Abstract
Wnt signaling maintains homeostasis in the bone marrow cavity: if Wnt signaling is inhibited then bone volume and density would decline. In this study, we identified a population of Wnt-responsive cells as osteoprogenitor in the intact trabecular bone region, which were responsible for bone development and turnover. If an implant was placed into the long bone, this Wnt-responsive population and their progeny contributed to osseointegration. We employed Axin2CreCreERT2/+;R26mTmG/+ transgenic mouse strain in which Axin2-positive, Wnt-responsive cells, and their progeny are permanently labeled by GFP upon exposure to tamoxifen. Each mouse received femoral implants placed into a site prepared solely by drilling, and a single-dose liposomal WNT3A protein was used in the treatment group. A lineage tracing strategy design allowed us to identify cells actively expressing Axin2 in response to Wnt signaling pathway. These tools demonstrated that Wnt-responsive cells and their progeny comprise a quiescent population residing in the trabecular region. In response to an implant placed, this population becomes mitotically active: cells migrated into the peri-implant region, up-regulated the expression of osteogenic proteins. Ultimately, those cells gave rise to osteoblasts that produced significantly more new bone in the peri-implant region. Wnt-responsive cells directly contributed to implant osseointegration. Using a liposomal WNT3A protein therapeutic, we showed that a single application at the time of implant placed was sufficient to accelerate osseointegration. The Wnt-responsive cell population in trabecular bone, activated by injury, ultimately contributes to implant osseointegration. Liposomal WNT3A protein therapeutic accelerates implant osseointegration in the long bone.
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References
Davila Castrodad IM et al (2019) Rehabilitation protocols following total knee arthroplasty: a review of study designs and outcome measures. Ann Transl Med 7(Suppl 7):S255
Branemark PI (1983) Osseointegration and its experimental background. J Prosthet Dent 50(3):399–410
Adell R et al (1981) A 15-year study of osseointegrated implants in the treatment of the edentulous jaw. Int J Oral Surg 10(6):387–416
Heinecke M et al (2018) The proximal and distal femoral canal geometry influences cementless stem anchorage and revision hip and knee implant stability. Orthopedics 41(3):e369–e375
Pilliar RM, Lee JM, Maniatopoulos C (1986) Observations on the effect of movement on bone ingrowth into porous-surfaced implants. Clin Orthop Relat Res 208:108–113
Malak TT et al (2016) Surrogate markers of long-term outcome in primary total hip arthroplasty: a systematic review. Bone Jt Res 5(6):206–214
Ramamurti BS et al (1997) Factors influencing stability at the interface between a porous surface and cancellous bone: a finite element analysis of a canine in vivo micromotion experiment. J Biomed Mater Res 36(2):274–280
Liu Y et al (2019) WNT3A accelerates delayed alveolar bone repair in ovariectomized mice. Osteoporos Int 30:1873–1885
Rodari G et al (2018) Progressive bone impairment with age and pubertal development in neurofibromatosis type I. Arch Osteoporos 13(1):93
Alghamdi HS, van den Beucken JJ, Jansen JA (2014) Osteoporotic rat models for evaluation of osseointegration of bone implants. Tissue Eng C 20(6):493–505
He YX et al (2011) Impaired bone healing pattern in mice with ovariectomy-induced osteoporosis: a drill-hole defect model. Bone 48(6):1388–1400
Song L et al (2012) Loss of wnt/beta-catenin signaling causes cell fate shift of preosteoblasts from osteoblasts to adipocytes. J Bone Miner Res 27(11):2344–2358
Zhang X et al (2018) Global transcriptome analysis to identify critical genes involved in the pathology of osteoarthritis. Bone Jt Res 7(4):298–307
Boyden LM et al (2002) High bone density due to a mutation in LDL-receptor-related protein 5. N Engl J Med 346(20):1513–1521
Lewiecki EM et al (2019) One year of romosozumab followed by two years of denosumab maintains fracture risk reductions: results of the FRAME Extension Study. J Bone Miner Res 34(3):419–428
Sovak G, Weiss A, Gotman I (2000) Osseointegration of Ti6Al4V alloy implants coated with titanium nitride by a new method. J Bone Jt Surg Br 82(2):290–296
Workman P et al (2010) Guidelines for the welfare and use of animals in cancer research. Br J Cancer 102(11):1555–1577
