200825232 九、發明說明： 【务明所屬之技術領域】 且特別是關 本發明是有關於一種不織布之製備方法 於一種止血不織布的製備方法及其應用。 【先前技術】 。乡原虫白（c〇llagen)為一種動物性高分子，當血管壁受 知後’♦原蛋白會立刻與血小板膜醣蛋白結合，進而促使 血小板：附於傷口上，以形成血拴，達到止血的功能。 儘官膠原蛋白已逐漸地應用在止血敷料上，但由於其 生物分解性極佳’於實際應用時會有分解過快等缺點。因 此多將海藻酸鈉與膠原蛋白複合，以束乾技術製造成多孔 體或稱海綿體(spGnge)的形式，強化其機械性。然而，此種 多孔體止血材，其吸液速度較慢，因此容易產生回滲現象， 無法使傷口保持乾燥，易造成感染。 儘管市面上尚有其他種類之止血敷料，但亦有諸多缺 點尚待改善。舉例而言，以海藻酸鹽所製成的海藻片，其 材料為天然植物性高分子，故與動物細胞之生物相容性有 待加強。而以氧化纖維素所製成的止血紗布，則是無法有 效促進凝血，以於短時間内迅速達到止血功效。 5 200825232 除了具有良好的生 ，同時可促進凝血 因此，仍待開發—種新的止血材， 物相合I·生外，並能提供較佳之吸液速度 速度，以保持傷口乾燥。 L贫明門答』 因此本發明的目的— „ 就疋在铨供一種止血不織布， 用以k供較佳之吸液性，並促進凝血速度。 冲 根據本發明之上述目的， 法。首先，分別配IMyu/ T織布的製備方 十 衣、、方、、、糸原液及金屬鹽成型液，並中钫絲 原液“驗金族海藻酸鹽、膠原蛋白與白复:，二、 濕式紡絲，使上述之纺/者’進行 維。然後利用交聯劑使複4二=形成複合纖 W丨义復口緘維父聯聚合，最 聚合後的複合纖維製成不織布。 、父汁 本發明的另-目的，則是提出—種止血不織布，此不 布至少包含複數個複合纖維 … 外，其成分包含海萍_ “ 義、准除了父聯成型 疋么A甘 鈣白复以及膠原蛋白，其中膠焉 虫 與海藻酸約係均勻混合，二一: 與膠原蛋白之混合重量比係 /、 她剩 主u丨尔冷n9:1，而旦 鹼土族海藻酸鹽與膠原蛋白 里糸為 y史臼重里總和之0.01-1.0%。 與傳統止血敷料相較， .,^ π 这之止血不織布除了具有較佳 之血液吸附性外，亦可於 m土 傷口癒合。 才門内達到政血效果，有助於 【實施方式】 6 200825232 於本發明之一實施例中 係提出一種止血不織布之fJ 備方：，並於製備的過程中，加入膠原蛋白與常用之中華 速度铜etlllastnata)，以增進其生物相容性，並促進凝金 睛茶照第1圖，：M:孫絡—丄々 ，、係〜不本發明一實施例中，止血 織布的製備概略流程。首先，制人 + 配衣έ有驗金族海藻酸鹽、 白芨與膠原蛋白之紡絲原液（步 ^ y使（步知102)。另外，配製金屬鹽 成型液（步驟104)。然後，推γ斗、&从 後進仃濕式紡絲，使紡絲原液及金 屬風成型液形成複合纖維(步驟106)。接著，將複合_ 泡於交聯劑中，以使複合纖維交聯（⑽㈣nkmg)(步ς 1〇8)。最後，將交聯後的複合纖維製成不織布（步驟“ο), 此過程以實施例為例詳述如下。 f施例一止血不今1《製備 麥照第2圖，其麵示本發明—實_之止*不織布之 製備流程。首先，將5公克海藻酸鈉、G.7公克的白芨與2 公克膠原蛋白加入140毫升蒸餾水中，以配製成紡絲原液 (步驟202)。同時，將50克的氯化鈣加入1〇〇〇毫升體積百 分比為50%的乙醇溶液中，以配製成一金屬鹽成型液(二 2〇4)。接著，進行濕式紡絲’將上述配製好的紡絲原^經 纺口擠入金屬鹽成型液中(步驟2〇6)。由於，海藻酸鈉中： 鈉離子會與金屬鹽成型液中的鈣離子發生離子交換，使一 價鈣離子與海藻酸鍵結形成海藻酸鈣，進而與膠原蛋白 白芨形成複合纖維。 接著，將複合纖維浸泡於重量百分率濃度為〇·5%的戊 二醛（glutaraldehyde)中，以使所形成的複合纖維進行六聯 200825232 反應，使複合纖維形成交聯架橋結構（步驟2〇8)。最後，將 父聯聚合後的複合纖維剪成短纖後，以水抄法製成不織布 (步驟210)。 於本發明之另-實施例中，紡絲原液中的海藻酸納，以 可用其他驗金族海蕩酸鹽代替’ &於纺絲原液中的重量百 分率濃度係為(U-10%。而膠原蛋白之重量百分率濃度係為 o.i-io%，白芨之重量百分率濃度則為〇 〇ι ι 〇%。此外， 用來製備纺絲原液之溶劑亦可為其他親水性溶劑，例如甲 醇或乙醇。再則’金屬鹽成型液中之所使用的金屬鹽可為 鹼土金屬鹽，其重量百分率濃度係為0.01-15%，另外可使 用：醇：乙醇或丙酮溶液作為金屬成型液之溶劑，其重量 百/刀率濃度係為3G.7G%。當驗金族海藻酸鹽與驗土金屬離 子相接觸日寸’會發生離子交換，進而形成驗土族海藻酸鹽200825232 IX. INSTRUCTIONS: [Technical field to which the invention belongs] and especially the present invention relates to a method for preparing a non-woven fabric and a method for preparing the same. [Prior Art]. C〇llagen is an animal polymer. When the blood vessel wall is known, the original protein will immediately bind to the platelet membrane glycoprotein, which will promote platelets: attach to the wound to form blood stasis and achieve hemostasis. The function. Collagen has been gradually applied to hemostatic dressings, but due to its excellent biodegradability, it has the disadvantage of being too fast to decompose in practical applications. Therefore, sodium alginate is often compounded with collagen, and it is produced into a porous body or spongy body (spGnge) by a bundle drying technique to enhance its mechanical properties. However, such a porous hemostatic material has a slow liquid absorption rate, and thus is prone to back osmosis, and it is impossible to keep the wound dry and