JP6213964B2 - Method and apparatus for producing low-degradable fibrin gel membrane for regenerative treatment - Google Patents
Method and apparatus for producing low-degradable fibrin gel membrane for regenerative treatment Download PDFInfo
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Description
本発明は、再生治療に使用可能な低分解性フィブリンゲル膜の製造方法及び製造装置に関するものである。 The present invention relates to a method and an apparatus for producing a low-degradable fibrin gel membrane that can be used for regenerative treatment.
（フィブリンゲル及びその用途）
近年、患者の血液から調製されたゲル状の多血小板フィブリン（ＰＲＦ）が、歯周病等の再生治療、人工インプラントの埋入や美容外科などの分野で、組織再生促進材料として利用されつつある。本明細書ではこのＰＲＦを「フィブリンゲル」とも呼ぶ。
(Fibrin gel and its use)
In recent years, gel-like platelet-rich fibrin (PRF) prepared from patient's blood is being used as a tissue regeneration promoting material in fields such as regenerative treatment of periodontal disease, implantation of artificial implants and cosmetic surgery. . In the present specification, this PRF is also referred to as “fibrin gel”.
（フィブリンゲルの長所）
フィブリンゲルは、（１）治療後の痛みを少なくしかつ傷の治りを早くする点、（２）血小板や増殖因子が豊富に含まれた血液由来の材料であるため、骨の少ない所に使用するとその再生が早くなり、ひいては、治癒期間が短縮する点といった優れた長所を有する。
(Advantages of fibrin gel)
Fibrin gels are (1) less pain after treatment and faster healing of wounds, and (2) blood-derived materials rich in platelets and growth factors, so they are used where bones are scarce. As a result, the regeneration is accelerated, and as a result, the healing period is shortened.
（先行技術：フィブリンゲル圧延装置）
本発明者らは、このフィブリンゲルを、内在する増殖因子や血漿等の栄養分の喪失やその表面に付着した血小板の損傷を引き起こすことなく、フィブリンゲルを均一な厚みに圧延することができる器具を既に提案している（特許文献１を参照）。すなわち、特許文献１に開示の圧延器具は、「組織再生促進材料」としてのフィブリンゲルの本来の機能・長所を保持したままでこれをシート状に形成することを目的として創出された器具である。
(Prior art: Fibrin gel rolling equipment)
The inventors have developed a device that can roll the fibrin gel to a uniform thickness without causing loss of nutrients such as endogenous growth factors and plasma and damage to the platelets adhering to the surface. It has already been proposed (see Patent Document 1). That is, the rolling instrument disclosed in Patent Document 1 is an instrument created for the purpose of forming a fibrin gel as a “tissue regeneration promoting material” in the form of a sheet while retaining the original functions and advantages. .
（遮断膜の必要性）
ところで、歯周病等の外科手術後の創傷治癒においては、歯肉粘膜上皮細胞の治癒の速度が結合組織の線維芽細胞や骨芽細胞より早いので、上皮細胞の深行増殖（down growth）が起こり、骨や歯周組織の再生／新生が得られにくくなることが問題となっている。この深行増殖の問題を解決するために、歯周組織誘導再生（ＧＴＲ）法や骨組織誘導再生（ＧＢＲ）法が提案・実施されている。これらの方法は、具体的には、遮断膜（barrier membrane）を用いて上皮細胞の患部への侵入を防ぎ、線維芽細胞や骨芽細胞の誘導を助けることによって歯周組織や歯槽骨の再生／新生を図るものである（例えば、特許文献２を参照）。
(Necessity of barrier film)
By the way, in wound healing after surgery such as periodontal disease, the healing speed of gingival mucosal epithelial cells is faster than that of connective tissue fibroblasts and osteoblasts, so epithelial cell down growth It is a problem that it is difficult to obtain regeneration / neogenesis of bone and periodontal tissue. In order to solve this deep growth problem, a periodontal tissue guided regeneration (GTR) method and a bone tissue guided regeneration (GBR) method have been proposed and implemented. Specifically, these methods use a barrier membrane to prevent epithelial cells from entering the affected area and help induce fibroblasts and osteoblasts to regenerate periodontal tissue and alveolar bone. / A new life is intended (see, for example, Patent Document 2).
遮断膜には、非吸収性のものと吸収性のものとに大別される。非吸収性の遮断膜には、ポリテトラフルオロエチレン（ＰＴＦＥ）膜などが挙げられるが、生体内で非吸収性のため術後に取り除く必要があり、生体組織（歯肉骨膜弁等）と癒着しないので、術後縫合部の開列が起こり易いという欠点が指摘されている。 The barrier film is roughly classified into a non-absorbing type and an absorbing type. Non-absorbable barrier membranes include polytetrafluoroethylene (PTFE) membranes, but they need to be removed after surgery because they are non-absorbable in vivo and do not adhere to living tissue (gingival periosteal flaps, etc.) Therefore, it has been pointed out that the postoperative suture part is likely to be opened.
一方、吸収性の遮断膜には、コラーゲン膜、ＰＬＡ／ＰＧＡ（ポリ乳酸／ポリグリコール酸共重合体）膜、等の生体分解性膜が挙げられる。しかしながら、コラーゲン等の架橋を架橋剤により化学的に行っていることから生体内でのこれらの膜の分解は遅々としたものであり、また過度の炎症を引き起こすなどの有害事象も報告されている。また、ＰＬＡ／ＰＧＡ膜は分解時に強烈な炎症を引き起こすことも報告されている。 On the other hand, the absorbable barrier membrane includes biodegradable membranes such as a collagen membrane and a PLA / PGA (polylactic acid / polyglycolic acid copolymer) membrane. However, since these membranes are chemically cross-linked with a cross-linking agent, degradation of these membranes in vivo is slow, and adverse events such as excessive inflammation have been reported. Yes. It has also been reported that PLA / PGA membranes cause intense inflammation upon degradation.
このような欠点に依るのか、近年、遮断膜の市販品の入手が困難となっており、主たる需要者である開業歯科医は困惑している。 Whether it depends on such a defect, in recent years, it is difficult to obtain a commercial product of the barrier film, and the practicing dentist who is the main consumer is confused.
