Fabrication of an rhBMP-2 loaded porous β-TCP microsphere-hyaluronic acid-based powder gel composite and evaluation of implant osseointegration

Jae Hyup Lee, Jungju Kim, Hae-Ri Baek, Kyung Mee Lee, Jun-Hyuk Seo, Hyun-Kyung Lee, A-Young Lee, Guang Bin Zheng, Bong-Soon Chang, Choon-Ki Lee, Jae Hyup Lee, Jungju Kim, Hae-Ri Baek, Kyung Mee Lee, Jun-Hyuk Seo, Hyun-Kyung Lee, A-Young Lee, Guang Bin Zheng, Bong-Soon Chang, Choon-Ki Lee

Abstract

Methods to improve osseointegration that include implantation of rhBMP-2 with various kinds of carriers are currently of considerable interest. The present study was conducted to evaluate if the rhBMP-2 loaded β-TCP microsphere-hyaluronic acid-based powder-like hydrogel composite (powder gel) can act as an effective rhBMP-2 carrier for implantation in host bone with a bone defect or poor bone quality. The release pattern for rhBMP-2 was then evaluated against an rhBMP-2-loaded collagen sponge as a control group. Dental implants were also inserted into the tibias of three groups of rabbits: an rhBMP-2 (200 µg) loaded powder gel composite implanted group, an implant only group, and a powder gel implanted group. Micro-CT and histology of the implanted areas were carried out four weeks later. The rhBMP-2 powder gel released less rhBMP-2 than the collagen sponge, but it continued a slow release for more than 7 days. The rhBMP-2 powder gel composite improved osseointegration of the dental implant by increasing the amount of new bone formation in the implant pitch and it improved the bone quality and bone quantity of new bone. The histology results indicated that the rhBMP-2 powder gel composite improved the osseointegration in the cortical bone as well as the marrow space along the fixture. The bone-to-implant contact ratio of the rhBMP-2 (200 µg) loaded powder gel composite implanted group was significantly higher than those of the implant only group and the powder gel implanted group. The powder gel appeared to be a good carrier and could release rhBMP-2 slowly to promote the formation of new bone following implantation in a bone defect, thereby improving implant osseointegration.

Figures

Fig. 1
Fig. 1
SEM images of β-TCP microspheres generated by the spray-dry method after sintering at 1,250 °C
Fig. 2
Fig. 2
Surgical procedures. a Tibial diaphysis in the tibial medial aspect was exposed and two, 4 mm diameter holes were generated; b A composite of rhBMP-2 loaded porous β-TCP microspheres and hyaluronic acid powder-gel was injected; c A dental implant fixture was inserted
Fig. 3
Fig. 3
a SEM images of cross-linked and freeze-dried HA-based hydrogel; b Images of the hydrated HA-based powder-gel; c Swelling ratio of the HA-based powder-gel
Fig. 4
Fig. 4
In vitro release of BMP-2 (A: Hyaluronic acid (HA)-based powder-gel; Control: collagen sponge)
Fig. 5
Fig. 5
Plane radiographs. H: A composite of the β-TCP microspheres and the hyaluronic acid powder gel was injected and a dental implant fixture was inserted (hydrogel group); B: A composite of the rhBMP-2 loaded porous β-TCP microspheres and the hyaluronic acid powder gel was injected and then a dental implant fixture was inserted (BMP-2 group). I: Only a dental implant fixture was inserted (implant group)
Fig. 6
Fig. 6
Micro-CT results. a Only a dental implant fixture was implanted (implant group); b A composite of the β-TCP microspheres and the hyaluronic acid powder gel was injected and a dental implant fixture was inserted (hydrogel group); c A composite of the rhBMP-2 loaded β-TCP microspheres and the hyaluronic acid powder gel was injected and a dental implant fixture was inserted (BMP-2 group). In panel a, new bone formation was insignificant around the implant fixture but some new bone was shown in panel b, while extensive new bone formation was observed in panel c (arrows: newly formed bony tissue)
Fig. 7
Fig. 7
Undecalcified histology. The image magnification on the left and right was ×12.5 and ×40, respectively. a Only a dental implant fixture was implanted (implant group). Bone coupling with cortical bone was stable, but the osseous tissue associated with the fixture screws in the marrow space was insignificant; b A composite of the β-TCP microspheres and the hyaluronic acid powder gel was injected and a dental implant fixture was inserted (hydrogel group). Bone bonding in the cortical bone was also robust, but new bone formation in the marrow space was insignificant even in comparison to the implant group. The tissue within the pitch is composed of fibrous tissue and exists as intervening tissue between the bony tissues of the marrow space; c A composite of the rhBMP-2 loaded β-TCP microspheres and the hyaluronic acid powder gel was injected and a dental implant fixture was inserted (BMP-2 group). The bone bonding in the cortical bone was stable and osseous tissue was connected from the cortical bone to the marrow space. Newly formed bone was evident around the fixture within the marrow space, and newly formed bone was fused with the fixture