easily cause infection. Although there are other types of hemostatic dressings on the market, there are still many shortcomings that need to be improved. For example, a seaweed tablet made of alginate is a natural plant polymer, so the biocompatibility with animal cells needs to be strengthened. The hemostatic gauze made of oxidized cellulose is not effective in promoting coagulation, so that hemostasis can be quickly achieved in a short time. 5 200825232 In addition to having a good life, it can also promote blood clotting. Therefore, it is still to be developed - a new hemostatic material, which is combined with I. It can provide better aspiration speed to keep the wound dry. Therefore, the object of the present invention is to provide a hemostatic non-woven fabric for better liquid absorption and to promote the rate of blood clotting. According to the above object of the present invention, the method firstly With the preparation of IMyu/T weaving, weigh the clothes, the square, the sputum, the sputum and the metal salt molding liquid, and the sputum silk liquid "the gold alginate, collagen and white complex:, two, wet spinning Silk, so that the above spinning / person's dimension. Then, the cross-linking agent is used to form a composite fiber, and the composite fiber is formed into a non-woven fabric. The other purpose of the present invention is to propose a hemostatic non-woven fabric, which does not contain at least a plurality of composite fibers... In addition, the composition thereof contains Haiping _ "Yi, quasi-except for the father's joint molding, A calcium calcium white Complex and collagen, in which the aphid and alginic acid are evenly mixed, 21: the weight ratio of the mixture with collagen /, her remaining main uur cold n9:1, and the alkaline alginate and collagen The protein is 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与 与. The effect of political blood is helpful [Embodiment] 6 200825232 In an embodiment of the present invention, a fJ preparation for hemostatic non-woven fabric is proposed: and in the process of preparation, collagen is added with common Chinese speed copper etlllastnata) In order to enhance its biocompatibility, and to promote the condensed eye tea photo 1st: M: Sunluo, 系, 系~ In one embodiment of the invention, the outline process of preparation of the hemostatic woven fabric. First, Manufacture + dressing έ There is a spinning stock solution of gold alginate, chalk and collagen (step y (step 102). In addition, a metal salt molding liquid is prepared (step 104). Then, γ bucket, & Wet spinning, the spinning dope and the metal wind forming liquid form a composite fiber (step 106). Next, the composite _ is bubbled in the crosslinking agent to crosslink the composite fiber ((10) (four) nkmg) (step 〇 1 〇 8) Finally, the crosslinked composite fiber is made into a non-woven fabric (step "o", and the process is described in detail by way of example below. f. Example 1 Stop bleeding without