200526279 九、發明說明： I：發明戶斤屬之技術領域3 發明領域 本發明涉及醫療領域中用於修復骨缺損部的多孔質骨 5 修復材料及其製造方法。 L先前技術U 發明背景 作為針對腫瘤或壞死等骨病變的骨組織切除手術、或 針對外傷引起的骨組織缺損部位的治療方法，近年來使用 10 再生醫療的手法。迄今為止，針對骨的缺損部位，填充金 屬等非生物體吸收性材料，只是單純修復缺損部的形態， 沒有考慮骨組織本身的再生。再生醫療的手法根據組織學 的概念、結合構成生物體組織的細胞和固定材料以及衍生 細胞分化的因數(信號物質等）而使失去的生物體組織（此處 15 是指骨組織）再生的醫療手法。 作為腳手架材料，要求具有對生物體的危害性低且在 一定時間内可被代謝吸收的特性，在有機材料中，正在嘗 試使用膠原、明膠、殼聚糖、聚乳酸、聚乙醇酸等。另外 ，特別是在骨組織或牙齒再生中，正在應用無機材料如磷 20 酸三鈣等磷酸鈣類材料。 羧曱基幾丁質（下面稱為CM幾丁質）與殼聚糖一樣，是 構成螃蟹等甲殼類的外骨骼的甲殼質的衍生物，通過適當 的交聯處理而具有腳手架材料的特性。 例如，對混入了磷酸鈣類化合物粉末的幾丁質（含有幾 200526279 丁質、设聚糖以及㈣的衍生物)誠實施真空熱乾燥等， 父聯狀悲的幾丁質形成承载有磷酸鈣類化合物粒子的複合 體，進而通過未交聯的幾丁質使該複合體表面形成被膜， 該生物體移植材料和生理鹽水等液體進行捏煉而具有適當 5的粘合性（參照專利文獻1)。 根據專利文獻1，通過該手法，由未交聯的幾丁質構成 的被膜具有水溶性，所以和生理鹽水等液體進行捏練而具 有適當的东占度，進而因為幾丁質進行交聯形成的承载磷酸 躺化合物粒子的複合體不溶於水，所以在複合體内牢固 地承載上述粒子並填充在骨的缺損部，且沒有出現移動、 脫落，骨絡在骨缺損部早期出現再生增殖，從而能夠提供 具有很大治療效果的生物體移植材料。 另外，也公開了由熱交聯後的幾丁質或其衍生物構成 的吸收性生物體材料、以及以實施真空熱交聯處理為主要 特欲的吸收性生物體材料的製造方法(參照專利文獻2)。 根據專利文獻2,相對容易地提供作為生物體内的安全 收1±材料的生物體材料’同時能夠通過使用該材料而 提供促進骨生成的環境。 $外’為了改善在吸收性生物體材料中修正(trimmi⑽ 〇時的強度以及生物體内環境下的強度，充分減緩在生物體 的吸收速度’也有由通過環氧化合物交聯的衍生物構成 的、且麵的吸收性生物體材料（參照專利文獻3)。 另外，作為促進生物體骨形成的材料，示出由包括下 述工序的方法形成的骨形成材料，即，通過有機酸溶解幾 200526279200526279 IX. Description of the invention: I: Technical field of inventors 3 Field of the invention The present invention relates to porous bone 5 for repairing bone defects in the medical field. 5 A repair material and a method for manufacturing the same. L Prior art U Background of the invention In recent years, regenerative medicine has been used as a treatment method for bone tissue resection surgery for bone lesions such as tumors or necrosis, or for bone tissue defects caused by trauma. Hitherto, non-bioresorbable materials such as metal have been used to fill bone defect sites, and the morphology of the defect sites has been simply repaired without considering the regeneration of bone tissue itself. Regenerative medicine is a medical technique that regenerates lost biological tissues (here, 15 refers to bone tissues) based on the concept of histology, combining cells and fixed materials that make up biological tissues and factors (derived signals, etc.) derived from cell differentiation. . As a scaffolding material, it is required to have the characteristics of being less harmful to living organisms and being metabolized within a certain period of time. Among organic materials, collagen, gelatin, chitosan, polylactic acid, polyglycolic acid, etc. are being tried. In addition, especially in the regeneration of bone tissue or teeth, inorganic materials such as calcium phosphate materials such as tricalcium phosphate 20 are being used. Carboxamidine chitin (hereinafter referred to as CM chitin), like chitosan, is a chitin derivative that constitutes the exoskeleton of crustaceans such as crabs, and has the characteristics of scaffolding material by appropriate cross-linking treatment. For example, chitin (containing chitin 200526279 chitin, chitosan, and derivatives of osmium) mixed with calcium phosphate powders is subjected to vacuum heat drying, etc., and chitin is formed to bear calcium phosphate. A composite of compound-like particles, and further formed a coating on the surface of the composite with uncrosslinked chitin, and the biotransplant material and a liquid such as physiological saline are kneaded to have an appropriate adhesiveness of 5 (see Patent Document 1) ). According to Patent Document 1, by this method, a film made of non-crosslinked chitin is water-soluble, so it is kneaded with a liquid such as physiological saline to have an appropriate eastern occupation, and is further formed because chitin is crosslinked. The complex bearing the phosphate compound