{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T10:59:29Z","timestamp":1788260369786,"version":"build-2803163510"},"reference-count":49,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2023,10,6]],"date-time":"2023-10-06T00:00:00Z","timestamp":1696550400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Medicina"],"abstract":"<jats:p>Objectives: We aimed to investigate the therapeutic effects of thymoquinone (TMQ) treatment in osteonecrotic rats by evaluating protein levels, osteonecrosis (ON) levels, fatty acid degeneration, oxidative status, and plasma levels of Urotensin-II (U-II) and transforming growth factor-beta (TGF-\u03b21). Materials and Methods: 40 weight-matched adult male Wistar rats were grouped as control (n = 10), methylprednisolone acetate (MPA) (n = 10), thymoquinone (TMQ) (n = 10), and MPA + TMQ (n = 10). To induce ON, 15-week-old animals were subcutaneously injected with MPA at a dose of 15 mg\/kg twice weekly for 2 weeks. TMQ was injected into 15-week-old rats via gastric gavage at a dose of 80 mg\/kg per day for 4 weeks. The rats in the MPA + TMQ group were administered TMQ 2 weeks before the MPA injection. At the end of the treatments, cardiac blood samples and femur samples were collected for biochemical and histological evaluations. Results: In the control and TMQ groups, no ON pattern was observed. However, in tissues exposed to MPA, TMQ treatment resulted in significantly decreased ON levels compared to the MPA group. The number of cells that were positive for 8-OHdG and 4-HNE was significantly lower in the MPA + TMQ group than in the MPA group (p &lt; 0.05). In terms of TGF-\u03b21 and U-II levels, we observed that both TGF-\u03b21 (367.40 \u00b1 23.01 pg\/mL vs. 248.9 \u00b1 20.12 pg\/mL) and U-II protein levels (259.5 \u00b1 6.0 ng\/mL vs. 168.20 \u00b1 7.90 ng\/mL) increased significantly in the MPA group compared to the control group (p &lt; 0.001). Furthermore, TGF-\u03b21 (293.50 \u00b1 14.18 pg\/mL) and U-II (174.80 \u00b1 4.2 ng\/mL) protein levels were significantly decreased in the MPA + TMQ group compared to the MPA group (p &lt; 0.05 and p &lt; 0.01, respectively). There was a statistically positive correlation (p &lt; 0.05) between the TGF-\u03b21 and U-II protein levels in all groups (p = 0.002, rcontrol = 0.890; p = 0.02, rTMQ = 0.861; p = 0.024, rMPA+TMQ = 0.868) except for the MPA group (p &lt; 0.03, rMedrol = \u22120.870). Conclusions: As far as we know, this is the first study to demonstrate the curative functions of TMQ on ON by causing a correlated decrease in the expression of U-II and TGF-\u03b21 in the femoral heads of rats.<\/jats:p>","DOI":"10.3390\/medicina59101781","type":"journal-article","created":{"date-parts":[[2023,10,6]],"date-time":"2023-10-06T07:49:29Z","timestamp":1696578569000},"page":"1781","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Effects of Thymoquinone on Urotensin-II and TGF-\u03b21 Levels in Model of Osteonecrosis in Rats"],"prefix":"10.3390","volume":"59","author":[{"given":"Mehmet","family":"Yilmaz","sequence":"first","affiliation":[{"name":"Department of Orthopedic Surgery, 25 Aralik State Hospital, Gaziantep 27090, Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Recep","family":"Dokuyucu","sequence":"additional","affiliation":[{"name":"Department of Physiology, Private Fizyoclinic Wellness Center, Gaziantep 27560, Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Microsurgery Department of the Orthopedics Branch of the Chinese Medical Doctor Association, Group from the Osteonecrosis and Bone Defect Branch of the Chinese Association of Reparative and Reconstructive Surgery, and Microsurgery and Reconstructive Surgery Group of the Orthopedics Branch of the Chinese Medical Association (2017). Chinese Guideline for the Diagnosis and Treatment of Osteonecrosis of the Femoral Head in Adults. Orthop. Surg., 9, 3\u201312.","DOI":"10.1111\/os.12302"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1016\/j.bone.2009.06.030","article-title":"Glucocorticoids, osteocytes, and skeletal fragility: The role of bone vascularity","volume":"46","author":"Weinstein","year":"2010","journal-title":"Bone"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1016\/j.sdentj.2022.01.003","article-title":"Medication-Related Osteonecrosis of the Jaw (MRONJ): A Review of Pathophysiology, Risk Factors, Preventive Measures and Treatment Strategies","volume":"34","author":"AlRowis","year":"2022","journal-title":"Saudi Dent. J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1080\/10641960802251883","article-title":"Osteonecrosis of femoral head in the stroke-prone spontaneously hypertensive rats, especially old rats","volume":"30","author":"Suzuki","year":"2008","journal-title":"Clin. Exp. Hypertens."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1259","DOI":"10.1007\/s43465-022-00635-2","article-title":"Aggressive Presentation and Rapid Progression of Osteonecrosis of the Femoral Head After COVID-19","volume":"56","author":"Dhanasekararaja","year":"2022","journal-title":"Indian J. Orthop."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e22.00036","DOI":"10.2106\/JBJS.RVW.22.00013","article-title":"What Do We Need to Know About Musculoskeletal Manifestations of COVID-19?: A Systematic Review","volume":"10","author":"Pires","year":"2022","journal-title":"JBJS Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.2106\/JBJS.20.00847","article-title":"Musculoskeletal Consequences of COVID-19","volume":"102","author":"Disser","year":"2020","journal-title":"J. Bone Jt. Surg. Am."},{"key":"ref_8","first-page":"7404","article-title":"Alpha-lipoic acid could be a promising treatment in steroid-induced osteonecrosis","volume":"26","author":"Koca","year":"2022","journal-title":"Eur. Rev. Med. Pharmacol. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Koca, I., Dokuyucu, R., Tas, Z., Gokce, H., and Ozcan, O. (2020). Effects of Alpha-Lipoic Acid on TGF-beta 1 and Urotensin-II Levels in Glucocorticoid-Induced Osteonecrosis in Rats. Iran. Red Crescent Med. J., 22.","DOI":"10.5812\/ircmj.97279"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"237","DOI":"10.2485\/jhtb.27.237","article-title":"The Effect of Cape on Steroid Induced Osteonecrosis of the Femoral Head in Rat Model","volume":"27","author":"Uruc","year":"2018","journal-title":"J. Hard Tissue Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.phrs.2015.03.011","article-title":"Thymoquinone and its therapeutic potentials","volume":"95\u201396","author":"Darakhshan","year":"2015","journal-title":"Pharmacol. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Hannan, M.A., Rahman, M.A., Sohag, A.A.M., Uddin, M.J., Dash, R., Sikder, M.H., Rahman, M.S., Timalsina, B., Munni, Y.A., and Sarker, P.P. (2021). Black Cumin (Nigella sativa L.): A Comprehensive Review on Phytochemistry, Health Benefits, Molecular Pharmacology, and Safety. Nutrients, 13.","DOI":"10.3390\/nu13061784"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"113213","DOI":"10.1016\/j.phytochem.2022.113213","article-title":"Therapeutic implications and clinical manifestations of thymoquinone","volume":"200","author":"Alam","year":"2022","journal-title":"Phytochemistry"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Ali, M.Y., Akter, Z., Mei, Z., Zheng, M., Tania, M., and Khan, M.A. (2021). Thymoquinone in autoimmune diseases: Therapeutic potential and molecular mechanisms. Biomed. Pharmacother., 134.","DOI":"10.1016\/j.biopha.2020.111157"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"154768","DOI":"10.1016\/j.phymed.2023.154768","article-title":"Therapeutic application of natural products: NAD(+) metabolism as potential target","volume":"114","author":"Guo","year":"2023","journal-title":"Phytomedicine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1111\/andr.12268","article-title":"Comparison of combined antioxidants and thymoquinone in the prevention of testis ischemia\u2014Reperfusion injury","volume":"5","author":"Erol","year":"2017","journal-title":"Andrology"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Pal, R.R., Rajpal, V., Singh, P., and Saraf, S.A. (2021). Recent Findings on Thymoquinone and Its Applications as a Nanocarrier for the Treatment of Cancer and Rheumatoid Arthritis. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13060775"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"553","DOI":"10.52312\/jdrs.2022.752","article-title":"The effects of thymoquinone on steroid-induced femoral head osteonecrosis: An experimental study in rats","volume":"33","author":"Dasci","year":"2022","journal-title":"Jt. Dis. Relat. Surg."},{"key":"ref_19","first-page":"111","article-title":"Effects of sustained delivery of thymoqiunone on bone healing of male rats","volume":"40","author":"Kirui","year":"2004","journal-title":"Biomed. Sci. Instrum."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Kalamegam, G., Alfakeeh, S.M., Bahmaid, A.O., AlHuwait, E.A., Gari, M.A., Abbas, M.M., Ahmed, F., Abu-Elmagd, M., and Pushparaj, P.N. (2020). In vitro Evaluation of the Anti-inflammatory Effects of Thymoquinone in Osteoarthritis and in silico Analysis of Inter-Related Pathways in Age-Related Degenerative Diseases. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.00646"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1042\/CBI20090104","article-title":"Urotensin-2 promotes collagen synthesis via ERK1\/2-dependent and ERK1\/2-independent TGF-beta1 in neonatal cardiac fibroblasts","volume":"35","author":"Dai","year":"2011","journal-title":"Cell Biol. Int."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.regpep.2012.12.006","article-title":"Urotensin II-induced collagen synthesis in cultured smooth muscle cells from rat aortic media and a possible involvement of transforming growth factor-beta1\/Smad2\/3 signaling pathway","volume":"182","author":"Zhao","year":"2013","journal-title":"Regul. Pept."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/j.peptides.2014.01.010","article-title":"Urotensin II (U-II), a novel cyclic peptide, possibly associated with the pathophysiology of osteoarthritis","volume":"54","author":"Gogebakan","year":"2014","journal-title":"Peptides"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"3442","DOI":"10.1002\/art.11328","article-title":"Reduction of osteophyte formation and synovial thickening by adenoviral overexpression of transforming growth factor beta\/bone morphogenetic protein inhibitors during experimental osteoarthritis","volume":"48","author":"Scharstuhl","year":"2003","journal-title":"Arthritis Rheum."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"R65","DOI":"10.1186\/ar1931","article-title":"TGF beta-induced cartilage repair is maintained but fibrosis is blocked in the presence of Smad7","volume":"8","author":"Vitters","year":"2006","journal-title":"Arthritis Res. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"511","DOI":"10.3109\/10641963.2010.496511","article-title":"Pentosan reduces osteonecrosis of femoral head in SHRSP","volume":"32","author":"Miyata","year":"2010","journal-title":"Clin. Exp. Hypertens."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.cbi.2018.02.006","article-title":"Improvement of antihyperglycemic activity of nano-thymoquinone in rat model of type-2 diabetes","volume":"295","author":"Rani","year":"2018","journal-title":"Chem. Biol. Interact."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"87","DOI":"10.3109\/17453674.2011.641103","article-title":"Pravastatin reduces steroid-induced osteonecrosis of the femoral head in SHRSP rats","volume":"83","author":"Nozaki","year":"2012","journal-title":"Acta Orthop."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1136\/bmj.288.6419.741","article-title":"Early diagnosis and treatment of steroid induced avascular necrosis of bone","volume":"288","author":"Nixon","year":"1984","journal-title":"Br. Med. J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.1002\/art.1780401119","article-title":"Effects of pulse methylprednisolone on bone and marrow tissues: Corticosteroid-induced osteonecrosis in rabbits","volume":"40","author":"Yamamoto","year":"1997","journal-title":"Arthritis Rheum."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1302\/0301-620X.83B2.0830274","article-title":"Selective reduction of bone blood flow by short-term treatment with high-dose methylprednisolone. An experimental study in pigs","volume":"83","author":"Drescher","year":"2001","journal-title":"J. Bone Jt. Surg. Br. Vol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1093\/rheumatology\/40.2.196","article-title":"A high low-density lipoprotein cholesterol to high-density lipoprotein cholesterol ratio as a potential risk factor for corticosteroid-induced osteonecrosis in rabbits","volume":"40","author":"Miyanishi","year":"2001","journal-title":"Rheumatology"},{"key":"ref_33","first-page":"453","article-title":"Effect of methylprednisolone on coagulation","volume":"91","author":"Pandit","year":"1999","journal-title":"J. Natl. Med. Assoc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1007\/s00776-010-1452-6","article-title":"Role of hypercoagulability in steroid-induced femoral head necrosis in rabbits","volume":"15","author":"Guan","year":"2010","journal-title":"J. Orthop. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2483","DOI":"10.1111\/j.1538-7836.2010.04034.x","article-title":"Systematic review on the effect of glucocorticoid use on procoagulant, anti-coagulant and fibrinolytic factors","volume":"8","author":"Nur","year":"2010","journal-title":"J. Thromb. Haemost. JTH"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3387","DOI":"10.1002\/art.20517","article-title":"Combined effects of an anticoagulant and a lipid-lowering agent on the prevention of steroid-induced osteonecrosis in rabbits","volume":"50","author":"Motomura","year":"2004","journal-title":"Arthritis Rheum."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1111\/j.1365-2613.2010.00705.x","article-title":"Effects of an anticoagulant and a lipid-lowering agent on the prevention of steroid-induced osteonecrosis in rabbits","volume":"91","author":"Kang","year":"2010","journal-title":"Int. J. Exp. Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1177\/1535370213519215","article-title":"Pravastatin prevents steroid-induced osteonecrosis in rats by suppressing PPARgamma expression and activating Wnt signaling pathway","volume":"239","author":"Jiang","year":"2014","journal-title":"Exp. Biol. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1093\/rheumatology\/keh518","article-title":"DNA oxidation injury in bone early after steroid administration is involved in the pathogenesis of steroid-induced osteonecrosis","volume":"44","author":"Ichiseki","year":"2005","journal-title":"Rheumatology"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.cbi.2013.11.004","article-title":"Effect of glucocorticoid-induced oxidative stress on the expression of Cbfa1","volume":"207","author":"Feng","year":"2014","journal-title":"Chem. Biol. Interact."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1007\/s00776-014-0654-8","article-title":"Beneficial effect of grape seed proanthocyanidin extract in rabbits with steroid-induced osteonecrosis via protecting against oxidative stress and apoptosis","volume":"20","author":"Song","year":"2015","journal-title":"J. Orthop. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1007\/s00296-011-1846-6","article-title":"Lipoic acid prevents steroid-induced osteonecrosis in rabbits","volume":"32","author":"Lu","year":"2012","journal-title":"Rheumatol. Int."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Kohandel, Z., Farkhondeh, T., Aschner, M., and Samarghandian, S. (2021). Anti-inflammatory effects of thymoquinone and its protective effects against several diseases. Biomed. Pharmacother., 138.","DOI":"10.1016\/j.biopha.2021.111492"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.ijporl.2017.03.011","article-title":"The effect of caffeic acid phenethyl ester and thymoquinone on otitis media with effusion in rats","volume":"96","author":"Gulmez","year":"2017","journal-title":"Int. J. Pediatr. Otorhinolaryngol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"285","DOI":"10.5152\/iao.2017.4213","article-title":"The Role of Topical Thymoquinone in the Treatment of Acute Otitis Externa; an Experimental Study in Rats","volume":"14","author":"Demirel","year":"2018","journal-title":"J. Int. Adv. Otol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1831","DOI":"10.3892\/ijmm.2016.2789","article-title":"Role of urotensin II in advanced glycation end product-induced extracellular matrix synthesis in rat proximal tubular epithelial cells","volume":"38","author":"Tian","year":"2016","journal-title":"Int. J. Mol. Med."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1186\/1479-5876-9-102","article-title":"Bisphosphonate-associated osteonecrosis of the jaw is linked to suppressed TGFbeta1-signaling and increased Galectin-3 expression: A histological study on biopsies","volume":"9","author":"Wehrhan","year":"2011","journal-title":"J. Transl. Med."},{"key":"ref_48","first-page":"1161","article-title":"Roles of TGF-beta 1 signaling in the development of osteoarthritis","volume":"31","author":"Fang","year":"2016","journal-title":"Histol. Histopathol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/j.bbadis.2008.01.006","article-title":"Role of transforming growth factor-beta superfamily signaling pathways in human disease","volume":"1782","author":"Gordon","year":"2008","journal-title":"Biochim. Biophys. Acta"}],"container-title":["Medicina"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1648-9144\/59\/10\/1781\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:02:02Z","timestamp":1760130122000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1648-9144\/59\/10\/1781"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,6]]},"references-count":49,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["medicina59101781"],"URL":"https:\/\/doi.org\/10.3390\/medicina59101781","relation":{},"ISSN":["1648-9144"],"issn-type":[{"value":"1648-9144","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,6]]}}}