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Published in: Clinical Orthopaedics and Related Research® 8/2013

01-08-2013 | Symposium: Nanoscience in Musculoskeletal Medicine

pHEMA-nHA Encapsulation and Delivery of Vancomycin and rhBMP-2 Enhances its Role as a Bone Graft Substitute

Authors: Xinning Li, MD, Jianwen Xu, PhD, Tera M. Filion, PhD, David C. Ayers, MD, Jie Song, PhD

Published in: Clinical Orthopaedics and Related Research® | Issue 8/2013

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Abstract

Background

Bone grafts are widely used in orthopaedic procedures. Autografts are limited by donor site morbidity while allografts are known for considerable infection and failure rates. A synthetic composite bone graft substitute poly(2-hydroxyethyl methacrylate)-nanocrystalline hydroxyapatite (pHEMA-nHA) was previously developed to stably press-fit in and functionally repair critical-sized rat femoral segmental defects when it was preabsorbed with a single low dose of 300 ng recombinant human bone morphogenetic protein-2/7 (rhBMP-2/7).

Questions/purposes

To facilitate clinical translation of pHEMA-nHA as a synthetic structural bone graft substitute, we examined its ability to encapsulate and release rhBMP-2 and the antibiotic vancomycin.

Methods

We analyzed the compressive behavior and microstructure of pHEMA-nHA as a function of vancomycin incorporation doses using a dynamic mechanical analyzer and a scanning electron microscope. In vitro release of vancomycin was monitored by ultraviolet-visible spectroscopy. Release of rhBMP-2 from pHEMA-nHA-vancomycin was determined by ELISA. Bioactivity of the released vancomycin and rhBMP-2 was examined by bacterial inhibition and osteogenic transdifferentiation capabilities in cell culture, respectively.

Results

Up to 4.8 wt% of vancomycin was incorporated into pHEMA-nHA without compromising its structural integrity and compressive modulus. Encapsulated vancomycin was released in a dose-dependent and sustained manner in phosphate-buffered saline over 2 weeks, and the released vancomycin inhibited Escherichia coli culture. The pHEMA-nHA-vancomycin composite released preabsorbed rhBMP-2 in a sustained manner over 8 days and locally induced osteogenic transdifferentiation of C2C12 cells in culture.

Conclusions

pHEMA-nHA can encapsulate and deliver vancomycin and rhBMP-2 in a sustained and localized manner with reduced loading doses.