particles is insoluble in water, so the particles are firmly supported in the complex and filled in the bone defect, and there is no movement or shedding, and the bone network appears to regenerate and proliferate in the bone defect in the early stage. It can provide biological transplant material with great therapeutic effect. Also disclosed are absorbent biomaterials composed of chitin or its derivatives after thermal cross-linking, and methods for producing absorbent biomaterials that are primarily intended to be subjected to vacuum thermal cross-linking treatment (see patents). Reference 2). According to Patent Document 2, it is relatively easy to provide a biological material ', which is a safe material to be received within a living body, and it is possible to provide an environment that promotes osteogenesis by using this material. $ 外 'In order to improve the correction in absorbent biomaterials (trimmi〇 〇 strength and strength in the living environment of the body to fully slow down the rate of absorption in the organism' there is also a derivative crosslinked by epoxy compounds And absorptive biomaterial (see Patent Document 3). As a material for promoting bone formation in a living body, a bone-forming material formed by a method including the following steps is shown, that is, an organic acid is used to dissolve the material 200526279.
丁質·殼聚糖以成為幾丁質·殼聚糖溶膠的工序、在該幾 丁質·殼聚糖溶膠中混合骨形成用蛋白質製作含有該骨形 成用蛋白質的幾丁質·殼聚糖溶膠的工序、把混合了上述 骨形成用蛋白質的上述幾丁質·殼聚糖溶膠與羥基磷灰石 5 、a - TCP、/3 - TCP、CaC03、CaO、ZnO、CaSi03、MgO 等的粉末進行捏煉而得到捏煉物的工序（參照專利文獻4)。 專利文獻1 :特開平6 — 105901號公報， 專利文獻2 :特開平7—116241號公報， 專利文獻3 : 2002 — 11090號公報， 10 專利文獻4 :特開平11 — 347112號公報。 如前所述，再生醫療的手法是根據組織學的概念，組 合構成生物體組織的細胞和成為腳手架的材料以及衍生細 胞分化的因數(信號物質等）而使失去的生物體組織（此處是 指骨組織)再生的醫療手法。當在生物體内只埋置材料時， 15 生物體内預先存在的細胞以及因數進入到材料内部，由此 在該處(in situ)期望出現組織的再生。從這種觀點出發，如 果研究現有技術需要考慮下述問題。 首先，專利文獻1所述的生物體移植材料，是用生物體 吸收性較慢的幾丁質或其衍生物覆蓋磷酸鈣類化合物顆粒 20 ，從而減慢成骨細胞與磷酸鈣類顆粒的接觸。 另外，對於專利文獻2、3所述的材料，由於通過溶液 的凍結乾燥形成多孔體，因此得到的成形體的微細結構， 成為用溶質凝聚形成的（此時是指幾丁質衍生物）的隔膜遮 蔽冰的晶體昇華而殘留的空洞的形狀(所謂蜂窩狀結構）。當 200526279 為這種結構時，因為各氣孔沒有形成完全的連續氣孔，所 以減慢再生骨組織的成骨細胞等細胞群進入到材料内部。 當為專利文獻3所述的材料時，由於進一步地用環氧化人物 父聯幾丁質衍生物，所以也擔心有交聯劑殘留。 5 此外，專利文獻4中所述的骨形成促進材料，採用在生 物體内或生物體外固化捏煉物的方法。在該手法中，固化 物只含有數微米以下的氣孔，實質上阻斷了再生骨組織的 細胞。為此，骨組織只在材料被生物體吸收之後才出現再 生。 1〇 綜上所述，現有手法的缺點都在於，減慢磷酸鈣類粒 子與對成骨細胞等骨進行再構建的細胞群之間的接觸。 另外’從幾丁質和磷酸鈣顆粒的複合體形成方法的觀 點來看，存在下述的課題。 上述的現有技術都使用在幾丁質溶液中混合磷酸鈣顆 15粒的手法◦通常情況下，幾丁質溶液的密度稍超過1，磷酸 鈣顆粒的密度約為3。為此，密度差在成形過程中引起顆粒 的沈殿，製作的複合體成為磷酸鈣下部的配合比較高的不 均勻物質。為了避免這種問題，必須採用在溶液冰點以下 的溫度下進行顆粒的攪拌混合以及成形等繁瑣的手法。 20 【明内】 發明概要 馨於現有技術的這種問題，本發明的目的在於，通過 防止碟酸辑顆粒分佈的偏差而使骨均勻進入其内部，通過 使骨修復材料内部的氣孔處於基本連通狀態而使骨大量進 200526279 入其内部，增大磷酸鈣顆粒在骨修復材料内部的氣孔内的 露出面積，從而促進骨進入其内部。 為瞭解決上述課題，本發明者進行了潛心研究，其結 果發現，當使用由幾丁質或其衍生物構成的纖維形成集合體 5 時，能夠構成基本上完全連續的氣泡，其實質上能在集合體 纖維之間均勻地承載磷酸鈣顆粒，從而完成了本發明。 即，本發明提供一種多孔質骨修復材料，在含有由幾 丁質或其衍生物構成的腳手架材料和由磷酸鈣構成的細胞 分化衍生材料的骨修復材料中，上述幾丁質或其衍生物形 10 成纖維狀並成為多孔集合體，同時磷酸鈣成為顆粒狀並在 上述集合體的纖維間被承載。 另外，本發明在製造由幾丁質或其衍生物的纖維集合 體和其纖維間所承載的鱗酸#5顆粒構成的多孔質骨修復材 料時，提供一種多孔質骨修復材料的製造方法，包括：幾 15 丁質或其衍生物形成纖維的工序、在減壓條件下交聯該纖 維的工序、把該交聯纖維與磷酸鈣顆粒以及該交聯纖維的 保水量以下的水相混合而形成大致均勻的混合物的工序、 凍結乾燥該混合物而成為多孔性材料的工序。 在本發明的多孔質骨修復材料中，幾丁質或其衍生物 20 在再生醫療手法中是作為腳手架的材料，磷酸鈣顆粒是衍 生細胞分化的材料。 在本發明中，使幾丁質或其衍生物成為纖維的集合胃 有下述理由。 即，通過形成在纖維集合體的纖維之間承載碟酸句顆 200526279 粒的結構，能夠防止在製造骨修復材料時出現顆粒沈澱且 顆粒刀佈出現偏差。另外，纖維集合體具有基本上完全連 、哀的氣孔’所以骨細胞能夠大量進入到其内部。而且，能 夠成為磷酸鈣顆粒在連續氣孔内露出的狀態，所以能夠很 5好地衍生細胞分化。 作為幾丁質或其衍生物，在製造工序上優選可以溶解 於中性水的羧甲基幾丁質（鈉鹽）或水溶性殼聚糖。另外，也 可以使用羥乙基幾丁質（乙二醇幾丁質）或乙二醇殼聚糖等 水溶性幾丁質衍生物。 10 這裡，衍生物是指對天然存在的幾丁質進行了化學修 飾後的物質。 ’ 作為磷酸鈣，也可能是磷灰石、磷酸三鈣、磷酸八鈣 等各種材料。 作為磷酸鈣顆粒的尺寸，只要為2000微米以下就能夠 15使用，根據應用部位的骨種類（皮質骨、海綿骨）或基於部位 不同而產生的骨反應性的變化，可以選擇最佳尺寸。 在本發明的製造方法中，作為形成幾丁質或其衍生物 的纖維的方法，當為水溶性幾丁質衍生物時，把該材料的 水溶液與乙醇或丙酮等水溶性有機溶劑相混合，通過快速 2〇㈣而析出短纖維狀的沈殿物，收集、清洗該沈搬物之後 ’乾燥並除去溶劑，由此能夠得到纖維。