References

    1. Sakka S, Coulthard P. 2009 Bone quality: a reality for the process of osseointegration. Implant dent. 2009;18:480–485. doi: 10.1097/ID.0b013e3181bb840d.
    1. Lambert PM, Morris HF, Ochi S. The influence of smoking on 3-year clinical success of osseointegrated dental implants. Ann Periodontol. 2000;5:579–589. doi: 10.1902/annals.2000.5.1.79.
    1. Carini F, Pisapia V, Monai D, Barbano L, Porcaro G. Implant rehabilitation in patients irradiated for head and neck cancer: role of intensity-moduled radiotherapy (IMRT) in planning the insertion site. Ann Stomatol (Roma) 2012;3:8–20.
    1. Mellado-Valero A, Ferrer-Garcia JC, Calvo-Catala J, Labaig-Rueda C. Implant treatment in patients with osteoporosis. Med Oral Patol Oral Cir Bucal. 2010;15:e52–e57.
    1. Lee JH, Ryu MY, Baek HR, Lee KM, Seo JH, Lee HK, Ryu HS. Effects of porous beta-tricalcium phosphate-based ceramics used as an E. coli-derived rhBMP-2 carrier for bone regeneration. J Mater Sci Mater Med. 2013;24:2117–2127.
    1. Barnes B, Boden SD, Louis-Ugbo J, Tomak PR, Park JS, Park MS, Minamide A. Lower dose of rhBMP-2 achieves spine fusion when combined with an osteoconductive bulking agent in non-human primates. Spine. 2005;15:1127–1133. doi: 10.1097/01.brs.0000162623.48058.8c.
    1. Fei Z, Hu Y, Wu D, Wu H, Lu R, Bai J, Song H. Preparation and property of a novel bone graft composite consisting of rhBMP-2 loaded PLGA microspheres and calcium phosphate cement. J Mater Sci Mater Med. 2008;19:1109–1116.
    1. Laurent TC, Laurent UB, Fraser JR. Functions of hyaluronan. Ann Rheum Dis. 1995;54(429–3):2.
    1. Karvinen S, Pasonen-Seppanen S, Hyttinen JM, Pienimaki JP, Torronen K, Jokela TA, Tammi MI, Tammi R. Keratinocyte growth factor stimulates migration and hyaluronan synthesis in the epidermis by activation of keratinocyte hyaluronan synthases 2 and 3. J Biol Chem. 2003;278:49495–49504. doi: 10.1074/jbc.M310445200.
    1. Bastow ER, Byers S, Golub SB, Clarkin CE, Pitsillides AA, Fosang AJ. Hyaluronan synthesis and degradation in cartilage and bone. Cell Mol Life Sci. 2008;65:395–413. doi: 10.1007/s00018-007-7360-z.
    1. West DC, Hampson IN, Arnold F, Kumar S. Angiogenesis induced by degradation products of hyaluronic acid. Science. 1985;228:1324–1326. doi: 10.1126/science.2408340.
    1. Itano N, Atsumi F, Sawai T, Yamada Y, Miyaishi O, Senga T, Hamaguchi M, Kimata K. Abnormal accumulation of hyaluronan matrix diminishes contact inhibition of cell growth and promotes cell migration. Proc Natl Acad Sci U S A. 2002;99:3609–3614. doi: 10.1073/pnas.052026799.
    1. Peng L, Bian WG, Liang FH, Xu HZ. Implanting hydroxyapatite-coated porous titanium with bone morphogenetic protein-2 and hyaluronic acid into distal femoral metaphysis of rabbits. Chin J Traumatol. 2008;11:179–185. doi: 10.1016/S1008-1275(08)60038-3.
