Angiogenesis and bone regeneration by mesenchymal stem cell transplantation with danshen in a rabbit model of avascular necrotic femoral head

Wu,Yungang; Zhang,Chunwu; Wu,Jianjing; Han,Yan; Wu,Chunlei

doi:10.3892/etm.2019.7556

Angiogenesis and bone regeneration by mesenchymal stem cell transplantation with danshen in a rabbit model of avascular necrotic femoral head

Authors:
- Yungang Wu
- Chunwu Zhang
- Jianjing Wu
- Yan Han
- Chunlei Wu
View Affiliations

Affiliations: Department of Orthopedics, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China
Published online on: May 8, 2019 https://doi.org/10.3892/etm.2019.7556
Pages: 163-171
Copyright: © Wu 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

The present study aimed to explore the potential of combined treatment with mesenchymal stem cells (MSCs) and danshen for angiogenesis and bone regeneration in a rabbit model of avascular necrosis of femoral head (ANFH). A rabbit model of ANFH was established using the Shwartzman reaction with methylprednisolone and Escherichia coli endotoxin injection. Magnetic resonance imaging (MRI) and histopathological examination were used to evaluate the rabbit model of ANFH. The rabbits were randomly divided into the danshen group, the MSCs group, the danshen combined with MSCs group and the model group (treated with physiological saline). The expression level of monocyte chemoattractant protein‑1 (MCP‑1) and stromal cell‑derived factor‑1 (SDF‑1) were determined by reverse transcription polymerase chain reaction (RT‑PCR). The expression level of bone morphogenetic protein‑2 (BMP‑2) and vascular endothelial growth factor (VEGF) were detected by immunofluorescence and the mRNA expression of BMP‑2 and VEGF were detected by RT‑PCR. Typical osteonecrosis occurred in the rabbit model of ANFH, which indicated that the model was successfully established. MCP‑1 and SDF‑1 were significantly increased in the model group compared with the normal group (P<0.05). Following the administration of MSCs and Salvia miltiorrhiza (danshen), MSCs labeled with 5‑bromo‑2‑deoxyuridine were observed to be gathered in the necrotic area. The increased migration of MSCs to the necrotic area may be due to the upregulated expression of the chemokines MCP‑1 and SDF‑1. ANFH treated with danshen combined with MSCs may promote revascularization by increasing the expression of VEGF and BMP‑2 in the femoral head, promoting re‑ossification and revascularization. Danshen combined with the transplantation of MSCs may be regarded as a novel therapy for the treatment of ANFH in a clinical setting.