另 、 乃外，用下述的 方法也能夠得到纖維。即，使用鹼性溶劑使幾丁質溶液化 並將其從細孔(數十微米）的噴嘴中高壓擠出到熱水中，^此 使其凝固而成為纖維狀。 200526279 另外’關於上述水溶性幾丁質衍生物，進行交聯處理 以使纖維在體内不容易被溶出。 父聯處理是通過在2kPa左右的減壓情況下12CTC至180 勺力”、、清況下進行熱交聯而貫現的。與上述的石粦酸詞顆 5粒相同，能夠根據應用部位選擇最佳溫度。 另外’當混合交聯纖維和磷酸鈣顆粒時，添加交聯纖 維的保水量以下的水。這裡，交聯纖維的保水量是指當纖 維集合體浸潰於大量水中後取出來時，纖維集合體攝入至 纖維間以及纖維内並保持的水量。這是因為當添加保水量 10以上的水時，在和磷酸鈣顆粒進行混合時，由於剩餘的水 具有過度的流動性，所以出現因重力造成的顆粒沈澱或凝 聚，造成其分散不均勻。當為保水量以下時，纖維集合體 所保持的水分難以流動，抑製磷酸鈣顆粒的凝聚或沈澱， 其結果是形成均勻的複合體。 15 交聯纖維和磷酸鈣顆粒的混合比（重量）在1 : 0.1至1 : 10之間是可以製作的。 為了使該混合物能夠成可使用狀態，進行混合物的;東 結乾燥。凍結乾燥是在醫藥品、乾燥食品的製造中等通常 進行的工序，通過使凍結乾燥機内部的溫度保持在混合物 20中所含的水分的凝固點以下(〇°c以下），使用真空泵等進行 減壓而使水分昇華蒸發而實現的。作為凍結乾燥的條件， 對溫度、減壓程度、乾燥時間等沒有特別限製。 根據本發明的多孔質骨修復材料，通過形成纖雉集合 體的纖維間承載鱗酸鈣顆粒的結構，能夠防止在製造骨修 200526279 復材料時出現顆粒沈澱以及顆粒分佈出現偏差。因此，能 夠實現骨細胞沒有偏差地進入内部。另外，因為纖維集合 體具備基本上完全連續的氣孔，所以骨細胞能夠大量進入 内部。而且，成為磷酸鈣顆粒在連續氣孔内露出的狀態， 5 所以能夠大量衍生細胞分化。 另外，根據本發明的製造方法，通過下述工序能夠製 造上述多孔質骨修復材料，即，幾丁質或其衍生物形成纖 維的工序、在真空中交聯該纖維的工序、把該交聯纖維與 磷酸鈣顆粒以及該交聯纖維的保水量以下的水相混合而形 10 成大致均勻的混合物的工序、凍結乾燥該混合物而成為多 孔性材料的工序。 圖式簡單說明 第1圖是表示實施例的多孔質骨修復材料表面的SEM 圖像。 15 第2圖是表示實施例的多孔質骨修復材料斷面的S Ε Μ 圖像(反射電子圖像）。 L實施方式3 較佳實施例之詳細說明 下面，表示上述發明的優選實施例，不過本發明如上 20 所述，交聯處理能夠在2kPa左右的減壓情況下120°C至180 °C的加熱情況下進行熱交聯，與上述的磷酸鈣顆粒相同， 能夠根據應用部位選擇最佳溫度。 另外，當混合交聯纖維和磷酸鈣顆粒時，添加交聯纖 維保水量以下的水而可以實現。另外，交聯纖維和磷酸鈣 12 200526279 顆粒的混合比（重量）幻：G.H : 1G之間是可以製作的。 因此，本發明並不限於下面的實施例。 (優選的製造方法） 精密稱取CM幾丁質原料3g，連同純水(mil 〇水)72§一 5起添加到玻璃製容器中，使用電磁式授掉器進行授摔、溶 解’製成偏％的CM幾丁質水溶液。為了加快復幾丁質的 溶解，水浴中加熱容器至30〜5〇t ’攪拌3〜5小時，得到 目視情況下均質的溶液。 把溶液倒入内裝15 〇 m丨乙醇的燒杯中並用玻璃棒快速 1〇攪拌。通過這-操作，析出短纖維狀的復幾丁質沈殿物。 把沈殿物轉移到其他容H巾，接著細乙醇⑽⑹清洗數次 。在盡可月b除去乙醇之後，把該沈殿物放置在4〇ι的乾燥 器中12小時以上，使乙醇以及殘留的水分蒸發。然後，在 真空熱處理裝置在140°C減壓條件下（約2kpa以下）處理12小 15 時。 以重蓋比1 · 5 · 32的比例稱量上述的CM幾丁質以及冷 TCP顆粒（粒搜50〜150微米）以及純水放在容器中，用刮勺 攪拌混合。把該混合物填充到尺寸適於以後的試驗的聚笨 乙稀製圓间中，用PET薄膜封閉圓筒的兩端，然後迅速放置 20到— 的冷凍庫中。經過1小時以上且混合物完全凍結之 後，去除雨端的PET薄膜並移至凍結乾燥器中，在凍結狀態 下直接蒸發水分。然後，從聚苯乙烯製圓筒中取出試驗體 ’供於下述試驗。 [微細結構觀察] ‘ 13 200526279 使用掃描型電子顯微鏡[SEM]實施。對試樣進行汽化鍍 金之後，使用35倍的倍率、以2維電子圖像進行觀察（參照 第1圖）。觀察到纖維狀的CM幾丁質（纖維集合體）中的纖維 間承載有TCP顆粒的狀態。氣孔的尺寸為數十〜數百μιη。 5 為了觀察TCP顆粒的分散狀態，把試驗體包埋於環氧 樹脂中之後，切斷並研磨切斷面，平整化之後實施相同的 SEM觀察。此時，為了只清楚地看到tcp顆粒，採用反射 電子圖像進行觀察（參照第2圖）。在所觀察的整個斷面（約 4x8mm)上，觀察到大致均勻分散有Tcp顆粒，沒有觀察到 10 顆粒因沈澱造成的偏載。 [向骨缺損部的埋置試驗] 將本材料埋置到形成在家兔下頜骨上的直徑為4mm的 骨缺損處，術後4、8周後與周圍組織一起回收，調查骨組 織的再生狀況。針對在規定時間後回收的組織，按照〜船 15方法製作非脫灰樹脂包埋組織標本，計算各標本中再生骨 組織佔骨缺損部位的面積比。作為比較，只形成骨缺損且 不埋置任何材料的例子（空白例）、埋置有通過現有技術即上 述專利文獻2(特開平7— 116241號公報）中所述的手法製造 的试驗體的例子，以供於相同的試驗。把空白例的骨体復 20面積作為1，比較研究各種材料埋置例子的面積比的值。 為現有技術樣本的製作如下所示。在冷卻浴（—1 > 中攪拌CM幾丁質4.24wt%的CM幾丁質水溶液，同時使其 束結’製成微細的晶體冰和溶液混合的狀態(fr〇zen)。添加 規定量的TCP顆粒(粒徑50〜150μηι)並攪拌，使其均勻分1 200526279 在尺寸適於w相試驗的聚苯乙稀製圓筒中填充該束結 物，用PET薄膜封閉圓筒兩端之後，迅速放到一⑽。C的冷康 j後使其4結乾燥。除掉圓筒，使用真空熱處理裝 在C或160 C的減壓條件下（2kPa以下）加熱處理24小 5 ^力到海綿狀體。常溫冷卻後，用體積是成形體表觀體 積2(H口以上的純水清洗3小日夺，把液體部分置換成乙醇，在 8〇 C的乾爍态中乾燥。製作的試驗體經由25kGy的伽馬射線 進行滅菌處理，供於試驗。 其中’只使用通過現有技術的手法，則出現TCP顆粒 10的偏載，但是，通過與如上所述在溶液冰點以下的溫度下 對顆粒進行攪拌混合以及成形等的手法並用，製作顆粒大 致均質分散的試驗體以用於試驗。 其試驗結果匯總如表1所示。 表1 埋置材料 缺損部的骨佔有率 4周後 8周後 本發明 1.3 1.2 現有技術 0.8 1 空白 1 1 15 由表1可知，和空白例相比，當埋置本發明製成的材料 時’在4周後，8周後的任何時點都有很好的骨修復。通過 現有技術製造的材料，其在4周後骨修復稍微差於空白例， 在8周後則相同。 20 【圖式簡單說明】 15 200526279 第1圖是表示實施例的多孔質骨修復材料表面的SEM 圖像。 第2圖是表示實施例的多孔質骨修復材料斷面的SEM 圖像(反射電子圖像）。 5 【主要元件符號說明】 (無） 16Chitin · chitosan is a step of forming chitin · chitosan sol, and chitin · chitosan sol is mixed with a protein for bone formation to produce chitin · chitosan containing the protein for bone formation A sol process, and powders of the chitin · chitosan sol mixed with the above-mentioned bone-forming protein and hydroxyapatite 5, a-TCP, / 3-TCP, CaC03, CaO, ZnO, CaSi03, MgO, etc. A step of kneading to obtain a kneaded product (see Patent Document 4). Patent Document 1: Japanese Patent Application Laid-Open No. 6-105901, Patent Document 2: Japanese Patent Application Laid-Open No. 