicine, due to its high bioactivity and biocompatibility. According to the method applied for HAp preparation, various mechanical properties bioactivity level, and dissolution behavior in the biological environment is expected [24]. It has been shown that HAp exerts anti-cancer effects via increasing drug release and consequently greater growth inhibition properties. Taken together, HAp enhances the chemotherapeutic efficacy of various agents including Cisplatin, Methotrexate, and Adriamycin. This could be because HAp mediates drug penetration into the tumor and improves drug delivery [25] improvement. It has been shown that the green synthesis of NPs has opened up new possibilities in material development, and there is research conducted on the green synthesis of HAp [26].

A different source of flavonoids including, catechin, epicatechin, gallocatechin, epigallocatechin, kaempferol, quercetin, luteolin, isorhamnetin, and isorhamnetin-3–0-β-D-galactopyranoside have been isolated from EA [27]. Furthermore, various phenolic components such as 4- hydroxybenzoic acid and caffeic acid have also been found in EA [28]. It has been shown that flavonoids contain phenolic hydroxyl groups, they may play a role in metals chelating efficiency, lowering lipid peroxidation process, and increasing antioxidant and free radical scavenging capacity [29, 30]. According to these findings, in the current study, nano hydroxyapatite (nHAp) was synthesized with the EA extract, which shows reducing/stabilizing effects, as well as capping properties. This synthesis method is inexpensive and simple, has long-lasting stability, and is appropriate for macro-scale procedures [7, 31,32,33].

Therefore, the current study aimed to synthesize and evaluate the anticancer effects of hydroalcoholic EA extract (HEAE), nHAp, and nHAp synthesized trough EA (nHA-EA) in MCF-7 breast cancer cell line. The synthesized nanoparticles were characterized, and their structural, morphological, and optical properties were determined via different analytical tools, including x-ray diffraction (XRD), fourier-transform infrared (FT-IR), scanning electron microscopy (SEM), and transmission electron microscopy (TEM).

Materials & methods

In the present study, all chemicals and reagents used to synthesize nHAp and nHA-EA were of analytical grade. Calcium nitrate tetrahydrate [Ca (NO3)2·4H2O], diammonium hydrogen phosphate [(NH4)2 HPO4], sodium hydroxide (NaOH), and were purchased from Sigma Aldrich. All chemicals solution was prepared using deionized water.

Plant material and extract preparation

The Russian Olive was obtained from South Khorasan, Birjand, Iran. It was identified as Elaeagnus Angustifolia L. by Dr. F. Askari (Assistant Professor of Traditional Pharmacy, School of Pharmacy). The voucher specimen was deposited in the Herbarium Center of the School of Pharmacy, Birjand University of Medical Sciences (221).

Hydroalcoholic EA extract preparation & green synthesis of nHA-EA

We have discussed synthesis of nHAp and nHA-EA in the previous studies [29, 34]. Briefly, EA pulps were obtained from fresh fruits and HEAE was prepared using the maceration procedure as below. 40 gr of dried EA pulp powder with 320 mL methanol and 80 ml distilled water were mixed. Finally, the extracts were filtered and concentrated using a rotating vacuum and HEAE was stored at 4 °C.

nHA-EAs were synthesized using the sol–gel technique, using calcium nitrate tetrahydrate and diammonium hydrogen phosphate, (molar ratio: 1.67). V/V HEAE (10 ml, 10%), diammonium hydrogen phosphate (5 ml, 0.3 M), and calcium nitrate tetrahydrate (15 ml, 0.3 M) were dissolved in ionized water. The Calcium nitrate tetrahydrate solution was introduced to the HEAE solution, then stirred upon slow warming to 50 °C, for 0.5 h. Then, diammonium hydrogen phosphate solution was added to the above-mixed solution at the flow rate of 1 ml/min. NaOH solution was used for pH adjustment (pH = 11). At 50 °C for 90 min, the suspension was agitated (Fig. 1). The resulting solution was subjected to the centrifuge and then gently rinsed with deionized water and ethanol (nHA-EA). Finally, nHAp solution without HEAE was synthesized under the same situations described for comparison.

