1287978 九、發明說明： 【發明所屬之技術領域】 本發明是有關於一種生物高分子複合纖維，且特別是 有關於一種由海藻酸鹽及膠原蛋白交聯所形成的複合海 藻酸鹽纖維。 【先前技術】 常見的天然高分子材料有來自動物的膠原蛋白 (collagen)、明膠（gelatin)、透明質酸（hyaluronic acid)、幾 丁質（chitin)、幾丁聚醣（chitosan)及其衍生物等；以及來自 於植物的海藻酸（alginate)、纖維素（ceiiui〇se)及其衍生物 等。海藻酸鹽與膠原蛋白皆為生物可吸收性材料，目前廣 泛應用於醫療用敷材、美容保養用品及食品添加物上。 過去曾利用海藻酸鹽以濕式紡絲方式製成海蒸酸鹽 纖維’其纖維之機械強度有限以致受到應用上的限制，若 將海藻酸鹽纖維織成不織布則可具有相當強度，故目前以 製成不織布形式的海藻酸鹽纖維應用較廣。海藤酸鹽纖維 為天然植物高分子材料，雖然具有優異之生物安全性，但 由於創傷敷材的應用範圍以動物為主，細胞實驗顯示，海 藻酸鹽纖維與動物細胞之細胞相容性有待加強。 膠原蛋白為動物性高分子，其具有優異的生物安全性 及細胞相容性，但由於其生物分解性極佳，實際應用於創 傷表面時，其膠原蛋白分解過快，以致增加換藥成本及感 染機率。另外，尚有利用海藻酸鈉與膠原蛋白複合，以来1287978 IX. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD The present invention relates to a biopolymer composite fiber, and more particularly to a composite alginate fiber formed by cross-linking of alginate and collagen. [Prior Art] Common natural polymer materials include collagen, gelatin, hyaluronic acid, chitin, chitosan, and derivatives derived from animals. And the like; and alginate, cellulose (ceiiui〇se) and derivatives thereof derived from plants. Alginate and collagen are bioabsorbable materials and are widely used in medical dressings, beauty care products and food additives. In the past, alginate was used to make seawater-steamed fiber by wet spinning. The mechanical strength of the fiber is limited, so that it is limited by application. If the alginate fiber is woven into a non-woven fabric, it can have considerable strength. Alginate fibers in the form of non-woven fabrics are widely used. Calcium silicate fiber is a natural plant polymer material. Although it has excellent bio-safety, the application range of wound dressing is mainly based on animals. Cell experiments show that the cell compatibility of alginate fiber and animal cells needs to be strengthened. . Collagen is an animal polymer with excellent biosafety and cytocompatibility. However, due to its excellent biodegradability, collagen is decomposed too quickly when applied to the wound surface, which increases the cost of dressing change. The probability of infection. In addition, there is still a combination of sodium alginate and collagen.
1287978 乾技術製造之創傷敷材或創傷植入體，其產生之多孔體創 傷敷材具有良好之吸液性，但敷材之強度及加工性仍有待 加強。 因此，需要一種結合海藻酸鹽與膠原蛋白之特性，以 提高生物相容性及生物可分解性，並具有高機械強度及適 用各種加工應用方式之生物高分子材料。 【發明内容】 因此，本發明之主要目的就是在提供一種生物高分子 材料，結合海藻酸鹽及膠原蛋白之特性，經由纖維共成型 技術，產生高生物相容性及生物可分解性的海藻酸鹽複合 纖維’可具有高強度及適用各種加工應用料，用來改善 傳統海藻酸鹽纖維材料之缺點，並增加海㈣鹽纖維應用 之領域。 根據本發明之上述目的，提出一種生物高分子複合纖 維，係以«酸鹽為基礎，以膠原蛋白作為添加劑，藉由 膠：蛋白與海藻酸鹽間之交聯作用均句混合成海藻酸鹽 =口纖維、。依照本發明—較佳實施例，海藻酸鹽纖維與膠 約為1〇的:比例可為19 # 9:1之間’此複合纖維直徑 而二 卡’其機械強度可隨著添加的膠原蛋白濃度增加 而提高。 傷敷發明之目的，提出一種生物高分子複合纖維創 或料纟___複合纖維剪成短纖後，以水抄法 5 製成"'海藻酸鹽複合纖維不織布敷^依照本發 6 1287978 明一較佳實施例，海藻酸鹽複合纖維不織布敷材具有高細 胞相容性及生物分解性，較其他敷材具有更好的傷口促癒 效果。 根據本發明之目的，提出一種海藻酸鹽複合纖維培養 基材，可於體外供纖維母細胞生長。依照本發明一較佳實 施例’纖維母細胞於本發明之海藻酸鹽複合纖維培養基材 上生長良好。海藻酸鹽複合纖維培養基材結合動物性生物 高分子膠原蛋白之高細胞相容特性，可進一步應用於製作 創傷植入體，具有高生物安全性、生物分解性及創傷促癒 效果’亦可應用於醫療實驗用途之細胞培養基材，可有效 促進細胞生長並降低成本。 根據本發明之另一目的，提出一種海藻酸鹽複合纖維 製造方法。首先配製一海藻酸鹽與膠原蛋白混合紡絲原液 及一金屬鹽類親水性成型液，接著將紡絲原液經由紡口擠 入親水性成型液中，以形成一海藻酸鹽複合纖維，並利用 交聯劑使海藻酸鹽複合纖維形成交聯架橋結構，最後以一 水洗程序將交聯的海藻酸鹽複合纖維表面之鹽類及殘留之 父聯劑去除。其中，海藻酸銅濃度可介於0· 1 %到1 〇%之 間’膠原蛋白濃度可介於01%到10%之間。 因此，本發明提供之海藻酸鹽複合纖維具有高細胞相 容性、生物分解性及較高之纖維強度以提供廣泛之可加工 特性： 1 ·以海藻酸鹽複合纖維作為醫療敷材具有高細胞相容 性及生物分解性，並可達到優異之傷口促癒效果，更兼具 1287978 耐用性及降低醫療成本的優點； 2.