Journal of the Korean Association of Oral and Maxillofacial Surgeons

  • 제35권1호
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  • Pages.7-12
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  • 2009
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  • 2234-7550(pISSN)
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  • 2234-5930(eISSN)
대한구강악안면외과학회 (The Korean Association of Oral and Maxillofacial Surgeons)
권호 표지

Osteogenic differentiation of bone marrow derived stem cells in gelatin-hydroxyapatite nanocomposite

  • 발행 : 2009.02.28
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초록

Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores ($20-40{\mu}m$). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.

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참고문헌

  1. Zerwekh JE, Kourosh S, Schienbergt R.: Fibrillar collagen.biphasic calcium phosphate composite as a bone graft substitute for spinal fusion. J Orthop Res 1992;10: 562-72 https://doi.org/10.1002/jor.1100100411
  2. Liu L, Zhang L, Ren B, Wang F, Zhang Q. : Preparation and charac-terization of collagen-hydroxyapatite composite used for bone tissue engineering scaffold. Artif Cells Blood Substit ImmobilBiotechnol 2003; 31:435-48 https://doi.org/10.1081/BIO-120025414
  3. Sachlos E, Gotora D, Czernuszka JT.: Collagen scaffolds reinforced with biomimetic composite nano-sized carbonate-substituted hydroxyapatite crystals and shaped by rapid prototyping to contain internal microchannels. Tissue Eng 2006; 12:2479-87 https://doi.org/10.1089/ten.2006.12.2479.
  4. Chang MC, Ikoma T, Tanaka J.: Cross-linkage of hydroxyapatitegelatin nanocomposite using EDGE. J Mater Sci 2004; 39: 5547 - 5550 https://doi.org/10.1023/B:JMSC.0000039284.70028.fa
  5. Kim HW, Knowles JC, Kim HE.: Porous scaffolds of gelatin- hydroxyapatite nanocomposites obtained by biomimetic approach: Characterization and antibiotic drug release. J Biomed Mater Res B Appl Biomater 2005; 74: 686-698 https://doi.org/10.1002/jbm.b.30236
  6. Chang MC, Ko CC, Douglas WH.: Modification of hydroxyapatitegelatin composite by polyvinylalcohol. J Mater Sci 2005; 40: 505 - 509 https://doi.org/10.1007/s10853-005-6115-1
  7. Chang MC, Ko CC, Douglas WH.: Preparation of hydroxyapatite - gelatin nanocomposite. Biomaterials 2003; 24: 2853-2862 https://doi.org/10.1016/S0142-9612(03)00115-7
  8. Chang MC.: Molecular orbital calculation on the configuration of hydroxyl group in hexagonal hydroxyapatite. J Kor Ceramic Society 2005; 42: 304-307 https://doi.org/10.4191/KCERS.2005.42.5.304
  9. Chang MC, Douglas WH, Tanaka J.: Organic-inorganic interaction and the growth mechanism of hydroxyapatite crystals in gelatin matrics between 37 and 80${^{\circ}C}$. J Mater Sci:Mater Med 2006; 17: 387-396 https://doi.org/10.1007/s10856-006-8243-9
  10. Chang MC.: Morphology development of HAp crystallites in Gel Matrix. J Kor Ceramic Society 2007; 44: 133-136 https://doi.org/10.4191/KCERS.2007.44.3.133
  11. Chang MC, Ko CC, Douglas WH.: Conformational change of hydroxyaptite/gelatin nanocomposite by glutaradehyde. Biomaterials 2003; 23: 3087-3094 https://doi.org/10.1016/S0142-9612(03)00150-9
  12. Preece A: A manual for histologic technicians. Boston. MA.Little, Bowrn and Co, 1972: 145-147
  13. Lee RH, Kim BC, Choi IS: Characterizaiton and expression analysis of mesenchymal stem cells from human bone marrow and adipose Tissue. Cell Physiol Biochem 2004; 14: 311-324 https://doi.org/10.1159/000080341
  14. Kim HW, Kim HE, Salih V.: Stimulation of osteoblast responses to biomimetic nanocomposites of gelatin-hydroxyapatite for tissue enginnering scaffolds. Biomaterials 2005; 26: 5221-5230 https://doi.org/10.1016/j.biomaterials.2005.01.047
  15. Yaylaoglu MB, Korkusuz P, Ors U, Korkusuz F, Hasirci V.: Development of a calcium phosphate.gelatin composite as a bone substitute and its use in drug release. Biomaterials1999; 20:711-719 https://doi.org/10.1016/S0142-9612(98)00199-9
  16. Fukunaka Y, Iwanaga K, Morimoto K, Kakemi M, Tabata Y.: Controlled release of plasmid DNA from cationized gelatin hydrogels based on hydrogel degradation. J Control Rel 2002; 80:333-43 https://doi.org/10.1016/S0168-3659(02)00026-3
  17. Kim HW, Knowles JC, Kim HE.: Hydroxyapatite and gelatin composite foams processed via novel freeze-drying and crosslinking for use as temporary hard tissue scaffolds. J Biomed Mater Res A. 2005; 72:136-45 https://doi.org/10.1002/jbm.a.30168
  18. Shen FH, Zeng Q, Choi L: Osteogenic differentiation of adiposederived stromal cells treated with GDF-5 cultured on a novel threedimensional sintered microsphere matrix. Spine 2006; 6: 615-623 https://doi.org/10.1016/j.spinee.2006.03.006
  19. Donzelli E, Salvade A, Mimo P: Mesenchymal stem cells cultured on a collagen scaffold: in vitro osteogenic differentiation. Arch Oral Biol 2007; 52: 64-73 https://doi.org/10.1016/j.archoralbio.2006.07.007
  20. Kimura Y. Biodegradable polymers. In; Tsuruta T, editor: Biomedical applications in polymeric materials. Boca Raton: CRC press, 1993: 163-165
  21. Wei G, Ma PX.: Structure and properties of nanohydroxyapatite/ polymer composite scaffolds for bone tissue engineering. Biomaterials 2004; 25:4749-57 https://doi.org/10.1016/j.biomaterials.2003.12.005
  22. Kenley RA, Yim K, Abrams J, Ron E, Turek T, Marden LJ: Biotechnology and bone graft substitutes. Pharm Res 1993;10: 1393- 401 https://doi.org/10.1023/A:1018902720816
  23. Khan SN, Cammisa Jr FP, Sandhu HS, Diwan AD, Girardi FP, Lane JM.: The biology of bone grafting. J Am Acad Orthop Surg, 2005; 13: 77-86 https://doi.org/10.5435/00124635-200501000-00010
  24. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD: Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284:143-7 https://doi.org/10.1126/science.284.5411.143
  25. Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP.: Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 1997; 64: 295-312 https://doi.org/10.1002/(SICI)1097-4644(199702)64:2<295::AID-JCB12>3.0.CO;2-I
  26. Yang S, Leong K-F, Du Z, Chua C-K.: The design of scaffolds for use in tissue engineering. Part I.traditional factors. Tissue Eng 2001; 7:679-89
  27. El-Amin SF, Lu HH, Burems J, Mitchell J, Tuan RS, Laurencin CT.: Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering. Biomaterials 2003; 4:1213-21 https://doi.org/10.1016/S0142-9612(02)00451-9