Fig. 1
figure 1

A schematic representation of the nHA-EA preparation process


The phase composition, functional groups, and crystallin phase of nHA-EA and nHAp were determined using FTIR and XRD. The characteristics of synthesized nanoparticles including structural and morphological parameters were investigated using SEM and TEM techniques.

Cell culture

The human MCF-7 cells were purchased from the Pasteur institute (Tehran, Iran). The cells were cultured in dulbecco's modified eagle's medium (DMEM) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% streptomycin/ penicillin and maintained at 37 °C in a 5% CO2 atmosphere.


The MTT-assay (Dimethylthiazoldiphenyltetrazolium) was used to assess the in vitro cytotoxic and half maximal inhibitory concentration (IC50) of EA extract, nHAp, and nHA-EA in the MCF-7 breast cancer cell line. Cells were treated with different concentrations of EA extract, nHAp, and nHA-EA for 24 h. Then, the medium was removed from the wells and MTT substrate (Sigma, Germany) was added to each well, and the plate was incubated for 4 h. Then, the resulting crystals were dissolved in dimethylsulfoxide (DMSO) and incubated for 1 h. Finally, the absorbance was measured at 540 nm using a microplate reader. The experiments were performed in triplicate.

Scratch assay

Briefly, MCF-7 cells were seeded in a 12-well plate at a concentration of 4 × 105 cells /well. Then, a narrow scratch was created with a 100 μl sterile pipette tip through a monolayer of adherent cells growing on the bottom of a cell culture plate, 24 h after cell seeding. Then, cell debris was removed. Plates were treated and after incubation at different time points were imaged and analyzed by using Digimizer 5.4.9 software.

Quantitative reverse-transcriptase polymerase-chain-reaction (qRT-PCR)

In this study, a quantitative RT-PCR assay was used to measure transcript levels of different genes. First, total RNA was extracted from the cells using the Pars Tous kit, (Tehran, Iran) according to the manufacturer's instructions. Then, RNAs were converted to complementary DNA (cDNA) using the commercial cDNA synthesis kit (Parstous kit). Quantitative RT-PCR was performed with specific forward and reverse primers for target genes (Table 1). The cDNA amplification was performed by using the StepOne instrument (Applied Biosystems, Foster City, CA). The gene expression levels were normalized to a housekeeping control gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Table 1 Primers used for real-time polymerase chain reaction

Statistical analysis

All values are expressed as mean ± standard error of the mean. GraphPad Prism (version 9) was used to conduct the analyses. Statistical comparisons were determined using student’s t-test or one‐way analysis of variance (ANOVA) followed by tukey’s multiple comparison test. The differences were considered to be statistically significant at P < 0.05.



The FTIR technique in the middle infrared spectral (400–4000 cm−1) wavelength was used to evaluate the functional groups of HEAE, nHAp and nHA-EA (Fig. 2). In the HEAE spectra, the continued peak at 3367 cm−1 is defined to O–H vibration and peaks at 1060–1283 cm−1 are the C–O–C characteristic peaks. In the nHAp, the intense peak around at 564–601 and 1030 cm−1 indicated the PO43− bands. Moreover, the O–H vibrations on the water molecule were found around 3000 to 3600 cm−1. In the nHA-EA spectra, the extended peak around at 3443 shows water absorption in the synthesized nanoparticles. nHA-EA spectra analysis showed distinct bands of PO43− at 564, 604, and, 1029 cm−1. The extra peak at 870 showed the presence of C = O stretching of carboxylic acid, and clearly suggests the interaction between carboxyl group and nHAp. These FTIR functional groups findings clearly demonstrates formation of nHAp and its interaction with HEAE during synthesis process.

