JP5924610B2 - 人工骨膜の作製方法 - Google Patents
人工骨膜の作製方法 Download PDFInfo
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- JP5924610B2 JP5924610B2 JP2011113498A JP2011113498A JP5924610B2 JP 5924610 B2 JP5924610 B2 JP 5924610B2 JP 2011113498 A JP2011113498 A JP 2011113498A JP 2011113498 A JP2011113498 A JP 2011113498A JP 5924610 B2 JP5924610 B2 JP 5924610B2
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- bmp
- biodegradable polymer
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- artificial periosteum
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- 208000006735 Periostitis Diseases 0.000 title claims description 30
- 210000003460 periosteum Anatomy 0.000 title claims description 30
- 238000004519 manufacturing process Methods 0.000 title claims description 15
- 239000000463 material Substances 0.000 claims description 41
- 229920002988 biodegradable polymer Polymers 0.000 claims description 37
- 239000004621 biodegradable polymer Substances 0.000 claims description 37
- 241000588724 Escherichia coli Species 0.000 claims description 13
- 108010049931 Bone Morphogenetic Protein 2 Proteins 0.000 claims description 12
- 102100024506 Bone morphogenetic protein 2 Human genes 0.000 claims description 12
- 229920001577 copolymer Polymers 0.000 claims description 10
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 5
- 108010049870 Bone Morphogenetic Protein 7 Proteins 0.000 claims description 4
- 102100022544 Bone morphogenetic protein 7 Human genes 0.000 claims description 4
- 108010049955 Bone Morphogenetic Protein 4 Proteins 0.000 claims description 2
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- 102100022545 Bone morphogenetic protein 8B Human genes 0.000 claims description 2
- 108010007726 Bone Morphogenetic Proteins Proteins 0.000 description 25
- 102000007350 Bone Morphogenetic Proteins Human genes 0.000 description 25
- 229940112869 bone morphogenetic protein Drugs 0.000 description 25
- 210000000988 bone and bone Anatomy 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 9
- 230000007547 defect Effects 0.000 description 6
- 238000004108 freeze drying Methods 0.000 description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000035876 healing Effects 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 210000004962 mammalian cell Anatomy 0.000 description 4
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 108010010803 Gelatin Proteins 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000008273 gelatin Substances 0.000 description 3
- 229920000159 gelatin Polymers 0.000 description 3
- 235000019322 gelatine Nutrition 0.000 description 3
- 235000011852 gelatine desserts Nutrition 0.000 description 3
- 230000011164 ossification Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- 229920003232 aliphatic polyester Polymers 0.000 description 2
- 230000010478 bone regeneration Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
- 238000002513 implantation Methods 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
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- 210000000130 stem cell Anatomy 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- VSKXVGWORBZZDY-UHFFFAOYSA-N 2-hydroxypropanoic acid;oxepan-2-one Chemical compound CC(O)C(O)=O.O=C1CCCCCO1 VSKXVGWORBZZDY-UHFFFAOYSA-N 0.000 description 1
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
