Nanohydroxyapatite synthesized from kombucha SCOBY and its effect on ovariectomized-induced osteoporosis in rats

Document Type : Research Paper


Department of Biology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran



Objective(s): Calcium phosphates, particularly hydroxyapatite, are the main inorganic compounds of vertebrate bone.In this study, nanohydroxyapatite was prepared using kombucha Symbiotic Culture of Bacteria and Yeast (SCOBY), and its effect was investigated on osteoporosis in ovariectomized rats. 
Materials and Methods: Kombucha-nanohydroxyapatite was synthesized by adding phosphoric acid to the kambucha scoby. Characterization of the nanoparticle was performed through X-ray diffraction, X-ray fluorescence, and transmission electron microscopy techniques. Female rats were divided into 5 groups: control, ovariectomized groups, and three ovariectomized groups treated with concentrations of 25, 50, and 100 mg/kg of nanoparticles. At the end of the treatment period, the levels of calcium, phosphorus, parathyroid hormone, and activity of alkaline phosphatase were measured in the blood samples. Calcium and phosphorus levels were also measured in bone and liver. The bone was evaluated histopathologically.
Results: The synthesis of nanohydroxyapatite with particle size of 30 nm was confirmed through the use of X-ray diffraction (XRD) and TEM techniques. A significant increase in calcium and phosphorus levels in the femur bone was observed in the ovariectomized group, which received the highest nanoparticle concentration compared to the ovariectomized group. Parathyroid hormone and alkaline phosphatase activity inhibition were increased in ovariectomized rats following treatment with the highest nanoparticle concentration. In the mentioned group, bone trabeculae proliferation and increased lacuna-containing osteocytes were also observed.
Conclusion: This study suggested that the highest concentration of kombucha-nanohydroxyapatite could be partially absorbed into bone tissues and recover the bone-destructive changes caused by ovariectomy, although additional experiments are needed for confirmation.


