Spinal fusion remains the gold-standard treatment for several pathological spine conditions. Although, autologous Iliac Crest Bone Grafting is considered the goldstandard graft choice to promote spinal fusion; however...Spinal fusion remains the gold-standard treatment for several pathological spine conditions. Although, autologous Iliac Crest Bone Grafting is considered the goldstandard graft choice to promote spinal fusion; however, it is associated with significant donor site morbidity and a limited graft quantity. Therefore, several bone graft alternatives have been developed, to augment arthrodesis. The purpose of this review is to present the results of clinical studies concerning the use of demineralized bone matrix(DBM), alone or as a composite graft, in the spinal fusion. A critical review of the English-language literature was conducted on Pubmed, using key word "demineralized bone matrix", "DBM", "spinal fusion", and "scoliosis". Results had been restricted to clinical studies. The majority of clinical trials demonstrate satisfactory fusion rates when DBM is employed as a graft extender or a graft enhancer.Limited number of prospective randomized controlled trials(4 studies), have been performed comparing DBM to autologous iliac crest bone graft in spine fusion. The majority of the clinical trials demonstrate comparable efficacy of DBM when it used as a graft extender in combination with autograft, but there is no clinical evidence to support its use as a standalone graft material. Additionally, high level of evidence studies are required, in order to optimize and clarify the indications of its use and the appropriate patient population that will benefit from DBM in spine arthrodesis.展开更多
Autologous bone graft is considered as the gold standard for all indications for bone grafting procedures but the limited availability and complications in donor site resulted in seeking other options like allografts ...Autologous bone graft is considered as the gold standard for all indications for bone grafting procedures but the limited availability and complications in donor site resulted in seeking other options like allografts andbone graft substitutes. Demineralized bone matrix(DBM) is an allograft product with no quantity limitation. It is an osteoconductive material with osteoinductive capabilities, which vary among different products, depending on donor characteristics and differences in processing of the bone. The purpose of the present review is to provide a critical review of the existing literature concerning the use of DBM products in various procedures in the extremities. Clinical studies describing the use of DBM alone or in combination with other grafting material are available for only a few commercial products. The Level of Evidence of these studies and the resulting Grades of Recommendation are very low. In conclusion, further clinical studies of higher quality are required in order to improve the Recommendation Grades for or against the use of DBM products in bone grafting procedures.展开更多
AIM To assess use of demineralized bone matrix(DBM) use in anterior cervical discectomy and fusion(ACDF) in outpatient setting.METHODS One hundred and forty-five patients with prospectively collected data undergoing s...AIM To assess use of demineralized bone matrix(DBM) use in anterior cervical discectomy and fusion(ACDF) in outpatient setting.METHODS One hundred and forty-five patients with prospectively collected data undergoing single and two level ACDF with DBM packed within and anterior to polyetheretherketone(PEEK) cages. Two groups created, Group 1(75) outpatients and control Group 2(70) hospital