聚（3-羟基丁酸3-羟基戊酸3-羟基己酸）生物相容性的研究
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摘要
三聚物P(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate)(PHBVHHx)是通过重组菌Aeromonas hydrophila发酵获得的，其包括5.3 mol％3-hydroxyvalerate(3-HV)和10.2 mol％3-hydroxyhexanoate(3-HHx)。本文第一次在体外通过与poly(L-lactic acid)(PLA)，poly(3-hydroxybutyrate)(PHB)，poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(PHBV)and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)(PHBHHx)的对比研究三聚物PHBVHHx的生物相容性。扫描电镜结果表明只有在PHBVHHx上才具有螺纹状的形态结构。噻唑蓝检测证明三聚物PHBVHHx在促进小鼠成纤维细胞L929和成骨细胞MC3T3的粘附和繁殖方面都分别比其他四种多聚物材料强。兔子体内组织学研究表明PHBVHHx是一个温和无伤害的生物植入材料。因此，PHBVHHx其可塑的物化性质和其生物相容性都使它可能成为新一代的生物植入材料。
Terpolyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoat)(PHBVHHx) containing 5.3 tool% 3-hydroxyvalerate (3-HV) and 10.2 tool%3-hydroxyhexanoate (3-HHx) was obtained via microbial synthesis using recombinantAeromonas hydrophila. For the first time in vitro biocompatibility of the terpolyesterwas evaluated in comparison with poly(L-lactic acid) (PLA), poly (3-hydroxybutyrate)(PHB), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) andpoly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx). Scanning electronmicroscopy (SEM) showed whorls-like morphology only on PHBVHHx film surface.Methylthiazol tetrazolium (MTT) assay demonstrated that PHBVHHx was better thanthe above four materials in promoting cell attachment and proliferation of fibroblastcell line L929 and osteoblast cell line MC3T3, respectively. Histological study usingrabbits proved that PHBVHHx was a fairly harmless implantable biomaterial. Thus,PHBVHHx with an adjustable mechanical property, combined with itsbiocompatibility, are in the process of developing into a new generation of bioimplantmaterial.
Terpolyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoat)(PHBVHHx) containing 5.3 tool% 3-hydroxyvalerate (3-HV) and 10.2 tool%3-hydroxyhexanoate (3-HHx) was obtained via microbial synthesis using recombinantAeromonas hydrophila. For the first time in vitro biocompatibility of the terpolyesterwas evaluated in comparison with poly(L-lactic acid) (PLA), poly (3-hydroxybutyrate)(PHB), poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) andpoly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx). Scanning electronmicroscopy (SEM) showed whorls-like morphology only on PHBVHHx film surface.Methylthiazol tetrazolium (MTT) assay demonstrated that PHBVHHx was better thanthe above four materials in promoting cell attachment and proliferation of fibroblastcell line L929 and osteoblast cell line MC3T3, respectively. Histological study usingrabbits proved that PHBVHHx was a fairly harmless implantable biomaterial. Thus,PHBVHHx with an adjustable mechanical property, combined with itsbiocompatibility, are in the process of developing into a new generation of bioimplantmaterial.
引文
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[16] Hazenberg W, Witholt B. Efficient production of medium-chain-length poly(3-hydroxyalkanoate) from octane by Pseudomonas oleovorans: economical considerations. Appl. Microbiol. Biotechnol. 1997, 48: 588-596.
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[27] Beckstead B L, Pan S, Bhrany A D, Bratt-Leal AM, Ratner B D, Giachelli C M. Esophageal epithelial cell interaction with synthetic and natural scaffolds for tissue engineering. Biomaterials. 2005, 26: 6217-28.
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[29] Qiu Q Q, Ducheyne P, Ayyaswamy P S. New bioactive, degradable composite microspheres as tissue engineering substrates. J. Biomed. Mater. Res. 2000, 52: 66-76.
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[33] Du C, Cui F Z, Feng Q L, zhu C D, de Groot K. Tissue response to nano-hydroxyapatite/collagen composite implants in marrow cavity. J Biomed Mater Res. 1998, 42: 540-548
[34] Zinn, M., Witholt, B. and Egli, T. Occurrence, synthesis and medical application of bacterial polyhydroxyalkanoates. Adv. Drug Delivery Rev. 2001, 53: 5-21.
