Deficiency of PDK1 in osteoclasts delays fracture healing and repair

Xiao,Dongliang; Zhou,Quan; Bai,Yiguang; Cao,Baichuan; Zhang,Qiong; Zeng,Gaofeng; Zong,Shaohui

doi:10.3892/mmr.2020.11209

Deficiency of PDK1 in osteoclasts delays fracture healing and repair

Authors:
- Dongliang Xiao
- Quan Zhou
- Yiguang Bai
- Baichuan Cao
- Qiong Zhang
- Gaofeng Zeng
- Shaohui Zong
View Affiliations

Affiliations: Department of Spine Osteopathia, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Collaborative Innovation Center of Guangxi Biological Medicine, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, College of Public Hygiene, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
Published online on: June 3, 2020 https://doi.org/10.3892/mmr.2020.11209
Pages: 1536-1546
Copyright: © Xiao 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

Bone fractures are common traumatic injuries of the musculoskeletal system. However, delayed union and non‑union fractures are a major clinical problem that present significant socioeconomic burden to patients and the public health sector. The bone‑resorbing osteoclasts and bone‑forming osteoblasts serve important roles in the fracture repair/healing process. Osteoclast deficiency or decreased osteoblast activity negatively impacts fracture healing. We previously demonstrated that the specific deletion of the serine/threonine kinase 3‑phosphoinositide‑dependent protein kinase 1 (PDK1) in osteoclasts leads to abrogated osteoclast formation and bone resorption in response to receptor activator of nuclear factor‑κB in vitro and protected mice against ovariectomized‑induced bone loss and lipopolysaccharide‑induced osteolysis in vivo. Given the importance of osteoclasts in fracture repair, we hypothesized that the specific loss of PDK1 in osteoclasts will alter the fracture healing process. Mice of tibial fracture were constructed, and tibial specimens were sampled at 7‑, 14‑, 21‑ and 28‑days post‑fracture to observe the effect of PDK1 gene regulated osteoclasts on fracture healing process by X‑ray radiography, microcomputed tomography scanning, histomorphological staining and biomechanical testing. The present study revealed, using the tibial fracture model, that the specific deletion of the PDK1 gene in osteoclasts impeded the fracture healing process by delaying the resorption of the cartilaginous callus and subsequent remodeling of immature woven bone to structurally and mechanically ensure lamellar bone is stronger. No effect on osteoblast bone formation and osteogenesis was observed, thus indicating that delayed fracture healing is primarily due to defective osteoclast activity. These results provide important clinical implications for the use of anti‑resorptive agents, such as bisphosphonates, for the treatment of osteolytic conditions. Such anti‑resorptive therapies may detrimentally delay fracture healing and repair.