Fig. 2
figure 2

FTIR spectra of HEAE, nHAp and nHA-EA


The X-ray diffraction patterns results of the synthesized nanoparticles are presented at Fig. 3, nHA-EA and nHAp showed similar XRD pattern. There was no peak related to the calcium hydroxide and calcium phosphates, which indicates synthesized nanomaterials purity. The seven peaks observed at 26◦, 32◦, 39◦, 46◦, 49◦,53◦ and 64◦, were corresponded to the plane (002), (112), (310), (222), (123), (004), and (233), respectively, and related to the hydroxyapatite, which find to be matched with ICDD: 01–084-1998.

Fig. 3
figure 3

XRD patterns of nHA and nHAEA


Data obtained from SEM and TEM images revealed nanorod particles (Figs. 4 and 5). Notably, nHAps have 17–29 nm width and 62–89 nm length, whereas nHA-EA nanorods have 17–23 nm width and 93–146 nm length.

Fig. 4
figure 4

SEM images of nHAp and nHA-EA

Fig. 5
figure 5

TEM images of nHAp and nHA-EA

Cell proliferation

In the current study, an MTT assay was performed to evaluate the in vitro cytotoxic and IC50 of EA, nHAp, and nHAEA extract in the MCF-7 breast cancer cell line. Our results indicated that the average IC50 of EA, nHAp, and nHAEA extract after treatment of cancer cells for 24 h was 400 µg/mL, 200 µg/mL, and 100 µg/mL, respectively (Fig. 6).

Fig. 6
figure 6

MTT assay images show cytotoxic effects and IC50 values of EA, nHAp and nHA-EA

Scratch assay

Regarding the potential effects of EA nanoparticles on breast cancer cell migration, in the present study, we examined the breast cancer cells invasiveness inhibitory effects of nHA-EA using the scratch assay. Our results revealed that nHA-EA significantly reduced the migration and invasion of the MCF-7 cells, in comparison to the nHAp and EA extract (Fig. 7).

Fig. 7
figure 7

The effects of EA, nHAp and nHA-EA on the migration of the MCF-7 cells after 24 h


To determine the anti-cancer effects of EA extract, nHAp, and a combination of EA extract and nHAp in the MCF-7 cells, we evaluated the expression level of Bax/Bcl2 and p53 as key markers in the carcinogenesis process. Our results revealed that the level of Bax/Bcl2 and p53 are significantly higher in the nHA-EA extract group in comparison to the EA extract and nHAp alone (Fig. 8).

Fig. 8
figure 8

EA, nHAp and nHA-EA triggered MCF-7 cells apoptosis genes as showed by qRT-PCR


In the past few years, there have been reports of nanoscale HAp that exhibit superior biocompatibility compared to regular HAp. Furthermore, these nanoscale HAp particles have shown anti-cancer properties and play a crucial role in controlling the behavior of breast cancer cells [35]. There are multiple methods for synthesizing nHAp, such as co-precipitation, hydrothermal, sol–gel, and others. Additionally, the utilization of metal nanoparticles extracted from plants is considered environmentally friendly. As a result, the use of natural materials in the production of nHAp has captured the interest of numerous researchers, contributing to the development of a significant field of study within the realm of nanotechnology science [29].

A wide range of natural plant-derived products and extracts possess anti-tumoral properties, presenting potential value in the creation and advancement of novel anti-cancer nanoparticles. There are growing body of evidences showing that natural products have an important role in treatment of wide variety of human disease, including types of cancer [30, 36]. EA, as an herbal medicine plant with different properties, have been used extensively for a long time to treat different disorders [13, 33]. There are various bioactive constituent in EA, including phenolic acids and flavonoids, which exerts a critical role in cancer development and progression inhibition [13, 27]. It has been shown that bioactive compounds of EA could modulate different biological processes of cells including cell cycle progression, apoptosis and DNA repair [28, 37]. In recent years, EA has received considerable attention for cancer therapy because of its promising effectiveness and potential therapeutic effects as chemo preventive and antitumor natural product. It has been shown that flavonoids with different signaling pathways which are correlated with carcinogenesis, including cellular proliferation, apoptosis, angiogenesis, and metastasis. Moreover, apigenin, as a phytoestrogen aglycone has shown to suppress apoptosis, cell cycle and invasion in malignant cells, alone or in combination with other chemotherapeutic agents [38].