Clinical Relevance

The elasticity, osteoconductivity, and rhBMP-2/vancomycin delivery characteristics of pHEMA-nHA may benefit orthopaedic reconstructions or fusions with enhanced safety and efficiency and reduced infection risk.
Literature
1.
go back to reference Abrarrategi A, Moreno-Vincent C, Ramos V, Aranaz L, Sanz Casado JV, Lopez-Lacomba JL. Improvement of porous beta-TCP scaffolds with rhBMP-2 choitosan carrier film for bone tissure application. Tissue Eng Part A. 2008;14:1305–1319.CrossRef Abrarrategi A, Moreno-Vincent C, Ramos V, Aranaz L, Sanz Casado JV, Lopez-Lacomba JL. Improvement of porous beta-TCP scaffolds with rhBMP-2 choitosan carrier film for bone tissure application. Tissue Eng Part A. 2008;14:1305–1319.CrossRef
2.
go back to reference Anderson EM, Noble ML, Garty S, Ma HY, Bryers JD, Shen TT, Ratner BD. Sustained release of antibiotic from poly(2-hydroxyethyl methacrylate) to prevent blinding infections after cataract surgery. Biomaterials. 2009;30:5675–5681.PubMedCrossRef Anderson EM, Noble ML, Garty S, Ma HY, Bryers JD, Shen TT, Ratner BD. Sustained release of antibiotic from poly(2-hydroxyethyl methacrylate) to prevent blinding infections after cataract surgery. Biomaterials. 2009;30:5675–5681.PubMedCrossRef
3.
go back to reference Arrington ED, Smith WJ, Chambers HG, Bucknell AL, Davino NA. Complications of iliac crest bone graft. Clin Orthop Relat Res. 1996;329:300–309.PubMedCrossRef Arrington ED, Smith WJ, Chambers HG, Bucknell AL, Davino NA. Complications of iliac crest bone graft. Clin Orthop Relat Res. 1996;329:300–309.PubMedCrossRef
4.
go back to reference Bostrom MP, Camacho NP. Potential role of bone morphogenetic proteins in fracture healing. Clin Orthop Relat Res. 1998;355(suppl):S274–S282.PubMedCrossRef Bostrom MP, Camacho NP. Potential role of bone morphogenetic proteins in fracture healing. Clin Orthop Relat Res. 1998;355(suppl):S274–S282.PubMedCrossRef
5.
go back to reference Bostrom MP, Seigerman DA. The clinical use of allografts, demineralized bone matricies, synthetic bone graft substitutes and osteoinductive growth factors: a survey study. HSS J. 2005;1:9–18.PubMedCrossRef Bostrom MP, Seigerman DA. The clinical use of allografts, demineralized bone matricies, synthetic bone graft substitutes and osteoinductive growth factors: a survey study. HSS J. 2005;1:9–18.PubMedCrossRef
6.
go back to reference Bourque WT, Gross M, Hall BK. A reproducible method for producing and quantifying the stages of fracture repair. Lab Anim Sci. 1992;42:369–374.PubMed Bourque WT, Gross M, Hall BK. A reproducible method for producing and quantifying the stages of fracture repair. Lab Anim Sci. 1992;42:369–374.PubMed
7.
go back to reference Carragee EJ, Hurwitz EL, Weiner BK. A critical review of recombinant human bone morphogenetic protein-2 trials in spinal surgery: emerging safety concerns and lessons learned. Spine J. 2011;11:471–491.PubMedCrossRef Carragee EJ, Hurwitz EL, Weiner BK. A critical review of recombinant human bone morphogenetic protein-2 trials in spinal surgery: emerging safety concerns and lessons learned. Spine J. 2011;11:471–491.PubMedCrossRef
8.
go back to reference Clellan J, Mulconrey D, Forbes R, Fullmer N. Vertebral bone resorption after transforaminal lumbar interbody fusion with bone morphogenetic protein (rhBMP-2). J Spinal Disord Tech. 2006;19:483–486.CrossRef Clellan J, Mulconrey D, Forbes R, Fullmer N. Vertebral bone resorption after transforaminal lumbar interbody fusion with bone morphogenetic protein (rhBMP-2). J Spinal Disord Tech. 2006;19:483–486.CrossRef
9.