本発明者らは、上記遮断膜を取り巻く事情を鑑み、患者の血液由来のフィブリンゲルを、本来の用途たる組織再生促進材料としてではなく、遮断膜としても利用できないかと思案した。 In view of the circumstances surrounding the barrier film, the present inventors wondered whether fibrin gel derived from a patient's blood could be used as a barrier film, not as a tissue regeneration promoting material, which is the original use.
しかしながら、従来のフィブリンゲルは治癒期間を短縮する目的で作製されているため、数日から１０日程度の極めて短い期間で生体内に完全に分解してしまう。遮断膜として使用するには、術後、骨芽細胞が十分に生成するまで所定の空隙を確保できるように、フィブリンゲルが６〜１２週間程度は分解されずに残存する必要がある。 However, since the conventional fibrin gel is produced for the purpose of shortening the healing period, it is completely decomposed in the living body in a very short period of several days to 10 days. In order to use it as a blocking membrane, it is necessary that fibrin gel remains without being decomposed for about 6 to 12 weeks so that a predetermined void can be secured after the operation until osteoblasts are sufficiently generated.
そこで、本発明では、低分解性でかつ膜状（シート状或いはフィルム状）のフィブリンゲルを容易に製造可能な方法及び装置を提供することを目的とする。 Accordingly, an object of the present invention is to provide a method and an apparatus capable of easily producing a fibrin gel having a low decomposability and a film shape (sheet shape or film shape).
本発明者らは、鋭意検討の末、フィブリンゲルに所定の前処理を行い、所定の熱を付与しながら圧延することにより、遮断膜にも利用可能な低分解性のフィブリンゲル膜を見出し、本発明を完成するに至った。 The present inventors, after earnest study, perform a predetermined pretreatment on the fibrin gel, find a low-degradable fibrin gel film that can also be used as a barrier film by rolling while applying a predetermined heat, The present invention has been completed.
すなわち本発明は、例えば、以下の構成・特徴を備えるものである。
（態様１）
フィブリンゲル内の増殖因子及び血小板の一部を除去する予備除去工程と、
前記フィブリンゲルに熱を付与する加熱工程と、
前記フィブリンゲルを圧延する圧延工程と、
を含み、かつ、
前記加熱工程と、前記圧延工程と、を同時に行い、
前記加熱工程では、８０〜１００℃の温度に維持された少なくとも一対の伝熱性部材を用いて、前記フィブリンゲルを１〜１０秒間、加熱しながら均一厚さに圧延することを特徴とする再生治療用低分解性フィブリンゲル膜の製造方法。
（態様２）
前記伝熱性部材が金属部材で構成されていることを特徴とする態様１に記載の製造方法。
（態様３）
前記加熱工程では、前記伝熱性部材の表面の少なくとも一部をセラミックス又は生体親和性を有した油で予め被覆するか、又は、前記フィブリンゲルをフィルム状シートで予め包むことを特徴とする態様１又は２に記載の製造方法。
（態様４）
前記予備除去工程では、前記フィブリンゲルをガーゼで包んで予備押圧することを特徴とする態様１〜３のいずれかに記載の製造方法。
（態様５）
フィブリンゲルを挟持可能な少なくとも一対の伝熱性部材と、
前記伝熱性部材を介して前記フィブリンゲルに熱を供給可能な熱源が設けられた本体と、
前記伝熱性部材の温度を検知する第１センサと、
前記第１センサから得られた情報を取得し、該情報を基に前記熱源での熱の供給を調節して、前記伝熱性部材の温度を８０〜１００℃の範囲に設定可能な制御部と、
を備えたことを特徴とする再生治療用低分解性フィブリンゲル膜の製造装置。
（態様６）
前記伝熱性部材によって前記フィブリンゲルが挟持されているかどうかを検知する第２センサをさらに備え、かつ、前記制御部は、前記第２センサから得られた情報も取得し、該情報を基に前記フィブリンゲルの加熱時間を計算可能にすることを特徴とする態様５に記載の製造装置。
（態様７）
前記制御部は、前記フィブリンゲルの加熱時間を１〜１０秒間の範囲に設定可能であることを特徴とする態様６に記載の製造装置。
（態様８）
前記伝熱性部材は前記本体に着脱自在であることを特徴とする態様５〜７のいずれかに記載の製造装置。
（態様９）
前記伝熱性部材が金属部材で構成されていることを特徴とする態様５〜８のいずれかに記載の製造装置。
（態様１０）
前記伝熱性部材は、セラミックスで被覆された表面を含んでいることを特徴とする態様５〜９のいずれかに記載の製造装置。
（態様１１）
前記伝熱性部材の目標温度及び目標加熱時間の少なくとも一つを受け付けかつこれらの情報を前記制御部に伝達可能な入力装置を備えたことを特徴とする態様５〜１０のいずれかに記載の製造装置。
（態様１２）
前記伝熱性部材の温度及び加熱時間の少なくとも一つを表示可能な表示装置、或いは、前記伝熱性部材の温度及び加熱時間の少なくとも一つが前記制御部で決定した設定範囲に出入りする際に警報を発することが可能な警報装置、又は、該表示装置及び該警報装置をさらに備えたことを特徴とする態様５〜１１のいずれかに記載の製造装置。
That is, the present invention includes, for example, the following configurations and features.
(Aspect 1)
A preliminary removal step of removing a part of growth factors and platelets in the fibrin gel;
A heating step of applying heat to the fibrin gel;
Rolling process for rolling the fibrin gel;
Including, and
Performing the heating step and the rolling step simultaneously,
In the heating step, using at least a pair of heat conductive members maintained at a temperature of 80 to 100 ° C., the fibrin gel is rolled to a uniform thickness while heating for 1 to 10 seconds. Of producing a low-degradable fibrin gel membrane for medical use.
(Aspect 2)
The manufacturing method according to aspect 1, wherein the heat conductive member is formed of a metal member.
(Aspect 3)
In the heating step, at least a part of the surface of the heat conductive member is previously coated with ceramics or biocompatible oil, or the fibrin gel is previously wrapped with a film-like sheet. Or the manufacturing method of 2.
(Aspect 4)
In the preliminary removal step, the fibrin gel is preliminarily pressed by wrapping with gauze.