7-116241, Patent Literature 3: Japanese Patent Application No. 2002-11090, 10 Japanese Patent Application No. 4: Japanese Patent Application Laid-Open No. 11-347112. As mentioned earlier, the technique of regenerative medicine is based on the concept of histology, combining the cells that make up biological tissues, the materials that become scaffolds, and the factors that induce cell differentiation (signal substances, etc.) to make the lost biological tissues (here is Finger bone tissue) regeneration. When only the material is embedded in the living body, the pre-existing cells and factors in the living body enter the inside of the material, and thus tissue regeneration is expected to occur in the situ. From this point of view, the following issues need to be considered when studying the prior art. First, the biotransplantation material described in Patent Document 1 covers the calcium phosphate-based compound particles 20 with chitin or a derivative thereof which is slower in the body, thereby slowing the contact between osteoblasts and calcium phosphate-based particles. . In addition, for the materials described in Patent Documents 2 and 3, the porous body is formed by freeze-drying the solution. Therefore, the fine structure of the obtained molded body is formed by solute aggregation (in this case, a chitin derivative). The shape of the cavity (a so-called honeycomb structure) where the crystals of the ice sublimate and remain. When 200526279 has this structure, because each stomata does not form a complete continuous stomata, it slows down the cell population such as osteoblasts that regenerate bone tissue into the material. In the case of the material described in Patent Document 3, since the chitin derivative is further epoxidized with the epoxidized character, there is also concern that a crosslinking agent may remain. 5 In addition, the bone formation promoting material described in Patent Document 4 employs a method of solidifying a kneaded material in or out of a living body. In this method, the solidified material contains stomata of a few microns or less, which substantially blocks the cells that regenerate bone tissue. For this reason, bone tissue regenerates only after the material is absorbed by the organism. 10 In summary, the disadvantages of the existing methods are that they slow down the contact between calcium phosphate particles and the cell population that reconstructs bones such as osteoblasts. In addition, from the viewpoint of a method for forming a complex of chitin and calcium phosphate particles, there are the following problems. In the above-mentioned prior art, a method of mixing 15 calcium phosphate particles in a chitin solution is used. ◦ Generally, the density of the chitin solution is slightly over 1, and the density of the calcium phosphate particles is about 3. For this reason, the density difference causes Shen Dian of the particles during the forming process, and the produced composite becomes a heterogeneous substance with a relatively high blending of the lower portion of calcium phosphate. In order to avoid this problem, complicated methods such as stirring and mixing the granules at a temperature below the freezing point of the solution must be adopted. 