go back to reference Decoster TA, Gehlert RJ, Mikola EA, Pirela-Cruz MA. Management of posttraumatic segmental bone defects. J Am Acad Orthop Surg. 2004;12:28–38.PubMed Decoster TA, Gehlert RJ, Mikola EA, Pirela-Cruz MA. Management of posttraumatic segmental bone defects. J Am Acad Orthop Surg. 2004;12:28–38.PubMed
10.
go back to reference Delloye C, Cornu O, Druez V, Barbier O. Bone allografts: what they can offer and what they cannot. J Bone Joint Surg Br. 2007;89:574–580.PubMedCrossRef Delloye C, Cornu O, Druez V, Barbier O. Bone allografts: what they can offer and what they cannot. J Bone Joint Surg Br. 2007;89:574–580.PubMedCrossRef
11.
go back to reference Filion TM, Li X, Mason-Savas A, Kreider JM, Goldstein SA, Ayers DC, Song J. Elasteromeric osteoconductive synthetic scaffolds with acquired osteoinductivity expedite the repair of critical femoral defects in rats. Tissue Eng Part A. 2011;17:503–511.PubMedCrossRef Filion TM, Li X, Mason-Savas A, Kreider JM, Goldstein SA, Ayers DC, Song J. Elasteromeric osteoconductive synthetic scaffolds with acquired osteoinductivity expedite the repair of critical femoral defects in rats. Tissue Eng Part A. 2011;17:503–511.PubMedCrossRef
12.
go back to reference Gilbert M, Shaw WJ, Long JR, Nelson K, Drobny GP, Giachelli CM, Stayton PS. Chimeric peptides of statherin and osteopontin that bind hydroxyapatite and mediate cell adhesion. J Biol Chem. 2000;275:16213–16218.PubMedCrossRef Gilbert M, Shaw WJ, Long JR, Nelson K, Drobny GP, Giachelli CM, Stayton PS. Chimeric peptides of statherin and osteopontin that bind hydroxyapatite and mediate cell adhesion. J Biol Chem. 2000;275:16213–16218.PubMedCrossRef
13.
go back to reference Glassman SD, Howard J, Dimar J, Sweet A, Wilson G, Carreon L. Complications with rhBMP-2 in posterolateral spine fusion: a consecutive series of one thousand thirty-seven cases. Spine (Phila Pa 1976). 2011;36:1849–1854. Glassman SD, Howard J, Dimar J, Sweet A, Wilson G, Carreon L. Complications with rhBMP-2 in posterolateral spine fusion: a consecutive series of one thousand thirty-seven cases. Spine (Phila Pa 1976). 2011;36:1849–1854.
14.
go back to reference Hanssen AD, Spangehl MJ. Practical applications of antibiotic-loaded bone cement for treatment of infected joint replacement. Clin Orthop Relat Res. 2004;427:79–85.PubMedCrossRef Hanssen AD, Spangehl MJ. Practical applications of antibiotic-loaded bone cement for treatment of infected joint replacement. Clin Orthop Relat Res. 2004;427:79–85.PubMedCrossRef
15.
go back to reference Jeon O, Song SJ, Kang SW, Putnam AJ, Kim BS. Enhancement of ectopic bone formation by bone morphogenetic protein-2 released from a herparin conjugated poly(l-lactic-co-glycolic acid) scaffold. Biomaterials. 2007;28:2763–2771.PubMedCrossRef Jeon O, Song SJ, Kang SW, Putnam AJ, Kim BS. Enhancement of ectopic bone formation by bone morphogenetic protein-2 released from a herparin conjugated poly(l-lactic-co-glycolic acid) scaffold. Biomaterials. 2007;28:2763–2771.PubMedCrossRef
16.
go back to reference Jones AL. Recombinant human bone morphogenic protein-2 in fracture care. J Orthop Trauma. 2005;19(10 suppl):S23–S25.PubMedCrossRef Jones AL. Recombinant human bone morphogenic protein-2 in fracture care. J Orthop Trauma. 2005;19(10 suppl):S23–S25.PubMedCrossRef
17.
go back to reference Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahashi N, Ikeda T, Rosen V, Wozney JM, Fujisawa-Sehara A, Suda T. Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage. J Cell Biol. 1994;127:1755–1766.PubMedCrossRef Katagiri T, Yamaguchi A, Komaki M, Abe E, Takahashi N, Ikeda T, Rosen V, Wozney JM, Fujisawa-Sehara A, Suda T. Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage. J Cell Biol. 1994;127:1755–1766.PubMedCrossRef