(Aspect 5)
At least a pair of heat conductive members capable of sandwiching fibrin gel;
A main body provided with a heat source capable of supplying heat to the fibrin gel via the heat conductive member;
A first sensor for detecting a temperature of the heat conductive member;
A controller capable of acquiring information obtained from the first sensor, adjusting a supply of heat from the heat source based on the information, and setting a temperature of the heat transfer member in a range of 80 to 100 ° C; ,
An apparatus for producing a low-degradable fibrin gel membrane for regenerative treatment, comprising:
(Aspect 6)
Further comprising a second sensor for detecting whether or not the fibrin gel is sandwiched by the heat conductive member, and the control unit also acquires information obtained from the second sensor, based on the information The manufacturing apparatus according to aspect 5, wherein the heating time of the fibrin gel can be calculated.
(Aspect 7)
The said control part can set the heating time of the said fibrin gel to the range of 1 to 10 second, The manufacturing apparatus of the aspect 6 characterized by the above-mentioned.
(Aspect 8)
The manufacturing apparatus according to any one of aspects 5 to 7, wherein the heat conductive member is detachable from the main body.
(Aspect 9)
The manufacturing apparatus according to any one of aspects 5 to 8, wherein the heat conductive member is formed of a metal member.
(Aspect 10)
The manufacturing apparatus according to any one of aspects 5 to 9, wherein the heat conductive member includes a surface coated with ceramics.
(Aspect 11)
The manufacturing according to any one of aspects 5 to 10, further comprising an input device that receives at least one of a target temperature and a target heating time of the heat transfer member and can transmit the information to the control unit. apparatus.
(Aspect 12)
A display device capable of displaying at least one of the temperature and heating time of the heat transfer member, or an alarm when at least one of the temperature and heating time of the heat transfer member enters and exits a set range determined by the control unit. The manufacturing apparatus according to any one of aspects 5 to 11, further comprising an alarm device capable of emitting, or the display device and the alarm device.
本発明によれば、生体内での分解速度（吸収速度）が低いフィブリンゲルを製造することができ、例えば、これを遮断膜として提供することができる。従来のフィブリンゲルは１０日程度で生体内に完全に吸収されるが、本発明により製造されたフィブリンゲル膜は、後述するように、分解が著しく遅延し、３〜４週間経過しても完全に吸収されることは無い。また、この遮断膜は血液由来のフィブリンゲルを利用している為、生体内での炎症を招くこともない。 According to the present invention, a fibrin gel having a low degradation rate (absorption rate) in a living body can be produced, and for example, it can be provided as a blocking film. Conventional fibrin gel is completely absorbed in vivo in about 10 days. However, as will be described later, the fibrin gel membrane produced according to the present invention is significantly delayed in decomposition, and will remain completely after 3 to 4 weeks. Will not be absorbed. Further, since this barrier film uses blood-derived fibrin gel, it does not cause inflammation in the living body.
以下、本考案を図面に示す実施の形態に基づき説明するが、本考案は、下記の具体的な
実施形態に何等限定されるものではない。
Hereinafter, although the present invention will be described based on the embodiments shown in the drawings, the present invention is not limited to the following specific embodiments.
（実施形態１）
（再生治療用低分解性フィブリンゲル膜の製造方法）
本発明は、再生治療を目的として、患者の血液から調整されたフィブリンゲルを原料として、低分解性を有しかつフィルム状（つまり膜状）を成すフィブリンゲル（以下、単に「フィブリンゲル膜」又は「フィブリンゲル」とも呼ぶ。）を製造する方法を提供する。先ず、この製造方法について、図１を参照しながら、説明する。なお、以下詳述する各工程を実施する前に、血液由来のフィブリンゲルを調整・用意する工程Ｓ０を含むことは言うまでも無い。
(Embodiment 1)
(Method for producing low-degradable fibrin gel membrane for regenerative treatment)
For the purpose of regenerative treatment, the present invention uses a fibrin gel prepared from a patient's blood as a raw material and has a fibrin gel having a low degradability and forming a film (that is, a film) (hereinafter simply referred to as “fibrin gel film”). Or a “fibrin gel”). First, this manufacturing method will be described with reference to FIG. In addition, before implementing each process explained in full detail below, it cannot be overemphasized that process S0 which prepares and prepares blood-derived fibrin gel is included.
（予備除去工程Ｓ１）
この製造方法は、フィブリンゲル内の増殖因子及び血小板の一部を除去する予備除去工程Ｓ１を含む。この予備除去工程Ｓ１は、ガーゼの利用が挙げられ、フィブリンゲルをガーゼで包んで予備押圧することが好ましい。これにより、フィブリンゲルが遮断膜として生体内に取り込まれた際に、フィブリンゲルの分解速度を高める要因となり得る増殖因子及び血小板を減らすことが可能となる。
(Preliminary removal step S1)
This manufacturing method includes a preliminary removal step S1 for removing a part of growth factors and platelets in the fibrin gel. This preliminary removal step S1 includes the use of gauze, and it is preferable to pre-press the fibrin gel wrapped with gauze. Thereby, when fibrin gel is taken into the living body as a blocking film, it is possible to reduce growth factors and platelets that can increase the rate of fibrin gel degradation.
また、本発明者らが既に提案しているフィブリンゲル圧延器具（特許文献１）や公知の圧延器具を用いて予備的に押圧してもよい。また、本発明の製造装置に設けられた後述する伝熱性部材（例えば、金属部材）を用いて、フィブリンゲルを加熱しない状態で予備的に押圧するようにしてもよい。以上の公知の圧延器具や本製造装置の金属部材を利用した場合は、ガーゼを利用した場合に比べ、増殖因子の数を削減しにくいものの血小板等の液体成分を迅速かつ手軽に排出することが可能となる。 Moreover, you may press preliminarily using the fibrin gel rolling tool (patent document 1) which the present inventors have already proposed, or a well-known rolling tool. Moreover, you may make it preliminarily press in the state which does not heat fibrin gel using the heat conductive member (for example, metal member) mentioned later provided in the manufacturing apparatus of this invention. When using the above-mentioned known rolling tools and metal members of this production apparatus, it is possible to quickly and easily discharge liquid components such as platelets, although it is difficult to reduce the number of growth factors compared to the case of using gauze. It becomes possible.