blood 1 "Preparation of wheat photo 2, which shows Invention - Realization * Preparation process of non-woven fabric. First, 5 g of sodium alginate, G. 7 g of white peony and 2 g of collagen were added to 140 ml of distilled water to prepare a spinning dope (step 202). At the same time, 50 g of calcium chloride is added to 1 ml of a 50% by volume ethanol solution to prepare a metal salt forming solution (2 2 4). Then, wet spinning is carried out. The prepared spinning original spinning port is extruded into a metal salt forming liquid (step 2〇6). Because, sodium alginate Medium: The sodium ion will ion exchange with the calcium ion in the metal salt forming solution, and the monovalent calcium ion and the alginic acid bond to form calcium alginate, and then form a composite fiber with the collagen white ash. Then, the composite fiber is immersed in the weight. The percentage concentration is 〇·5% of glutaraldehyde, so that the formed composite fiber undergoes a six-200825232 reaction, so that the composite fiber forms a cross-linked bridging structure (step 2〇8). Finally, the parent is polymerized. After the composite fiber is cut into staple fibers, the nonwoven fabric is formed into a non-woven fabric by water (step 210). In another embodiment of the present invention, the sodium alginate in the spinning dope can be used for other gold-recognition The weight percent concentration of the salt instead of ' & in the spinning dope is (U-10%). The concentration percentage of collagen is o.i-io%, and the weight percentage of white peony is 〇 〇ι ι 〇%. Further, the solvent used to prepare the spinning dope may be other hydrophilic solvents such as methanol or ethanol. Further, the metal salt used in the metal salt molding liquid may be an alkaline earth metal salt having a concentration percentage of 0.01 to 15%, and an alcohol: ethanol or acetone solution may be used as a solvent for the metal molding liquid, and the weight thereof. The 100/knife rate concentration is 3G.7G%. When the gold test alginate is in contact with the soil metal ion, ion exchange occurs, and the soil alginate is formed.
之複合纖維析出。 I 於本务明之另一實施例中，作為交聯劑的戊二酸亦可使 用綠栀子素（g吻ln)代替。而交聯劑的重量百分率濃度係為 Ο力i Ο·5/”另外，交聨聚合後的複合纖維則利用針紮法製 成不織布。 、 、 種m明之另—實施例中，係、提供—種止血不織布。此 含複數個複合纖維。該些複合纖維除了交 中膠原务/成分包含海藻_、白芨以及膠原蛋白，其 多”、白芨與海藻酸鈣係均勾合， 藻酸鹽與膠屌疋a /、甲鹼土私海 旦儀纽…、之混合重量比係為ι:9-9:ι，而白芨之重 里双土族海藻酸鹽與膠原蛋白重量總和 200825232 為了測減所製備出的止血不織布是否具有良好的生物 相容性，因此依據國際標準化組織（Internati〇nai Organization for Standardization)所制定之 ISOH)993-PART4方法，進行测試。 於此測試方法中，係將測試樣品與兔子血液相混合， 若測試樣品之生物相容性不#，則會造成溶血現象，使紅 血球破裂，進而釋放出血紅素。而利用所釋放出血紅素的 里，則可以下式（1)計算出測試樣品的溶血指數。 溶血指數=積所釋放出血紅素的量（毫克/毫升) 單位體積原血紅素的量（毫克/毫升） Xl〇〇/° 最後’再將所測得的溶血指數與is〇1〇993_PART4方法 中，所制定的標準溶血指標進行比對。而上述實施例之止 血不織布的測試結果，如表一所示。 1 一生物相> 性測試結果 _ 標準溶, ik指標 — — -----—.— 止血不織布測試結果 溶血程度 溶血指數 ----——____Η 測試樣品 :~^ 〉谷血指數 益 0-2% 1------!__——一 輕微 3-10% 陰性對照組 0.01% 中等 11-20% 顯著 21-40% 測試組 1.08% ------- ——-—— 嚴重 大於40% 有效陽性對照組之測試值為> 850/0 有效陽性對照組之測試值為< 2〇/〇 陽性對照組 85.2% 9 200825232 旦表-左側列出了標準溶血指標，係作為測試結果的評 1基準，可知隨著溶血指數的， J 开/合血私度也隨之增加。 