patients. Prevertebral soft tissue swelling(PVSTS) was measured anterior to C2 and C6 on plain lateral cervical radiographs preoperatively and one week postoperatively and fusion assessed at two years. RESULTS There was no intergroup significance between preoperative and postoperative visual analogue scales(VAS)and neck disability index(NDI) scores between Group 1 and 2. Mean preoperative PVSTS in Group 1 was 4.7 ± 0.2 mm at C2 level and 11.1 ± 0.5 at C6 level compared to Group 2 mean PVSTS of 4.5 ± 0.5 mm and 12.8 ± 0.5, P = 0.172 and 0.127 respectively. There was no radiographic or clinical evidence of adverse reaction noted. In Group 1 mean postoperative PVSTS was 5.5 ± 0.4 mm at C2 and 14.9 ± 0.6 mm at C6 compared Group 2 mean PVSTS was 4.9 ± 0.3 mm at C2 and 14.8 ± 0.5 mm at C6, P = 0.212 and 0.946 respectively. No significant increase in prevertebral soft tissue space at C2 and C6 level demonstrated.CONCLUSION ACDF with adjunct DBM packed PEEK cages showed a statistical significant intragroup improvement in VAS neck pain scores and NDI scores(P = 0.001). There were no reported serious patient complications; post-operative radiographs demonstrated no significant difference in prevertebral space. We conclude that ACDF with DBMpacked PEEK cages can be safely done in an ASC with satisfactory outcomes.展开更多
Matrix stiffness has been demonstrated in many studies to adjust the biological behaviors of mesenchymal stem cells (MSCs). However, in the initial phase of bone restore, MSCs will encounter a hypoxic microenvironment...Matrix stiffness has been demonstrated in many studies to adjust the biological behaviors of mesenchymal stem cells (MSCs). However, in the initial phase of bone restore, MSCs will encounter a hypoxic microenvironment. Studying the connection existing between the matrix stiffness and biological behavior of MSCs under hypoxic condition can better simulate the microenvironment at the prime period of bone repairment. In this work, three-dimensional (3D) decalcified bone scaffolds with diverse stiffness (high stiffness (66.06 ± 27.83) MPa, medium stiffness (26.90 ± 13.16) MPa, and low stiffness (0.67 ± 0.14) MPa) but same microstructure have been prepared by controlling decalcification time. In addition, the decellularized bone scaffold was regard as control group and its stiffness was (230.93 ± 72.65) MPa. The viability, proliferation, infiltration, and osteogenic differentiation of MSCs seeded into these 3D demineralized bone scaffolds were systematically investigated under 100 μM CoCl2-simulated hypoxic and normoxic environments. The results showed that the viability, proliferation, and extracellular matrix (ECM) secretion of MSCs had no significant difference on scaffolds with diverse stiffness but the degree of collagen deposition of MSCs gradually increased with the increase of scaffold stiffness both under normoxia and hypoxia. Compared to normoxia, the viability, proliferation, ECM secretion, vascular endothelial growth factor (VEGF) expression, and osteogenis of MSCs on the scaffolds with the same stiffness were evidently inhibited by hypoxia. Additionally, under hypoxic condition, the expression of VEGF and hypoxia inducible factor 1α(HIF-1α) in MSCs on the low stiffness scaffold was markedly increased comparing to those on other groups. In summary, we found that the low stiffness scaffold can improved the proliferation and osteoginic differentiation of MSCs under hypoxic environment, which may help to explore efficient methods for bone defect repairing.展开更多