In this study, nHAp was synthesized with EA extract, which could act as a reducing, stabilizing, and capping agent. This synthesis is easy, cheap, durable, and suitable for largescale processing. To determine the anticancer effects of nHA-EA in MCF-7 cells were treated with nHA-EA.

According to result of this study, nHA-EA had more growth inhibitory properties on MCF-7 breast cancer cell line than other groups. Our results revealed that, nHA-EA decreased the viability and proliferation of MCF-7 cells and significantly increased the expression level of p53 and Bax/Bcl2 genes.

Flavonoids have been demonstrated to increase the expression of p53 and induce cell cycle arrest specifically in the G2/M phase in cancer cells. Additionally, they are recognized for their ability to inhibit the expression of Ras proteins and modulate heat-shock proteins in different types of cancers, particularly in leukemia and colorectal cancer.

The flavonoids in the EA extract surround the nHAp like a cap. As a result, they may increase the anti-cancerous properties of nHAp. Quercetin, a prominent flavonoid found in EA, is a significant anti-proliferative compound. In addition, it plays a role in enhancing tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by increasing the expression of Bax and suppressing the activity of Bcl2 protein [39].

These results are in accordance with previous studies showing the suppressive effects of EA on different type of cancer including glioblastoma and breast cancer [38, 40]. Although, the results of the current study are not consistent with other investigation, showing that EA extract did not exerted any significant antiproliferative activity against cancerous cells, which may be due to the different concentration of extract and its characteristics [41].

Uncontrolled cell division and growth and escape from apoptosis are well-known hallmarks of malignant cells and important mechanism involved in cancer treatment resistance; therefore, targeting specific cancer deregulated pathways could be an important way for providing novel therapeutics strategies [42, 43].

Moreover, we revealed that the ethanolic extract of EA could decrease migration of MCF-7 cells and induces apoptosis in this cell line. Our results are in agreement with other studies that confirmed EA exerts its anti-tumor impacts via upregulation of p53 and Bax/Bcl2.

The observed anti-proliferative effects of EA extract on breast cancer cells in our study could mediate via increasing the expression of p53 as a tumor suppressor gene. It has been shown that bioactive constituents of in EA, including flavonoids, triterpenoid, lignanoid, benzenoid, pro-anthocyanosides, polysaccharides and phenolic acids, plays an important role in preventing cancer development and progression. Investigations revealed that these constituents could regulate various biological pathways including apoptosis, DNA repair, inflammatory processes and cell cycle progression [13, 33, 44]. p53 regulates the expression of Bcl2, a proapoptotic factor, in which triggers apoptosis process [45]. Moreover, there are studies showing that EA extract induces apoptosis via different mechanisms including, human epidermal growth factor receptor 2 gene (HER2) and Jun N-terminal kinases (JNK) inhibition [39]. Thus, these results support the role for EA extract in the Bcl2-associated intrinsic pathway of apoptosis via p53 upregulation.

It is well known that invasion and migration, which are key underling mechanisms of metastasis plays an important role in breast cancer disease. Metastasis is a process in which malignant cells leaving primary tissue, disseminate, circulate and induces secondary tumors in distant sites [46].

In the present study, analysis of scratch assay revealed that, nHA-EA significantly inhibited migration and invasion in MCF-7 cell line. The anti-migratory and anti-metastatic potential of EA herbal plant has been confirmed in a study conducted by Jabeen showing that, EA extract inhibited the invasiveness of HER2-positive breast cancer cell lines. They showed that, this effect is associated with JNK signaling suppression and mesenchymal-epithelial transition (MET) inhibition. They also reported that suppression of JNK signaling pathway is associated with induction of apoptosis process [39]. In another study conducted by Saleh, it has been found that EA extract suppresses the invasiveness and migration of oral carcinoma cell lines. They reported that this extract inhibits cell invasion via increasing the amplification of E-cadherin, an important modulator of MET process. The also indicated that this reduction in migratory properties of oral carcinoma cell lines is mediated via inhibition of extracellular-signal-regulated kinases 1 and 2 (ERK1/ERK2) pathway [47].