以海藻酸鹽複合纖維作為細胞培養基材，其高細胞 相容特性可於體外供纖維母細胞生長，應用於製作創傷植 入體時’此細胞培養基材可根據傷口深淺形成特定形狀， 以利後續植入傷口，並具有促進傷口復原的效用；或應用 於醫療實驗用途之細胞培養，可提供有別於傳統細胞培養 基材之外的另一種選擇。 【實施方式】 以下提供數個較佳實施例用來幫助了解本發明之各 種特點與製造方法。 請參照第1圖，其繪示依照本發明一較佳實施例的一 種製造海藻酸鹽複合纖維的方法流程圖。如步驟1〇2所 示’首先配製海藻酸鹽與膠原蛋白混合紡絲原液，其中海 藻酸鈉濃度可介於〇·1%到10%之間，膠原蛋白濃度可介 於〇·1%到10%之間。根據本發明之較佳實施例，以5公 克海藻酸鈉與2公克之膠原蛋白溶於14〇毫升之蒸餾水.或 蒸餾水加親水性溶劑中，此親水性溶劑可至少包含甲醇、 乙醇、丙_或其組合之族群，接著攪拌使之溶解以配製5〇/〇 之海藻酸鹽與膠原蛋白混合紡絲原液。 ° 步驟104為配製金屬鹽類親水性成型液。使海藻酸鹽 纖維成型之金屬鹽類可至少包含鈹、n錄、鎖、銅二 銀、鐵及其組合之族群，例如氣化鈣（CaC12)。而膠原蛋白 之親水性成型液可至少包含甲醇、乙醇、丙酮或其組合之 1287978 圖所不之方法，並藉由調整複合纖維製中 添加之膠原蛋白+μ ^1287978 A wound dressing or wound implant made by dry technology has a good liquid absorbing property, but the strength and workability of the dressing need to be strengthened. Therefore, there is a need for a biopolymer material that combines the characteristics of alginate and collagen to improve biocompatibility and biodegradability, and has high mechanical strength and various processing applications. SUMMARY OF THE INVENTION Accordingly, the main object of the present invention is to provide a biopolymer material which combines the characteristics of alginate and collagen to produce high biocompatibility and biodegradable alginic acid via fiber co-molding technology. The salt composite fiber' can be of high strength and suitable for various processing applications, to improve the shortcomings of traditional alginate fiber materials, and to increase the application of sea (four) salt fiber. According to the above object of the present invention, a biopolymer composite fiber is proposed, which is based on an acid salt and uses collagen as an additive to form alginate by cross-linking between a gel: protein and alginate. = mouth fiber,. According to the present invention - a preferred embodiment, the alginate fiber and the gum are about 1 Torr: the ratio may be between 19 #9:1 'this composite fiber diameter and the second card' is mechanically strong with the added collagen The concentration increases and increases. For the purpose of the invention, a biopolymer composite fiber crepe or 纟___ composite fiber is cut into staple fibers, and then prepared by water splicing method 5 "'alginate composite fiber non-woven fabric ^ according to the hair 6 1287978 In a preferred embodiment, the alginate composite fiber nonwoven fabric has high cell compatibility and biodegradability, and has a better wound healing effect than other dressings. In accordance with the purpose of the present invention, an alginate composite fiber culture substrate is proposed which can be used to grow fibroblasts in vitro. According to a preferred embodiment of the present invention, fibroblasts grow well on the alginate composite fiber culture substrate of the present invention. The alginate composite fiber culture medium combines the high cell compatibility of animal biopolymer collagen, which can be further applied to the manufacture of wound implants, which has high biosafety, biodegradability and wound healing effect. The cell culture substrate used for medical experiments can effectively promote cell growth and reduce costs. According to another object of the present invention, a method for producing alginate composite fiber is proposed. First, a seaweed and collagen mixed spinning dope and a metal salt hydrophilic molding liquid are prepared, and