1. Zhao R, Shang T, Yuan B, Zhu X, Zhang X, Yang X. Osteoporotic bone recovery by a bamboo-structured bioceramic with controlled release of hydroxyapatite nanoparticles. Bioact Mater 2022; 17 (2022): 379–393.  
2.    Yatsyshyn R, Cherniuk N, Drohomeretska O, Kaminskyi V, Gerych P, Stoika I, et al. A silent epedamic of XXI century: Secondary forms. Arch Clin Med. 2022; 2(28):1-9. 
3.    Castelo-Branco C, Cancelo Hidalgo MJ, Palacios S, Ciria-Recasens M, Fernández-Pareja A. Carbonell-Abella C, et al. Efficacy and safety of ossein-hydroxyapatite complex versus calcium carbonate to prevent bone loss. Climacteric. 2019; 21(2019):1-7.  
4.    Cosman F, de Beur SJ, Le Boff MS. Clinician’s guide to prevention and treatment of osteoporosis. Osteoporos Int. 2014; 25: 2359–2381. 
5.    Compston J, Cooper A, Cooper C, Gittoes N,  Gregson N,   Harvey N, et al. UK clinical guideline for the prevention and treatment of osteoporosis. Arch Osteoporos. 2017;12(43): 1-24.
6.    Martiniakova M, Babikova M, Mondockova V, Blahova J, Kovacova V, Omelka R. The Role of Macronutrients, Micronutrients and Flavonoid Polyphenols in the Prevention and Treatment of Osteoporosis. Nutrients 2022; 14(523): 1-30. 
7.    Cengiz B, Gokce Y, Yildiz N, Aktas Z, Calimli A. Synthesis and characterization of hydroxyapatite nanoparticles. Colloids and Surfaces A: Physicochem Eng Aspects. 2008;  322(1-3): 29-33.
8.    Chandra A, Rajawat J. Skeletal Aging and Osteoporosis: Mechanisms and Therapeutics. Int J Mol Sci. 2021; 22(7): 1-25. 
9.    Maia ALC, Ferreira C de A, Barros ALB de,  Aline Teixeira Maciel e Silva ATM,  Ramaldes GA, Júnior A da SC, et al. Vincristine-loaded hydroxyapatite nanoparticles as a potential delivery system for bone cancer therapy. J Drug Target. 2018; 26(7): 592-603. 
10.    Kapp JM, Sumner W. Kombucha: A systematic review of the empirical evidence of human health benefit. Ann Epidem. 2019; 30: 66-70.
11.    Ledormand P, Desmasures N, Dalmasso M. Phage community involvement in fermented beverages, an open door to technological advances? Crit Rev Food Sci Nutr. 2020;61(17):2911-2920. 
12.    Jayabalan R, Malbasa RV,  Loncar ES, Vitas JS, Sathishkumar MA. Review on kombucha tea-microbiology, composition, fermentation, beneficial effects, toxicity, and tea fungus. Compr Rev Food Sci Food Saf. 2014; 13: 538-550.
13.    Kalaiappan K, Marimuthu S, Rengapillai S, Murugan R,  Premkumar T. Kombucha scoby-based carbon as a green scaffold for high-capacity cathode in lithium–sulfur batteries. Ionics. 2019; 25: 4637-4650.  
14.    KraeuterAK. Pre-Clinical Models: Techniques and Protocols, Methods in Molecular Biology, Methods Mol Biology Clifton N J. 1916; 105: 303-309. 
15.    Abdelhalim MAK, Jarrar BM. Histological alterations in the liver of rats induced by different gold nanoparticle sizes, doses and exposure duration. J Nanobiotechnology. 2012; 10(5): 1-9. 
16.    Fatemi M, Moshtaghian J, Ghaedi K, Jafari dinani N, Naderi GH. Effects of Silver Nanoparticle on the Developing Liver of Rat Pups after Maternal Exposure. Iranian J Pharmaceut Res. 2017;16 (2): 685-693.  
17.    Amiri GhR, Yousefi MH, Aboulhassani MR, Keshavarz MH, Shahbazi D, Fatahian S, et al. Radar absorption of Ni0.7Zn0.3Fe2O4 nanoparticles. Dig J Nanomater Biostructures. 2010; 5(3):719-725.
18.    Mansouri F, Amiri GH, Fatemi M. Synthesis and tissue distribution of CoFe2O4 Na. Nanomed J. 2016; 3(3):196-201. 
19.    Kattimani1 VS, Kondaka S, Lingamaneni KP. Hydroxyapatite Past, Present, and Future in Bone Regeneration. Bone Tissue Regen Insights. 2016;7: 9–19.
20.    Hanh NT, Bich PTN, Thao HTT. Acute and subchronic oral toxicity assessment of calcium hydroxyapatite-alginate in animals. Vietnam J Chem. 2019; 57(1): 16-20 . 
21.    Remya NS, Syama S, Sabareeswaran A, Mohanan PV. Investigation of chronic toxicity of hydroxyapatite nanoparticles administered orally for one year in wistar rats. Mater Sci Engi C. 76(1): 2017; 518–527. 
22.    Hanh NT, Bich PTN, Thao HTT.  Acute and subchronic oral toxicity assessment of calcium hydroxyapatite-alginate in animals. Vietnam J Chem. 2019; 57(1): 16-20.  
23.    Houa T, Zhangb L, Yangc X. Ferulic acid, a natural polyphenol, protects against osteoporosis by activating SIRT1 and NF-κB in neonatal rats with glucocorticoid-induced osteoporosis. Biomed  Pharmacother. 2019;120:1-7.  
24.    Fan H, Cao Y, Xi Z, Wang G, Zheng J. Protective Effect of Panax ginseng on osteoporosis in Ovariectomized Female Guinea-pigs.  Front Med Sci Res. 2021; 3( 5): 24-29. 
25.    Hurjui LL, Hurjui I, Delianu C, Tărniceriu CC, Mârțu AM, Balcoș C, et al. Biological markers importance in the  diagnosis of osteoporosis. Rom J Oral Rehabil. 2020; 2( 4): 181-189.  
26.    Elbahnasawy AS, Valeeva ER, El-Sayed EM, Rakhimov II. The Impact of Thyme and Rosemary on Prevention of Osteoporosis in Rats. J Nutr Metab. 2019; (2019):1-11. 
27.    Berlin T, Bjorkhem I. On the regulatory importance of 1,25-dihydroxyvitamin D3 and dietary calcium on serum levels of 25-hydroxyvitamin D3 in rats. Biochem Biophys Res Commun. (144): 1055–1058.
28.    Bhattarai T, Bhattacharya K, Chaudhuri P, Sengupta P. Correlation of common biochemical markers for bone turnover, serum calcium, and alkaline phosphatase in post-menopausal women. Malays J Med Sci. 2014; 21(1):58-61.
29.    Moore-Schiltz L, Albert JM, Singer ME, Swain J, Nock NL. Dietary intake of calcium and magnesium and the metabolic syndrome in the National Health and Nutrition Examination (NHANES) 2001–2010 data. Be J Nutr. 2015;114(6): 924-935.
30.    Lee KJ, Kim KS, Kim HN, Seo JA, Song SW. Association between dietary calcium and phosphorus intakes, dietary calcium/phosphorus ratio and bone mass in the Korean population. Nutr J. 2014; 13(114): 1-8.. 
31.    Kumar Bhattarai H, Shrestha S, Rokka K, Shakya R. Vitamin D, Calcium, Parathyroid Hormone, and Sex Steroids in Bone Health and Effects of Aging. J Osteoporosis. 2020; 2020: 1-10. 
32.    Ahmadzadeh E, Talebnia Rowshan F, Mashkour M. Enhancement of bone mineral density and body mass in newborn chickens by in ovo injection of ionic-hydroxyapatite nanoparticles of bacterial origin. J Mater Sci Mater Med. 2019; 30(16): 1-11.  
33.    Butera A, Gallo S, Pascadopoli M, Montasser MA, Abd El Latief MH, Modica GG, et al. Home Oral Care with Biomimetic Hydroxyapatite vs. Conventional Fluoridated Toothpastefor the Remineralization and Desensitizing of White Spot Lesions: Randomized Clinical Trial. Int J Environ Res. 2022; (19):1-10. 
34.    Lee AW, Cho SS. Association between Phosphorus Intake and Bone Health in the NHANES Population. Nutr J. 2015;14(28):1-7. 
35.    Sahana H, Kumar Khajuria D, Razdan R, Roy Mahapatra D, Bhat M R, Suresh S, et al. Improvement in Bone Properties by Using Risedronate Adsorbed Hydroxyapatite Novel Nanoparticle Based Formulation in a Rat Model of Osteoporosis. J Biomed Nanotechnol. 2013; 9(2): 193-201.
36.    Nanotechnol. 2013; 9(2): 193-201.