View Figures

View References

1	Kanis JA, Odén A, McCloskey EV, Johansson H, Wahl DA and Cooper C; IOF Working Group on Epidemiology and Quality of Life, : A systematic review of hip fracture incidence and probability of fracture worldwide. Osteoporos Int. 23:2239–2256. 2012. View Article : Google Scholar : PubMed/NCBI
2	Tzioupis C and Giannoudis PV: Prevalence of long-bone non-unions. Injury. 38 (Suppl 2):S3–S9. 2007. View Article : Google Scholar : PubMed/NCBI
3	Hak DJ, Fitzpatrick D, Bishop JA, Marsh JL, Tilp S, Schnettler R, Simpson H and Alt V: Delayed union and nonunions: Epidemiology, clinical issues, and financial aspects. Injury. 45 (Suppl 2):S3–S7. 2014. View Article : Google Scholar : PubMed/NCBI
4	Einhorn TA: The cell and molecular biology of fracture healing. Clin Orthop Relat Res. 355S (Suppl):S7–S21. 1998. View Article : Google Scholar
5	Claes L, Recknagel S and Ignatius A: Fracture healing under healthy and inflammatory conditions. Nat Rev Rheumatol. 8:133–143. 2012. View Article : Google Scholar : PubMed/NCBI
6	Schindeler A, McDonald MM, Bokko P and Little DG: Bone remodeling during fracture repair: The cellular picture. Semin Cell Dev Biol. 19:459–466. 2008. View Article : Google Scholar : PubMed/NCBI
7	Hadjiargyrou M, Lombardo F, Zhao S, Ahrens W, Joo J, Ahn H, Jurman M, White DW and Rubin CT: Transcriptional profiling of bone regeneration. Insight into the molecular complexity of wound repair. J Biol Chem. 277:30177–30182. 2002. View Article : Google Scholar : PubMed/NCBI
8	Gerstenfeld LC, Cullinane DM, Barnes GL, Graves DT and Einhorn TA: Fracture healing as a post-natal developmental process: Molecular, spatial, and temporal aspects of its regulation. J Cell Biochem. 88:873–884. 2003. View Article : Google Scholar : PubMed/NCBI
9	Marsell R and Einhorn TA: The biology of fracture healing. Injury. 42:551–555. 2011. View Article : Google Scholar : PubMed/NCBI
10	Einhorn TA and Gerstenfeld LC: Fracture healing: Mechanisms and interventions. Nat Rev Rheumatol. 11:45–54. 2015. View Article : Google Scholar : PubMed/NCBI
11	Ferguson C, Alpern E, Miclau T and Helms JA: Does adult fracture repair recapitulate embryonic skeletal formation? Mech Dev. 87:57–66. 1999. View Article : Google Scholar : PubMed/NCBI
12	Xin C, Zhi-Ming H, Yasuaki S, Tomoo T and Akira Y: Quantitative study of osteoclastic related factors in the process of bone reconstruction. Hua Xi Kou Qiang Yi Xue Za Zhi. 24:164–165. 2006.(In Chinese). PubMed/NCBI
13	Lin HN and OConnor JP: Osteoclast depletion with clodronate liposomes delays fracture healing in mice. J Orthop Res. 35:1699–1706. 2017. View Article : Google Scholar : PubMed/NCBI
14	Cao Y, Mori S, Mashiba T, Westmore MS, Ma L, Sato M, Akiyama T, Shi L, Komatsubara S, Miyamoto K, et al: Raloxifene, estrogen, and alendronate affect the processes of fracture repair differently in ovariectomized rats. J Bone Miner Res. 17:2237–2246. 2002. View Article : Google Scholar : PubMed/NCBI
15	Flick LM, Weaver JM, Ulrich-Vinther M, Abuzzahab F, Zhang X, Dougall WC, Anderson D, OKeefe RJ and Schwarz EM: Effects of receptor activator of NFkappaB (RANK) signaling blockade on fracture healing. J Orthop Res. 21:676–684. 2003. View Article : Google Scholar : PubMed/NCBI
16	Gerstenfeld LC, Sacks DJ, Pelis M, Mason ZD, Graves DT, Barrero M, Ominsky MS, Kostenuik PJ, Morgan EF and Einhorn TA: Comparison of effects of the bisphosphonate alendronate versus the RANKL inhibitor denosumab on murine fracture healing. J Bone Miner Res. 24:196–208. 2009. View Article : Google Scholar : PubMed/NCBI
17	Xiao D, Zhou Q, Gao Y, Cao B, Zhang Q, Zeng G and Zong S: PDK1 is important lipid kinase for RANKL-induced osteoclast formation and function via the regulation of the Akt-GSK3β-NFATc1 signaling cascade. J Cell Biochem. Feb 12–2020.(Epub ahead of print). View Article : Google Scholar