    1. Hulsart-Billstrom G, Bergman K, Andersson B, Hilborn J, Larsson S, Jonsson KB. A unicortical femoral defect model in the rat: evaluation using injectable hyaluronan hydrogel as a carrier for bone morphogenetic protein-2. J Tissue Eng Regen Med. 2012; Epub ahead of print.
    1. Young S, Wong M, Tabata Y, Mikos AG. Gelatin as a delivery vehicle for the controlled release of bioactive molecules. J Control Release. 2005;109:256–274. doi: 10.1016/j.jconrel.2005.09.023.
    1. Maus U, Andereya S, Gravius S, Ohnsorge JA, Niedhart C, Siebert CH. BMP-2 incorporated in a tricalcium phosphate bone substitute enhances bone remodeling in sheep. J Biomater Appl. 2008;22:559–576. doi: 10.1177/0885328207083311.
    1. Uy R, Wold F. 1,4-Butanediol diglycidyl ether coupling of carbohydrates to sepharose: affinity adsorbents for lectins and glycosidases. Anal Biochem. 1977;81:98–107. doi: 10.1016/0003-2697(77)90602-9.
    1. Fiszer-Szafarz B, Czartoryska B, Tylki-Szymanska A. Serum hyaluronidase aberrations in metabolic and morphogenetic disorders. Glycoconjugate. 2005;22:395–400. doi: 10.1007/s10719-005-1390-2.
    1. Rifas L, Halstead LR, Peck WA, Avioli LV, Welgus HG. Human osteoblasts in vitro secrete tissue inhibitor of metalloproteinases and gelatinase but not interstitial collagenase as major cellular products. J Clin Invest. 1989;84:686–694. doi: 10.1172/JCI114216.
    1. Simon M, Trisi P, Piattelli A, Ridge V. Augmentation using a membrane technique associated with osseointegrated implants. Int J Periodontics Restor Dent. 1994;14:497–511.
    1. Leknes KN, Yang J, Qahash M, Polimeni G, Susin C, Wikesjö UM. Alveolar ridge augmentation using implants coated with recombinant human bone morphogenetic protein-2: radiographic observations. Clin Oral Implant Res. 2008;19:1027–1033. doi: 10.1111/j.1600-0501.2008.01567.x.
    1. Gloria A, Borzacchiello A, Causa F, Ambrosio L. Rheological characterization of hyaluronic acid derivatives as injectable materials toward nucleus pulposus regeneration. J Biomater Appl. 2012;26:745–759. doi: 10.1177/0885328210387174.
    1. Yuan H, Fernandes H, Habibovic P, de Boer J, Barradas AMC, de Ruiter A, Walsh WR, van Blitterswijk CA, de Bruijn JD. Osteoinductive ceramics as a synthetic alternative to autologous bone grafting. Proc Natl Acad Sci. 2010;107:13614–13619. doi: 10.1073/pnas.1003600107.
    1. Barradas AM, Fernandes HA, Groen N, Chai YC, Schrooten J, van de Peppel J, van Leeuwen JP, van Blitterswijk CA, de Boer J. A calcium-induced signaling cascade leading to osteogenic differentiation of human bone marrow-derived mesenchymal stromal cells. Biomaterials. 2012;33:3205–3215. doi: 10.1016/j.biomaterials.2012.01.020.
    1. Hänseler P, Ehrbar M, Kruse A, Fischer E, Schibli R, Ghayor C, Weber FE. Delivery of BMP-2 by two clinically available apatitie materials: In vitro and in vivo comparison. J Biomed Mater Res A. 2014; E-pub ahead of print.
    1. Reddi AH. Role of morphogenetic proteins in skeletal tissue engineering and regeneration. Nat Biotechnol. 1998;16:247–252. doi: 10.1038/nbt0398-247.