而表-右側則列出了止血不織布的測試結果，其中陽 照組之測試樣品為注射S蒸館水，所測得的溶血指數為 \ 85.25%’對照左側之標準溶血指標，為溶血現象嚴重。此 係因為加入蒸德水後，血球細胞内外產生渗透麼差，而使 纟分參人血球内部。隨著血球細胞内部Μ力增大，最後導 致細胞膜的破裂，致使血紅素釋出，產生溶血覌象。 # 至於陰性對照組之測試樣品為高密度聚氯乙浠(HlghThe composite fiber is precipitated. I In another embodiment of the present invention, glutaric acid as a crosslinking agent may also be replaced by scorpionin (g kiss ln). The weight percentage of the cross-linking agent is Ο力 Ο·5/". In addition, the conjugated fiber after the cross-linking polymerization is made into a non-woven fabric by a needle-punching method, and the other is in the embodiment. - Hemostasis non-woven fabric. This contains a plurality of composite fibers. In addition to the intermediate collagen/components containing seaweed _, white peony and collagen, the conjugated fibers are multi-", white peony and calcium alginate are combined, alginate and The glue a /, the alkali-alkali soil Hainan instrument ..., the mixture weight ratio is ι: 9-9: ι, and the weight of the white sorghum double earth alginate and collagen weight sum 200825232 prepared for the reduction Whether the hemostatic non-woven fabric has good biocompatibility, the test was carried out according to the ISOH) 993-PART4 method established by the International Organization for Standardization (ISO). In this test method, the test sample is mixed with the blood of the rabbit. If the biocompatibility of the test sample is not #, the hemolysis phenomenon is caused, the red blood cell is broken, and the hemoglobin is released. In the case of using the released hemoglobin, the hemolysis index of the test sample can be calculated by the following formula (1). Hemolysis index = amount of hemoglobin released by the product (mg/ml) The amount of raw heme per unit volume (mg/ml) Xl〇〇/° Finally 're-measured hemolysis index and is〇1〇993_PART4 method In the middle, the standard hemolysis indicators were compared. The test results of the non-woven fabric of the above embodiment are shown in Table 1. 1 a biological phase > sex test results _ standard solution, ik index ---- ------.- hemostatic non-woven test results hemolysis degree hemolysis index ----- ____ Η test sample: ~ ^ 〉 谷血指数益0-2% 1------!__——A slight 3-10% Negative control group 0.01% Medium 11-20% Significant 21-40% Test group 1.08% ------- ——- —— Severely greater than 40% Effective positive control group test value> 850/0 Effective positive control group test value is < 2〇/〇 positive control group 85.2% 9 200825232 Dan table - left side lists standard hemolysis index As a benchmark for the test results, it can be seen that with the hemolysis index, the J open/combined blood is also increased. The table-right side lists the test results of hemostatic non-woven fabrics. The test sample of the Yangzhao group is injected with S steaming water. The measured hemolysis index is \ 85.25%' standard hemolysis index on the left side of the control, which is a serious hemolysis phenomenon. . This is because after the addition of steamed water, the infiltration of blood cells inside and outside the cell is poor, and the sputum is divided into the blood cells. As the internal force of the blood cells increases, the rupture of the cell membrane eventually leads to the release of hemoglobin, which produces hemolysis. # As for the negative control group, the test sample is high density polychloroethylene (Hlgh)
