Osteogenic differentiation of bone marrow derived stem cells in gelatinhydroxyapatite nanocomposite
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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 (20-40μm). 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.
Abstract
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 (20-40μm). 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.
참고문헌 (27)
- Zerwekh JE, "Fibrillar collagen.biphasic calcium phosphate composite as a bone graft substitute for spinal fusion", J Orthop Res, 10, pp.562~572 (1992)
- Liu L, "Preparation and charac-terization of collagen-hydroxyapatite composite used for bone tissue engineering scaffold", Artif Cells Blood Substit ImmobilBiotechnol, 31, pp.435~448 (2003)
- Sachlos E, "Collagen scaffolds reinforced with biomimetic composite nano-sized carbonate-substituted hydroxyapatite crystals and shaped by rapid prototyping to contain internal microchannels", Tissue Eng, 12, pp.2479~2487 (2006)
- Chang MC, "Cross-linkage of hydroxyapatitegelatin nanocomposite using EDGE", J Mater Sci, 39, pp.5547~5550 (2004)
- Kim HW, "Porous scaffolds of gelatin- hydroxyapatite nanocomposites obtained by biomimetic approach: Characterization and antibiotic drug release", J Biomed Mater Res B Appl Biomater, 74, pp.686~698 (2005)
- Chang MC, "Modification of hydroxyapatitegelatin composite by polyvinylalcohol", J Mater Sci, 40, pp.505~509 (2005)
- Chang MC, "Preparation of hydroxyapatite - gelatin nanocomposite", Biomaterials, 24, pp.2853~2862 (2003)
- 장명철, "Molecular Orbital Calculation on the Configuration of Hydroxyl Group in Hexagonal Hydroxyapatite", 한국세라믹학회지, 42(5), pp.15~30 (2005), 한국세라믹학회
- Chang MC, "Organic-inorganic interaction and the growth mechanism of hydroxyapatite crystals in gelatin matrics between 37 and 80℃", J Mater Sci:Mater Med, 17, pp.387~396 (2006)
- 장명철, "Morphology Development of HAp Crystallites in GEL Matrix", 한국세라믹학회지, 44(3), pp.133~136 (2007), 한국세라믹학회
- Chang MC, "Conformational change of hydroxyaptite", gelatin nanocomposite by glutaradehyde, 2003, pp.23, 3087~3094
- Preece A, "A manual for histologic technicians", pp.145~147 (1972), Bowrn and Co
- Lee RH, "Characterizaiton and expression analysis of mesenchymal stem cells from human bone marrow and adipose Tissue", Cell Physiol Biochem, 14, pp.311~324 (2004)
- Kim HW, "Stimulation of osteoblast responses to biomimetic nanocomposites of gelatin-hydroxyapatite for tissue enginnering scaffolds", Biomaterials, 26, pp.5221~5230 (2005)
- Yaylaoglu MB, "Development of a calcium phosphate.gelatin composite as a bone substitute and its use in drug release", Biomaterials, 20, pp.711~719 (1999)
- Fukunaka Y, "Controlled release of plasmid DNA from cationized gelatin hydrogels based on hydrogel degradation", J Control Rel, 80, pp.333~343 (2002)
- Kim HW, "Hydroxyapatite and gelatin composite foams processed via novel freeze-drying and crosslinking for use as temporary hard tissue scaffolds", J Biomed Mater Res A, 72, pp.136~145 (2005)
- Shen FH, "Osteogenic differentiation of adiposederived stromal cells treated with GDF-5 cultured on a novel threedimensional sintered microsphere matrix", Spine, 6, pp.615~623 (2006)
- Donzelli E, "Mesenchymal stem cells cultured on a collagen scaffold: in vitro osteogenic differentiation", Arch Oral Biol, 52, pp.64~73 (2007)
- Kimura Y, "Biodegradable polymers, in Biomedical applications in polymeric materials", pp.163~165 (1993), CRC press
- Wei G, "Structure and properties of nanohydroxyapatite", polymer composite scaffolds for bone tissue engineering, 2004, pp.14, 4749~4757
- Kenley RA, "Biotechnology and bone graft substitutes", Pharm Res, 10, pp.1393~1401 (1993)
- Khan SN, "The biology of bone grafting", J Am Acad Orthop Surg, 13, pp.77~86 (2005)
- Pittenger MF, "Multilineage potential of adult human mesenchymal stem cells", Science, 284, pp.143~147 (1999)
- Jaiswal N, "Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro", J Cell Biochem, 64, pp.295~312 (1997)
- Yang S, "The design of scaffolds for use in tissue engineering. Part I.traditional factors", Tissue Eng, 7, pp.679~689 (2001)
- El-Amin SF, "Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering", Biomaterials, 4, pp.1213~1221 (2003)