Demineralized bone matrix(DBM)has been widely used clinically for dental,craniofacial and skeletal bone repair,as an osteoinductive and osteoconductive material.3D printing(3DP)enables the creation of bone tissue engi...Demineralized bone matrix(DBM)has been widely used clinically for dental,craniofacial and skeletal bone repair,as an osteoinductive and osteoconductive material.3D printing(3DP)enables the creation of bone tissue engineering scaffolds with complex geometries and porosity.Photoreactive methacryloylated gelatin nanoparticles(GNP-MAs)3DP inks have been developed,which display gel-like behavior for high print fidelity and are capable of post-printing photocrosslinking for control of scaffold swelling and degradation.Here,novel DBM nanoparticles(DBM-NPs,∼400 nm)were fabricated and characterized prior to incorporation in 3DP inks.The objectives of this study were to determine how these DBM-NPs would influence the printability of composite colloidal 3DP inks,assess the impact of ultraviolet(UV)crosslinking on 3DP scaffold swelling and degradation and evaluate the osteogenic potential of DBM-NP-containing composite colloidal scaffolds.The addition of methacryloylated DBM-NPs(DBM-NP-MAs)to composite colloidal inks(100:0,95:5 and 75:25 GNP-MA:DBM-NP-MA)did not significantly impact the rheological properties associated with printability,such as viscosity and shear recovery or photocrosslinking.UV crosslinking with a UV dosage of 3 J/cm2 directly impacted the rate of 3DP scaffold swelling for all GNP-MA:DBM-NP-MA ratios with an∼40%greater increase in scaffold area and pore area in uncrosslinked versus photocrosslinked scaffolds over 21 days in phosphate-buffered saline(PBS).Likewise,degradation(hydrolytic and enzymatic)over 21 days for all DBM-NP-MA content groups was significantly decreased,∼45%less in PBS and collagenase-containing PBS,in UV-crosslinked versus uncrosslinked groups.The incorporation of DBM-NP-MAs into scaffolds decreased mass loss compared to GNP-MA-only scaffolds during collagenase degradation.An in vitro osteogenic study with bone marrow-derived mesenchymal stem cells demonstrated osteoconductive properties of 3DP scaffolds for the DBM-NP-MA contents examined.The creation of photoreactive DBM-NP-MAs and their application in 3DP provide a platform for the development of ECM-derived colloidal materials and tailored control of biochemical cue presentation with broad tissue engineering applications.展开更多
Bone protein extract is regarded as the new generation of demineralized bone matrix. The aim of this paper is to describe and characterize the properties of demineralized bone matrix and its new generation product in ...Bone protein extract is regarded as the new generation of demineralized bone matrix. The aim of this paper is to describe and characterize the properties of demineralized bone matrix and its new generation product in addition to its application in animal and human studies. Bone protein extract has features of osteoconductivity, osteoinductivity and osteogenicity, which originate from its unique and precise processing. It has exhibited powerful bone formation capacity both in animal experiments and in clinical trials by providing an optimal microenvironment for osteogenesis. Furthermore, not only does it have excellent bio- compatibility, it also has good compatibility with other implant materials, helping it bridge the host and implanted materials. Bone protein extract could be a promising alternative for demineralized bone matrix as a bone graft substitute.展开更多