View Figures

View References

1	Zhao D, Cui D, Wang B, Tian F, Guo L, Yang L, Liu B and Yu X: Treatment of early stage osteonecrosis of the femoral head with autologous implantation of bone marrow-derived and cultured mesenchymal stem cells. Bone. 50:325–330. 2012. View Article : Google Scholar : PubMed/NCBI
2	Ohzono K, Saito M, Takaoka K, Ono K, Saito S, Nishina T and Kadowaki T: Natural history of nontraumatic avascular necrosis of the femoral head. J Bone Joint Surg Br. 73:68–72. 1991. View Article : Google Scholar : PubMed/NCBI
3	Arlet J: Nontraumatic avascular necrosis of the femoral head. Past, present, and future. Clin Orthop Relat Res. 277:12–21. 1992.
4	Mankin HJ: Nontraumatic necrosis of bone (osteonecrosis). N Engl J Med. 326:1473–1479. 1992. View Article : Google Scholar : PubMed/NCBI
5	Roshan A and Ram S: The neglected femoral neck fracture in young adults: Review of a challenging problem. Clin Med Res. 6:33–39. 2008. View Article : Google Scholar : PubMed/NCBI
6	Ciapetti G, Granchi D, Fotia C, Savarino L, Dallari D, Del Piccolo N, Donati DM and Baldini N: Effects of hypoxia on osteogenic differentiation of mesenchymal stromal cells used as a cell therapy for avascular necrosis of the femoral head. Cytotherapy. 18:1087–1099. 2016. View Article : Google Scholar : PubMed/NCBI
7	Tabatabaee RM, Saberi S, Parvizi J, Mortazavi SM and Farzan M: Combining Concentrated autologous bone marrow stem cells injection with core decompression improves outcome for patients with early-stage osteonecrosis of the femoral head: A comparative study. J Arthroplasty. 30 (Suppl 9):S11–S15. 2015. View Article : Google Scholar
8	Song HM, Wei YC, Li N, Wu B, Xie N, Zhang KM, Wang SZ and Wang HM: Effects of Wenyangbushen formula on the expression of VEGF, OPG, RANK and RANKL in rabbits with steroid-induced femoral head avascular necrosis. Mol Med Rep. 12:8155–8161. 2015. View Article : Google Scholar : PubMed/NCBI
9	Rajpura A, Wright AC and Board TN: Medical management of osteonecrosis of the hip: A review. Hip Int. 21:385–392. 2011. View Article : Google Scholar : PubMed/NCBI
10	Zhai L, Sun N, Zhang B, Liu ST, Zhao Z, Jin HC, Ma XL and Xing GY: Effects of focused extracorporeal shock waves on bone marrow mesenchymal stem cells in patients with avascular necrosis of the femoral head. Ultrasound Med Biol. 42:753–762. 2016. View Article : Google Scholar : PubMed/NCBI
11	Kim N and Cho SG: Clinical applications of mesenchymal stem cells. Korean J intern Med. 28:387–402. 2013. View Article : Google Scholar : PubMed/NCBI
12	Wen Q, Zhou C, Luo W, Zhou M and Ma L: Pro-osteogenic effects of fibrin glue in treatment of avascular necrosis of the femoral head in vivo by hepatocyte growth factor-transgenic mesenchymal stem cells. J Transl Med. 12:1142014. View Article : Google Scholar : PubMed/NCBI
13	Ye YT, Zhong W, Sun P, Wang D, Wang C, Hu LM and Qian JQ: Apoptosis induced by the methanol extract of salvia miltiorrhiza bunge in non-small cell lung cancer through PTEN-mediated inhibition of PI3K/Akt pathway. J Ethnopharmacol. 200:107–116. 2017. View Article : Google Scholar : PubMed/NCBI
14	Park CH, Shin SH, Lee EK, Kim DH, Kim MJ, Roh SS, Yokozawa T and Chung HY: Magnesium lithospermate B from salvia miltiorrhiza bunge ameliorates aging-induced renal inflammation and senescence via NADPH oxidase-mediated reactive oxygen generation. Phytother Res. 31:721–728. 2017. View Article : Google Scholar : PubMed/NCBI
15	Zhou L, Zuo Z and Chow MS: Danshen: An overview of its chemistry, pharmacology, pharmacokinetics, and clinical use. J Clin Pharmacol. 45:1345–1359. 2005. View Article : Google Scholar : PubMed/NCBI
16	Argenbright LW and Barton RW: The Shwartzman response: A model of ICAM-1 dependent vasculitis. Agents Actions. 34:208–210. 1991. View Article : Google Scholar : PubMed/NCBI