    1. Qiao C, Zhang K, Jin H, Miao L, Shi C, Liu X, Yang A, Liu J, Li D, Zheng C, Zhang G, Li X, Yang B, Sun H. Using poly(lactic-co-glycolic acid) microspheres to encapsulate plasmid of bone morphogenetic protein 2/polyethylenimine nanoparticles to promote bone formation in vitro and in vivo. Int J Nanomedicine. 2013;8:2985–2995.
    1. Jung MR, Shim IK, Chung HJ, Lee HR, Park YJ, Lee MC, Yang YI, Do SH, Lee SJ. Local BMP-7 release from a PLGA scaffolding-matrix for the repair of osteochondral defects in rabbits. J Control Release. 2012;162:485–491. doi: 10.1016/j.jconrel.2012.07.040.
    1. Yang C, Unursaikhan O, Lee JS, Jung UW, Kim CS, Choi SH. Osteoconductivity and biodegradation of synthetic bone substitutes with different tricalcium phosphate contents in rabbits. J Biomed Mater Res B. 2014;102:80–88. doi: 10.1002/jbm.b.32984.
    1. Kondo N, Ogose A, Tokunaga K, Umezu H, Arai K, Kudo N, Hoshino M, Inoue H, Irie H, Kuroda K, Mera H, Endo N. Osteoinduction with highly purified beta-tricalcium phosphate in dog dorsal muscles and the proliferation of osteoclasts before heterotopic bone formation. Biomaterials. 2006;27:4419–4427. doi: 10.1016/j.biomaterials.2006.04.016.
    1. Suzuki K, Anada T, Miyazaki T, Miyatake N, Honda Y, Kishimoto KN, Hosaka M, Imaizumi H, Itoi E, Suzuki O. Effect of addition of hyaluronic acids on the osteoconductivity and biodegradability of synthetic octacalcium phosphate. Acta Biomater. 2014;10:531–543. doi: 10.1016/j.actbio.2013.09.005.
    1. Bhakta G, Rai B, Lim ZX, Hui JH, Stein GS, van Wijnen AJ, Nurcombe V, Prestwich GD, Cool SM. Hyaluronic acid-based hydrogels functionalized with heparin that support controlled release of bioactive BMP-2. Biomaterials. 2012;33:6113–6122. doi: 10.1016/j.biomaterials.2012.05.030.
    1. Kisiel M, Klar AS, Ventura M, Buijs J, Mafina MK, Cool SM, Hilborn J. Complexation and sequestration of BMP-2 from an ECM mimetic hyaluronan gel for improved bone formation. PLoS ONE. 2013;8:e78551. doi: 10.1371/journal.pone.0078551.
    1. Ikada Y, Tabata Y. Protein release from gelatin matrices. Adv Drug Deliv Rev. 1998;31:287–301. doi: 10.1016/S0169-409X(97)00125-7.
    1. Poldervaart MT, Wang H, van der Stok J, Weinans H, Leeuwenburgh SC, Oner FC, Dhert WJ, Alblas J. Sustained release of BMP-2 in bioprinted alginate for osteogenicity in mice and rats. PLoS ONE. 2013;8:e72610. doi: 10.1371/journal.pone.0072610.
    1. Cao L, Duan PG, Wang HR, Li XL, Yuan FL, Fan ZY, Li SM, Dong J. Degradation and osteogenic potential of a novel poly(lactic acid)/nano-sized beta-tricalcium phosphate scaffold. Int J Nanomedicine. 2012;7:5881–5888. doi: 10.2147/IJN.S38127.
    1. Yang HS, La WG, Bhang SH, Jeon JY, Lee JH, Kim BS. Heparin-conjugated fibrin as an injectable system for sustained delivery of bone morphogenetic protein-2. Tissue Eng Part A. 2010;16:1225–1233. doi: 10.1089/ten.tea.2009.0390.

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