then the spinning dope is extruded into a hydrophilic molding liquid through a spinning port to form an alginate composite fiber, and is utilized. The cross-linking agent forms a cross-linked bridging structure of the alginate composite fiber, and finally removes the salt of the surface of the cross-linked alginate composite fiber and the residual parent-linking agent by a water washing process. Among them, the concentration of copper alginate may range from 0.1% to 1%%. The collagen concentration may be between 01% and 10%. Therefore, the alginate composite fiber provided by the present invention has high cell compatibility, biodegradability and high fiber strength to provide a wide range of processability characteristics: 1. High-cell with alginate composite fiber as medical dressing material Compatibility and biodegradability, and can achieve excellent wound healing effect, and has the advantages of 1287978 durability and reduced medical cost; 2. High acellular compatibility with alginate composite fiber as cell culture substrate The characteristics can be used for the growth of fibroblasts in vitro. When used in the manufacture of wound implants, the cell culture medium can form a specific shape according to the depth of the wound, so as to facilitate subsequent implantation of the wound and have the effect of promoting wound healing; or Cell culture for medical laboratory use provides an alternative to traditional cell culture media. [Embodiment] Several preferred embodiments are provided below to assist in understanding various features and manufacturing methods of the present invention. Referring to Figure 1, there is shown a flow chart of a method of making alginate composite fibers in accordance with a preferred embodiment of the present invention. As shown in step 1〇2, first prepare the mixed alginate and collagen spinning solution, wherein the concentration of sodium alginate can be between %·1% and 10%, and the collagen concentration can be between 〇·1%. Between 10%. According to a preferred embodiment of the present invention, 5 g of sodium alginate and 2 g of collagen are dissolved in 14 ml of distilled water or distilled water in a hydrophilic solvent, and the hydrophilic solvent may contain at least methanol, ethanol, and c. Or a group of combinations thereof, followed by stirring to dissolve it to prepare a 5 〇/〇 alginate-collagen mixed spinning dope. ° Step 104 is to prepare a metal salt hydrophilic molding liquid. The metal salt forming the alginate fibers may comprise at least a group of ruthenium, n-record, lock, copper-silver, iron, and combinations thereof, such as calcium carbonate (CaC12). The hydrophilic molding liquid of collagen may contain at least a method of methanol, ethanol, acetone or a combination thereof, and the collagen added by the composite fiber is adjusted by μ +
震曰比例，試驗膠原蛋白添加比例對於纖維 度之影響。 啊、中WThe proportion of shock, the effect of the proportion of collagen added on the fiber. Ah, medium W
海藻酸鹽複合纖^^ 原蛋白Alginate composite fiber