DenSlty Polyethylene，HDpE)之萃取液其溶血指數為 0.01%，對照左側之標準溶血指標，屬於無溶血現象產生。 此係由於高密度聚氯乙烯不會造成金液滲透壓的改變，故 紅血球不會破裂，無溶血現象產生。至於測試組中，則為 止血不織布之萃取液與兔子血液，所測得的溶血指數為 咖/。，對照標準溶血指標可知，止血不織布並不會造成溶 血現象，據此，本發明之實施例的止血不織布具有良好的 Φ 生物相容性。 (二）血液吸附測訧 為了測試所製得的止血不織布是否具有較佳的血液吸 附性，因此藉由測定纖維素原（FlbrinogeI1)濃度，進行血液 吸附測試，並與海澡酸鈉與膠原蛋白所製成的多孔體止血 材，以及由海澡酸鹽所製成的海藻片進行比較，其測試方 法如下。 首先，取10毫克的樣品放入重量百分率為〇·3%的纖 維素原（Fibnnogen)溶液中，於37。(：下放置24小時，使纖 200825232 維原吸附在測試樣品上，再利用二辛可寧酸（bicinchoninic acid，BCA)法測試所吸附纖維素原的量。二辛可寧酸法是 利用血液中的蛋白質（即纖維素原）會將兩價銅離子還原成 一價的銅離子，所生成的一價銅離子又可與二辛可寧酸形 成錯合物而呈色，最後藉由吸收度的測試來決定所吸附蛋 白夤的置。由於纖維素原係為血漿中的一種蛋白質，據此， 若得知測試樣品對於纖維素原的吸附性，即相當於得知測 试樣品對於血液的吸附性。The extract of DenSlty Polyethylene, HDpE has a hemolysis index of 0.01%, and the standard hemolysis index on the left side of the control belongs to no hemolysis. This is because the high-density polyvinyl chloride does not cause a change in the osmotic pressure of the gold liquid, so the red blood cells do not rupture and no hemolysis occurs. As for the test group, the hemolysis non-woven extract and the rabbit blood were measured, and the measured hemolysis index was coffee/. According to the standard hemolysis index, it is known that the hemostatic non-woven fabric does not cause hemolysis, and accordingly, the hemostatic non-woven fabric of the embodiment of the present invention has good Φ biocompatibility. (2) Blood adsorption test In order to test whether the prepared hemostatic non-woven fabric has better blood adsorption property, blood adsorption test is carried out by measuring the concentration of cellulose raw material (Flbrinoge I1), and sodium and collagen with sea bath and collagen The prepared porous hemostatic material and the seaweed sheet made of sea bath salt were compared, and the test method was as follows. First, 10 mg of the sample was placed in a Fibnnogen solution having a weight percentage of 〇·3%, at 37. (: Place for 24 hours, so that the fiber 200825232 dimension is adsorbed on the test sample, and then the amount of adsorbed cellulose original is tested by the bicinchoninic acid (BCA) method. The dicinocinic acid method utilizes blood. The protein (ie, cellulose) reduces the divalent copper ions to monovalent copper ions, and the resulting monovalent copper ions form a complex with dioctocinic acid to form a color, and finally by absorbance. The test determines the position of the adsorbed peptone. Since the cellulose is a protein in plasma, according to this, if the adsorption of the test sample to the cellulose is known, it is equivalent to knowing the test sample for the blood. Adsorption.