18.
go back to reference Kempen DH, Kruyt MC, Lu L, Wilson CE, Florschutz AV, Creemers LB, Yaszemski MJ, Dhert WJ. Effect of autogous BMSCs seeding and BMP-2 delivery on ectopic bone formation in a microsphere/poly(propylene fumarate) composite. Tissue Eng Part A. 2008;15:587–594.CrossRef Kempen DH, Kruyt MC, Lu L, Wilson CE, Florschutz AV, Creemers LB, Yaszemski MJ, Dhert WJ. Effect of autogous BMSCs seeding and BMP-2 delivery on ectopic bone formation in a microsphere/poly(propylene fumarate) composite. Tissue Eng Part A. 2008;15:587–594.CrossRef
19.
go back to reference Kim SS, Gwak SJ, Sim BS. Orthotopic bone formation by implantation of apatite-coated poly(lactide-co-glycolide)/hydroxyapatite composite particulate and bone morphogenetic protein-2. J Biomed Mater Res A. 2008;87:245–253.PubMed Kim SS, Gwak SJ, Sim BS. Orthotopic bone formation by implantation of apatite-coated poly(lactide-co-glycolide)/hydroxyapatite composite particulate and bone morphogenetic protein-2. J Biomed Mater Res A. 2008;87:245–253.PubMed
20.
go back to reference Lee SH, Lee JE, Baek WY, Lim JO. Regional delivery of vancomycin using pluronic F-127 to inhibit methicillin resistant Staphylococcus aureus (MRSA) growth in chronic otitis media in vitro and in vivo. J Control Release. 2004;96:1–7.PubMedCrossRef Lee SH, Lee JE, Baek WY, Lim JO. Regional delivery of vancomycin using pluronic F-127 to inhibit methicillin resistant Staphylococcus aureus (MRSA) growth in chronic otitis media in vitro and in vivo. J Control Release. 2004;96:1–7.PubMedCrossRef
21.
go back to reference Le Nihouannen D, Hacking SA, Gbureck U, Komarova SV, Barralet JE. The use of RANKL-coated brushite cement to stimulate bone remodelling. Biomaterials. 2008;29:3253–3259.PubMedCrossRef Le Nihouannen D, Hacking SA, Gbureck U, Komarova SV, Barralet JE. The use of RANKL-coated brushite cement to stimulate bone remodelling. Biomaterials. 2008;29:3253–3259.PubMedCrossRef
22.
go back to reference Li B, Brown KV, Wenke JC, Guelcher SA. Sustained release of vancomycin from polyurethane scaffolds inhibits infection of bone wounds in a rat femoral segmental defect model. J Control Release. 2010;145:221–230.PubMedCrossRef Li B, Brown KV, Wenke JC, Guelcher SA. Sustained release of vancomycin from polyurethane scaffolds inhibits infection of bone wounds in a rat femoral segmental defect model. J Control Release. 2010;145:221–230.PubMedCrossRef
23.
go back to reference McCall TA, Brokaw DS, Jelen BA, Scheid KD, Scharfenberger AV, Maar DC, Green JM, Shipps MR, Stone MB, Musapatika D, Weber TG. Treatment of large segmental bone defects with reamer irrigator-aspirator bone graft: technique and case series. Orthop Clin North Am. 2010;41:63–73.PubMedCrossRef McCall TA, Brokaw DS, Jelen BA, Scheid KD, Scharfenberger AV, Maar DC, Green JM, Shipps MR, Stone MB, Musapatika D, Weber TG. Treatment of large segmental bone defects with reamer irrigator-aspirator bone graft: technique and case series. Orthop Clin North Am. 2010;41:63–73.PubMedCrossRef
24.
go back to reference McKay B, Sandhu HS. Use of recombinant human bone morphogenetic protein-2 in spinal fusion. Spine (Phila Pa 1976). 2002;27(16 suppl 1):S66–S85. McKay B, Sandhu HS. Use of recombinant human bone morphogenetic protein-2 in spinal fusion. Spine (Phila Pa 1976). 2002;27(16 suppl 1):S66–S85.
25.
go back to reference Montgomery DM, Aronson DD, Lee CL, LaMont RL. Posterior spinal fusion: allograft versus autograft bone. J Spinal Disord Tech. 1990;3:370–375. Montgomery DM, Aronson DD, Lee CL, LaMont RL. Posterior spinal fusion: allograft versus autograft bone. J Spinal Disord Tech. 1990;3:370–375.