Taken together, our results demonstrated that bioactive constituents of EA medicinal plant in form of nHA-EA particles, can effectively exerts potential anticancer and chemo preventive effect against breast cancer growth and can be proposed as a promising beneficial candidate for BC therapy. Moreover, we showed that nHA-EA triggers cell apoptosis in MCF-7 cells via increasing proapoptotic gene. Additionally, our findings indicate the role of nHA-EA as a natural product in targeting p53 as a key marker in BC treatment. However, further investigations are required to discover what bioactive compounds are responsible for the chemo preventive effect of this extract.

Availability of data and materials

The datasets used and/or analysed during the current study available from the corresponding author on reasonable request.



Elaeagnus Angustifolia


Hydroalcoholic EA extract


Nano hydroxyapatite


nHAp synthesized trough EA


Fourier-transform infrared


X-ray diffraction


Scanning electron microscopy


Transmission electron microscopy


Breast cancer



Ca (NO3)2·4H2O:

Calcium nitrate tetrahydrate


Diammonium hydrogen phosphate




Converted to complementary DNA


Glyceraldehyde-3-phosphate dehydrogenase


One‐way analysis of variance


Dulbecco's Modified Eagle's Medium


quantitative reverse-transcriptase polymerase-chain-reaction




Fetal bovine serum


Mesenchymal-epithelial transition


Human epidermal growth factor receptor 2 gene


Jun N-terminal kinases


Extracellular-signal-regulated kinases 1 and 2


tumor necrosis factor-related apoptosis-inducing ligand


Half maximal inhibitory concentration


  1. Binabaj MM, Hosseini SA, Khoshnazar AK, Asadi J. The simultaneous effect of valproic acid and gamma radiation on telomerase activity and Bax and Bcl-2 protein levels in MCF-7 breast cancer cell line. Jundishapur J Nat Pharm Prod. 2015;10(3):e22818.

  2. Khoshakhlagh M, Soleimani A, Binabaj MM, Avan A, Ferns GA, Khazaei M, et al. Therapeutic potential of pharmacological TGF-β signaling pathway inhibitors in the pathogenesis of breast cancer. Biochem Pharmacol. 2019;164:17–22.

    CAS  PubMed  Google Scholar 

  3. Housman G, Byler S, Heerboth S, Lapinska K, Longacre M, Snyder N, et al. Drug resistance in cancer: an overview. Cancers. 2014;6(3):1769–92.

    PubMed  PubMed Central  Google Scholar 

  4. Moza MK. Forsteronia refracta holds the key to breast cancer treatment. Curr Sci. 2005;88(8):1122.

    Google Scholar 

  5. Farzaei MH, Bahramsoltani R, Abbasabadi Z, Rahimi R. A comprehensive review on phytochemical and pharmacological aspects of Elaeagnus angustifolia L. J Pharm Pharmacol. 2015;67(11):1467–80.

    CAS  PubMed  Google Scholar 

  6. Fonia A, White I, White J. Allergic contact dermatitis to Elaeagnus plant (Oleaster). Contact Dermatitis. 2009;60(3):178–9.

    CAS  PubMed  Google Scholar 

  7. Hamidpour R, Hamidpour S, Hamidpour M, Shahlari M, Sohraby M, Shahlari N, et al. Russian olive (Elaeagnus angustifolia L.): From a variety of traditional medicinal applications to its novel roles as active antioxidant, anti-inflammatory, anti-mutagenic and analgesic agent. J Tradit Complement Med. 2017;7(1):24–9.

    PubMed  Google Scholar 

  8. Tehranizadeh ZA, Baratian A, Hosseinzadeh H. Russian olive (Elaeagnus angustifolia) as a herbal healer. BioImpacts: BI. 2016;6(3):155.