—第3圖則顯示了血液吸附測試之結果，樣品（A)為以海 藻酸鹽製成的海藻片（購自維康醫療用品器材公司，商品名 為kaltostat) ’樣品⑻則為多孔體止血材（購自維康醫療用 口口為材公司’商品名為sp〇nt〇stan)，而樣品⑷為實施例一 所製備之止血不織布。由第3圖可知，每毫克的止血不織 =則可吸㈣0.83笔克的纖維素原。然而，於同樣的單位重 里下U僅可吸附〇.58毫克的纖維素原，多孔性止血 材吸附性更差，只有G25毫克。由此可知，止血不織布與 /、他止血敷料相較的確具有較佳的血液吸附性。 ^了驗證上述之止血不織布，除了具有良好的血液吸 附Γ，亦可促進凝血，因此，使用凝血分析儀進行測試， 乳化纖維素所製成之紗布進行比較。同時，為了驗 If加之*芨對於凝血效果有極大助益，因&，亦設置 組以進行比較’其結果如表二所示。 200825232- Figure 3 shows the results of the blood adsorption test. Sample (A) is a seaweed tablet made of alginate (purchased from Wellcome Medical Supplies Equipment, trade name kaltostat). Sample (8) is a porous body to stop bleeding. The material (purchased from Weikang Medical Oral Products Co., Ltd.'s trade name sp〇nt〇stan), and the sample (4) was the hemostatic non-woven fabric prepared in Example 1. As can be seen from Fig. 3, every milligram of hemostasis can absorb (four) 0.83 grams of cellulose. However, in the same unit weight, U can only adsorb 58.58 mg of cellulose original, and the porous hemostatic material is more poorly adsorbed, only G25 mg. It can be seen that the hemostatic non-woven fabric does have better blood adsorption properties than the hemostatic dressing. ^ The above-mentioned hemostatic non-woven fabric was verified, and in addition to having good blood absorbing sputum, coagulation was also promoted. Therefore, a gauze prepared by emulsifying cellulose was used for comparison using a coagulation analyzer. At the same time, in order to check If plus * 芨 is very helpful for the coagulation effect, because &, also set the group for comparison', the results are shown in Table 2. 200825232
3 6.0 秒 可知’於相同的測試條 的平均凝血時間為521 y ”〜布 加白 ^ 3秒’而對照組為於製備過程中未添 至於添 此可 助益 芨之止血不織布，其平均凝血時間為39· 1秒 加白复之止血不織布的平均凝血時間僅需36.0秒，由 知’添加白I的止血不織布，對於凝血效果的確有顯著的 4可知本發明上述實施例之止血不織布，除了 ㈣良好的生物相容性外，亦能提供較佳之吸液速度，預 防回滲，並可有效促進凝血。 雖然本發明已以較佳實施例揭露如上，然其並非用以 限定本發明，任何熟習此技藝者，在不脫離本發明之精神 和範圍内’當可作各種之更動與潤飾，因此本發明之保護 範圍當視後附之申請專利範圍所界定者為準。 12 200825232 【圖式簡單說明】 ▲為讓本發明之上述和其他目的、特徵、優點與" 能更明顯易懂，所附圖式之詳細說明如下： 、也例 的製備 、第1目係緣示本發明-實施例中，纟血不織布 流程圖。3 6.0 seconds, it is known that the average clotting time of the same test strip is 521 y ”~ Bujia white ^ 3 seconds' and the control group is not added to the hemostatic non-woven fabric during the preparation process. The average clotting time of the time is 39. 1 second plus the white hemostasis non-woven fabric only takes 36.0 seconds. It is known that the hemostatic non-woven fabric of adding white I has a significant effect on the blood coagulation effect, and the hemostatic non-woven fabric of the above embodiment of the present invention is known. (4) In addition to good biocompatibility, it can also provide a better liquid absorption rate, prevent back osmosis, and can effectively promote blood coagulation. Although the present invention has been disclosed in the preferred embodiments as above, it is not intended to limit the invention, any A person skilled in the art will be able to make various changes and modifications without departing from the spirit and scope of the invention, and the scope of the invention is defined by the scope of the appended claims. 12 200825232 BRIEF DESCRIPTION OF THE DRAWINGS ▲ In order to make the above and other objects, features, advantages and advantages of the present invention more obvious, the detailed description of the drawings is as follows: The first edge line head of the present invention shown - embodiment, the non-woven Si blood flow chart.
第2圖騎示依照本發明—實施例中，止血不 製備流程圖。 、第3圖係繪不依照本發明一實施例中，止血不織布血 液吸附測試結果之長條圖。 【主要元件符號說明】 104 :步驟 108 :步驟 204 :步驟 208 =步驟 102 :步驟 106 :步驟 11 〇 :步驟 2()2 :步驟 206 :步驟 21 〇 :步驟 13Fig. 2 shows a flow chart for the preparation of hemostasis in accordance with the present invention. Fig. 3 is a bar graph showing the results of blood adsorption test of hemostatic non-woven fabric in accordance with an embodiment of the present invention. [Main component symbol description] 104: Step 108: Step 204: Step 208 = Step 102: Step 106: Step 11 〇: Step 2 () 2: Step 206: Step 21 〇: Step 13