17	Xia CS, Zuo AJ, Wang CY and Wang YZ: Isolation of rabbit bone marrow mesenchymal stem cells using density gradient centrifugation and adherence screening methods. Minerva Medica. 104:519–525. 2013.PubMed/NCBI
18	Yamamoto T, Hirano K, Tsutsui H, Sugioka Y and Sueishi K: Corticosteroid enhances the experimental induction of osteonecrosis on rabbits with Shwartzman reaction. Clin Orthop Relat Res. 316:235–243. 1995. View Article : Google Scholar
19	Saeed H, Abdallah BM, Ditzel N, Catala-Lehnen P, Qiu W, Amling M and Kassem M: Telomerase-deficiency-related bone loss is caused by intrinsic impairment of mesenchymal stem cell (MSC) functions and increased osteoclastogenesis due to pro-inflammatory micro-environment. Bone. 47:S392010. View Article : Google Scholar
20	Wang D, Wang G, Liu M, Sun L, Zong W, Jiang H, Zhang H, Li H, Gong J and Sun S: A novel animal model of osteonecrosis of the femoral head induced using a magnetic resonance imaging-guided argon-helium cryotherapy system. Exp Ther Med. 7:1525–1528. 2014. View Article : Google Scholar : PubMed/NCBI
21	Wen Q, Ma L, Chen YP, Yang L, Luo W and Wang XN: A rabbit model of hormone-induced early avascular necrosis of the femoral head. Biomed Environ Sci. 21:398–403. 2008. View Article : Google Scholar : PubMed/NCBI
22	Nowak DA and Yeung J: Steroid-induced osteonecrosis in dermatology: A review. J Cutan Med Surg. 19:358–360. 2015. View Article : Google Scholar : PubMed/NCBI
23	Li Z, Liao W, Zhao Q, Liu M, Xia W, Yang Y and Shao N: Angiogenesis and bone regeneration by allogeneic mesenchymal stem cell intravenous transplantation in rabbit model of avascular necrotic femoral head. J Surg Res. 183:193–203. 2013. View Article : Google Scholar : PubMed/NCBI
24	Bartonicek J, Vavra J and Bartoska R: Operative treatment of avascular necrosis of the femoral head after slipped capital femoral epiphysis. Arch Orthop Trauma Surg. 131:497–502. 2011. View Article : Google Scholar : PubMed/NCBI
25	Zlotnik A and Yoshie O: Chemokines: A new classification system and their role in immunity. Immunity. 12:121–127. 2000. View Article : Google Scholar : PubMed/NCBI
26	Fritz EA, Glant TT, Vermes C, Jacobs JJ and Roebuck KA: Chemokine gene activation in human bone marrow-derived osteoblasts following exposure to particulate wear debris. J Biomed Mater Res A. 77:192–201. 2006. View Article : Google Scholar : PubMed/NCBI
27	Marra F and Tacke F: Roles for chemokines in liver disease. Gastroenterology. 147:577–594, e571. 2014. View Article : Google Scholar : PubMed/NCBI
28	Wasmuth HE, Tacke F and Trautwein C: Chemokines in liver inflammation and fibrosis. Semin Liver Dis. 30:215–225. 2010. View Article : Google Scholar : PubMed/NCBI
29	Zhang C, Wang KZ, Qiang H, Tang YL, Li Q, Li M and Dang XQ: Angiopoiesis and bone regeneration via co-expression of the hVEGF and hBMP genes from an adeno-associated viral vector in vitro and in vivo. Acta Pharmacol Sin. 31:821–830. 2010. View Article : Google Scholar : PubMed/NCBI
30	Young S, Patel ZS, Kretlow JD, Murphy MB, Mountziaris PM, Baggett LS, Ueda H, Tabata Y, Jansen JA, Wong M and Mikos AG: Dose effect of dual delivery of vascular endothelial growth factor and bone morphogenetic protein-2 on bone regeneration in a rat critical-size defect model. Tissue Eng Part A. 15:2347–2362. 2009. View Article : Google Scholar : PubMed/NCBI
31	Zhang C, Ma J, Li M, Li XH, Dang XQ and Wang KZ: Repair effect of coexpression of the hVEGF and hBMP genes via an adeno-associated virus vector in a rabbit model of early steroid-induced avascular necrosis of the femoral head. Transl Res. 166:269–280. 2015. View Article : Google Scholar : PubMed/NCBI
