2004 Fiscal Year Final Research Report Summary
Development of bioresponsive material for controllong cell proliferation and exfoliation
| Project/Area Number |
15500334
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | Kinki University |
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Principal Investigator |
SUGIYAMA Kazuo Kinki University, School of Engineering, Professor, 工学部, 教授 (00088577)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRAISHI Kohei Kinki University, School of Engineering, Professor, 工学部, 教授 (10196602)
IHARA Tatsuhiko Kinki University, School of Engineering, Professor, 工学部, 教授 (50133541)
HOSOYA Hiroshi Hiroshima University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (90183102)
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| Project Period (FY) |
2003 – 2004
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| Keywords | 2-hydroxypropyl methacrylamide / methyl methacrylate / Sodium Copper-chlorophyllin / thermoresponsible polymer / light-responsible polymer / cell exfoliation / HeLa Cell / 3T3 Cell |
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| Research Abstract |
Novel thermoresponsible random copolymers, poly(HPMA-co-MMA), and light responsible random copolymers, poly(HPMA-co-MMA/Chlph) were prepared from biocompatible 2-hydroxypropylmethacrylamide(HPMA), methyl methacrylate (MMA), and sodium copper-chlorophyllin(Chlph) as a basic polymer backbone to give an artificial extra cellular matrix for controlling cell proliferation and exfoliation. This research results ranged from (I) to (V) as described below in basic and application examination.
(I)Graft-polymerization onto diamond like carbon (DLC) coating material : The graft-polymerization of vinyl monomer was developed onto DLC coated inorganic materials on three-dimensional pieces with complex shapes based on low temperature plasma irradiation/post-polymerization technique onto polymer material.
(II)Development of novel polymer material as an artificial extra cellular matrix : RGDS(cell attachment peptide) was immobilized on the thermoresponsible poly(HPMA-co-MMA) grafted polyethyleneterephthal
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ate(g-PET) as a substrate via two step reaction as follows : (1)N-methacryloylsuccinimide (NASu) was grafted onto g-PET by Ar plasma / post polymerization, (2)Synthetic RGDS was reacted with MASu grafted g-PET. The rate of HeLa cells proliferation until 48 hr was considerably increased comparing to non-RGDS treated poly(HPMA-co-MMA) under normal cell cultivative condition. The more than 80% of cultured HeLa cells were exfoliated by decreasing temperature (25℃) for 2hr while non RGDS treated poly(HPMA-co-MMA) and untreated PET was about 70% and 10%, respectively Platelets adhesion was suppressed on newly designed a zwitterionic poly(N^α-methacrylamide-L-lysiene) grafted film as a typical example among examined samples. It was found that the developed polymer surface modification enables us to control cell attachment and exfoliation. The evaluation of surface properties for designing appropriate chemical structure was newly proposed by using bacterial random peptide library method(FliTrx^<TM> : Invitrogen). The hydrophilic-hydrophobic change of poly(HPMA-co-MMA) grafted PET on heating was able to be shown by using hydrophobic parameters from selective amino acid sequences.
(III)Light responsible polymer material as a cell scaffold : Poly(HPMA-co-MMA/Chlph) showed a lower critical solution temperature(LCST) under visible light irradiation. For example, the maximum 68% of attached 3T3 cells was exfoliated after turning off the light irradiation for 2hr. The novel cell culture and exfoliation systems was newly developed by visible light controlling.
(IV)Microporous hydroxyapatide(HAP) and/or visible light responsible TiO_2 hybrid material : HAP and/or TiO_2 hybrid material as a scaffold was prepared as follows ; (1)Microspheres with phosphoric acid on the surface was allowed to stand at 80 ℃ in HAP forming and/or TiO_2 suspended solution to give HAP/TiO_2 coated microspheres(HAP/TiO_2 -MS). (2)The molded HAP/TiO_2-MS to like disk was calcinated at 600℃ to give a microprous HAP/TiO_2.The cell attachment and exfoliation is now under consideration.
(V)Thermo- and/or pH sensitive hydrogels : Molecularly imprinted thermosenstive and/or pH responsible hydrogels obtained from N-isopropylacrylamide and/or zwitterionic O-methacryloyl-L-serine were prepared far basic examinations for controlling and timing release of such as cell differentiation or proliferation factors. Less
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