背景:脊柱融合术中选择有效的植骨材料十分有必要,自体骨移植被视为脊柱融合术的“金标准”,但其存在取骨区疼痛等并发症。同种异体骨供骨量充足、可避免取骨区并发症,不同的同种异体骨材料在不同部位的脊柱融合中展现了良好的成骨能力...背景:脊柱融合术中选择有效的植骨材料十分有必要,自体骨移植被视为脊柱融合术的“金标准”,但其存在取骨区疼痛等并发症。同种异体骨供骨量充足、可避免取骨区并发症,不同的同种异体骨材料在不同部位的脊柱融合中展现了良好的成骨能力。目的:综述近年来同种异体骨移植材料在脊柱融合术中的应用现状。方法:检索1990-2022年间,在PubMed、Web of Science数据库和中国知网发表的相关中英文文献,英文检索词:“fresh frozen bone,autograft,allograft,freeze-dried bone,demineralized bone matrix,allograft spacer,spinal fusion”;中文检索词:“新鲜冰冻骨,自体骨,同种异体骨,冷冻干燥骨,脱矿骨基质,同种异体骨椎间融合器,脊柱融合”,经筛选后最终纳入95篇文献进行综述。结果与结论:新鲜冰冻骨由于其存在免疫排斥、传播疾病等风险临床运用较少;冷冻干燥骨移植虽然存在延迟愈合,但是其最终融合效果与自体骨相似;脱钙骨基质常作为自体骨填充剂或用于填充椎间融合器而最大发挥其融合能力,与硫酸钙或羟磷灰石等载体的联用使其可塑性和成骨活性提升明显;新型解剖型同种异体椎间融合器成骨活性好、临床融合率高、可有效维持椎间隙高度和椎间孔的正常形状。现有的大量临床研究结果表明同种异体骨材料在脊柱融合术中能达到与自体骨一样的融合效率,并且和自体骨一样安全,是外科医生良好的选择,在临床中得到了大量应用。展开更多
目的:建立骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)、脱钙骨基质和切断跟腱诱导的异位骨化动物模型,并初步探讨其形成机制,为异位骨化的研究奠定实验基础。方法:制备动物模型:(1)构建BMP4重组腺病毒,实验组裸鼠的一侧腓肠...目的:建立骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)、脱钙骨基质和切断跟腱诱导的异位骨化动物模型,并初步探讨其形成机制,为异位骨化的研究奠定实验基础。方法:制备动物模型:(1)构建BMP4重组腺病毒,实验组裸鼠的一侧腓肠肌内注入50μl1×107pfu BMP4重组腺病毒液,对照组腓肠肌内注入50μl1×107pfu空病毒液。4周后行X线和组织学检查。(2)无菌条件下,股后外侧入路,将50mg脱钙骨基质植入裸鼠股后肌群内,4周后行X线和组织学检查。(3)20只小鼠于跟腱中点行跟腱切断术,10周后行X线和组织学检查。结果:X线和组织学检查显示4周后,注射BMP4重组腺病毒的动物均出现异位骨,注射空病毒组未见异位骨形成。4周后,植入脱钙骨基质的动物均出现异位骨。10周后,行跟腱切断术的动物均在跟腱部位出现异位骨。结论:BMP4、脱钙骨基质和切断跟腱可有效诱导异位骨化,结果稳定可靠。展开更多
目的:制备犬脱钙骨基质(demineralized bone matrix,DBM),研究DBM作为骨组织工程支架材料的可行性。方法:杂种犬的股骨行脱脂、脱钙、脱非胶原蛋白、冷冻干燥、灭菌制成DBM,与骨髓间充质干细胞(mesenchymal stemcells,MSCs)复合,应用倒...目的:制备犬脱钙骨基质(demineralized bone matrix,DBM),研究DBM作为骨组织工程支架材料的可行性。方法:杂种犬的股骨行脱脂、脱钙、脱非胶原蛋白、冷冻干燥、灭菌制成DBM,与骨髓间充质干细胞(mesenchymal stemcells,MSCs)复合,应用倒置相差显微镜、扫描电镜观察脱钙骨基质的结构及细胞在材料表面的生长情况。结果:犬脱钙骨基质具有三维立体网孔结构,平均孔隙直径为(254.39±88.71)μm,孔隙率约为70%。与MSCs复合培养后,细胞黏附其上,上架率高,生长状态良好,增殖迅速,并分泌大量细胞外基质。结论:DBM具有良好的生物相容性。展开更多
目的研究经多聚左旋赖氨酸修饰的山羊脱钙骨基质(demineralized bone matrix decorated with poly-L-lysine,PLL-DBM)的性能,为选择性细胞滞留技术(selective cell retention,SCR)提供一种对骨髓干细胞黏附性能良好的富集基质材料。方...目的研究经多聚左旋赖氨酸修饰的山羊脱钙骨基质(demineralized bone matrix decorated with poly-L-lysine,PLL-DBM)的性能,为选择性细胞滞留技术(selective cell retention,SCR)提供一种对骨髓干细胞黏附性能良好的富集基质材料。方法将取材于山羊股骨近端的松质骨去皮质理化处理制成脱钙骨基质(demineralized bonem atrix,DBM),用多聚左旋赖氨酸(poly-L-lysine,PLL)对其进行修饰。扫描电镜分析其显微结构;氨基酸成分分析检测PLL与DBM的复合情况;通过纤维母细胞集落形成单位(CFU-F)计数,检测山羊PLL-DBM对骨髓干细胞的富集效果;在山羊横突间融合模型中,通过X线片及组织学评价其成骨能力。结果PLL-DBM具有天然网状孔隙结构系统,空隙内修饰有PLL形成的蜘蛛网样结构;PLL与DBM能较好地复合;山羊PLL-DBM对骨髓干细胞富集效果明显优于空白DBM(P<0.01)。PLL-DBM的成骨能力与自体髂骨相当。结论山羊PLL-DBM对骨髓干细胞有较好的富集效果,成骨能力强,是一种理想的富集基质材料。展开更多
目的 探索表面有纳米级沟槽的冻干脱钙骨基质（freeze-dried demineralized bone matrix with nanoscale topography，nFDBM）的制备方法，并探讨其构建组织工程骨的可行性。方法取犬趾皮质骨按改良Urist法制备FDBM，经掺钕钇铝石榴石...目的 探索表面有纳米级沟槽的冻干脱钙骨基质（freeze-dried demineralized bone matrix with nanoscale topography，nFDBM）的制备方法，并探讨其构建组织工程骨的可行性。方法取犬趾皮质骨按改良Urist法制备FDBM，经掺钕钇铝石榴石（Nd：YAG）激光处理后，与犬自体骨髓间充质干细胞诱导分化来的成骨细胞体外构建nFDBM-细胞复合物，原子力显微镜及扫描电镜观察材料表面不同形态特征对细胞行为的影响;同时将片状nFDBM复合物植入右侧犬背深筋膜袋内（实验组A），短管状nFDBM植入右侧趾骨节段性缺损处（实验组C），以片状FDBM-成骨细胞复合物（对照组B）及短管状FDBM（对照组D）植入同一犬体内左侧相对应位置作为对照，术后第4、8、12周行X线片、组织学及扫描电镜观察。结果FDBM经Nd：YAG激光处理后表面形成了规则的纳米级沟槽状三维结构（深150nm，宽600-800nm）。成骨细胞在nFDBM上的黏附密度及分泌基质量均高于FDBM。实验组复合物植入体内12周时，HE染色、扫描电镜发现材料纳米沟槽一侧、材料内有新生骨组织，X线片检查呈部分钙化，而对照组几无成骨表现，其中X线片表现评分差异有统计学意义（P〈0．05）。结论Nd：YAG激光可在FDBM表面形成纳米沟槽结构，该结构有利于成骨细胞的黏附、生长和基质分泌。以组织工程方法结合纳米技术构建的nFDBM，成骨细胞复合物植入动物模型体内表现出一定的成骨能力。展开更多
文摘Spinal fusion remains the gold-standard treatment for several pathological spine conditions. Although, autologous Iliac Crest Bone Grafting is considered the goldstandard graft choice to promote spinal fusion; however, it is associated with significant donor site morbidity and a limited graft quantity. Therefore, several bone graft alternatives have been developed, to augment arthrodesis. The purpose of this review is to present the results of clinical studies concerning the use of demineralized bone matrix(DBM), alone or as a composite graft, in the spinal fusion. A critical review of the English-language literature was conducted on Pubmed, using