The Effect of Bone Morphogenic Protein-2 (BMP-2)-Immobilizing Heparinized-Chitosan Scaffolds for Enhanced Osteoblast Activity

다국어 초록 (Multilingual Abstract)

The aim of orthopedic and dental tissue engineering is to generate synthetic bone-graft tissue substitutes. It is generally comprised through the combination of viable cells, a scaffolding material, and sometimes the inclusion of bone morphogenic proteins. The object of this study is to develop novel bone-grafting scaffolds that enhance osteoblast activity. We were fabricated the chitosan scaffolds with channel-shaped and spherically shaped pore morphologies.
Also, Bone morphogenic protein-2 (BMP-2) was sequentially immobilized to the heparinized-chitosan (Hep-chitosan)scaffolds. Osteoblast activities of all chitosan scaffolds were investigated by a cell proliferation assay, alkaline phosphatase (ALP) activity, calcium deposition, and the expression of osteogenic markers. The results showed that BMP-2-immobilizing heparinized-chitosan (BMP-2/Hep-chitosan) scaffolds significantly enhanced ALP activity and calcium deposition of the osteoblast cells when compared with chitosan scaffolds only. Also, mRNA expressions of osteocalcin and osteopontin of osteoblast cells cultured on BMP-2 (100 ng)/Hep-chitosan scaffolds were increased versus chitosan scaffolds only. Taken together, BMP-2 (100 ng)/Hep-chitosan scaffolds could achieve the functions of excellent osteoblast promotion. Therefore, osteoinductive protein-functionalizing scaffold substrates such as BMP-2/Hepchitosan scaffolds are a promising material for the enhanced osteoblast activity in orthopedic and dental fields

참고문헌 (Reference)

1 RS Sellers, "The effect of recombinanthuman bone morphogenetic protein-2 (rhBMP-2) on the healingof full-thickness defects of articular cartilage" 79 : 1452-, 1997

2 SE Kim, "The effect of immobilizationof heparin and bone morphogenic protein-2 (BMP-2) to titaniumsurfaces on inflammation and osteoblast function" 32 : 366-, 2011

3 H Lin, "The effect of crosslinking heparinto demineralized bone matrix on mechanical strength and specific binding to human bone morphogenetic protein-2" 29 : 1189-, 2008

4 S Yang, "The design of scaffolds for usein tissue engineering. Part I. traditional factors" 7 : 679-, 2001

5 Z Shi, "Surface functionalization oftitanium with carboxymethyl chitosan and immobilized bonemorphogenetic protein-2 for enhanced osseointegration" 10 : 1603-, 2009

6 GN King, "Recombinant humanbone morphogenetic protein-2 promotes wound healing in ratperiodontal fenestration defects" 76 : 1460-, 1997

7 MC Wake, "Pore morphology effectson the fibrovascular tissue growth in porous polymer substrates" 3 : 339-, 1994

8 ST Choi, "Osteopontin might beinvolved in bone remodelling rather than in inflammation inankylosing spondylitis" 47 : 1775-, 2008

9 R Sasisekharan, "On the regulation offibroblast growth factor activity by heparin-like glycosaminoglycans" 1 : 45-, 1997

10 JM Wozney, "Novel regulators ofbone formation: molecular clones and activities" 242 : 1528-, 1988

11 GC Steffens, "Modulation of angiogenicpotential of collagen matrices by covalent incorporation ofheparin and loading with vascular endothelial growth factor" 10 : 1502-, 2004

12 A Khademhosseini, "Microscaletechnologies for tissue engineering and biology" 103 : 2480-, 2006

13 BG Amsden, "Methacrylated glycolchitosan as a photopolymerizable biomaterial" 8 : 3758-, 2007

14 F Deschaseaux, "Mechanisms of bonerepair and regeneration" 15 : 417-, 2009

15 Y Sheng, "Long-circulating polymericnanoparticles bearing a combinatorial coating of PEG andwater-soluble chitosan" 30 : 2340-, 2009

16 K Turksen, "Isolation of monoclonalantibodies recognizing rat bone-associated molecules in vitro andin vivo" 40 : 1339-, 1992

17 GK Tan, "Interactions betweenmeniscal cells and a self assembled biomimetic surface composedof hyaluronic acid, chitosan and meniscal extracellular matrixmolecules" 31 : 6104-, 2010

18 ST Nillesen, "Increased angiogenesisand blood vessel maturation in acellular collageneheparin scaffoldscontaining both FGF2 and VEGF" 28 : 1123-, 2007