（加熱工程Ｓ２及び圧延工程Ｓ３）
この製造方法は、さらに、フィブリンゲルに熱を付与する加熱工程Ｓ２と、このフィブリンゲルを圧延する圧延工程Ｓ３と、を含む。なお、後述する実施例のように、加熱工程Ｓ２と圧延工程Ｓ３とを同時に行うことが好ましい。
(Heating step S2 and rolling step S3)
The manufacturing method further includes a heating step S2 for applying heat to the fibrin gel and a rolling step S3 for rolling the fibrin gel. In addition, it is preferable to perform heating process S2 and rolling process S3 simultaneously like the Example mentioned later.
（加熱条件）
この加熱工程Ｓ２では、８０〜１００℃の周囲温度かつ１〜１０秒間、前記フィブリンゲルを加熱する。好ましくは、周囲温度は８０〜９０℃で、加熱時間は１〜５秒間である。要求されるフィブリンゲル膜の厚さが薄い場合には、加熱時間は１〜３秒であることが好ましい。周囲温度や加熱時間が上限を超えてしまうとフィブリンゲル内の蛋白質が過度に変性（場合によっては、フィブリンゲルの反り返り（curled）等、その形状が変形）してしまう。一方、下限を下回るとフィブリンゲルに熱を付与してもその内部のフィブリン繊維同士を効率的又は効果的に熱架橋させることができなくなる恐れがある。上記好ましい温度でフィブリンゲルを加熱できれば、フィブリン繊維間に効率的又は効果的な熱架橋が施され、生体内の分解酵素に対して抵抗性を増加させることができる。なお、一対の伝熱性部材を用いて上下（又は左右）からフィブリンゲルを短時間に加熱することにより、外側は比較的硬く、内側は比較的柔らかいフィブリンゲル膜を形成することができる。
(Heating conditions)
In this heating step S2, the fibrin gel is heated at an ambient temperature of 80 to 100 ° C. and for 1 to 10 seconds. Preferably, the ambient temperature is 80-90 ° C. and the heating time is 1-5 seconds. When the required fibrin gel film is thin, the heating time is preferably 1 to 3 seconds. If the ambient temperature or the heating time exceeds the upper limit, the protein in the fibrin gel is excessively denatured (in some cases, the shape of the fibrin gel is curled or the like is deformed). On the other hand, if the temperature is below the lower limit, even if heat is applied to the fibrin gel, there is a possibility that the fibrin fibers in the inside cannot be efficiently or effectively thermally crosslinked. If the fibrin gel can be heated at the above-mentioned preferable temperature, efficient or effective thermal cross-linking can be performed between the fibrin fibers, and resistance to degrading enzymes in the living body can be increased. In addition, by heating the fibrin gel from above and below (or left and right) using a pair of heat conductive members in a short time, a fibrin gel film that is relatively hard on the outside and relatively soft on the inside can be formed.
（同時処理）
なお、圧延工程Ｓ３が加熱工程Ｓ２と同時に行われる場合、前述した加熱温度に維持された少なくとも一対の伝熱性部材でフィブリンゲルを加熱しながら均一厚さに圧延することが望ましい。遮断膜に利用できるよう、その厚みを０．１ｍｍ〜２ｍｍ程度に設定することが好ましい。
(Simultaneous processing)
When the rolling step S3 is performed simultaneously with the heating step S2, it is desirable to roll the fibrin gel to a uniform thickness while heating the fibrin gel with at least a pair of heat conductive members maintained at the heating temperature described above. The thickness is preferably set to about 0.1 mm to 2 mm so that it can be used as a barrier film.
なお、伝熱性部材として、金属部材、又は、伝熱性材料で外表面を被覆した樹脂、など伝熱性を有する材料の使用が挙げられ、好ましくは、フィブリンゲルを圧延可能な面を有したシート状の金属部材であり、さらに好ましくは、後述の実施形態２に示すようにフィブリンゲルを把持・収容が可能な匙状の金属部材である。なお、樹脂を利用する場合、上述した加熱温度にも耐えられる耐熱性を有する材料を選択する必要がある。 Examples of the heat transfer member include use of a metal member or a material having heat transfer properties such as a resin whose outer surface is coated with a heat transfer material, and preferably a sheet-like shape having a surface on which fibrin gel can be rolled. More preferably, the metal member is a bowl-shaped metal member capable of gripping and containing fibrin gel as shown in a second embodiment described later. In addition, when using resin, it is necessary to select the material which has heat resistance which can endure the heating temperature mentioned above.
（フィブリンゲルの付着防止処理）
また、加熱工程Ｓ２では、伝熱性部材の表面の少なくとも一部をセラミックス又は生体親和性を有した油で予め被覆するか、又は、フィブリンゲルをフィルム状シートで予め包んでおくこと（図１では、付着防止処理工程Ｓ２Ａ）が好ましい。この処理工程Ｓ２Ａにより、加熱後のフィブリンゲルが、伝熱性部材の表面にこびりつかずに難無く採取することができるようになる。なお、フィルム状シートには、市販の食品包装用フィルムや公知の医療用フィルムが挙げられる。
(Fibrin gel adhesion prevention treatment)
In addition, in the heating step S2, at least a part of the surface of the heat transfer member is previously coated with ceramics or biocompatible oil, or fibrin gel is pre-wrapped with a film sheet (in FIG. 1). Adhesion prevention treatment step S2A) is preferred. By this treatment step S2A, the heated fibrin gel can be collected without difficulty without sticking to the surface of the heat conductive member. Examples of the film-like sheet include commercially available food packaging films and known medical films.
（実施形態２）
（再生治療用低分解性フィブリンゲル膜の製造装置）
次に、上述した方法の加熱工程Ｓ２及び圧延工程Ｓ３を実現可能な器具（製造装置）１について、図２を参照しながら説明する。なお、上記加熱工程Ｓ２は、下記の製造装置１に限らず、他の装置を用いても実現可能であり、例えば、高温の液体や気体を収容した容器（図示せず）を用意し、フィブリンゲルを所定時間、この容器内に配置することでも達成可能であろう。
(Embodiment 2)
(Production equipment for low-degradable fibrin gel membrane for regenerative treatment)
Next, an instrument (manufacturing apparatus) 1 capable of realizing the heating step S2 and the rolling step S3 of the above-described method will be described with reference to FIG. The heating step S2 is not limited to the manufacturing apparatus 1 described below, and can be realized by using other apparatuses. For example, a container (not shown) containing a high-temperature liquid or gas is prepared and fibrin is prepared. It may also be achieved by placing the gel in this container for a predetermined time.