key word "demineralized bone matrix", "DBM", "spinal fusion", and "scoliosis". Results had been restricted to clinical studies. The majority of clinical trials demonstrate satisfactory fusion rates when DBM is employed as a graft extender or a graft enhancer.Limited number of prospective randomized controlled trials(4 studies), have been performed comparing DBM to autologous iliac crest bone graft in spine fusion. The majority of the clinical trials demonstrate comparable efficacy of DBM when it used as a graft extender in combination with autograft, but there is no clinical evidence to support its use as a standalone graft material. Additionally, high level of evidence studies are required, in order to optimize and clarify the indications of its use and the appropriate patient population that will benefit from DBM in spine arthrodesis.
文摘Autologous bone graft is considered as the gold standard for all indications for bone grafting procedures but the limited availability and complications in donor site resulted in seeking other options like allografts andbone graft substitutes. Demineralized bone matrix(DBM) is an allograft product with no quantity limitation. It is an osteoconductive material with osteoinductive capabilities, which vary among different products, depending on donor characteristics and differences in processing of the bone. The purpose of the present review is to provide a critical review of the existing literature concerning the use of DBM products in various procedures in the extremities. Clinical studies describing the use of DBM alone or in combination with other grafting material are available for only a few commercial products. The Level of Evidence of these studies and the resulting Grades of Recommendation are very low. In conclusion, further clinical studies of higher quality are required in order to improve the Recommendation Grades for or against the use of DBM products in bone grafting procedures.
文摘AIM To assess use of demineralized bone matrix(DBM) use in anterior cervical discectomy and fusion(ACDF) in outpatient setting.METHODS One hundred and forty-five patients with prospectively collected data undergoing single and two level ACDF with DBM packed within and anterior to polyetheretherketone(PEEK) cages. Two groups created, Group 1(75) outpatients and control Group 2(70) hospital patients. Prevertebral soft tissue swelling(PVSTS) was measured anterior to C2 and C6 on plain lateral cervical radiographs preoperatively and one week postoperatively and fusion assessed at two years. RESULTS There was no intergroup significance between preoperative and postoperative visual analogue scales(VAS)and neck disability index(NDI) scores between Group 1 and 2. Mean preoperative PVSTS in Group 1 was 4.7 ± 0.2 mm at C2 level and 11.1 ± 0.5 at C6 level compared to Group 2 mean PVSTS of 4.5 ± 0.5 mm and 12.8 ± 0.5, P = 0.172 and 0.127 respectively. There was no radiographic or clinical evidence of adverse reaction noted. In Group 1 mean postoperative PVSTS was 5.5 ± 0.4 mm at C2 and 14.9 ± 0.6 mm at C6 compared Group 2 mean PVSTS was 4.9 ± 0.3 mm at C2 and 14.8 ± 0.5 mm at C6, P = 0.212 and 0.946 respectively. No significant increase in prevertebral soft tissue space at C2 and C6 level demonstrated.CONCLUSION ACDF with adjunct DBM packed PEEK cages showed a statistical significant intragroup improvement in VAS neck pain scores and NDI scores(P = 0.001). There were no reported serious patient complications; post-operative radiographs demonstrated no significant difference in prevertebral space. We conclude that ACDF with DBMpacked PEEK cages can be safely done in an ASC with satisfactory outcomes.