18	Morshed S: Current options for determining fracture union. Adv Med. 2014.doi:10.1155/2014/708574. View Article : Google Scholar : PubMed/NCBI
19	Mi M, Jin H, Wang B, Yukata K, Sheu TJ, Ke QH, Tong P, Im HJ, Xiao G and Chen D: Chondrocyte BMP2 signaling plays an essential role in bone fracture healing. Gene. 512:211–218. 2013. View Article : Google Scholar : PubMed/NCBI
20	van t Hof RJ, Rose L, Bassonga E and Daroszewska A: Open source software for semi-automated histomorphometry of bone resorption and formation parameters. Bone. 99:69–79. 2017. View Article : Google Scholar : PubMed/NCBI
21	Abstracts of the Academy of Pediatric Physical Therapy Poster Presentations at the Combined Sections Meeting. Pediatr Phys Ther. 31:e31–e69. 2019. View Article : Google Scholar
22	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 : PubMed/NCBI
23	Inada M, Matsumoto C and Miyaura C: Animal models for bone and joint disease. Ovariectomized and orchidectomized animals. Clin Calcium. 21:164–170. 2011.(In Japanese). PubMed/NCBI
24	Frost HM and Jee WS: On the rat model of human osteopenias and osteoporoses. Bone Miner. 18:227–236. 1992. View Article : Google Scholar : PubMed/NCBI
25	Ghiasi MS, Chen J, Vaziri A, Rodriguez EK and Nazarian A: Bone fracture healing in mechanobiological modeling: A review of principles and methods. Bone Rep. 6:87–100. 2017. View Article : Google Scholar : PubMed/NCBI
26	Loi F, Córdova LA, Pajarinen J, Lin TH, Yao Z and Goodman SB: Inflammation, fracture and bone repair. Bone. 86:119–130. 2016. View Article : Google Scholar : PubMed/NCBI
27	Kamimura M, Mori Y, Sugahara-Tobinai A, Takai T and Itoi E: Impaired fracture healing caused by deficiency of the immunoreceptor adaptor protein DAP12. PLoS One. 10:e01282102015. View Article : Google Scholar : PubMed/NCBI
28	Shefelbine SJ, Augat P, Claes L and Beck A: Intact fibula improves fracture healing in a rat tibia osteotomy model. J Orthop Res. 23:489–493. 2005. View Article : Google Scholar : PubMed/NCBI
29	He LH, Liu M, He Y, Xiao E, Zhao L, Zhang T, Yang HQ and Zhang Y: TRPV1 deletion impaired fracture healing and inhibited osteoclast and osteoblast differentiation. Sci Rep. 7:423852017. View Article : Google Scholar : PubMed/NCBI
30	McDonald MM, Dulai S, Godfrey C, Amanat N, Sztynda T and Little DG: Bolus or weekly zoledronic acid administration does not delay endochondral fracture repair but weekly dosing enhances delays in hard callus remodeling. Bone. 43:653–662. 2008. View Article : Google Scholar : PubMed/NCBI
31	Ota N, Takaishi H, Kosaki N, Takito J, Yoda M, Tohmonda T, Kimura T, Okada Y, Yasuda H, Kawaguchi H, et al: Accelerated cartilage resorption by chondroclasts during bone fracture healing in osteoprotegerin-deficient mice. Endocrinology. 150:4823–4834. 2009. View Article : Google Scholar : PubMed/NCBI
32	Takahara M, Naruse T, Takagi M, Orui H and Ogino T: Matrix metalloproteinase-9 expression, tartrate-resistant acid phosphatase activity, and DNA fragmentation in vascular and cellular invasion into cartilage preceding primary endochondral ossification in long bones. J Orthop Res. 22:1050–1057. 2004. View Article : Google Scholar : PubMed/NCBI
33	Colnot C, Thompson Z, Miclau T, Werb Z and Helms JA: Altered fracture repair in the absence of MMP9. Development. 130:4123–4133. 2003. View Article : Google Scholar : PubMed/NCBI
34	Li J, Mori S, Kaji Y, Kawanishi J, Akiyama T and Norimatsu H: Concentration of bisphosphonate (incadronate) in callus area and its effects on fracture healing in rats. J Bone Miner Res. 15:2042–2051. 2000. View Article : Google Scholar : PubMed/NCBI