Green Extraction of Bioactive Compounds from Solanum nigrum L Fruit Using Ultrasound-Assisted Technology: Antioxidant Properties and Proliferative Effects on NIH-3T3 Fibroblasts
DOI:
https://doi.org/10.48048/tis.2026.12682Keywords:
Antioxidant, Cosmetic, Fibroblasts, Proliferation, NIH-3T3, Skin agingAbstract
Skin aging is a process that involves a gradual decline in the function of fibroblasts in the dermis layer. Fibroblasts in the dermis play a role in controlling the structure of the extracellular matrix, supporting wound healing, and maintaining skin tissue stability. When fibroblast function declines, it results in damage to the skin structure, prolonged inflammation, and a reduction in the skin’s ability to regenerate. This study aimed to evaluate the antiaging potential of the extract of Solanum nigrum L fruit from Majenang, Central Java, Indonesia. The effects of solvent type and extraction duration on the bioactive content and biological activities of Solanum nigrum L fruit extracts obtained by Ultrasound-Assisted Extraction (UAE). Proliferation activity was carried out on NIH-3T3 fibroblast cell lines, while antioxidant activity was done with DPPH methods. We also analyzed the content of phenolic and flavonoid compounds. The research method was experimental. Extraction was performed using the Ultrasonic-Assisted Extraction (UAE) method. Phytochemical analysis included the determination of total phenol and total flavonoid content. The activity of the extract was evaluated through antioxidant testing using the DPPH method and testing of the proliferation effect on NIH 3T3 cells. The results indicated that solvent selection and extraction time had a significant influence on both extraction yield and biological activity. Optimal conditions were achieved using 96% ethanol as the solvent and an extraction time of 30 min, yielding a total phenolic content of 61.18 mg GAE/g, flavonoids of 73.99 mg Rutin/g, antioxidant activity of 87.164 ppm, and an EC50 value for NIH-3T3 fibroblast cell proliferation of 12.48 ppm. These proved that Solanum nigrum L fruit has antioxidant activity and can stimulate the proliferation of NIH-3T3 fibroblast cells, making it a promising candidate for further development in the pharmaceutical and cosmetic industries as an active ingredient that supports cell regeneration.
HIGHLIGHTS
- Fresh Solanum nigrum fruit was extracted using Ultrasound-Assisted Technology (UAE)
- Evaluating the effect of solvent variation and extraction time on the extraction efficiency of Solanum nigrum fruit using UAE technology.
- Reporting that 96% ethanol is the best solvent compared to other solvents in increasing the extraction yield and determining flavonoid and phenol content.
- Confirming that an extraction duration of 30 minutes is the optimal condition for the highest yield and quality of
- Reporting an EC₅₀ value of 12.48 ppm in the NIH-3T3 fibroblast cell proliferation test, indicating strong bioactive potential for skin cell regeneration.
GRAPHICAL ABSTRACT
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J Zhang, H Yu, MQ Man and L Hu. Aging in the dermis: Fibroblast senescence and its significance. Aging Cell 2023; 23(2), 14054.
Y Lu, G Pan, Z Wei, Y Li and X Pan. Role of fibroblast autophagy and proliferation in skin anti-aging. Experimental Gerontology 2024; 196, 112559.
L Nan, P Guo, W Hui, F Xia and C Yi. Recent advances in dermal fibroblast senescence and skin aging: Unraveling mechanisms and pioneering therapeutic strategies. Frontiers in Pharmacology 2025; 16, 1592596.
M Wlaschek, P Maity, E Makrantonaki and K Scharffetter-Kochanek. Connective tissue and fibroblast senescence in Skin Aging. Journal of Investigative Dermatology 2021;141(4), 985-992.
X Xiong, W Yang, G Huang and H Huang. Ultrasonic-assisted extraction , characteristics and activity of Ipomoea batatas polysaccharide. Ultrasonics Sonochemistry 2023; 96, 106420.
J Yang, N Li, C Wang, T Chang and H Jiang. Ultrasound-homogenization-assisted extraction of polyphenols from coconut mesocarp: Optimization study. Ultrasonics Sonochemistry 2021; 78, 105739.
Z Xu, Y Cai, Q Ma, Z Zhao, D Yang and X Xu. Optimization of extraction of bioactive compounds from Baphicacanthus cusia leaves by hydrophobic deep eutectic solvents. Molecules 2021; 26(6), 1729.
CP Mungwari, CK King’Ondu, P Sigauke and BA Obadele. Conventional and modern techniques for bioactive compounds recovery from plants: Review. Scientific African 2024; 27, 02509.
PA Uwineza and A Waśkiewicz. Recent advances in supercritical fluid extraction of natural bioactive compounds from natural plant materials. Molecule 2025; 25(17), 3847.