[35] Anderson A J, Dawes E A. Occurrence, metabolism, metabolic role and industrial uses of bacterial polyhydroxyalkanoates. Microbiol. Rev. 1990, 54: 450-472
[36] Korsatko W. Retard implantation tablets on a matrix base with Eudragit as a structural substance. Pharmazie 1982,37: 272-3
[37] Laffferty R M, Korsatko B, Korsatko W. Microbial production of poly-3-hydroxybutyric acid. Biotechnology: A comprehensive treatise, special microbial processes, VCH Verlagsgesellschaft, Weinhheim.1988,6: 135-176
[38] 王常勇，袁晓辉 刘爽等．聚羟基丁酸酯载体人工软骨体内培育的实验研究．中华外科杂志，2000，38：269-271．
[39] Sevastianov V I, Perova N V, Shishatskaya E I, Kalacheva G S, Volova T G. Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood. J. Biomater. Sci. Polym. Ed. 2003,14: 1029-42
[40] Mosahebi A, Fuller P, Wiberg M, Terenghi G. Effect of allergenic Schwann cell transplantation on peripheral nerve regeneration. Exp. Neurol. 2002, 173:213-23
[41] Gerlach K L and Pesch H J. In vivo evaluation of a copolymer of polyhydroxybutyrate and polyhydroxyvalerate. Abstracts of the 8th European Conference on Biomaterials. Heidelberg. 1989, B3-1:79.
[42] Leenstra T S, Kuijpers-Jagtman A M, Maltha J C. The healing process of palatal tissues after palatal surgery with and without implantation of membranes: an experimental study in dogs. J. Mater. Sci. Mater. Med. 1998, 9:249-55
[43] Rivard C H, Chaput C, Rhalmis. Bioabsorbable synthetic polyesters and tissue regeneration. A study of three-dimensional proliferation of ovin chondrocytes and psteoblasts. Ann. Chir. 1996, 50: 651-658.
[44] Sevastianov V I, Perova N V, Shishatskaya E I, Kalacheva G S, Volova T G. Production of purified polyhydroxyalkanoates (PHAs) for applications in contact with blood. J. Biomater. Sci. Polym. Ed. 2003, 14:1029-42.
[45] Shishatskaya E I, Volova T G. A comparative investigation of biodegradable polyhydroxyalkanoate films as matrices for in vitro cell cultures. J. Mater. Sc.i. Mater. Med. 2004, 15: 915-923
[46] Williams S F, Martin. Therapeutic uses of polymers and oligomers comprising gamma-hydroxybutyrate. US Patent Appl. 661948, 2000.
[47] Engelmayr G C, Hildebrand D K, Sutherland F W H. A novel bioreactor for the dynamic flexural stimulation of tissue engineered heart valve biomaterials. Bimaterials 2003, 24: 2523-2532.
[48] Sodian R, Loebe M, Hein A, Martin D P, Hoerstrup S P, Potapov E V, Hausmann H, Lueth T, Hetzer R. Application of stereo lithography for scaffold fabrication for tissue engineered heart valves. Asaio. J. 2002, 48: 12-6.
[49] Opitz F, Schenke-Layland K, Cohnert T U, Starcher B, Halbhuber K J, Martin D P, Stock U A. Tissue engineering of aortic tissue: dire consequence of suboptimal elastic fiber synthesis in vivo. Cardiovasc. Res. 2004, 63: 719-30.
[50] Sodian R, Hoerstrup S P, Sperling J S. Early in vivo experience with tissue-engineered trileaflet heart valves. Circulation. 2000, 102: 22-29.
[51] Sodian R, Hoerstrup S P, Sperling J S. Tissue engineering of heart valves: In vitro experiences. Ann. Thorac. Surg. 2000, 70: 140-144.
[52] Perry T E, Kaushal S, Sutherland F W H. Bone marrow as a cell source for tissue engineering heart valves. Ann. Thorac. Surg. 2003, 75: 761-767.
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