    CAS  Google Scholar 

  9. Faramarz S, Dehghan G, Jahanban-Esfahlan A. Antioxidants in different parts of oleaster as a function of genotype. BioImpacts: BI. 2015;5(2):79.

    PubMed  PubMed Central  Google Scholar 

  10. Panahi Y, Alishiri GH, Bayat N, Hosseini SM, Sahebkar A. Efficacy of Elaeagnus Angustifolia extract in the treatment of knee osteoarthritis: a randomized controlled trial. EXCLI J. 2016;15:203.

    PubMed  PubMed Central  Google Scholar 

  11. Boudraa S, Hambaba L, Zidani S, Boudraa H. Mineral and vitamin composition of fruits of five underexploited species in Algeria: Celtis australis L., Crataegus azarolus L., Crataegus monogyna Jacq., Elaeagnus angustifolia L. and Zizyphus lotus L. Fruits. 2010;65(2):75–84.

    CAS  Google Scholar 

  12. Taheri JB, Anbari F, Maleki Z, Boostani S, Zarghi A, Pouralibaba F. Efficacy of Elaeagnus angustifolia topical gel in the treatment of symptomatic oral lichen planus. J Dent Res Dent Clin Dent Prospects. 2010;4(1):29.

    Google Scholar 

  13. Saboonchian F, Jamei R, Sarghein SH. Phenolic and flavonoid content of Elaeagnus angustifolia L.(leaf and flower). Avicenna J Phytomed. 2014;4(4):231.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Niknam F, Azadi A, Barzegar A, Faridi P, Tanideh N, Zarshenas MM. Phytochemistry and Phytotherapeutic Aspects of Elaeagnus angustifolia L. Current Drug Discov Technol. 2016;13(4):199–210.

    CAS  Google Scholar 

  15. Torbati M, Asnaashari S, Afshar FH. Essential oil from flowers and leaves of Elaeagnus angustifolia (Elaeagnaceae): Composition, radical scavenging and general toxicity activities. Adv Pharm Bull. 2016;6(2):163.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Kurdali F, Al-Shamma’a M. Natural abundances of 15N and 13C in leaves of some N2-fixing and non-N2-fixing trees and shrubs in Syria. Isot Environ Health Stud. 2009;45(3):198–207.

    CAS  Google Scholar 

  17. Murakami A, Ashida H, Terao J. Multitargeted cancer prevention by quercetin. Cancer Lett. 2008;269(2):315–25.

    CAS  PubMed  Google Scholar 

  18. Wang X, Liu L-H, Ramström O, Yan M. Engineering nanomaterial surfaces for biomedical applications. Exp Biol Med. 2009;234(10):1128–39.

    CAS  Google Scholar 

  19. Tanaka T, Hirose M, Kotobuki N, Ohgushi H, Furuzono T, Sato J. Nano-scaled hydroxyapatite/silk fibroin sheets support osteogenic differentiation of rat bone marrow mesenchymal cells. Mater Sci Eng C. 2007;27(4):817–23.

    CAS  Google Scholar 

  20. Takemoto S, Kusudo Y, Tsuru K, Hayakawa S, Osaka A, Takashima S. Selective protein adsorption and blood compatibility of hydroxy-carbonate apatites. J Biomed Mater Res A. 2004;69(3):544–51.

    PubMed  Google Scholar 

  21. Iqbal J, Abbasi BA, Ahmad R, Shahbaz A, Zahra SA, Kanwal S, et al. Biogenic synthesis of green and cost effective iron nanoparticles and evaluation of their potential biomedical properties. J Mol Struct. 2020;1199:126979.

    CAS  Google Scholar 

  22. Hameed S, Iqbal J, Ali M, Khalil AT, Abbasi BA, Numan M, et al. Green synthesis of zinc nanoparticles through plant extracts: establishing a novel era in cancer theranostics. MRX. 2019;6(10):102005.