M Saifullah, R McCullum, A McCluskey and Q Vuong. Comparison of conventional extraction technique with ultrasound assisted extraction on recovery of phenolic compounds from lemon scented tea tree (Leptospermum petersonii) leaves. Heliyon 2020; 6(4), 03666.
L Shen, S Pang, M Zhong, Y Sun, A Qayum, Y Liu, A Rashid, B Xu, Q Liang, H Ma and X Ren. A comprehensive review of ultrasonic assisted extraction (UAE) for bioactive components: Principles, advantages, equipment, and combined technologies. Ultrasonics Sonochemistry 2023; 101, 106646.
A Carreira-Casais, P Otero, P Garcia-Perez, P Garcia-Oliveira, AG Pereira, M Carpena, A Soria-Lopez, J Simal-Gandara and MA Prieto. Benefits and drawbacks of ultrasound-assisted extraction for the recovery of bioactive compounds from marine algae. International Journal of Environmental Research and Public Health 2021; 18(17), 9153.
I Călinescu, M Vinatoru, D Ghimpețeanu, V Lavric and TJ Mason. A new reactor for process intensification involving the simultaneous application of adjustable ultrasound and microwave radiation. Ultrasonics Sonochemistry 2021; 77, 105701.
RS Hussein, SBin Dayel, O Abahussein and AA El-Sherbiny. Influences on skin and intrinsic aging: Biological, environmental, and therapeutic insights. Journal of Cosmetic Drmatology 2025; 24(2), 16688.
X Chen, X Dai, Y Liu, Y Yang, L Yuan, X He and G Gong. Solanum nigrum Linn.: An insight into current research on traditional uses, phytochemistry, and pharmacology. Frontiers in Pharmacology 2022; 13, 918071.
A Aouadi, DH Saoud, A Rebiai, MH Ibrahim, M Messaoudi, K Alia, H Zidane, AV Atoki and FMA El-Mordy. Chemical composition’s effect on Solanum nigrum Linn.’s antioxidant capacity and erythrocyte protection: Bioactive components and molecular docking analysis. Open Life Sciences 2024; 19(1), 20220944.
A Campisi, R Acquaviva, G Raciti, A Duro, M Rizzo and NA Santagati. Antioxidant activities of Solanum nigrum L. Leaf extracts determined in in vitro cellular models. Foods 2019; 8(2), 63.
A Zorina, V Zorin, A Isaev, D Kudlay, M Vasileva and P Kopnin. Dermal fibroblasts as the main target for skin anti-age correction using a combination of regenerative medicine methods. Current Issues in Molecular Biology 2023; 45(5), 3829-3847.
J Hao, Z Wang, Y Jia, L Sun, Z Fu, M Zhao, Y Li, N Yuan, B Cong, L Zhao and G Ge. Optimization of ultrasonic-assisted extraction of flavonoids from Lactuca indica L. cv. Mengzao and their antioxidant properties. Frontiers in Nutrition 2023; 10, 1065662.
A El-Baakili, M Fadil and NE Es-Safi. Ultrasonic-assisted extraction for phenolic compounds and antioxidant activity of Moroccan Retama sphaerocarpa L. leaves: Simultaneous optimization by response surface methodology and characterization by HPLC/ESI-MS analysis. Heliyon 2023; 9(6), 17168.
S Baliyan, R Mukherjee, A Priyadarshini, A Vibhuti, A Gupta, RP Pandey and CM Chang. Determination of antioxidants by DPPH radical scavenging activity and quantitative phytochemical analysis of Ficus religiosa. Molecules 2022; 27(4), 1326.
V Buranasudja, D Rani, A Malla, K Kobtrakul and S Vimolmangkang. Insights into antioxidant activities and anti-skin-aging potential of callus extract from Centella asiatica (L.). Scientific Reports 2021; 11, 13459.
AM Cruz, MC Gonçalves, MS Marques, F Veiga, AC Paiva-Santos and PC Pires. In vitro models for antiaging efficacy assessment: A critical update in dermocosmetic research. Cosmetics 2023; 10(2), 66.
A Nurkhasanah, T Fardad, C Carrera, W Setyaningsih and M Palma. Ultrasound-assisted anthocyanins extraction from pigmented corn: Optimization using response surface methodology. Methods Protocols 2023; 6(4), 69.
K Kumar, S Srivastav and VS Sharanagat. Ultrasound assisted extraction (UAE) of bioactive compounds from fruit and vegetable processing by-products: A review. Ultrasonics Sonochemistry 2021; 70, 105325.
G Psakis, F Lia, VP Valdramidis and R Gatt. Exploring hydrodynamic cavitation for citrus waste valorisation in Malta: From beverage enhancement to potato sprouting suppression and water remediation. Frontiers in Chemistry 2024; 12, 1411727.