(ASTM D3822-1996，夾距 10mm，速度：6mm/min) 此纖維強度測試顯示，單纖斷裂強度隨著膠原蛋白添 加比例增加而有較高之強度，相較於未加入膠原蛋白之實 驗組’於海藻酸鹽纖維中添加膠原蛋白作為添加劑確實可 提升其機械強度，因此本發明提供之海藻酸鹽複合纖維比 起傳統單一成型之海藻酸鹽纖維更具有良好之可加工 性，增加海藻酸鹽纖維的應用範圍。 動物體内實驗 第3圖係繪示以海藻酸鹽複合纖維不織布作為創傷敷 材，對動物外科傷口之復原及促癒效果與傳統敷材的比 較。實驗使用之大白鼠（SD-rat)體重約為250公克至300 公克，利用乙醚作麻醉手術後，將背部剃毛並以10%水溶 性優蛾（aqueous betadine)及70%酒精消毒。接著以外科手 術刀造成一個深達panniculus carnosis的3x3公分傷口， 分別以本發明之海藻酸鹽複合纖維不織布、市售膠原蛋白 敷材及市售海藻酸鹽敷材（商品名：Kaltostat)平整的覆蓋 1287978 於肌肉層上’敷材尺寸需大”口。再以市售防水透明谬 布（商品名：Tegaderm®，6x7cm; 3M)將敷材固定在傷口之 上，並以自黏彈性繃帶輕輕纏繞於實驗鼠之胸腔外固定， 防止敷材脫落及傷口感染。上述實驗鼠於手術後需個別單 獨飼養，自由飲食。本實驗以天為紀錄單位，並分別於手 ㈣3天' 7天、Η天及21天對實驗鼠進行傷口復原程 度評估。 請參照第3圖，縱軸代表傷口癒合比例，橫軸為實驗 天數。其中，由於膠原蛋白之生物分解性極佳，故於實驗 第3天即觀察到膠原蛋白敷材之膠原蛋白成份已被分解殆 盡，其後續實驗期間之傷口癒合程度皆為實驗動物本身之 自癒能力所達成，因此可視使用膠原蛋白敷材之實驗鼠為 對照組。值得注意的是，第〇到3天為傷口之發炎反應期， 此階段傷口尚来開始癒合。第4到7天之傷口癒合雖不明 顯，但仍可發現使用本發明之海藻酸鹽複合纖維不織布敷 材之實驗鼠傷口癒合程度較佳；而使用市售之海藻酸鹽敷 材之實驗鼠，由於其敷材為植物性生物高分子，與動物細 胞之相谷性較差，故傷口復原程度遠低於本發明之海藻酸 鹽複合纖維不織布敷材。第7到14天進入傷口癒合期， 可明顯觀察到海藻酸鹽複合纖維不織布敷材較其他敷材 具有更好之傷口促癒效果。因此本發明之海藻酸鹽複合纖 維不織布敷材具有高細胞相容性及生物分解性，可達到優 異之傷口促癒效果，且其以海藻酸鹽為主、膠原蛋白為辅 的組成特性，更兼具耐用性及降低醫療成本的優點。 11 1287978 盘藻酸鹽複合嬝維表面爐錐母細胞培巷 第4圖係利用本發明之海藻酸鹽複合纖維作為細胞培 • 養基材，以纖維母細胞L929 (細胞株：L929)培養於此基材 ' 上的生長情形之電子顯微鏡照片，顯示纖維母細胞L929 ^ 於本發明之海藻酸鹽複合纖維培養基材上生長良好。已知 纖維母細胞L929無法於一般市售海藻酸鹽上生長，係由 φ 於海藻酸鹽為植物性生物高分子，其與動物細胞之相容性 差，且無促進動物細胞生長之功能。本發明之海藻酸鹽複 合纖維培養基材結合動物性生物高分子膠原蛋白之高細 胞相容特性，可於體外供纖維母細胞生長，因此可進一步 應用於製作創傷植入體，此細胞培養基材可根據傷口深淺 形成特定形狀，以利後續植入傷口，具有高生物安全性及 達到創傷促癒之效果；或可應用於醫療實驗用途之細胞培 養，提供有別於傳統細胞培養基材之外的另一種選擇。 結論 優點 由上述本發明較佳實施例可知，應用本發明具有下列 本發明之一特性為於海藻酸鹽纖維中添加膠原蛋白作 為添加劑’藉由兩種纖維間之交聯架橋所形成之海藻酸越 複合纖維而具有更高之機械強度。因此比起傳統單：成； 之海藻酸鹽纖較具有良狀可加卫性，增 維的應用範圍。 孤纖 12 1287978 本發明之另一特性為以濕式紡絲技術與水抄法或針 紮法織成之海藻酸鹽複合纖維不織布，動物體内實驗顯 示，以此海藻酸鹽複合纖維織不織布作為醫療敷材，具有 高細胞相容性及生物分解性，並可達到優異之傷口促癒效 果’且其以海蒸㉟鹽為主、膠原蛋白為輔的組成特性，更 兼具耐用性及降低醫療成本的優點。 本發明之又一特性為可利用海藻酸鹽複合纖維作為 細胞培養基材，結合動物性生物高分子膠原蛋白之高細胞 相容特性，可於體外供纖維母細胞生長，因此可進一步應 用於裝作創傷植人體，具有高生物安全性及達賴傷促癒 之效果，或可應用於醫療實驗用途之細胞培養基材，提供 有別於傳統細胞培養基材之外的另一種選擇。 以上所展示或描述之較佳實施例中之特定時間、濃度 等數據’僅疋幫助讀者了解本發明的原理及提供技術上之 概念，並非用以限^本發明。熟習此技藝者根據本發明原 理所作之任何細節或型態上的變化、或任意組合上述各較 佳實施例之方法，均不脫離本發明之精神及範圍，因此本 發明之保護範圍當視後附之申請專利摩請所界定者為準。 【圖式簡單說明】 為讓本發明之上述和其他目的、特徵、優點與實施例 能更明顯易懂，所附圖式之詳細說明如下： 第1圖疋依照本發明之製備海藻酸鹽複合纖維步驟流 13 1287978 程圖。 第2圖係本發明一較佳實施例的一種海藻酸鹽複合纖 維之截面圖。 第3圖係繪示依照本發明另一較佳實施例的一種海藻 酸鹽複合纖維敷料與單一成型纖維敷料之傷口促癒效果 比較之折線圖。 第4圖係本發明一較佳實施例的一種複合纖維培養基 材上細胞生長之電子顯微鏡照片。 【主要元件符號說明】 102 :步驟 104 ·•步驟 106 :步驟 108 :步驟 110 :步驟(ASTM D3822-1996, 10 mm pitch, speed: 6 mm/min) This fiber strength test showed that the breaking strength of the single fiber increased with the increase of the collagen addition ratio, compared to the experimental group without collagen. Adding collagen to the alginate fiber as an additive does improve its mechanical strength. Therefore, the alginate composite fiber provided by the present invention has better processability than the conventional single-molded alginate fiber, and increases alginic acid. The application range of salt fiber. In Vivo Animal Experiments Figure 3 shows the comparison of the recovery and healing effects of animal surgical wounds with traditional dressings using alginate composite fiber nonwoven fabric as a wound dressing. The rats used in the experiment (SD-rat) weighed about 250 to 300 grams. After anesthesia with ether, the back was shaved and disinfected with 10% aqueous betadine and 70% alcohol. A 3x3 cm wound of panniculus carnosis was then created with a surgical scalpel, which was flattened with the alginate conjugate fiber nonwoven fabric of the present invention, a commercially available collagen dressing, and a commercially available alginate dressing (trade name: Kaltostat). Cover 1287978 on the muscle layer 'the size of the dressing needs to be large'. Then use the commercially available waterproof transparent crepe (trade name: Tegaderm®, 6x7cm; 3M) to fix the dressing on the wound and lighten it with self-adhesive elastic bandage. Lightly entangled in the external thoracic cavity of the experimental rats to prevent the dressing from falling off and wound infection. The above-mentioned rats need to be individually raised and eat freely after surgery. The experiment is based on the day and is in the hands (4) 3 days ' 7 days, The degree of wound healing was evaluated in the rats and the 21st day. Please refer to Figure 3, the vertical axis represents the proportion of wound healing, and the horizontal axis is the number of experimental days. Among them, because the biodegradability of collagen is excellent, the experiment is the third. It was observed that the collagen component of the collagen dressing had been decomposed, and the degree of wound healing during the subsequent experiments was the self-healing ability of the experimental animals themselves. Therefore, it is obvious that the experimental rats using collagen dressings are used as the control group. It is worth noting that the third day is the inflammatory reaction period of the wound, and the wound is still healing at this stage. The wound healing on the 4th to 7th day is It is not obvious, but it can still be found that the test rats using the alginate composite fiber non-woven fabric of the present invention have better wound healing degree; and the experimental rats using the commercially available alginate dressing material are plant organisms because of their application materials. The polymer has poor phase contrast with animal cells, so the degree of wound recovery is much lower than that of the alginate composite fiber non-woven fabric of the present invention. On the 7th to 14th day, the alginate composite fiber can be observed. The non-woven fabric has better wound healing effect than other materials. Therefore, the alginate composite fiber non-woven fabric of the present invention has high cell compatibility and biodegradability, and can achieve excellent wound healing effect, and It is characterized by alginate and collagen, which combines the advantages of durability and lowering medical costs. 11 1287978 Alginate composite 袅Wei surface furnace cone Fig. 4 is a photomicrograph showing the growth of the alginate composite fiber of the present invention as a cell culture substrate and the growth of the fibroblast L929 (cell strain: L929) on the substrate. It is shown that the fibroblast L929 ^ grows well on the alginate composite fiber culture material of the present invention. It is known that the fibroblast L929 cannot grow on the commonly marketed alginate, and is based on φ on alginate as a plant organism. The polymer has poor compatibility with animal cells and has no function of promoting animal cell growth. The alginate composite fiber culture material of the present invention combines the high cell compatibility property of animal biopolymer collagen, and can be externally For the growth of fibroblasts, it can be further applied to the manufacture of wound implants. The cell culture substrate