    CAS  Google Scholar 

  23. Abbasi BA, Iqbal J, Mahmood T, Ahmad R, Kanwal S, Afridi S. Plant-mediated synthesis of nickel oxide nanoparticles (NiO) via Geranium wallichianum: Characterization and different biological applications. MRX. 2019;6(8):0850a7.

    CAS  Google Scholar 

  24. Riman RE, Suchanek WL, Byrappa K, Chen C-W, Shuk P, Oakes CS. Solution synthesis of hydroxyapatite designer particulates. Solid State Ionics. 2002;151(1–4):393–402.

    CAS  Google Scholar 

  25. Munir MU, Salman S, Javed I, Bukhari SNA, Ahmad N, Shad NA, et al. Nano-hydroxyapatite as a delivery system: overview and advancements. Artif Cells Nanomed Biotechnol. 2021;49(1):717–27.

    CAS  PubMed  Google Scholar 

  26. Kumar GS, Muthu D, Karunakaran G, Karthi S, Girija EK, Kuznetsov D. Curcuma longa tuber extract mediated synthesis of hydroxyapatite nanorods using biowaste as a calcium source for the treatment of bone infections. J Sol-Gel Sci Technol. 2018;86(3):610–6.

    CAS  Google Scholar 

  27. Abizov E, Tolkachev O, Mal’Tsev S, Abizova E. Composition of biologically active substances isolated from the fruits of Russian olive (Elaeagnus angustifolia) introduced in the European part of Russia. Pharm Chem J. 2008;42(12):696–8.

    CAS  Google Scholar 

  28. Hui C, Qi X, Qianyong Z, Xiaoli P, Jundong Z, Mantian M. Flavonoids, flavonoid subclasses and breast cancer risk: a meta-analysis of epidemiologic studies. PLoS One. 2013;8(1):e54318.

    PubMed  PubMed Central  Google Scholar 

  29. Azaryan E, Mortazavi-Derazkola S, Alemzadeh E, Emadian Razavi F, Yousefi M, Hanafi-Bojd MY, et al. Effects of hydroxyapatite nanorods prepared through Elaeagnus Angustifolia extract on modulating immunomodulatory/dentin–pulp regeneration genes in DPSCs. Odontology. 2023;111(2):461–73.

    CAS  PubMed  Google Scholar 

  30. Atanasov AG, Waltenberger B, Pferschy-Wenzig E-M, Linder T, Wawrosch C, Uhrin P, et al. Discovery and resupply of pharmacologically active plant-derived natural products: A review. Biotechnol Adv. 2015;33(8):1582–614.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Kalaiselvi V, Mathammal R, Vijayakumar S, Vaseeharan B. Microwave assisted green synthesis of Hydroxyapatite nanorods using Moringa oleifera flower extract and its antimicrobial applications. Int J Vet Sci Med. 2018;6(2):286–95.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Hamidpour R, Hamidpour S, Doostmohamadi P. Chemistry, pharmacology and medicinal property of Russian olive (Elaeagnus angustifolia L.). Cancer Sci Res. 2019;6:1–7.

    Google Scholar 

  33. Ayaz FA, Bertoft E. Sugar and phenolic acid composition of stored commercial oleaster fruits. J Food Compos Anal. 2001;14(5):505–11.

    CAS  Google Scholar 

  34. Azaryan E, Hanafi-Bojd MY, Alemzadeh E, Emadian Razavi F, Naseri M. Effect of PCL/nHAEA nanocomposite to osteo/odontogenic differentiation of dental pulp stem cells. BMC Oral Health. 2022;22(1):1–13.

    Google Scholar 

  35. Meena R, Kesari KK, Rani M, Paulraj R. Effects of hydroxyapatite nanoparticles on proliferation and apoptosis of human breast cancer cells (MCF-7). J Nanopart Res. 2012;14:1–11.

    Google Scholar 

  36. Atanasov AG, Zotchev SB, Dirsch VM, Supuran CT. Natural products in drug discovery: advances and opportunities. Nat Rev Drug Discovery. 2021;20(3):200–16.