基金the National Natural Science Foundation of China (Grants 11702043 and 11672051 and 11702044)Fundamental Research Funds for the Central Universities (Grant 2018CDQYSG0015).
文摘Matrix stiffness has been demonstrated in many studies to adjust the biological behaviors of mesenchymal stem cells (MSCs). However, in the initial phase of bone restore, MSCs will encounter a hypoxic microenvironment. Studying the connection existing between the matrix stiffness and biological behavior of MSCs under hypoxic condition can better simulate the microenvironment at the prime period of bone repairment. In this work, three-dimensional (3D) decalcified bone scaffolds with diverse stiffness (high stiffness (66.06 ± 27.83) MPa, medium stiffness (26.90 ± 13.16) MPa, and low stiffness (0.67 ± 0.14) MPa) but same microstructure have been prepared by controlling decalcification time. In addition, the decellularized bone scaffold was regard as control group and its stiffness was (230.93 ± 72.65) MPa. The viability, proliferation, infiltration, and osteogenic differentiation of MSCs seeded into these 3D demineralized bone scaffolds were systematically investigated under 100 μM CoCl2-simulated hypoxic and normoxic environments. The results showed that the viability, proliferation, and extracellular matrix (ECM) secretion of MSCs had no significant difference on scaffolds with diverse stiffness but the degree of collagen deposition of MSCs gradually increased with the increase of scaffold stiffness both under normoxia and hypoxia. Compared to normoxia, the viability, proliferation, ECM secretion, vascular endothelial growth factor (VEGF) expression, and osteogenis of MSCs on the scaffolds with the same stiffness were evidently inhibited by hypoxia. Additionally, under hypoxic condition, the expression of VEGF and hypoxia inducible factor 1α(HIF-1α) in MSCs on the low stiffness scaffold was markedly increased comparing to those on other groups. In summary, we found that the low stiffness scaffold can improved the proliferation and osteoginic differentiation of MSCs under hypoxic environment, which may help to explore efficient methods for bone defect repairing.
基金support from a National Science Foundation Graduate Research Fellowship(M.R.P.,E.J.,E.Y.J.)the National Institutes of Health(F31 DE030333,K.J.H.+3 种基金P41 EB023833,A.G.M.)the Baylor College of Medicine Medical Science Training Program(K.J.H.),the Scientific and Technological Research Council of Turkey International Research Fellowship Programme for PhD Students(H.O.)a Rubicon Postdoctoral Fellowship from the Dutch Research Council(NWOProject No.019.182 EN.004)(M.D.).
文摘Demineralized bone matrix(DBM)has been widely used clinically for dental,craniofacial and skeletal bone repair,as an osteoinductive and osteoconductive material.3D printing(3DP)enables the creation of bone tissue engineering scaffolds with complex geometries and porosity.Photoreactive methacryloylated gelatin nanoparticles(GNP-MAs)3DP inks have been developed,which display gel-like behavior for high print fidelity and are capable of post-printing photocrosslinking for control of scaffold swelling and degradation.Here,novel DBM nanoparticles(DBM-NPs,∼400 nm)were fabricated and characterized prior to incorporation in 3DP inks.The objectives of this study were to determine how these DBM-NPs would influence the printability of composite colloidal 3DP inks,assess the impact of ultraviolet(UV)crosslinking on 3DP scaffold swelling and degradation and evaluate the osteogenic potential of DBM-NP-containing composite colloidal scaffolds.The addition of methacryloylated DBM-NPs(DBM-NP-MAs)to composite colloidal inks(100:0,95:5 and 75:25 GNP-MA:DBM-NP-MA)did not significantly impact the rheological properties associated with printability,such as viscosity and shear recovery or photocrosslinking.UV crosslinking with a UV dosage of 3 J/cm2 directly impacted the rate of 3DP scaffold swelling for all GNP-MA:DBM-NP-MA ratios with an∼40%greater increase in scaffold area and pore area in uncrosslinked versus photocrosslinked scaffolds over 21 days in phosphate-buffered saline(PBS).Likewise,degradation(hydrolytic and enzymatic)over 21 days for all DBM-NP-MA content groups was significantly decreased,∼45%less in PBS and collagenase-containing PBS,in UV-crosslinked versus uncrosslinked groups.The incorporation of DBM-NP-MAs into scaffolds decreased mass loss compared to GNP-MA-only scaffolds during collagenase degradation.An in vitro osteogenic study with bone marrow-derived mesenchymal stem cells demonstrated osteoconductive properties of 3DP scaffolds for the DBM-NP-MA contents examined.The creation of photoreactive DBM-NP-MAs and their application in 3DP provide a platform for the development of ECM-derived colloidal materials and tailored control of biochemical cue presentation with broad tissue engineering applications.