この製造装置１は、フィブリンゲル（図２では符号Ｘ）を挟持可能な少なくとも一対の伝熱性部材２（図２では、一対の匙状の金属部材２ａ，２ｂ）と、伝熱性部材２を介してフィブリンゲルＸに熱を供給可能な熱源３（図２では、破線で示したコイル３ａ，３ｂ）が設けられた本体４と、を少なくとも備える。この形態を有した本体４は、ヒンジ部５にて一定距離だけ回動可能に接続された一対の柄部６（６ａ，６ｂ）内に熱源３ａ，３ｂが内蔵されている。また、本体４には、熱源３ａ，３ｂに電力を供給するための電源プラグ７も設けられている。さらに、ヒンジ部５の近傍には、加熱を開始又は終了させるためのスイッチ９が設けられていても良い。 The manufacturing apparatus 1 includes at least a pair of heat conductive members 2 (in FIG. 2, a pair of bowl-shaped metal members 2 a and 2 b) capable of sandwiching fibrin gel (symbol X in FIG. 2) and a heat conductive member 2. And at least a main body 4 provided with a heat source 3 (coils 3a and 3b shown by broken lines in FIG. 2) capable of supplying heat to the fibrin gel X. The main body 4 having this form includes heat sources 3a and 3b in a pair of handle portions 6 (6a and 6b) connected to be able to rotate by a fixed distance by a hinge portion 5. The main body 4 is also provided with a power plug 7 for supplying power to the heat sources 3a and 3b. Further, a switch 9 for starting or ending heating may be provided in the vicinity of the hinge portion 5.
また、伝熱性部材２は本体４に着脱自在であることが好ましい。図２の例では、図２（Ｂ）に示すように、それぞれの伝熱性部材２ａ，２ｂには延長部２１（２１ａ，２１ｂ）が接続されている一方、それぞれの柄部６ａ，６ｂの先端には、延長部２１ａ，２１ｂを熱源３ａ，３ｂへ導くための差し込み口８（８ａ，８ｂ）が設けられている。 Further, the heat conductive member 2 is preferably detachable from the main body 4. In the example of FIG. 2, as shown in FIG. 2B, the extension portions 21 (21 a, 21 b) are connected to the respective heat transfer members 2 a, 2 b, while the tips of the respective handle portions 6 a, 6 b are connected. Is provided with an insertion port 8 (8a, 8b) for guiding the extension portions 21a, 21b to the heat sources 3a, 3b.
伝熱性部材２ａ，２ｂの材料として、例えば、ステンレス、チタン、アルミニウム等の金属部材が好ましい。これらの材料は、良好な伝熱特性（熱伝導率及び熱伝達率）を有する他、耐圧性、耐熱性、及び耐腐食性を有する材料でもあり、オートクレーブ滅菌処理や超音波洗浄を適用することで再利用可能となる。また、フィブリンゲルＸと直接接触し最も汚れやすい伝熱性部材２ａ，２ｂを着脱自在とすることで、この製造装置１のうち当該部材２ａ，２ｂのみを一回限りの使い捨て（ディスポーザル）とすることもできる。 As a material of the heat conductive members 2a and 2b, for example, a metal member such as stainless steel, titanium, or aluminum is preferable. These materials have good heat transfer characteristics (thermal conductivity and heat transfer rate), and also have pressure resistance, heat resistance, and corrosion resistance. Apply autoclave sterilization and ultrasonic cleaning. Can be reused. In addition, by making the heat conductive members 2a and 2b that come in direct contact with the fibrin gel X and are most likely to be soiled freely attachable / detachable, only the members 2a and 2b of the manufacturing apparatus 1 are disposable (disposal). You can also
また、伝熱性部材２ａ，２ｂは、図示しないセラミックスで被覆された表面を含んでいることがさらに好ましい。例えば、フィブリンゲルＸを収容・挟持する各面２２ａ，２２ｂにこのセラミックスコーティングが施されていることが好ましい。これにより、加熱後のフィルム状のフィブリンゲル（図示では符号Ｙ）が、伝熱性部材２ａ，２ｂの表面２２ａ，２２ｂにこびりつかずに難無く採取することができるようになる。 Further, it is more preferable that the heat conductive members 2a and 2b include a surface coated with ceramics (not shown). For example, it is preferable that the ceramic coating is applied to each of the surfaces 22a and 22b that contain and sandwich the fibrin gel X. Thereby, the heated film-like fibrin gel (symbol Y in the drawing) can be collected without difficulty without sticking to the surfaces 22a, 22b of the heat conductive members 2a, 2b.
また、伝熱性部材２ａ，２ｂの形状としては、種々の形状を選択することができるが、当該金属部材が急速に加熱されて一定の温度を保持できかつフィブリンゲルＸを均一な薄い厚さで圧延できる性能が得られるようにするには、適度に薄くて扁平な板状に構成されていることが好ましい。なお、図示の例の伝熱性部材２ａ，２ｂは、フィブリンゲルＸを効率良く収容・挟持するため、匙状に形成されている。また、伝熱性部材２ａ，２ｂを最も閉じた状態では、これらの部材２ａ，２ｂの対向する面２２ａ，２２ｂが、フィブリンゲル膜Ｙの所望の厚み（０．１ｍｍ〜２ｍｍ程度）に応じた離間距離を有するように設定されていることが好ましい。 In addition, various shapes can be selected as the shape of the heat conductive members 2a and 2b. However, the metal member can be rapidly heated to maintain a constant temperature, and the fibrin gel X can be uniformly thin. In order to obtain the performance that can be rolled, it is preferable that the plate is configured to be a moderately thin and flat plate. In addition, the heat conductive members 2a and 2b in the illustrated example are formed in a bowl shape in order to efficiently store and sandwich the fibrin gel X. In the state where the heat conductive members 2a and 2b are most closed, the opposing surfaces 22a and 22b of the members 2a and 2b are separated according to the desired thickness (about 0.1 mm to 2 mm) of the fibrin gel film Y. It is preferable that the distance is set.