20 [Mei Nai] Summary of the invention: This problem is related to the prior art. The purpose of the present invention is to prevent the deviation of the distribution of the particles in the dish acid from entering the bone uniformly, and to make the pores in the bone repairing material basically communicate with each other. State, the bone enters 200526279 into its interior, increasing the exposed area of calcium phosphate particles in the pores inside the bone repair material, thereby promoting the entry of bone into its interior. In order to solve the above-mentioned problems, the present inventors conducted intensive studies, and as a result, found that when the aggregate 5 is formed using fibers composed of chitin or a derivative thereof, it is possible to form substantially completely continuous air bubbles, which is substantially effective. The calcium phosphate particles were uniformly carried between the aggregate fibers, thereby completing the present invention. That is, the present invention provides a porous bone repair material. Among the bone repair materials containing a scaffold material composed of chitin or a derivative thereof and a cell differentiation derived material composed of calcium phosphate, the chitin or a derivative thereof The shape 10 is fibrous and becomes a porous aggregate, and at the same time, calcium phosphate becomes granular and is carried between the fibers of the aggregate. In addition, the present invention provides a method for producing a porous bone repair material when producing a porous bone repair material composed of a fiber aggregate of chitin or a derivative thereof and scaly acid # 5 particles carried between the fibers. Including: a step of forming a fiber with several 15 butyrin or a derivative thereof, a step of crosslinking the fiber under reduced pressure, mixing the crosslinked fiber with calcium phosphate particles and water having a water retention capacity below the crosslinked fiber, and A step of forming a substantially uniform mixture, and a step of freeze-drying the mixture to form a porous material. In the porous bone repair material of the present invention, chitin or a derivative thereof 20 is used as a scaffolding material in a regenerative medical technique, and calcium phosphate particles are a material derived from cell differentiation. In the present invention, there are the following reasons for making chitin or its derivative into a collective stomach of fibers. That is, by forming a structure that supports 200526279 grains of disc acid particles between the fibers of the fiber assembly, it is possible to prevent the precipitation of particles and the deviation of the granular knife cloth during the manufacture of the bone repair material. In addition, the fibrous aggregate has substantially completely connected, sad stomata 'so that bone cells can enter a large number of them. Furthermore, since the calcium phosphate particles can be exposed in the continuous stomata, cell differentiation can be well derived. As the chitin or its derivative, carboxymethylchitin (sodium salt) or water-soluble chitosan which can be dissolved in neutral water is preferred in the production process. Alternatively, a water-soluble chitin derivative such as hydroxyethylchitin (ethylene chitin) or ethylene glycol chitosan may be used. 10 Here, derivatives are substances that have been chemically modified with naturally occurring chitin. As the calcium phosphate, various materials such as apatite, tricalcium phosphate, and octacalcium phosphate may be used. The size of the calcium phosphate particles can be used as long as it is 2000 microns or less. The optimal size can be selected according to the type of bone (cortical bone, cavernous bone) in the application site or changes in osteoresponsiveness depending on the site. In the manufacturing method of the present invention, as a method for forming a fiber of chitin or a derivative thereof, when the chitin derivative is