基金supported by the National Basic Research Program of China (Grant No. 2012CB619105)the National Natural Science Foundation of China (Grant Nos. 30571892, 81071512 and u0732001)the Fabrikant Mads Clausens Foundation of Denmark
文摘Bone protein extract is regarded as the new generation of demineralized bone matrix. The aim of this paper is to describe and characterize the properties of demineralized bone matrix and its new generation product in addition to its application in animal and human studies. Bone protein extract has features of osteoconductivity, osteoinductivity and osteogenicity, which originate from its unique and precise processing. It has exhibited powerful bone formation capacity both in animal experiments and in clinical trials by providing an optimal microenvironment for osteogenesis. Furthermore, not only does it have excellent bio- compatibility, it also has good compatibility with other implant materials, helping it bridge the host and implanted materials. Bone protein extract could be a promising alternative for demineralized bone matrix as a bone graft substitute.
文摘背景:脊柱融合术中选择有效的植骨材料十分有必要,自体骨移植被视为脊柱融合术的“金标准”,但其存在取骨区疼痛等并发症。同种异体骨供骨量充足、可避免取骨区并发症,不同的同种异体骨材料在不同部位的脊柱融合中展现了良好的成骨能力。目的:综述近年来同种异体骨移植材料在脊柱融合术中的应用现状。方法:检索1990-2022年间,在PubMed、Web of Science数据库和中国知网发表的相关中英文文献,英文检索词:“fresh frozen bone,autograft,allograft,freeze-dried bone,demineralized bone matrix,allograft spacer,spinal fusion”;中文检索词:“新鲜冰冻骨,自体骨,同种异体骨,冷冻干燥骨,脱矿骨基质,同种异体骨椎间融合器,脊柱融合”,经筛选后最终纳入95篇文献进行综述。结果与结论:新鲜冰冻骨由于其存在免疫排斥、传播疾病等风险临床运用较少;冷冻干燥骨移植虽然存在延迟愈合,但是其最终融合效果与自体骨相似;脱钙骨基质常作为自体骨填充剂或用于填充椎间融合器而最大发挥其融合能力,与硫酸钙或羟磷灰石等载体的联用使其可塑性和成骨活性提升明显;新型解剖型同种异体椎间融合器成骨活性好、临床融合率高、可有效维持椎间隙高度和椎间孔的正常形状。现有的大量临床研究结果表明同种异体骨材料在脊柱融合术中能达到与自体骨一样的融合效率,并且和自体骨一样安全,是外科医生良好的选择,在临床中得到了大量应用。