本発明の製造装置１は、さらに、伝熱性部材２ａ，２ｂの温度を検知する第１センサ（温度センサ）３０と、第１センサ３０から得られた情報を取得し、当該情報を基に熱源３ａ，３ｂでの熱の供給を調節して伝熱性部材２ａ，２ｂの温度を８０〜１００℃の範囲に設定可能な制御部４０と、を備える。これらの構成要素３０，４０を設けることにより、伝熱性部材２ａ，２ｂの表面の現在温度を実測し、その実測値に応じて熱源での熱の供給量を制御すること（場合によっては供給を遮断すること）によって、フィブリンゲルＸにおける繊維間の熱架橋を行うのに適切な熱量を伝熱性部材２ａ，２ｂに供給し、伝熱性部材２ａ，２ｂを所望の温度に維持することができるようになる。 The manufacturing apparatus 1 of the present invention further acquires a first sensor (temperature sensor) 30 that detects the temperature of the heat conductive members 2a and 2b, and information obtained from the first sensor 30, and based on the information, a heat source And a control unit 40 capable of adjusting the temperature of the heat transfer members 2a and 2b within a range of 80 to 100 ° C. by adjusting the supply of heat at 3a and 3b. By providing these components 30 and 40, the current temperatures of the surfaces of the heat transfer members 2a and 2b are measured, and the amount of heat supplied from the heat source is controlled according to the measured values (in some cases, the supply is performed). By cutting off), it is possible to supply the heat transfer members 2a and 2b with an appropriate amount of heat for performing thermal crosslinking between fibers in the fibrin gel X, and to maintain the heat transfer members 2a and 2b at a desired temperature. become.
この製造装置１は、さらに、伝熱性部材２ａ，２ｂによってフィブリンゲルＸが挟持されているかどうか（つまり、挟持の有無）を検知する第２センサ（接触検知センサ）５０を備え、かつ、制御部４０は、第２センサ５０から得られた情報も取得し、当該情報を基にフィブリンゲルＸの加熱時間を計算可能にすることが好ましい。 The manufacturing apparatus 1 further includes a second sensor (contact detection sensor) 50 that detects whether or not the fibrin gel X is sandwiched between the heat conductive members 2a and 2b (that is, whether or not the fibrin gel X is sandwiched), and a control unit. 40 also preferably obtains information obtained from the second sensor 50 and makes it possible to calculate the heating time of the fibrin gel X based on the information.
また、この製造装置１は、伝熱性部材２ａ，２ｂの目標温度及び目標加熱時間を受け付けかつこれらの情報を制御部４０に伝達可能な入力装置６１をさらに備えることが好ましい。このような入力装置６１を設けることで、目標とする温度や加熱時間を任意に設定したり、微調整したりすることが可能となる。 In addition, it is preferable that the manufacturing apparatus 1 further includes an input device 61 that can receive the target temperature and the target heating time of the heat conductive members 2 a and 2 b and can transmit these information to the control unit 40. By providing such an input device 61, it is possible to arbitrarily set or finely adjust the target temperature and heating time.
また、この製造装置１は、第１センサ３０で実測された温度、制御部４０で計算された加熱時間、及び、入力装置６１で受け付けられた目標温度若しくは目標時間の少なくとも一つを表示する表示装置６２や、伝熱性部材２ａ，２ｂの温度及び加熱時間が制御部４０で決定した設定範囲に出入りする際に警報を発する警報装置６３をさらに備えることが好ましい。このような表示装置６２や警報装置６３を設けることで、適切な加熱条件を順守したままフィブリンゲルＸを加熱及び圧延することができるため、熟練した者で無くともこの製造装置１を難なく適切に操作することが可能となる。 The manufacturing apparatus 1 also displays at least one of the temperature actually measured by the first sensor 30, the heating time calculated by the control unit 40, and the target temperature or target time received by the input device 61. It is preferable to further include an alarm device 63 that issues an alarm when the device 62 and the temperature and heating time of the heat conductive members 2a and 2b enter and exit the set range determined by the control unit 40. By providing such a display device 62 and an alarm device 63, the fibrin gel X can be heated and rolled while adhering to appropriate heating conditions. It becomes possible to operate.
なお、上述の制御部４０、入力装置６１、表示装置６２、及び警報装置６３の少なくとも一つは、公知のパーソナルコンピュータ６０を利用することができる。 A known personal computer 60 can be used as at least one of the control unit 40, the input device 61, the display device 62, and the alarm device 63 described above.
また、本発明の製造装置１には種々の変形・改良が可能である。例えば、図２において本発明の製造装置１の外部に現われている構成要素（例えば、第１センサ３０、制御部４０、入力装置６１、表示装置６２、及び警報装置６３）は、この製造装置１に一体となるよう内蔵されていてもよい。また、図２に示す製造装置１では電源プラグ７が設けられているが、これに代えて、例えば、使い捨て電池や充電式電池等を利用してもよい。 Moreover, various deformation | transformation and improvement are possible for the manufacturing apparatus 1 of this invention. For example, the components (for example, the first sensor 30, the control unit 40, the input device 61, the display device 62, and the alarm device 63) appearing outside the manufacturing apparatus 1 of the present invention in FIG. It may be built in so as to be integrated. Moreover, although the power supply plug 7 is provided in the manufacturing apparatus 1 shown in FIG. 2, it may replace with this and may utilize a disposable battery, a rechargeable battery, etc., for example.
（試験管を用いたインビトロでの実証試験）
健康な成人の新鮮な血液（末梢血）を遠心操作して調整し、ＰＢＳ液で洗浄したフィブリンゲルＸを用意した。これを乾いたガーゼで軽く押し潰して、内部の増殖因子及び血小板の一部を除去した（予備除去工程Ｓ１）。
(In vitro verification test using test tubes)
Fibrin gel X prepared by centrifuging fresh blood (peripheral blood) of healthy adults and washed with PBS solution was prepared. This was lightly crushed with dry gauze to remove some of the internal growth factors and platelets (preliminary removal step S1).