32	Aldridge JM III and Urbaniak JR: Avascular necrosis of the femoral head: Role of vascularized bone grafts. Orthop Clin North Am. 3813–22. (v)2007. View Article : Google Scholar : PubMed/NCBI
33	Fairbank AC, Bhatia D, Jinnah RH and Hungerford DS: Long-term results of core decompression for ischaemic necrosis of the femoral head. J Bone Joint Surg Br. 77:42–49. 1995. View Article : Google Scholar : PubMed/NCBI
34	Koo KH, Kim R, Ko GH, Song HR, Jeong ST and Cho SH: Preventing collapse in early osteonecrosis of the femoral head. A randomised clinical trial of core decompression. J Bone Joint Surg Br. 77:870–874. 1995. View Article : Google Scholar : PubMed/NCBI
35	Mont MA, Carbone JJ and Fairbank AC: Core decompression versus nonoperative management for osteonecrosis of the hip. Clin Orthop Relat Res. 324:169–178. 1996. View Article : Google Scholar
36	Aaron RK, Lennox D, Bunce GE and Ebert T: The conservative treatment of osteonecrosis of the femoral head. A comparison of core decompression and pulsing electromagnetic fields. Clin Orthop Relat Res. 249:209–218. 1989.
37	Bassett CA, Schink-Ascani M and Lewis SM: Effects of pulsed electromagnetic fields on Steinberg ratings of femoral head osteonecrosis. Clin Orthop Relat Res. 246:172–185. 1989.
38	Sugioka Y, Hotokebuchi T and Tsutsui H: Transtrochanteric anterior rotational osteotomy for idiopathic and steroid-induced necrosis of the femoral head. Indications and long-term results. Clin Orthop Relat Res. 277:111–120. 1992.
39	Boettcher WG, Bonfiglio M and Smith K: Non-traumatic necrosis of the femoral head. II. Experiences in treatment. J Bone Joint Surg Am. 52:322–329. 1970. View Article : Google Scholar : PubMed/NCBI
40	Buckley PD, Gearen PF and Petty RW: Structural bone-grafting for early atraumatic avascular necrosis of the femoral head. J Bone Joint Surg Am. 73:1357–1364. 1991. View Article : Google Scholar : PubMed/NCBI
41	Kirschenbaum IH, Vernace JV, Booth RE Jr, Balderston RA and Rothman RH: Total hip arthroplasty for osteonecrosis. Semin Arthroplasty. 2:234–240. 1991.PubMed/NCBI
42	Katz RL, Bourne RB, Rorabeck CH and McGee H: Total hip arthroplasty in patients with avascular necrosis of the hip. Follow-up observations on cementless and cemented operations. Clin Orthop Relat Res. 281:145–151. 1992.
43	Plakseychuk AY, Kim SY, Park BC, Varitimidis SE, Rubash HE and Sotereanos DG: Vascularized compared with nonvascularized fibular grafting for the treatment of osteonecrosis of the femoral head. J Bone Joint Surg Am 85-A. 589–596. 2003. View Article : Google Scholar
44	González Della Valle A, Bates J, Di Carlo E and Salvati EA: Failure of free vascularized fibular graft for osteonecrosis of the femoral head: A histopathologic study of 6 cases. J Arthroplasty. 20:331–336. 2005. View Article : Google Scholar : PubMed/NCBI
45	Duran JM, Makarewich CA, Sharp TE, Starosta T, Zhu F, Hoffman NE, Chiba Y, Madesh M, Berretta RM, Kubo H and Houser SR: Bone-derived stem cells repair the heart after myocardial infarction through transdifferentiation and paracrine signaling mechanisms. Circ Res. 113:539–552. 2013. View Article : Google Scholar : PubMed/NCBI
46	Zhang HX, Zhang XP, Xiao GY, Hou Y, Cheng L, Si M, Wang SS, Li YH and Nie L: In vitro and in vivo evaluation of calcium phosphate composite scaffolds containing BMP-VEGF loaded PLGA microspheres for the treatment of avascular necrosis of the femoral head. Mater Sci Eng C Mater Biol Appl. 60:298–307. 2016. View Article : Google Scholar : PubMed/NCBI
47	Street J, Bao M, deGuzman L, Bunting S, Peale FV Jr, Ferrara N, Steinmetz H, Hoeffel J, Cleland JL, Daugherty A, et al: Vascular endothelial growth factor stimulates bone repair by promoting angiogene-sis and bone turnover. Proc Natl Acad Sci USA. 99:9656–9661. 2002. View Article : Google Scholar : PubMed/NCBI