water-soluble, an aqueous solution of the material is mixed with a water-soluble organic solvent such as ethanol or acetone. A short fibrous sinker was precipitated quickly by 20 ° C, and the sinker was collected and washed, and the solvent was dried and removed to obtain fibers. In addition, fibers can also be obtained by the following methods. That is, the chitin is solutionized with an alkaline solvent and extruded into a hot water at a high pressure from a nozzle having pores (tens of micrometers), so that it is solidified to become fibrous. 200526279 In addition, the above-mentioned water-soluble chitin derivative is subjected to a cross-linking treatment so that the fiber is not easily dissolved in the body. The paternity treatment is carried out through thermal cross-linking under the condition of 12CTC to 180 spoons under decompression of about 2kPa. It is the same as the 5 capsules of carbamic acid described above, which can be selected according to the application site. Optimum temperature. In addition, when the crosslinked fiber and calcium phosphate particles are mixed, water with a water retention capacity of the crosslinked fiber is added. Here, the water retention capacity of the crosslinked fiber means that the fiber assembly is taken out after being immersed in a large amount of water. The amount of water that the fiber aggregate ingests into and between the fibers and maintains. This is because when the water holding capacity of 10 or more is added, the remaining water has excessive fluidity when mixed with calcium phosphate particles. Therefore, the precipitation or agglomeration of particles due to gravity causes uneven dispersion. When it is below the water retention capacity, the water held by the fiber assembly is difficult to flow, which inhibits the agglomeration or precipitation of calcium phosphate particles. As a result, a uniform composite is formed. 15 The mixing ratio (weight) of the crosslinked fiber and calcium phosphate particles can be made between 1: 0.1 and 1:10. In order to make the mixture into a usable state The drying of the mixture is carried out. The freeze drying is a process generally performed in the manufacture of pharmaceuticals and dried foods. The temperature inside the freeze dryer is kept below the freezing point of the water contained in the mixture 20 (0 ° c or less). ), Which is achieved by depressurizing using a vacuum pump or the like to sublimate and evaporate water. As conditions for freeze-drying, there are no particular restrictions on temperature, degree of decompression, and drying time. According to the porous bone repair material of the present invention, fibers The structure of the calcium aggregates carrying the calcium squamate particles between the fibers of the aggregates can prevent the precipitation of particles and deviations in the distribution of the particles during the manufacture of bone repair 200526279 composite materials. Therefore, bone cells can enter the interior without deviation. In addition, because of the fiber assembly The body has substantially completely continuous stomata, so that a large number of bone cells can enter the interior. Moreover, calcium phosphate particles are exposed in the continuous stomata, so that a large number of cell differentiation can be derived. In addition, according to the manufacturing method of the present invention, Said process can manufacture said porous bone repair material Material, that is, the step of forming a fiber with chitin or a derivative thereof, the step of cross-linking the fiber in a vacuum, mixing the cross-linked fiber with calcium phosphate particles and water having a water retention capacity below the cross-linked fiber, and forming 10 A step of forming a substantially homogeneous mixture, and a step of freeze-drying the mixture to form a porous material. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a SEM image showing the surface of a porous bone repair material of an example. 15 FIG. 2 is a view showing S EM image (reflected electron image) of a section of a porous bone repair material according to an embodiment. L Embodiment 3 Detailed description of a preferred embodiment The following shows a preferred embodiment of the present invention, but the present invention is as described above 20 It is stated that the cross-linking treatment can be thermally cross-linked under heating at 120 ° C to 180 ° C under a reduced pressure of about 2 kPa. Similar to the above-mentioned calcium phosphate particles, the optimal temperature can be selected according to the application site. In addition, when the crosslinked fiber and calcium phosphate particles are mixed, it can be achieved by adding water having a water retention capacity of less than the crosslinked fiber. In addition, the mixing ratio (weight) of crosslinked fiber and calcium phosphate 12 200526279 particles can be made between G.H: 1G. Therefore, the present invention is not limited to the following embodiments. (Preferred manufacturing method) Precisely weigh 3g of CM chitin raw material, add it to a glass container together with pure water (mil 0 water) 72§-5, and use an electromagnetic dispenser to drop and dissolve. Partial CM chitin aqueous solution. In order to accelerate the dissolution of the chitin, the container was heated in a water bath to 30 to 50 t 'and stirred for 3 to 5 hours to obtain a visually homogeneous solution. Pour the solution into a 150-m ethanol beaker and stir quickly with a glass rod for 10 minutes. Through this operation, short fibrous complex chitin sinks. Transfer Shen Dianwu to the other towels, and then wash with ethanol for several times. After removing the ethanol as much as possible, the Shen Dianwu was placed in a 40 m dryer for more than 12 hours to evaporate the ethanol and residual moisture. Then, it is processed in a vacuum heat treatment apparatus under a reduced pressure of 140 ° C (below about 2kpa) for 12 hours and 15 hours. Weigh the above CM chitin and cold TCP granules (50 ~ 150 microns) and pure water in a container with a weight ratio of 1 · 5 · 32 and mix in a container with a spatula. The mixture was filled into a polyethylene round cell of a size suitable for future experiments, and both ends of the cylinder were closed with PET film, and then quickly placed in a freezer of 20 to-. After more than 1 hour and the mixture is completely frozen, the PET film at the rain end is removed and moved to a freeze dryer to evaporate water directly in the frozen state. Then, the test body was taken out from a polystyrene cylinder and subjected to the following test. [Observation of fine structure] ‘13 200526279 This was performed using a scanning electron microscope [SEM]. After the sample was vapor-plated, it was observed with a two-dimensional electronic image at a magnification of 35 times (see Fig. 1). A state where TCP particles were carried between fibers in the fibrous CM chitin (fiber aggregate) was observed. The size of the pores is tens to hundreds of μm. 5 In order to observe the dispersed state of the TCP particles, the test body was embedded in an epoxy resin, the cut surface was cut and ground, and the same SEM observation was performed after flattening. At this