can form a specific shape according to the depth of the wound, so as to facilitate subsequent implantation of the wound, and has high biosafety and wound healing effect; Alternatively, it can be applied to cell culture for medical laboratory applications, providing an alternative to traditional cell culture media. Conclusion Advantages It is apparent from the above-described preferred embodiments of the present invention that the application of the present invention has the following characteristics of the present invention: the addition of collagen to alginate fibers as an additive 'alginic acid formed by cross-linking between two fibers The more composite fibers have higher mechanical strength. Therefore, compared with the traditional single: into; the alginate fiber has a good shape to enhance the scope of application. Solitary fiber 12 1287978 Another characteristic of the present invention is alginate composite fiber non-woven fabric woven by wet spinning technique and water-jet method or needle-punching method, and animal experiments show that the alginate composite fiber woven non-woven fabric As a medical dressing material, it has high cell compatibility and biodegradability, and can achieve excellent wound healing effect, and it is characterized by sea steamed 35 salt and collagen as a supplement, and is more durable. The advantage of reducing medical costs. Another feature of the present invention is that the alginate composite fiber can be used as a cell culture substrate, and the high cell compatibility property of the animal biopolymer collagen can be used for the growth of the fibroblast in vitro, so that it can be further applied to the device. It is a traumatic implanted human body with high biosafety and sturdy wound healing effect, or a cell culture substrate that can be used for medical experiments, providing an alternative to traditional cell culture materials. The specific time, concentration, and the like in the preferred embodiments shown or described herein are merely intended to assist the reader in understanding the principles of the present invention and the technical concept. The details of the present invention are not limited to the spirit and scope of the present invention, and the scope of protection of the present invention is apparent. The patent application is subject to the definition of patent application. BRIEF DESCRIPTION OF THE DRAWINGS In order to make the above and other objects, features, advantages and embodiments of the present invention more obvious, the detailed description of the drawings is as follows: FIG. 1 is a preparation of alginate composite according to the present invention. Fiber Step Flow 13 1287978. Fig. 2 is a cross-sectional view showing a sea alginate composite fiber according to a preferred embodiment of the present invention. Fig. 3 is a line graph showing a comparison of wound healing effects of an alginate composite fiber dressing and a single shaped fiber dressing according to another preferred embodiment of the present invention. Fig. 4 is an electron micrograph of cell growth on a composite fiber medium of a preferred embodiment of the present invention. [Description of main component symbols] 102: Step 104 ·•Step 106: Step 108: Step 110: Step