次に、伝熱性部材２ａ，２ｂを加熱してこれを約１００℃の温度に保たれるようにした。実証実験では、伝熱性部材２ａ，２ｂ及び熱源３ａ，３ｂとして毛髪用アイロン（テスコム電機株式会社製）と、伝熱性部材２ａ，２ｂの温度を検知する第１センサ３０やこの温度を表示する表示装置６２として放射温度計（株式会社エー・アンド・デイ製）とを用いて、フィブリンゲルＸの加熱工程Ｓ２及び圧延工程Ｓ３を同時に実行した。加熱時間は約１０秒間とした。なお、加熱・圧延工程Ｓ２，Ｓ３の前に、フィブリンゲルＸを食品包装用フィルムで包んでおいた（付着防止処理工程Ｓ２Ａ）。具体的には、食品包装用フィルムの表面をエタノールで拭いて紫外線（ＵＶ）で滅菌した後、フィブリンゲルＸを包むようにした。これにより、フィブリンゲルＸを無菌的に操作できただけでなく、伝熱性部材２ａ，２ｂの加熱面へのフィブリンゲルＸのこびりつきを防止することができた。 Next, the heat conductive members 2a and 2b were heated so as to be maintained at a temperature of about 100 ° C. In the demonstration experiment, a hair iron (manufactured by Tescom Electric Co., Ltd.) as the heat conductive members 2a and 2b and the heat sources 3a and 3b, a first sensor 30 for detecting the temperature of the heat conductive members 2a and 2b, and a display for displaying the temperature. Using a radiation thermometer (manufactured by A & D Co., Ltd.) as the device 62, the heating step S2 and the rolling step S3 of the fibrin gel X were simultaneously performed. The heating time was about 10 seconds. In addition, before heating and rolling process S2, S3, the fibrin gel X was wrapped with the film for food packaging (adhesion prevention process process S2A). Specifically, the surface of the food packaging film was wiped with ethanol and sterilized with ultraviolet rays (UV), and then fibrin gel X was wrapped. As a result, not only the fibrin gel X could be operated aseptically, but also the fibrin gel X could be prevented from sticking to the heating surfaces of the heat conductive members 2a and 2b.
その後、加熱・圧延されたフィブリンゲル膜Ｙにパンチ器具を押し当てパンチして、直径８ｍｍの円盤状の試験片を作製した（実施例１）。なお、予備除去工程Ｓ１のみを実行したフィブリンゲルＸを用いて同一寸法・形状の試験片を作製し、これを比較例１とした。 Thereafter, a punch device was pressed against the heated / rolled fibrin gel film Y to produce a disk-shaped test piece having a diameter of 8 mm (Example 1). In addition, the test piece of the same dimension and shape was produced using the fibrin gel X which performed only preliminary removal process S1, and this was made into the comparative example 1. FIG.
次に、２４−ｗｅｌｌ ｐｌａｔｅに上記試験片を挿入し、プラスミンを２μｇ／ｍＬ濃度で注入したハンクス液（ＨＢＳＳ）で充たし、ＣＯ２インキュベータ内で培養した。なお、ＨＢＳＳは２日おきに新しいものに取り換えた。 Next, the test piece was inserted into a 24-well plate, filled with Hanks' solution (HBSS) into which plasmin was injected at a concentration of 2 μg / mL, and cultured in a CO 2 incubator. The HBSS was replaced with a new one every two days.
（試験管での結果）
図３に、試験片の分解の経時変化を示す。上段は比較例１の試験片（図中、符号Ｘを参照）の外観を示す一方、下段は実施例１の試験片（図中、符号Ｙを参照）の外観を示す。この図３からも分かるように、ガーゼで押しつぶしただけでの比較例１の試験片Ｘでは１０日程度でほぼ分解されてしまうのが観察された。一方、実施例１の試験片Ｙでは、僅かな分解は確認されたが、１０日経過してもほぼ分解されずに残存していることが観察された。なお、別の成人の血液から調整したフィブリンゲルを用いて同様の方法で試験を何度か行ったが、同様の結果を得た。
(Results in test tubes)
FIG. 3 shows the change over time in the decomposition of the test piece. The upper part shows the appearance of the test piece of Comparative Example 1 (see the reference symbol X in the figure), while the lower part shows the appearance of the test piece of Example 1 (see the reference sign Y in the drawing). As can be seen from FIG. 3, it was observed that the test piece X of Comparative Example 1 that was simply crushed with gauze was almost decomposed in about 10 days. On the other hand, in the test piece Y of Example 1, although slight decomposition was confirmed, it was observed that it remained substantially undecomposed even after 10 days. In addition, although the test was done several times by the same method using the fibrin gel prepared from another adult's blood, the same result was obtained.
（動物を用いたインビトロでの実証試験）
動物実験のサンプルとして生後６週間の雄のネズミを複数調達し、本試験前に試験場所で１週間以上保管・生育した。また、この動物実験に供するフィブリンゲルＸも、上述の試験管試験の実施例１及び比較例１の場合と同様に調整し、実施例２及び比較例２として用意した。
(In vitro demonstration test using animals)
A plurality of 6-week-old male mice were procured as samples for animal experiments, and were stored and grown at the test site for more than one week before the main test. In addition, fibrin gel X to be subjected to this animal experiment was prepared in the same manner as in the above-described test tube test of Example 1 and Comparative Example 1, and prepared as Example 2 and Comparative Example 2.
これらの動物サンプルの背部を消毒した後、正中線皮膚切開を実施し、その皮下組織に、実施例２或いは比較例２の試験片を移植して、その後の試験片の分解・吸収の様子を観察した。試験片とこの周辺の結合組織は、これらをヘマトキシリン、エオシン、又はマッソン・トリクロームを用いて染色することで可視化された。 After disinfecting the back of these animal samples, a midline skin incision was performed, and the test piece of Example 2 or Comparative Example 2 was transplanted into the subcutaneous tissue, and the state of subsequent decomposition and absorption of the test piece was observed. Observed. The specimen and the surrounding connective tissue were visualized by staining them with hematoxylin, eosin, or Masson trichrome.