Szot GL, Koudria P, Bluestone JA (2007) Transplantation of pancreatic islets into the kidney capsule of diabetic mice. J Vis Exp. https://doi.org/10.3791/404
Movat HZ (1955) Demonstration of all connective tissue elements in a single section; pentachrome stains. AMA Arch Pathol 60(3):289–295
Leucht P et al (2007) Accelerated bone repair after plasma laser corticotomies. Ann Surg 246(1):140–150
Minear S et al (2010) Wnt proteins promote bone regeneration. Sci Transl Med 2(29):29ra30
Kawamoto T, Kawamoto K (2014) Preparation of thin frozen sections from nonfixed and undecalcified hard tissues using Kawamot’s film method (2012). Methods Mol Biol 1130:149–164
Yuan X et al (2018) Biomechanics of immediate postextraction implant osseointegration. J Dent Res 97(9):987–994
Sun Q et al (2019) Improving intraoperative storage conditions for autologous bone grafts: an experimental investigation in mice. J Tissue Eng Regen Med 13(12):2169–2180
Jing W et al (2015) Reengineering autologous bone grafts with the stem cell activator WNT3A. Biomaterials 47:29–40
Popelut A et al (2010) The acceleration of implant osseointegration by liposomal Wnt3a. Biomaterials 31(35):9173–9181
Dhamdhere GR et al (2014) Drugging a stem cell compartment using Wnt3a protein as a therapeutic. PLoS ONE 9(1):e83650
Morrell NT et al (2008) Liposomal packaging generates Wnt protein with in vivo biological activity. PLoS ONE 3(8):e2930
Hoyte DAN (1966) Experimental investigations of skull morphology and growth W.J.L. Felts and R.J. Harrison (eds). Int Rev Gen Exp Zool 2:345–407
Labek G et al (2011) Revision rates after total joint replacement: cumulative results from worldwide joint register datasets. J Bone Jt Surg Br 93(3):293–297
Sadoghi P et al (2013) Revision surgery after total joint arthroplasty: a complication-based analysis using worldwide arthroplasty registers. J Arthroplast 28(8):1329–1332
Apostu D et al (2018) Current methods of preventing aseptic loosening and improving osseointegration of titanium implants in cementless total hip arthroplasty: a review. J Int Med Res 46(6):2104–2119
Lam YF et al (2016) A review of the clinical approach to persistent pain following total hip replacement. Hong Kong Med J 22(6):600–607
Piscitelli P et al (2013) Painful prosthesis: approaching the patient with persistent pain following total hip and knee arthroplasty. Clin Cases Miner Bone Metab 10(2):97–110
Abu-Amer Y, Darwech I, Clohisy JC (2007) Aseptic loosening of total joint replacements: mechanisms underlying osteolysis and potential therapies. Arthritis Res Ther 9(Suppl 1):S6
Janssen D et al (2010) Computational assessment of press-fit acetabular implant fixation: the effect of implant design, interference fit, bone quality, and frictional properties. Proc Inst Mech Eng H 224(1):67–75
Soballe K et al (1992) Tissue ingrowth into titanium and hydroxyapatite-coated implants during stable and unstable mechanical conditions. J Orthop Res 10(2):285–299
Nazemi SM et al (2017) Optimizing finite element predictions of local subchondral bone structural stiffness using neural network-derived density–modulus relationships for proximal tibial subchondral cortical and trabecular bone. Clin Biomech (Bristol Avon) 41:1–8
Goltzman D (2019) The aging skeleton. Adv Exp Med Biol 1164:153–160
Salmon B et al (2017) WNT-activated bone grafts repair osteonecrotic lesions in aged animals. Sci Rep 7(1):14254
Virdi AS et al (2015) Sclerostin antibody treatment improves implant fixation in a model of severe osteoporosis. J Bone Jt Surg Am 97(2):133–140
Pei X et al (2017) Contribution of the PDL to osteotomy repair and implant osseointegration. J Dent Res 96(8):909–916
Li Z et al (2020) Effects of condensation and compressive strain on implant primary stability: a longitudinal, in vivo, multiscale study in mice. Bone Jt Res 9(2):60–70
Acknowledgements
We thank Dr. Yindong Liu, Bo Liu, and Dr. Giuseppe Salvi for their contributions to this manuscript. This work was supported by a Grant from the NIH (R01 DE024000-12) to JAH.
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Li, Z., Yuan, X., Arioka, M. et al. Pro-osteogenic Effects of WNT in a Mouse Model of Bone Formation Around Femoral Implants. Calcif Tissue Int 108, 240–251 (2021). https://doi.org/10.1007/s00223-020-00757-5
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DOI: https://doi.org/10.1007/s00223-020-00757-5