19 MJ Wissink, "Improvedendothelialization of vascular grafts by local release of growthfactor from heparinized collagen matrices" 64 : 103-, 2000

20 G Zellin, "Importance of delivery systems for growthstimulatoryfactors in combination with osteopromotivemembranes. an experimental study using rhBMP-2 in ratmandibular defects" 35 : 181-, 1997

21 E Khor, "Implantable applications of chitin andchitosan" 24 : 2339-, 2003

22 YJ Park, "Immobilization of bonemorphogenetic protein-2 on a nanofibrous chitosan membrane forenhanced guided bone regeneration" 43 : 17-, 2006

23 "ISO document 10993. Biological compatibility of medicaldevices* Part 5. Tests for cytotoxicity: in vitro methods"

24 N Bordenave, "Hydrophobization andantimicrobial activity of chitosan and paper-based packagingmaterial" 11 : 88-, 2010

25 BS Vladimirov, "Growth factors--importance andpossibilities for enhancement of the healing process in bonefractures" 46 : 11-, 2004

26 DW Lee, "Gentamicin and bonemorphogenic protein-2 (BMP-2)-delivering heparinizedtitaniumimplant with enhanced antibacterial activity andosteointegration" 50 : 974-, 2012

27 JJ van den Beucken, "Functionalization of multilayered DNA-coatings with bonemorphogenetic protein 2" 113 : 63-, 2006

28 S Hirota, "Expression of mRNAof murine bone-related proteins in ectopic bone induced bymurine bone morphogenetic protein-4" 277 : 27-, 1994

29 A Perets, "Enhancing thevascularization of three-dimensional porous alginate scaffolds byincorporating controlled release basic fibroblast growth factormicrospheres" 65 : 489-, 2003

30 K Whang, "Engineering boneregeneration with bioabsorbable scaffolds with novelmicroarchitecture" 5 : 35-, 1999

31 MJ Wissink, "Endothelialcell seeding of (heparinized) collagen matrices: effects of bFGFpre-loading on proliferation (after low density seeding) and procoagulantfactors" 67 : 141-, 2000

32 NK Lee, "Endocrine regulation ofenergy metabolism by the skeleton" 130 : 456-, 2007

33 QP Pham, "Electrospun poly(epsiloncaprolactone)microfiber and multilayer nanofiber/microfiberscaffolds: characterization of scaffolds and measurement ofcellular infiltration" 7 : 2796-, 2006

34 HJ Moon, "Effect of heparin andalendronate coating on titanium surfaces on inhibition ofosteoclast and enhancement of osteoblast function" 413 : 194-, 2011

35 VK Mourya, "Chitosan-modifications and applications:opportunities galore" 68 : 1013-, 2008

36 CKS Pillai, "Chitin and chitosan polymers:chemistry, solubility and fiber formation" 34 : 641-, 2009

37 J Kim, "Characterization of low-molecularweighthyaluronic acid-based hydrogel and differential stem cellresponses in the hydrogel microenvironments" 88 : 967-, 2009

38 D Depan, "Cell proliferation andcontrolled drug release studies of nanohybrids based on chitosang-lactic acid and montmorillonite" 5 : 93-, 2009

39 J Becerra, "Boneregeneration, cell therapy and tissue engineering" 116 : 23-, 2001

40 MR Urist, "Bone: formation by autoinduction" 150 : 893-, 1965

41 J Kim, "Bone regeneration usinghyaluronic acid-based hydrogel with bone morphogenicprotein-2 and human mesenchymal stem cells" 28 : 1830-, 2007

42 BE Bax, "Bone morphogeneticprotein-2 increases the rate of callus formation after fracture ofthe rabbit tibia" 65 : 83-, 1999

43 N Sykaras, "Bone morphogenetic proteins(BMPs): how do they function and what can they offer theclinician" 45 : 57-, 2003

44 T Ishibe, "Bone formation on apatitecoatedtitanium with incorporated BMP-2/heparin in vivo" 108 : 867-, 2009

45 YY Liang, "Bioconjugation of papain onsuperparamagnetic nanoparticles decorated with carboxymethylatedchitosan" 8 : 1480-, 2007

46 AC Larkin, "Biocompatible,detachable, and free-standing polyelectrolyte multilayer films" 11 : 2788-, 2010

47 RH Christenson, "Biochemical markers of bone metabolism: anoverview" 30 : 573-, 1997