文摘目的:建立骨形态发生蛋白4(bone morphogenetic protein 4,BMP4)、脱钙骨基质和切断跟腱诱导的异位骨化动物模型,并初步探讨其形成机制,为异位骨化的研究奠定实验基础。方法:制备动物模型:(1)构建BMP4重组腺病毒,实验组裸鼠的一侧腓肠肌内注入50μl1×107pfu BMP4重组腺病毒液,对照组腓肠肌内注入50μl1×107pfu空病毒液。4周后行X线和组织学检查。(2)无菌条件下,股后外侧入路,将50mg脱钙骨基质植入裸鼠股后肌群内,4周后行X线和组织学检查。(3)20只小鼠于跟腱中点行跟腱切断术,10周后行X线和组织学检查。结果:X线和组织学检查显示4周后,注射BMP4重组腺病毒的动物均出现异位骨,注射空病毒组未见异位骨形成。4周后,植入脱钙骨基质的动物均出现异位骨。10周后,行跟腱切断术的动物均在跟腱部位出现异位骨。结论:BMP4、脱钙骨基质和切断跟腱可有效诱导异位骨化,结果稳定可靠。
文摘目的:制备犬脱钙骨基质(demineralized bone matrix,DBM),研究DBM作为骨组织工程支架材料的可行性。方法:杂种犬的股骨行脱脂、脱钙、脱非胶原蛋白、冷冻干燥、灭菌制成DBM,与骨髓间充质干细胞(mesenchymal stemcells,MSCs)复合,应用倒置相差显微镜、扫描电镜观察脱钙骨基质的结构及细胞在材料表面的生长情况。结果:犬脱钙骨基质具有三维立体网孔结构,平均孔隙直径为(254.39±88.71)μm,孔隙率约为70%。与MSCs复合培养后,细胞黏附其上,上架率高,生长状态良好,增殖迅速,并分泌大量细胞外基质。结论:DBM具有良好的生物相容性。
文摘目的研究经多聚左旋赖氨酸修饰的山羊脱钙骨基质(demineralized bone matrix decorated with poly-L-lysine,PLL-DBM)的性能,为选择性细胞滞留技术(selective cell retention,SCR)提供一种对骨髓干细胞黏附性能良好的富集基质材料。方法将取材于山羊股骨近端的松质骨去皮质理化处理制成脱钙骨基质(demineralized bonem atrix,DBM),用多聚左旋赖氨酸(poly-L-lysine,PLL)对其进行修饰。扫描电镜分析其显微结构;氨基酸成分分析检测PLL与DBM的复合情况;通过纤维母细胞集落形成单位(CFU-F)计数,检测山羊PLL-DBM对骨髓干细胞的富集效果;在山羊横突间融合模型中,通过X线片及组织学评价其成骨能力。结果PLL-DBM具有天然网状孔隙结构系统,空隙内修饰有PLL形成的蜘蛛网样结构;PLL与DBM能较好地复合;山羊PLL-DBM对骨髓干细胞富集效果明显优于空白DBM(P<0.01)。PLL-DBM的成骨能力与自体髂骨相当。结论山羊PLL-DBM对骨髓干细胞有较好的富集效果,成骨能力强,是一种理想的富集基质材料。
文摘目的 探索表面有纳米级沟槽的冻干脱钙骨基质（freeze-dried demineralized bone matrix with nanoscale topography，nFDBM）的制备方法，并探讨其构建组织工程骨的可行性。方法取犬趾皮质骨按改良Urist法制备FDBM，经掺钕钇铝石榴石（Nd：YAG）激光处理后，与犬自体骨髓间充质干细胞诱导分化来的成骨细胞体外构建nFDBM-细胞复合物，原子力显微镜及扫描电镜观察材料表面不同形态特征对细胞行为的影响;同时将片状nFDBM复合物植入右侧犬背深筋膜袋内（实验组A），短管状nFDBM植入右侧趾骨节段性缺损处（实验组C），以片状FDBM-成骨细胞复合物（对照组B）及短管状FDBM（对照组D）植入同一犬体内左侧相对应位置作为对照，术后第4、8、12周行X线片、组织学及扫描电镜观察。结果FDBM经Nd：YAG激光处理后表面形成了规则的纳米级沟槽状三维结构（深150nm，宽600-800nm）。成骨细胞在nFDBM上的黏附密度及分泌基质量均高于FDBM。实验组复合物植入体内12周时，HE染色、扫描电镜发现材料纳米沟槽一侧、材料内有新生骨组织，X线片检查呈部分钙化，而对照组几无成骨表现，其中X线片表现评分差异有统计学意义（P〈0．05）。结论Nd：YAG激光可在FDBM表面形成纳米沟槽结构，该结构有利于成骨细胞的黏附、生长和基质分泌。以组织工程方法结合纳米技术构建的nFDBM，成骨细胞复合物植入动物模型体内表现出一定的成骨能力。