26.
27.
go back to reference Muscolo D, Ayerza M, Aponte-Tinao L. Massive allograft use in orthopedic oncology. Orthop Clinic North Am. 2006;37:65–74.CrossRef Muscolo D, Ayerza M, Aponte-Tinao L. Massive allograft use in orthopedic oncology. Orthop Clinic North Am. 2006;37:65–74.CrossRef
28.
go back to reference Naraghi F, Wolfer L, Lewis M, Palma C. BMP-2 dose correlates with increase postoperative edema and swallowing complications after cervical fusion. Spine J. 2006;6:7s–8s.CrossRef Naraghi F, Wolfer L, Lewis M, Palma C. BMP-2 dose correlates with increase postoperative edema and swallowing complications after cervical fusion. Spine J. 2006;6:7s–8s.CrossRef
29.
go back to reference Ozaki T, Hillmann A, Bettin D, Wuisman P, Winkelmann W. Intramedullary, antibiotic-loaded cemented, massive allografts for skeletal reconstruction: 26 cases compared with 19 uncemented allografts. Acta Orthop Scand. 1997;68:387–391.PubMedCrossRef Ozaki T, Hillmann A, Bettin D, Wuisman P, Winkelmann W. Intramedullary, antibiotic-loaded cemented, massive allografts for skeletal reconstruction: 26 cases compared with 19 uncemented allografts. Acta Orthop Scand. 1997;68:387–391.PubMedCrossRef
30.
go back to reference Penner MJ, Masri BA, Duncan CP. Elution characteristics of vancomycin and tobramycin combined in acrylic bone cement. J Arthroplasty. 1996;11:939–944.PubMedCrossRef Penner MJ, Masri BA, Duncan CP. Elution characteristics of vancomycin and tobramycin combined in acrylic bone cement. J Arthroplasty. 1996;11:939–944.PubMedCrossRef
31.
go back to reference Prokuski L. Prophylactic antibiotics in orthopaedic surgery. J Am Acad Orthop Surg. 2008;16:283–293.PubMed Prokuski L. Prophylactic antibiotics in orthopaedic surgery. J Am Acad Orthop Surg. 2008;16:283–293.PubMed
32.
go back to reference Raiche AT, Puleo DA. Cell responses to BMP-2 and IGF-I release with different time-dependent profiles. J Biomed Mater Res A. 2004;69:342–350.PubMedCrossRef Raiche AT, Puleo DA. Cell responses to BMP-2 and IGF-I release with different time-dependent profiles. J Biomed Mater Res A. 2004;69:342–350.PubMedCrossRef
33.
go back to reference Rauschmann MA, Wichelhaus TA, Stirnal V, Dingeldein E, Zichner L, Schnettler R, Volker A. Nanocrystalline hydroxyapatite and calcium sulphate as biodegradable composite carrier material for local delivery of antibiotics in bone infections. Biomaterials. 2005;26:2677–2684.PubMedCrossRef Rauschmann MA, Wichelhaus TA, Stirnal V, Dingeldein E, Zichner L, Schnettler R, Volker A. Nanocrystalline hydroxyapatite and calcium sulphate as biodegradable composite carrier material for local delivery of antibiotics in bone infections. Biomaterials. 2005;26:2677–2684.PubMedCrossRef
34.
go back to reference Shah RK, Moncayo VM, Smitson RD, Pierre-Jerome C, Terk MR. Recombinant human bone morphogenetic protein 2 induced heterotopic ossification of the retroperitoneum, psoas muscle, pelvis and abdominal wall following lumbar spinal fusion. Skeletal Radiol. 2010;39:501–504.PubMedCrossRef Shah RK, Moncayo VM, Smitson RD, Pierre-Jerome C, Terk MR. Recombinant human bone morphogenetic protein 2 induced heterotopic ossification of the retroperitoneum, psoas muscle, pelvis and abdominal wall following lumbar spinal fusion. Skeletal Radiol. 2010;39:501–504.PubMedCrossRef
35.