K Papoutsis, P Pristijono, JB Golding, CE Stathopoulos, MC Bowyer, CJ Scarlett and QV Vuong. Optimizing a sustainable ultrasound-assisted extraction method for the recovery of polyphenols from lemon by-products: Comparison with hot water and organic solvent extractions. European Food Research and Technology 2018; 244, 1353-1365.
NVT Nguyen, CV Nguyen, NT Duong, XTT Dai, KT Nguyen and CTT Le. Optimization of ultrasound-assisted extraction using response surface methodology and quantification of polyphenol compounds in Avicennia officinalis L. from Vietnam. Pharmacia 2024; 71, 115528.
AC Andrade, FT Borsoi, ASMC Saliba, SM de Alencar, GM Pastore and HS Arruda. Optimization of Ultrasonic-assisted extraction of phenolic compounds and antioxidant activity from araticum peel using response surface methodology. Plants 2024; 13(18), 2560.
M Sukma, AWR Akili1, A Hardianto, J Latip and T Herlina. Ultrasound-assisted extraction of flavonoids from Erythrina crista-galli twigs using natural deep eutectic solvents: Process optimization and antioxidant evaluation. Trends in Sciences 2025; 22(12), 11099.
E Herrera-Pool, AL Ramos-Díaz, MA Lizardi-Jiménez, S Pech-Cohuo, T Ayora-Talavera, JC Cuevas-Bernardino, U García-Cruz and N Pacheco. Effect of solvent polarity on the Ultrasound Assisted extraction and antioxidant activity of phenolic compounds from habanero pepper leaves (Capsicum chinense) and its identification by UPLC-PDA-ESI-MS/MS. Ultrasonics Sonochemistry 2021; 76, 105658.
JDC Gomes, FR Procópio, AC Mendonça, LM Vieira, ÍT Perrone, FAR de Barros and PC Stringhet. Optimization of ultrasound-assisted extraction of phenolic compounds from jussara (Euterpe edulis M.) and blueberry (Vaccinium myrtillus) fruits. Food Science and Technology 2018; 38(1), 36316.
M Pérez, I Dominguez-Lopez and RM Lamuela-Raventos. The chemistry behind the folin-ciocalteu method for the estimation of (poly)phenol content in food: Total phenolic intake in a mediterranean dietary pattern. Journal of Agricultural and Food Chem 2023; 71(46), 17543-17553.
M Zugazua-Ganado, A Bordagaray, J Ezenarro, R Garcia-Arrona, M Ostra and M Vidal. Adaptation of the Folin-ciocalteu and fast blue BB spectrophotometric methods to digital image analysis for the determination of total phenolic content: Reduction of reaction time, interferences and sample analysis. LWT 2023; 193, 115756.
H Bouakline, S Bouknana, M Merzouki, I Ziani, A Challioui, M Bnouham, A Tahani and AEL Bachiri. The Phenolic content of pistacia lentiscus leaf extract and its antioxidant and antidiabetic properties. Scientific World Journal 2024; 2024, 1998870.
B Poojar, B Ommurugan, S Adiga, H Thomas, RK Sori, B Poojar, N Hodlur, A Tilak, R Korde and P Gandigawad. Methodology used in the study. Asian Journal Pharmaceutical Clinical Research 2017; 7(10), 4103.
SK Zaidi, MN Hoda, S Tabrez, SA Ansari, MA Jafri, MS Khan, S Hasan, MH Alqahtani, AM Abuzenadah and N Banu. Protective effect of Solanum nigrum leaves extract on immobilization stress induced changes in rat’s brain. Evidence-Based Complementarary and Alternative Medicine 2014; 2014, 912450.
Y Desmiaty, FC Saputri, M Hanafi, R Prastiwi and B Elya. Anti-elastase, anti-tyrosinase, and anti-oxidant of Rubus fraxinifolius stem methanolic extract. Pharmacognosy Journal 2020; 12(2), 271-275.
M Veerapagu, KR Jeya, A Sankaranarayanan and A Rathika. In vitro antioxidant properties of methanolic extract of Solanum nigrum L fruit. The Pharma Innovation Journal 2018; 7(5), 371-374.
H Brem, MS Golinko, O Stojadinovic, A Kodra, RF Diegelmann, S Vukelic, H Entero, DL Coppock and M Tomic-Canic. Primary cultured fibroblasts derived from patients with chronic wounds : A methodology to produce human cell lines and test putative growth factor therapy such as GMCSF. Journal of Translational Medicine 2008; 6, 75.
H Moravvej, H Memariani, M Memariani, M Kabir-salmani, A Shoae-hassani and F Abdollahimajd. Evaluation of fibroblast viability seeded on acellular human amniotic membrane. Biomed Research Internasional 2021; 2021, 5597758.
