生物高分子PHBV之性質分析與孔洞基材製備及其細胞相容性之研究

Studies on properties and cell compatibility of porous PHBV Scaffolds

陶雨榮(Yu-Rong Tao)

指導教授 ： 陳志成

大同大學/工程學院/生物工程學系所/碩士(2007年)

摘要

聚羥基烷酯類 (polyhydroxyalkanoates, PHA) 具有與石化塑膠相似的物化特性並且又具有生物分解性和生物吸收性。應用醱酵技術生產之生物高分子聚羥基丁酯戊酯共聚酯(poly (3-hydroxybutyric acid-co-3-hydroxyvaleric acid);PHBV)可應用於供細胞生長之用的孔洞基材。本研究為評估 PHBV經熱壓鹽析法所生產之多孔基材，其材料性質和生物相容性。經由紅外線光譜法(FT-IR)發現，市售之ENMAT-PHBV(enPHBV)與耐鹽菌Haloferax mediterranei醱酵所生產之PHBV(hmPHBV)，具有相似的化學結構，經核磁共振光譜法(NMR)，發現兩者HV含量不同，enPHBV為3.61% 而hmPHBV為12.3% 。微分掃描式熱卡計(DSC)分析發現兩種PHBV之熔點(Tm)均有雙峰之情況，但因HV含量不同造成兩者Tm之差異；水接觸角分析發現熱壓製程溫度對於多孔性基材表面之水分吸附影響甚大；經由X-射線粉體繞射儀(XRD)發現以170℃製程溫度所製成之樣品其結晶性較佳，但高於170℃會造成材料結晶性的變化。而在鹽析法中利用不同鹽重量比例製備之多孔性基材，其鹽添加量的上升會造成孔隙度(Porosity)之上升、基材密度(Scaffold density)之下降與殘餘鹽量的降低。而在生物相容性方面，enPHBV多孔基材上進行人類皮膚纖維母細胞(CCD966SK)之培養，由掃描式電子顯微鏡SEM觀測，發現以熱壓法所生產之多孔性基材，其內孔分佈平均且孔徑均勻，且觀測到細胞於基材上生長並進行分化；經MTT (3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrasodium bromide)檢測活細胞數量後，發現160 ℃與170 ℃之製程溫度所生產之基材對於細胞培養的影響差異不大，但發現培養於enPHBV上之人類皮膚纖維母細胞，其活性與培養於無多孔enPHBV基材之環境下來比較可維持較長之時間。

關鍵字

生物高分子

並列摘要

Polyhydroxyalkanoates (PHA) have some characteristics similar to the petrochemical plastics and have bio-decomposition and bio-absorbability. In this study, PHBV was manufactured to the porous scaffold and used for cell culture. The PHBV porous scaffold was made by a heat-compressing method. By the infrared spectrum (FT-IR), it is found that the commercial ENMAT-PHBV (enPHBV) and PHBV (hmPHBV) have similar chemical structure. But by the analysis of the nuclear magnetic resonance (NMR), it was found that the HV content of two kinds of PHBV is different. The enPHBV has HV content of 3.61% and hmPHBV of 12.3%. By the differential scanning calorimetry (DSC) analysis, it was found that the melting point (Tm) of two kinds of PHBV all has a double peak phenomenon. The different HV content causes the difference of Tm in two type of PHBV. By water contact angle analysis, it was found that temperature of heat-compressing process had effect on the water-adsorbability of porous PHBV. By the X-ray diffraction (XRD) analysis, it was found that high temperature over 170℃ will cause the significant variation of crystallinity of material. It was found that higher weight ratio of salt in preparing the scaffold would increase the porosity and decrease the scaffold density. When human fibroblast cell (CCD966SK) was seeded in the enPHBV porous scaffold, we observed the cells growth on the scaffold by MTT (3- (4, 5 - dimethylthiazol-2-yl) -2,5-diphenyl tetrasodium bromide) method to quantify the living cells number. Different temperature of 160℃ and 170℃ to make the scaffold made no difference on cell culture. We found that the human fibroblast cell growth on enPHBV, compared with no porous scaffold, could maintain a longer time.

並列關鍵字

PHBV

參考文獻

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