time, in order to clearly see the TCP particles, the reflected electron image was used for observation (see Fig. 2). Throughout the observed section (approximately 4x8mm), Tcp particles were observed to be approximately uniformly dispersed, and no eccentric loading of 10 particles due to precipitation was observed. [Implantation test to bone defect] The material was embedded in a bone defect with a diameter of 4 mm formed on the mandible of rabbits, and recovered with surrounding tissues 4 and 8 weeks after surgery to investigate the regeneration of bone tissue situation. For tissues recovered after a predetermined period of time, non-delimed resin-embedded tissue specimens were prepared according to the ~ 15 method, and the area ratio of regenerated bone tissue to bone defect sites in each specimen was calculated. For comparison, an example (blank example) in which only a bone defect is formed and no material is embedded, and a test body manufactured by a method described in the above-mentioned Patent Document 2 (Japanese Patent Application Laid-Open No. 7-116241) is embedded. Example for the same experiment. The area of the bone complex 20 in the blank example was taken as 1, and the value of the area ratio of the examples in which the various materials were embedded was compared. The production of a prior art sample is shown below. In a cooling bath (—1 >), the CM chitin 4.24 wt% CM chitin aqueous solution was stirred, and at the same time, it was bound to form a state in which fine crystal ice and a solution were mixed (frOzen). A predetermined amount was added TCP particles (particle size 50 ~ 150μηι) and stirred to make them evenly divided 20052005279 Fill the knot in a polystyrene cylinder with a size suitable for the w-phase test, and close the two ends of the cylinder with PET film. Quickly put it in a stack. After the cold Kang of C, make the 4 knots dry. Remove the cylinder, and use a vacuum heat treatment under the reduced pressure of C or 160 C (less than 2kPa) and heat treatment for 24 small 5 ^ force to the sponge After cooling at room temperature, the volume is the apparent volume of the molded body 2 (washed with pure water above H mouth for 3 hours, the liquid part is replaced with ethanol, and dried in a dry state at 80 ° C. Production test The body was sterilized by 25 kGy gamma rays for testing. Among them, 'only using the method of the prior art, the TCP particles 10 were eccentrically loaded, but the particles were subjected to a temperature below the freezing point of the solution as described above. Use mixing methods such as stirring, mixing and forming to make A test body with substantially homogeneous and dispersed particles was used for the test. The test results are summarized in Table 1. Table 1 Bone occupancy at the defect site of the embedded material 4 weeks and 8 weeks later The present invention 1.3 1.2 The prior art 0.8 1 Blank 1 1 15 As can be seen from Table 1, when compared with the blank example, when the material made by the present invention is embedded, there is a good bone repair after 4 weeks and at any time after 8 weeks. The materials manufactured by the existing technology have After 4 weeks, the bone repair was slightly worse than the blank case, and it was the same after 8 weeks. 20 [Brief Description of the Drawings] 15 200526279 Figure 1 is an SEM image showing the surface of the porous bone repair material of the example. Figure 2 This is an SEM image (reflected electron image) showing a cross section of the porous bone repair material of the example. 5 [Explanation of Symbols of Major Components] (None) 16