go back to reference Shields LB, Raque GH, Glassman SD, Campbell M, Vitaz T, Harpring J, Shields CB. Adverse effects associated with high-dose recombinant human bone morphogenetic protein-2 use in anterior cervical spine fusion. Spine (Phila Pa 1976). 2006;31:542–547. Shields LB, Raque GH, Glassman SD, Campbell M, Vitaz T, Harpring J, Shields CB. Adverse effects associated with high-dose recombinant human bone morphogenetic protein-2 use in anterior cervical spine fusion. Spine (Phila Pa 1976). 2006;31:542–547.
36.
go back to reference Song J, Xu J, Filion T, Saiz E, Tomsia AP, Lian J, Stein GS, Ayers D, Bertozzi CR. Elastomeric high-mineral content hydrogel-hydroxyapatite composites for orthopedic applications. J Biomed Mater Res A. 2009;89:1098–1107.PubMed Song J, Xu J, Filion T, Saiz E, Tomsia AP, Lian J, Stein GS, Ayers D, Bertozzi CR. Elastomeric high-mineral content hydrogel-hydroxyapatite composites for orthopedic applications. J Biomed Mater Res A. 2009;89:1098–1107.PubMed
37.
go back to reference Stubbs JT 3rd, Mintz KP, Eanes ED, Torchia DA, Fisher LW. Characterization of native and recombinant bone sialoprotein: delineation of the mineral-binding and cell adhesion domains and structural analysis of the RGD domain. J Bone Miner Res. 1997;12:1210–1222.PubMedCrossRef Stubbs JT 3rd, Mintz KP, Eanes ED, Torchia DA, Fisher LW. Characterization of native and recombinant bone sialoprotein: delineation of the mineral-binding and cell adhesion domains and structural analysis of the RGD domain. J Bone Miner Res. 1997;12:1210–1222.PubMedCrossRef
38.
go back to reference Su A, Chen W, Chen C, Chen T. Innovative trident fixation technique for allograft knee arthrodesis for high-grade osteosarcoma around the knee. Jpn J Clin Oncol. 2009;39:739–744.PubMedCrossRef Su A, Chen W, Chen C, Chen T. Innovative trident fixation technique for allograft knee arthrodesis for high-grade osteosarcoma around the knee. Jpn J Clin Oncol. 2009;39:739–744.PubMedCrossRef
39.
go back to reference Whitehouse MR, Lankester BJ, Winson IG, Hepple S. Bone graft harvest from the proximal tibia in foot and ankle arthrodesis surgery. Foot Ankle Int. 2006;27:913–916.PubMed Whitehouse MR, Lankester BJ, Winson IG, Hepple S. Bone graft harvest from the proximal tibia in foot and ankle arthrodesis surgery. Foot Ankle Int. 2006;27:913–916.PubMed
40.
go back to reference Wong D, Kumar A, Jatana S, Ghiselli G, Wong K. Neurologic impairment from ectopic bone in the lumbar canal: a potential complications of off-label PLIF/TLIF use of bone morphogenetic protein-2 (BMP-2). Spine J. 2008;8:1011–1018.PubMedCrossRef Wong D, Kumar A, Jatana S, Ghiselli G, Wong K. Neurologic impairment from ectopic bone in the lumbar canal: a potential complications of off-label PLIF/TLIF use of bone morphogenetic protein-2 (BMP-2). Spine J. 2008;8:1011–1018.PubMedCrossRef
41.
42.
go back to reference Xu J, Li X, Lian J, Ayers DC, Song J. Sustained and localized in vitro release of BMP-2/7, RANKL, and tetracycline from FlexBone, an elastomeric osteoconductive bone substitute. J Orthop Res. 2009;27:1306–1311.PubMedCrossRef Xu J, Li X, Lian J, Ayers DC, Song J. Sustained and localized in vitro release of BMP-2/7, RANKL, and tetracycline from FlexBone, an elastomeric osteoconductive bone substitute. J Orthop Res. 2009;27:1306–1311.PubMedCrossRef
Metadata
Title
pHEMA-nHA Encapsulation and Delivery of Vancomycin and rhBMP-2 Enhances its Role as a Bone Graft Substitute
Authors
Xinning Li, MD
Jianwen Xu, PhD
Tera M. Filion, PhD
David C. Ayers, MD
Jie Song, PhD
Publication date
01-08-2013
Publisher
Springer US
Published in
Clinical Orthopaedics and Related Research® / Issue 8/2013
Print ISSN: 0009-921X
Electronic ISSN: 1528-1132
DOI
https://doi.org/10.1007/s11999-012-2644-5

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