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Title: 利用生物分子架構新型骨再生基質之研究
A Study for a Novel Bone Regeneration Matrix by Biomacromolecules
Authors: 許富銀
Contributors: NTOU:Institute of Bioscience and Biotechnology
Keywords: 膠原蛋白;明膠;二級結構;生物活性奈米纖維玻璃
collagen;gelatin;secondary structure;bioactive glass nanofiber
Date: 2009-08
Issue Date: 2011-06-28T07:18:01Z
Publisher: 行政院國家科學委員會
Abstract: 摘要:骨組織主要是由第一型膠原蛋白與生物磷灰石所組成的,所以骨組織為一硬且具有彈性特性之組織結構。由於膠原蛋白在體外的製程中常能保有其特定之生理及物化特性,以致於常被用來當作細胞培養時之培養支架,以提供較佳的臨床使用性。但是利用電氣紡織製備膠原蛋白纖維結構時,常使用氟代醇類之溶劑(例如1,1,1,3,3,3-hexafluoro-2-propanol 或 2,2,2-trifluoroethanol) 而造成所製得之膠原蛋白纖微喪失膠原蛋白特定之周期性微結構(D-banding structure)。過去我們已建立電氣紡織技術,並且成功的製備出膠原蛋白/透明質酸奈米複合纖維。所以本研究計畫第一年主要在製備並探討經電氣紡織製得之明膠及膠原蛋白纖維的生物分子二級結構,並探討不同二級結構之生物分子電氣紡織纖維對MG63類骨母細胞(osteoblasts)之增生與分化的影響。另一方面,所以我們將探討在不同溶劑條件下經電氣紡織所製備之膠原蛋白纖維的二級結構與四級結構,以製備出具有D-banding結構之膠原蛋白電氣紡織纖維。並探討具D-banding結構之膠原蛋白電氣紡織纖維對MG63類骨母細胞之增生與分化的影響。 過去我們也曾經製備出含生物分子及生物活性無機質之複合基材,以應用在骨組織再生。在體外實驗發現含生物活性無機質會有較佳的骨生成特性的表現(如osteocalcin 與alkaline phosphatase的表現)。因此,在第二年將利用電氣紡織技術製備具有生物活性玻璃奈米纖維強化型之順向性生物分子複合基質,並探討其對MG63類骨母細胞之增生與分化的影響。此外,將複合基質浸泡於模擬生物體體液中,探討生物活性奈米纖維玻璃對基質表面形成氫氧基磷灰石的影響,並探討不同形狀及大小的氫氧基磷灰石其對MG63類骨母細胞的影響。 在第三年則利用電氣紡織技術製備具有中空柱狀的玻璃奈米纖維強化型之順向性複合基質。此外,並將過去所開發的擬骨微粒載體,在體外培養骨髓幹細胞,並進一步將含骨髓幹細胞之微粒載體與platelet-rich plasma與凝血酶 (thrombi)混合,可得到含細胞、生長因子與支架之骨填補複合物充填於中空柱狀之複合基質內,最後以動物實驗驗證此新型骨再生基質對骨癒合之促進能力。
Abstract:Bone is both hard and resilient, mainly consisting of Type I collagen and bioapatite. Scaffolds manufactured from collagen promise better clinical functionality because its signature biological and physico-chemical properties are retained in in vitro preparations. However, numerous studies reveal collagen lack the unique ultra-structural axial periodicity when it is electro-spun into nano-fibres out of fluoroalcohols such as 1,1,1,3,3,3-hexafluoro-2-propanol or 2,2,2-trifluoroethanol. According the previous NSC project practiced, we had succeeded to fabricate an electrospun nanofibrous matrix composed of hyaluronic acid and collagen. Therefore, one of the aims of the project at the first year is to study the influence of the secondary structure of collagen and gelatin of electrospun nanofibrous cell-cultured matrix on the phenotype, proliferation, and maturation of osteoblasts. In the other hand, we will fabricate a collagen electrospun nanofibrous matrix with a characteristic D-pattern by adjusting solvent or other processing parameters, and evaluate the effects of specific structures on cellular behaviors. In the past, we fabricated a composite containing biomolecule and bioactive inorganics, for bone tissue regeneration. When compared to the biomolecule individual, its composite with bioactive inorganic has shown better osteogenic properties in vitro, such as the expression of alkaline phosphatase and osteocalcin. Along the conclusions, at 2nd year, we will focus on to fabricate a bioactive glass nanofiber-reinforced biomolecule fibrous matrix, and evaluate the effect of this composite on cellular behaviors. Besides, we will investigate of surface structure changes by immersing the electrospun scaffold in the simulated body fluids for various time periods, and assess the effects of size and shape of nanoapatite on cellular behaviors. At 3rd year, we will devote to fabricate a hollow cylinder of aligned fibers and reinforced with bioactive glass nanofiber. We had developed bone-like microcarriers, and it supported the attachment and proliferation of MG63 osteoblast-cells at the past NSC project. Thus, consequently, we will culture the bone marrow stem cells onto the bone-like microcarriers to expand the number of bone marrow stem cells. Moreover, we will mix platelet rich plasma, thrombin with microcarriers containing bone marrow stem cell to form a bone graft gel, and fill it into hollow cylinder. Finally, we will evaluate the performance of this novel bone regeneration matrix on bone regeneration.
Relation: NSC98-2221-E019-004
Appears in Collections:[Department of Bioscience and Biotechnology ] Research Reports

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