    CAS  PubMed  Google Scholar 

  37. Zhang H-W, Hu J-J, Fu R-Q, Liu X, Zhang Y-H, Li J, et al. Flavonoids inhibit cell proliferation and induce apoptosis and autophagy through downregulation of PI3Kγ mediated PI3K/AKT/mTOR/p70S6K/ULK signaling pathway in human breast cancer cells. Sci Rep. 2018;8(1):1–13.

    Google Scholar 

  38. Arab S, Bahraminasab M, Yazdani A, Abdolshahi A. Effects of whole fruit extract of Elaeagnus angustifolia L. on glioblastoma cell lines. J Microbiol Biotechnol Food Sci. 2022;11(5):e4314-e.

    Google Scholar 

  39. Jabeen A, Sharma A, Gupta I, Kheraldine H, Vranic S, Al Moustafa A-E, et al. Elaeagnus angustifolia plant extract inhibits epithelial-mesenchymal transition and induces apoptosis via HER2 inactivation and JNK pathway in HER2-positive breast cancer cells. Molecules. 2020;25(18):4240.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Fouzat A, Hussein OJ, Gupta I, Al-Farsi HF, Khalil A, Al Moustafa A-E. Elaeagnus angustifolia plant extract induces apoptosis via P53 and signal transducer and activator of transcription 3 signaling pathways in triple-negative breast cancer cells. Front Nutr 2022;9:418.

  41. Abu-Dahab R, Afifi F. Antiproliferative activity of selected medicinal plants of Jordan against a breast adenocarcinoma cell line (MCF7). Sci Pharm. 2007;75(3):121–46.

    Google Scholar 

  42. Hanahan D. Hallmarks of cancer: new dimensions. Cancer Discov. 2022;12(1):31–46.

    CAS  PubMed  Google Scholar 

  43. Plati J, Bucur O, Khosravi-Far R. Dysregulation of apoptotic signaling in cancer: molecular mechanisms and therapeutic opportunities. J Cell Biochem. 2008;104(4):1124–49.

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Carradori S, Cairone F, Garzoli S, Fabrizi G, Iazzetti A, Giusti AM, et al. Phytocomplex characterization and biological evaluation of powdered fruits and leaves from Elaeagnus angustifolia. Molecules. 2020;25(9):2021.

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Yan H, Huang W, Rao J, Yuan J. miR-21 regulates ischemic neuronal injury via the p53/Bcl-2/Bax signaling pathway. Aging (Albany NY). 2021;13(18):22242.

    CAS  PubMed  Google Scholar 

  46. Fidler IJ. Critical factors in the biology of human cancer metastasis: twenty-eighth G.H.A. Clowes memorial award lecture. Cancer Res. 1990;50(19):6130–8.

    CAS  PubMed  Google Scholar 

  47. Saleh AI, Mohamed I, Mohamed AA, Abdelkader M, Yalcin HC, Aboulkassim T, et al. Elaeagnus angustifolia plant extract inhibits angiogenesis and downgrades cell invasion of human oral cancer cells via Erk1/Erk2 inactivation. Nutr Cancer. 2018;70(2):297–305.

    CAS  PubMed  Google Scholar 

Download references


All the authors of the study would like to thank the Birjand University of Medical Science.


This work was supported by Birjand University of Medical Science (BUMS) (Grant No. 4277), Iran.

Author information

Authors and Affiliations



SK, EA contributed to research planning, case examination, and editing the manuscript. MMB contributed to examination and writing. AZ and MN contributed to statistical analysis, research planning, collect and enter data.

Corresponding authors

Correspondence to Samira Karbasi or Mohsen Naseri.

Ethics declarations

Ethics approval and consent to participate

Experimental research and field studies on plants including the collection of plant material are complying with relevant guidelines and regulation for plant ethics.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zarban, A., Azaryan, E., Binabaj, M.M. et al. Green synthesis of nanohydroxyapatite trough Elaeagnus angustifolia L. extract and evaluating its anti-tumor properties in MCF7 breast cancer cell line. BMC Complement Med Ther 23, 338 (2023).

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: