Dickkopf‑3 and β‑catenin play opposite roles in the Wnt/β‑catenin pathway during the abnormal subchondral bone formation of human knee osteoarthritis

Liang,Xuegang; Jin,Qunhua; Yang,Xiaochun; Jiang,Wenhui

doi:10.3892/ijmm.2022.5103

Dickkopf‑3 and β‑catenin play opposite roles in the Wnt/β‑catenin pathway during the abnormal subchondral bone formation of human knee osteoarthritis

Authors:
- Xuegang Liang
- Qunhua Jin
- Xiaochun Yang
- Wenhui Jiang
View Affiliations

Affiliations: Department of Orthopedics, Ningxia Medical University General Hospital, Ningxia Hui Autonomous Region 750000, P.R. China, Clinical Medical College, Xi'an Medical College, Xi'an, Shanxi 710000, P.R. China
Published online on: February 9, 2022 https://doi.org/10.3892/ijmm.2022.5103
Article Number: 48
Copyright: © Liang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Osteoarthritis (OA) is condition which poses a main concern to the aging population and its severity is expected to increase with the increasing life expectancy. In the future, several possible targets for OA treatment need to be defined. Dickkopf‑related protein 3 (DKK3) is an atypical member of the Wnt‑antagonistic dickkopf‑related protein (DKK) family. The availability of research into the role of DKK3 in the abnormal remodeling of subchondral bone in human knee joints is currently limited. Thus, the aim of the present study was the evaluation of DKK3 expression in the abnormal bone remodeling of subchondral bone in human knee OA in order to clarify the role of DKK3 in subchondral bone remodeling and to acknowledge its potential relevance to β‑catenin. In total, 38 specimens were collected from osteotomies of the medial tibial plateau of the human knee. The patient samples were then divided into the normal, mild, moderate and severe symptom groups, according to the Osteoarthritis Research Society International (OARSI) score. Following hematoxylin and eosin (H&E) and Safranin O‑fast green staining for alkaline phosphatase (AZO method), changes in the distribution and number of osteocytes in the subchondral bone and the degree of sclerosis of the subchondral bone were observed. Immunohistochemical staining, immunofluorescence, western blot analysis and reverse‑transcription quantitative PCR (RT‑qPCR) were used for the detection of DKK3 and β‑catenin expression level changes in osteoblasts in the subchondral bone of the medial tibial plateau. H&E and alkaline phosphatase staining revealed that the total number of osteocytes in the subchondral bone increased with the severity of the disease. The samples were also evaluated using Safranin O‑Fast Green staining and were attributed a score according to the OARSI scoring system: The scoring number and cartilage damage increased along with OA severity. Immunohistochemistry and immunofluorescence assays demonstrated that β‑catenin expression in osteocytes increased from mild to moderate, whereas DKK3 expression decreased with the development of arthritis from normal, mild to moderate. According to the results of western blot analysis, β‑catenin expression was higher in moderate OA and then decreased in severe OA. On the other hand, the DKK3 levels decreased along with the progression from normal, mild to moderate OA. The results of RT‑qPCR demonstrated that β‑catenin and DKK3 gene expression differed with the degree of OA. On the whole, the present study demonstrates that DKK3 and β‑catenin may play opposite roles in OA subchondral bone remodeling.

View Figures

View References

1	Qin HJ, Xu T, Wu HT, Yao ZL, Hou YL, Xie YH, Su JW, Cheng CY, Yang KF, Zhang XR, et al: SDF-1/CXCR4 axis coordinates crosstalk between subchondral bone and articular cartilage in osteoarthritis pathogenesis. Bone. 125:140–150. 2019. View Article : Google Scholar : PubMed/NCBI
2	Anderson-MacKenzie JM, Quasnichka HL, Starr RL, Lewis EJ, Billingham ME and Bailey AJ: Fundamental subchondral bone changes in spontaneous knee osteoarthritis. Int J Biochem Cell Biol. 37:224–236. 2005. View Article : Google Scholar
3	Omoumi P, Babel H, Jolles BM and Favre J: Relationships between cartilage thickness and subchondral bone mineral density in non-osteoarthritic and severely osteoarthritic knees: In vivo concomitant 3D analysis using CT arthrography. Osteoarthritis Cartilage. 27:621–629. 2019. View Article : Google Scholar : PubMed/NCBI
4	Neve A, Corrado A and Cantatore FP: Osteoblast physiology in normal and pathological conditions. Cell Tissue Res. 343:289–302. 2010. View Article : Google Scholar : PubMed/NCBI
5	Maruotti N, Corrado A and Cantatore FP: Osteoblast role in osteoarthritis pathogenesis. J Cell Physiol. 232:2957–2963. 2017. View Article : Google Scholar :
6	Di Pompo G, Errani C, Gillies R, Mercatali L, Ibrahim T, Tamanti J, Baldini N and Avnet S: Acid-induced inflammatory cytokines in osteoblasts: A guided path to osteolysis in bone metastasis. Front Cell Dev Biol. 9:6785322021. View Article : Google Scholar :
7	Wang Y, Fan X, Xing L and Tian F: Wnt signaling: A promising target for osteoarthritis therapy. Cell Commun Signal. 17:972019. View Article : Google Scholar : PubMed/NCBI
8	Lories RJ and Monteagudo S: Review article: Is Wnt signaling an attractive target for the treatment of osteoarthritis? Rheumatol Ther. 7:259–270. 2020. View Article : Google Scholar : PubMed/NCBI
9	Cherifi C, Monteagudo S and Lories RJ: Promising targets for therapy of osteoarthritis: A review on the Wnt and TGF-β signalling pathways. Ther Adv Musculoskel. 13:1759720X2110069592021.
10	Chen H, Tan XN, Hu S, Liu RQ, Peng LH, Li YM and Wu P: Molecular mechanisms of chondrocyte proliferation and differentiation. Front Cell Dev Biol. 9:6641682021. View Article : Google Scholar : PubMed/NCBI
11	Staines KA, Macrae VE and Farquharson C: Cartilage development and degeneration: A Wnt Wnt situation. Cell Biochem Funct. 30:633–642. 2012. View Article : Google Scholar
12	Hill TP, Später D, Taketo MM, Birchmeier W and Hartmann C: Canonical Wnt/beta-catenin signaling prevents osteoblasts from differentiating into chondrocytes. Dev Cell. 8:727–738. 2005. View Article : Google Scholar : PubMed/NCBI
13	Zhu M, Tang D, Wu Q, Hao S, Chen M, Xie C, Rosier RN, O'Keefe RJ, Zuscik M and Chen D: Activation of beta-catenin signaling in articular chondrocytes leads to osteoarthritis-like phenotype in adult beta-catenin conditional activation mice. J Bone Miner Res. 24:12–21. 2009. View Article : Google Scholar
14	Fjeld K, Kettunen P, Furmanek T, Kvinnsland IH and Luukko K: Dynamic expression of Wnt signaling-related Dickkopf1, -2, and -3 mRNAs in the developing mouse tooth. Dev Dyn. 233:161–166. 2005. View Article : Google Scholar : PubMed/NCBI
15	Chen J, Yang C, Yang Y, Liang Q, Xie K, Liu J and Tang Y: Targeting DKK1 prevents development of alcohol-induced osteonecrosis of the femoral head in rats. Am J Transl Res. 13:2320–2330. 2021.PubMed/NCBI
16	Baetta R and Banfi C: Dkk (Dickkopf) proteins. Arterioscler Thromb Vasc Biol. 39:1330–1342. 2019. View Article : Google Scholar : PubMed/NCBI
17	Lee EJ, Nguyen QTT and Lee M: Dickkopf-3 in human malignant tumours: A clinical viewpoint. Anticancer Res. 40:5969–5979. 2020. View Article : Google Scholar : PubMed/NCBI
18	Pritzker KP, Gay S, Jimenez SA, Ostergaard K, Pelletier JP, Revell PA, Salter D and van den Berg WB: Osteoarthritis cartilage histopathology: Grading and staging. Osteoarthritis Cartilage. 14:13–29. 2006. View Article : Google Scholar
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
20	Zhang L and Wen C: Osteocyte dysfunction in joint homeostasis and osteoarthritis. Int J Mol Sci. 22:65222021. View Article : Google Scholar : PubMed/NCBI
21	Tian J, Gao SG, Li YS, Cheng C, Deng ZH, Luo W and Zhang FJ: The β-catenin/TCF-4 pathway regulates the expression of OPN in human osteoarthritic chondrocytes. J Orthop Surg Res. 15:3442020. View Article : Google Scholar
22	Li W, Xiong Y, Chen W and Wu L: Wnt/β-catenin signaling may induce senescence of chondrocytes in osteoarthritis. Exp Ther Med. 20:2631–2638. 2020.PubMed/NCBI
23	Xuan F, Yano F, Mori D, Chijimatsu R, Maenohara Y, Nakamoto H, Mori Y, Makii Y, Oichi T, Taketo MM, et al: Wnt/β-catenin signaling contributes to articular cartilage homeostasis through lubricin induction in the superficial zone. Arthritis Res Ther. 21:2472019. View Article : Google Scholar
24	Yu H, Liu Y, Yang X, He J, Zhang F, Zhong Q and Guo X: Strontium ranelate promotes chondrogenesis through inhibition of the Wnt/β-catenin pathway. Stem Cell Res Ther. 12:2962021. View Article : Google Scholar
25	Le NH, Franken P and Fodde R: Tumour-stroma interactions in colorectal cancer: Converging on beta-catenin activation and cancer stemness. Brit J Cancer. 98:1886–1893. 2008. View Article : Google Scholar : PubMed/NCBI
26	Ren C, Gu X, Li H, Lei S, Wang Z, Wang J, Yin P, Zhang C, Wang F and Liu C: The role of DKK1 in Alzheimer's disease: A potential intervention point of brain damage prevention? Pharmacol Res. 144:331–335. 2019. View Article : Google Scholar : PubMed/NCBI
27	Uribe D, Cardona A, Esposti DD, Cros MP, Cuenin C, Herceg Z, Camargo M and Cortés-Mancera FM: Antiproliferative effects of epigenetic modifier drugs through E-cadherin up-regulation in liver cancer cell lines. Ann Hepatol. 17:444–460. 2018. View Article : Google Scholar : PubMed/NCBI
28	Suwa T, Chen M, Hawks CL and Hornsby PJ: Zonal expression of dickkopf-3 and components of the Wnt signalling pathways in the human adrenal cortex. J Endocrinol. 178:149–158. 2003. View Article : Google Scholar : PubMed/NCBI
29	Niehrs C: Function and biological roles of the Dickkopf family of Wnt modulators. Oncogene. 25:7469–7481. 2006. View Article : Google Scholar : PubMed/NCBI
30	Aslan H, Ravid-Amir O, Clancy BM, Rezvankhah S, Pittman D, Pelled G, Turgeman G, Zilberman Y, Gazit Z, Hoffmann A, et al: Advanced molecular profiling in vivo detects novel function of dickkopf-3 in the regulation of bone formation. J Bone Miner Res. 21:1935–1945. 2006. View Article : Google Scholar : PubMed/NCBI
31	De Palma A and Nalesso G: WNT signalling in osteoarthritis and its pharmacological targeting. Handb Exp Pharmacol. 269:337–356. 2021. View Article : Google Scholar : PubMed/NCBI
32	Funck-Brentano T, Bouaziz W, Marty C, Geoffroy V, Hay E and Cohen-Solal M: Dkk-1-mediated inhibition of Wnt signaling in bone ameliorates osteoarthritis in mice. Arthritis Rheumatol. 66:3028–3039. 2014. View Article : Google Scholar : PubMed/NCBI
33	Jiang A, Xu P, Sun S, Zhao Z, Tan Q, Li W, Song C and Leng H: Cellular alterations and crosstalk in the osteochondral joint in osteoarthritis and promising therapeutic strategies. Connect Tissue Res. 62:709–719. 2021. View Article : Google Scholar : PubMed/NCBI
34	Liao F, Hu X and Chen R: The effects of omarigliptin on promoting osteoblastic differentiation. Bioengineered. 12:11837–11846. 2021. View Article : Google Scholar : PubMed/NCBI
35	Miyamoto S, Yoshikawa H and Nakata K: Axial mechanical loading to ex vivo mouse long bone regulates endochondral ossification and endosteal mineralization through activation of the BMP-Smad pathway during postnatal growth. Bone Rep. 15:1010882021. View Article : Google Scholar
36	Vincent TL and Wann AKT: Mechanoadaptation: Articular cartilage through thick and thin. J Physiol. 597:1271–1281. 2019. View Article : Google Scholar :
37	Bhatla JL, Kroker A, Manske SL, Emery CA and Boyd SK: Differences in subchondral bone plate and cartilage thickness between women with anterior cruciate ligament reconstructions and uninjured controls. Osteoarthritis Cartilage. 26:929–939. 2018. View Article : Google Scholar : PubMed/NCBI
38	Hu W, Chen Y, Dou C and Dong S: Microenvironment in subchondral bone: Predominant regulator for the treatment of osteoarthritis. Ann Rheum Dis. 80:413–422. 2020. View Article : Google Scholar
39	Fell NLA, Lawless BM, Cox SC, Cooke ME, Eisenstein NM, Shepherd DET and Espino DM: The role of subchondral bone, and its histomorphology, on the dynamic viscoelasticity of cartilage, bone and osteochondral cores. Osteoarthritis Cartilage. 27:535–543. 2019. View Article : Google Scholar :
40	Smieszek A, Marcinkowska K, Pielok A, Sikora M, Valihrach L and Marycz K: The role of miR-21 in osteoblasts-osteoclasts coupling in vitro. Cells. 9:4792020. View Article : Google Scholar
41	Chu Y, Gao Y, Yang Y, Liu Y, Guo N, Wang L, Huang W, Wu L, Sun D and Gu W: β-Catenin mediates fluoride-induced aberrant osteoblasts activity and osteogenesis. Environ Pollut. 265:1147342020. View Article : Google Scholar
42	Huang Y, Jiang L, Yang H, Wu L, Xu N, Zhou X and Li J: Variations of Wnt/β-catenin pathway-related genes in susceptibility to knee osteoarthritis: A three-centre case-control study. J Cell Mol Med. 23:8246–8257. 2019. View Article : Google Scholar : PubMed/NCBI
43	Charlier E, Malaise O, Deroyer C, Zeddou M, Neuville S, Cobraiville G, Gillet P, Kurth W, de Seny D, Relic B and Malaise MG: Dickkopf 3 (DKK3) is increased along human hip OA chondrocytes dedifferentiation and can modulate Wnt/B-catenin and TGFβ Alk1/Smad1/5 signaling pathways, as well as leptin production. Osteoarthritis Cartilage. 24(Suppl 1): S1822016. View Article : Google Scholar