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- 230000037237 body shape Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000010030 laminating Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 230000002188 osteogenic effect Effects 0.000 description 1
- 230000002138 osteoinductive effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920000747 poly(lactic acid) Polymers 0.000 description 1
- 229920002463 poly(p-dioxanone) polymer Polymers 0.000 description 1
- 229920001610 polycaprolactone Polymers 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 239000004633 polyglycolic acid Substances 0.000 description 1
- 239000004626 polylactic acid Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
Description
即ち本発明は、大腸菌由来のBMPを含んだシート状の生分解性高分子材料からなる人工骨膜である。
1,4−ジオキサン中に乳酸−グリコール酸共重合体(乳酸:グリコール酸=75:25、分子量約250、000)を8重量%の濃度となるように入れ撹拌器で1時間撹拌させて溶解した。溶解液を100mm×100mm、隙間2mmの型に流し込み、24時間凍結後、48時間真空乾燥させて80mm×80mm、厚み約1mmの生分解性高分子材料を得た。得られた生分解性高分子材料を金属製のプレス型にて300kgf/cm2の条件にて厚さ約0.25mmにプレスした。作製されたシート状の生分解性高分子材料は、孔のサイズが50μm、気孔率50%、厚さが0.25mmであった。
その後、シート状生分解性高分子材料を10mm×10mmの大きさにカットし、濃度1,5,10μg/μLで調製した大腸菌由来BMP-2を溶解させたリン酸緩衝液0.1mL中に4℃で24時間浸漬させた後、取り出し、凍結乾燥させることで人工骨膜を作製した。なお、凍結乾燥前後の重量測定を行ったところ、本実施例の人工骨膜にはそれぞれ、約16,80,160μgの大腸菌由来BMP-2が含まれていた。
1,4−ジオキサン中に乳酸−グリコール酸共重合体(乳酸:グリコール酸=75:25、分子量約250、000)を8重量%の濃度となるように入れ撹拌器で1時間撹拌させて溶解した。溶解液を100mm×100mm、隙間2mmの型に流し込み、24時間凍結後、48時間真空乾燥させて80mm×80mm、厚み約1mmの生分解性高分子材料を得た。得られた生分解性高分子材料を隙間約0.25mmの金属製のプレス型にて300kgf/cm2の条件にて厚さ約0.25mmにプレスした。作製されたシート状の生分解性高分子材料は、孔のサイズが50μm、気孔率50%、厚さが0.25mmであった。
その後、シート状生分解性高分子材料を10mm×10mmの大きさにカットし、濃度1,5,10μg/μLで調製した哺乳動物細胞を利用したBMP-2を溶解させたリン酸緩衝液0.1mL中に4℃で24時間浸漬させた後、取り出し、凍結乾燥させることで人工骨膜を作製した。なお、凍結乾燥前後の重量測定を行ったところ、本実施例の人工骨膜にはそれぞれ、約16,80,160μgの哺乳動物細胞を利用したBMP-2が含まれていた。
哺乳動物由来BMP-2の製造コスト:¥10,000/mg
Claims (4)
- シート状の生分解性高分子材料からなる人工骨膜の作製方法であって、
宿主細胞として大腸菌を用いてBMPを生産し、前記BMPが溶解された水溶液に、孔のサイズが1〜2000μmφ、気孔率が5〜95%及び厚さが0.01〜2mmの多孔質形状であるシート状の生分解性高分子材料を浸漬させることを特徴とする人工骨膜の作製方法。 - 前記BMPが、BMP-2, BMP-4, BMP-5, BMP-6, BMP-7(OP-1),及びBMP-8(OP-2)からなる群より選択されるBMPの1種または2種以上である請求項1に記載の人工骨膜の作製方法。
- 前記生分解性高分子材料が、乳酸−グリコール酸共重合体である請求項1又は2に記載の人工骨膜の作製方法。
- 前記生分解性高分子材料を浸漬させた後、取り出して凍結乾燥させる請求項1〜3のいずれか記載の人工骨膜の作製方法。
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JP5924610B2 true JP5924610B2 (ja) | 2016-05-25 |
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CN112789034A (zh) * | 2018-09-14 | 2021-05-11 | 奥索赛尔有限公司 | 人造骨膜 |
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JPH053913A (ja) * | 1991-06-26 | 1993-01-14 | Sumitomo Cement Co Ltd | 人工生体複合材料 |
TW369414B (en) * | 1994-09-30 | 1999-09-11 | Yamanouchi Pharma Co Ltd | Bone formation transplant |
EP1053002A1 (en) * | 1998-02-10 | 2000-11-22 | Oregon Health Sciences University | Treatment of bony defects with osteoblast precursor cells |
JP3886834B2 (ja) * | 2002-03-25 | 2007-02-28 | オリンパス株式会社 | 骨再生シート |
US20060045902A1 (en) * | 2004-09-01 | 2006-03-02 | Serbousek Jon C | Polymeric wrap for in vivo delivery of osteoinductive formulations |
JP2007332106A (ja) * | 2006-06-16 | 2007-12-27 | Okayama Univ | 骨形成用徐放性医薬組成物 |
US8524265B2 (en) * | 2006-08-17 | 2013-09-03 | Warsaw Orthopedic, Inc. | Medical implant sheets useful for tissue regeneration |
JP2009045142A (ja) * | 2007-08-16 | 2009-03-05 | Okayama Univ | 生体内にて基材周囲に骨形成を誘導するための基材処理方法 |
EP2268330B1 (en) * | 2008-04-21 | 2012-02-29 | AO Technology AG | Biocompatible implant |
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