（動物実験の結果）
図４の各図に、上記動物実験の結果、具体的には、ヘマトキシリンで染色された試験片及びその周辺の結合組織の切断画像を示す。図４の（Ａ）及び（Ｂ）は、移植後１週間目及び２週間目の比較例２の試験片（図中の破線及び符号Ｘを参照）の状態を示す。一方、図４（Ｃ）、（Ｄ）及び（Ｅ）は、移植後１週間目、２週間目、及び３週間目の実施例２の試験片（図中の破線及び符号Ｙを参照）の状態を示す。なお、各画像の倍率・縮尺は同様である。
(Results of animal experiments)
Each figure of FIG. 4 shows the result of the animal experiment, specifically, a cut image of the test piece stained with hematoxylin and its surrounding connective tissue. 4 (A) and 4 (B) show the state of the test piece of Comparative Example 2 (see the broken line and symbol X in the figure) at 1 week and 2 weeks after transplantation. On the other hand, FIGS. 4C, 4D, and 4E show the test pieces of Example 2 (see broken line and symbol Y in the figure) at 1 week, 2 weeks, and 3 weeks after transplantation. Indicates the state. The magnification and scale of each image are the same.
これらの画像からも明らかなように、比較例２の試験片Ｘは、２週間を経過すると非常に薄くなっていることが観察される。なお、３週間目では完全に無くなってしまったため、図示していない。 As is apparent from these images, it is observed that the test piece X of Comparative Example 2 is very thin after two weeks. In addition, since it was completely lost in the 3rd week, it is not illustrated.
一方、この図４（Ｃ）、（Ｄ）及び（Ｅ）の各画像からも明らかなように、実施例２の試験片Ｙは、時間の経過とともに、縮小しているが、３週間を経過しても、確実に残存していることが観察された。 On the other hand, as is apparent from the images of FIGS. 4C, 4D, and 4E, the test piece Y of Example 2 is reduced with the passage of time, but 3 weeks have passed. Even then, it was observed that it remained reliably.
以上のように、試験管を用いた試験及び動物実験を用いた試験においても、実施例１，２及び比較例１，２において、生体分解酵素に対する分解速度において有意な差が観察された。 As described above, also in the test using the test tube and the test using the animal experiment, a significant difference was observed in the degradation rate for the biodegrading enzyme in Examples 1 and 2 and Comparative Examples 1 and 2.
上述したように、歯周病等の再生医療などの分野にて有害物質を含まない安全な遮断膜の需要が高いだけで無く、開業歯科医等が自ら手軽に遮断膜を作製したいとの要望も高い。従って、本発明は、これらの需要に応え、安心・安全な治療の発展に寄与するものであり、産業上の利用価値及び利用可能性は非常に高い。 As mentioned above, there is a high demand for safe barrier films that do not contain harmful substances in fields such as regenerative medicine such as periodontal disease, and there is a demand for practitioners to easily create barrier films themselves. Is also expensive. Therefore, the present invention responds to these demands and contributes to the development of safe and secure treatment, and its industrial utility value and applicability are very high.
Ｓ１ 予備除去工程
Ｓ２ 加熱工程
Ｓ２Ａ 付着防止処理工程
Ｓ３ 圧延工程
１ 再生治療用低分解性フィブリンゲル膜の製造装置
２（２ａ，２ｂ） 伝熱性部材（一対の金属部材）
３（３ａ，３ｂ） 熱源（一対の熱源）
４ 製造装置の本体
３０ 第１センサ（温度センサ）
４０ 制御部
５０ 第２センサ（接触検知センサ）
６１ 入力装置
６２ 表示装置
６３ 警報装置
Ｘ フィブリンゲル
Ｙ フィブリンゲル膜
S1 Preliminary removal step S2 Heating step S2A Adhesion prevention treatment step S3 Rolling step 1 Production device for low-degradable fibrin gel film for regenerative treatment 2 (2a, 2b) Heat transfer member (a pair of metal members)
3 (3a, 3b) Heat source (a pair of heat sources)
4 Manufacturing Equipment Main Body 30 First Sensor (Temperature Sensor)
40 control unit 50 second sensor (contact detection sensor)
61 Input device 62 Display device 63 Alarm device X Fibrin gel Y Fibrin gel membrane
Claims (12)
前記フィブリンゲルに熱を付与する加熱工程と、
前記フィブリンゲルを圧延する圧延工程と、
を含み、かつ、
前記加熱工程と、前記圧延工程と、を同時に行い、
前記加熱工程では、８０〜１００℃の温度に維持された少なくとも一対の伝熱性部材を用いて、前記フィブリンゲルを１〜１０秒間、加熱しながら均一厚さに圧延することを特徴とする再生治療用低分解性フィブリンゲル膜の製造方法。 A preliminary removal step of removing a part of growth factors and platelets in the fibrin gel;
A heating step of applying heat to the fibrin gel;
Rolling process for rolling the fibrin gel;
Including, and
Performing the heating step and the rolling step simultaneously,
In the heating step, using at least a pair of heat conductive members maintained at a temperature of 80 to 100 ° C., the fibrin gel is rolled to a uniform thickness while heating for 1 to 10 seconds. Of producing a low-degradable fibrin gel membrane for medical use.
前記伝熱性部材を介して前記フィブリンゲルに熱を供給可能な熱源が設けられた本体と、
前記伝熱性部材の温度を検知する第１センサと、
前記第１センサから得られた情報を取得し、該情報を基に前記熱源での熱の供給を調節して、前記伝熱性部材の温度を８０〜１００℃の範囲に設定可能な制御部と、
を備えたことを特徴とする再生治療用低分解性フィブリンゲル膜の製造装置。 At least a pair of heat conductive members capable of sandwiching fibrin gel;
A main body provided with a heat source capable of supplying heat to the fibrin gel via the heat conductive member;
A first sensor for detecting a temperature of the heat conductive member;
A controller capable of acquiring information obtained from the first sensor, adjusting a supply of heat from the heat source based on the information, and setting a temperature of the heat transfer member in a range of 80 to 100 ° C; ,
An apparatus for producing a low-degradable fibrin gel membrane for regenerative treatment, comprising:
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