A Nawaz, A Jamal, A Arif and Z Parveen. In vitro cytotoxic potential of Solanum nigrum against human cancer cell lines. Saudi Journal Biological Sciences 2021; 28(8), 4786-4792.
K Buranaamnuay. The MTT assay application to measure the viability of spermatozoa: A variety of the assay protocols. Open Veterinary Journal 2021; 11(2), 251-269.
M Ghasemi, T Turnbull, S Sebastian and I Kempson. The mtt assay: Utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. Internasional Journal Molecular Sciences 2021; 22(23), 12827.
RM da Gama, AA Ruggiero, GP Andrade, J Armando-Junior and MAS Pinhal. Optimizing bioactive compounds extraction and cell viability effect of Tithonia diversifolia (Hemsl.) A. Gray dry flower extracts. Journal of Pharmacy and Pharmacognosy Researchs 2025; 13(5), 1313-1326.
AB Achmad. In vitro cytotoxicity test of red okra (abelmoschus esculentus) fruit ethanolic extract on HeLa cells. Journal of Applied Veterinary Science and Technology 2022; 3(1), 22-26.
GA Lozano-Casabianca, SS Arango-Varela, J Aguillón-Osma, MA Llano-Ramírez and ME Maldonado-Celis. Inhibition of cell proliferation and induction of cell cycle arrest in colon cancer cells by lyophilized Mango (Mangifera indica L.) pulp extract. Preventive Nutrition and Food Science 2022; 27(4), 436-447.
R Bar-Shalom, M Bergman, S Grossman, N Azzam, L Sharvit and F Fares. Inula viscosa extract inhibits growth of colorectal cancer cells in vitro and in vivo through induction of apoptosis. Frontiers in Oncology 2019; 9, 227.
S Kumar, VK Maurya, SV Chitti, R Kabir, K Shanker, D Nayak, A Khurana, RK Manchanda, S Gadugu, V Kumar and SK Saxena. Wound healing activity of a novel formulation SKRIN via induction of cell cycle progression and inhibition of PCNA-p21 complex interaction leading to cell survival and proliferation. ACS Pharmacology and Translational Science 2021; 4(1), 352-364
S Manmuan, S Tubtimsri, N Chaothanaphat, N Issaro HM Tantisira and P Manmuan. Determination of the anticancer activity of standardized extract of Centella asiatica (ECa 233) on cell growth and metastatic behavior in oral cancer cells. Research in Pharmaceutical Sciences 2024; 19(2), 121-147.
NM Alfarafisa, AF Khairani, A Achadiyani, Y Chou, F Aulia and MB Firdaus. Assessment of alpha-tocopheryl acetate and metformin hydrochloride as independent agents on human dermal fibroblast viability: Findings from MTT Assay. Althea Medical Journal 2025; 12(2), 125-132.
S Hong, B Lee, JH Kim, EY Kim, M Kim, B Kwon HR Cho, Y Sohn and HS Jung. Solanum nigrum Linne improves DNCB ‑ induced atopic dermatitis ‑ like skin disease in BALB /c mice. Molecular Medicine Reports 2020; 22(4), 2878-2886.
CC Yang, CH Wu, TW Wong, FJ Lai, KC Wei, TK Lin, CS Lin, J Zhao, L Xiang, CH Kao, YJ Wang, CCE Lan and HM Sheu. Effects of Solanum undatum extract (SR-T100) on photocarcinogenesis and photoaging of actinic keratosis. The Journal of Dermatology 2021; 48(3), 344-352.
A Merecz-Sadowska, P Sitarek, E Kucharska, T Kowalczyk, K Zajdel, T Cegliński and R Zajdel. Antioxidant properties of plant-derived phenolic compounds and their effect on skin fibroblast cells. Antioxidants 2021; 10(5), 726.
YM Muflihah, G Gollavelli and YC Ling. Correlation study of antioxidant activity with phenolic and flavonoid compounds in 12 indonesian indigenous herbs. Antioxidants 2021; 10(10), 1530.
TY McCarty and CJ Kearney. Human dermal fibroblast senescence in response to single and recurring oxidative stress. Frontier in Aging 2025; 6, 1504977.
P Singh, SS Bhat, N Singh, BU Venkanna, R Mohamed and RP Rao. Cell-based model systems for validation of various efficacy-based claims for cosmetic ingredients. Cosmetics 2022; 9(5), 107.
F Boraldi, FD Lofaro, S Bonacorsi, A Mazzilli, M Garcia-fernandez and D Quaglino. The role of fibroblasts in skin homeostasis and repair. Biomedicines 2024; 12(7), 1586.
S Dierckx, M Patrizi, M Merino, S González, JL Mullor and R Nergiz-Unal. Collagen peptides affect collagen synthesis and the expression of collagen, elastin, and versican genes in cultured human dermal fibroblasts. Frontiers in Medicine 2024; 11, 1397517.
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