Angiogenic response induced by acellular femoral matrix in vivo

We investigated the angiogenic response induced by acellular femoral matrices implanted in vivo on to the chick embryo chorioallantoic membrane (CAM), a useful model for such investigation. The results showed that acellular matrices were able to induce a strong angiogenic response, comparable with that of fibroblast growth factor-2 (FGF-2), a well-known angiogenic cytokine. The angiogenic response was further increased when exogenous FGF-2 or transforming growth factor beta-1 (TGF-beta1) was added to the matrices and inhibited by the addition of anti-FGF-2 or anti-TGF-beta1 antibodies. The response may be considered to be dependent on a direct angiogenic effect exerted by the matrices, and also in part by the presence of FGF-2 and TGF-beta1 in the acellular matrices.     

即刻种植中异体脱细胞真皮基质引导的骨组织再生*

背景：脱细胞真皮基质可作为软组织支架使成纤维细胞和上皮细胞在其中增殖,并最终成为宿主组织的一部分。 目的：观察异体脱细胞真皮基质引导即刻植入种植体周围骨组织再生的效果。 方法：选择口腔修复行即刻种植患者15例,使用异体脱细胞真皮基质行骨组织引导再生,种植体植入后,测量术中、植入后6,12个月唇侧骨壁厚度,同时观察种植体周围软组织愈合情况。 结果与结论：6个月后均完成最终修复,无感染及明显并发症。异体脱细胞真皮基质有效关闭了即刻种植位点的软组织创口,6个月内逐渐与种植体周软组织发生整合。植入后6,12个月种植位点唇侧骨壁厚度与唇侧骨量高于术中测量值(P < 0.05),植入后6,12个月组间骨壁厚度及骨量变化不明显(P > 0.05)。说明异体脱细胞真皮基质作为屏障膜能有效引导骨组织再生,降低即刻种植术中关闭创口的难度,短期临床结果满意。关键词：异体脱细胞真皮基质;牙种植体;即刻种植;引导骨组织再生技术;组织膜 缩略语注释：GBR：guided bone regeneration,骨组织再生技术;ADM：acellular dermal matrix,异体脱细胞真皮基质 doi:10.3969/j.issn.1673-8225.2012.16.002     

灌注法制备全肾脏脱细胞基质支架的体内生物相容性

背景：应用灌注法制备的大鼠全肾脏脱细胞基质支架具有良好的体外细胞相容性,但其体内生物相容性尚不明确。 目的：应用灌注法制备大鼠全肾脏脱细胞基质支架,检测其体内生物相容性。 方法：应用灌注法制备Wistar大鼠全肾脏脱细胞基质支架,进行以下实验：①急性毒性实验：在小鼠腹腔分别注射全肾脏脱细胞基质支架浸提液、生理盐水及苯酚。②溶血实验：将抗凝新西兰兔血分别与全肾脏脱细胞基质支架浸提液、生理盐水及蒸馏水混合。③热源实验：向新西兰兔耳缘静脉注射全肾脏脱细胞基质支架浸提液。④内皮刺激实验：在新西兰兔皮下注射全肾脏脱细胞基质支架浸提液,观察有无皮肤刺激反应。⑤皮下植入实验：将全肾脏脱细胞基质支架埋入新西兰兔背部皮下。 结果与结论：全灌注法制备的Wistar大鼠全肾脏脱细胞基质支架无细胞残留,未引起全身毒性反应、急性溶血反应、热源反应及皮肤刺激反应,植入兔体内具有良好的组织相容性。说明大鼠全肾脏脱细胞基质材料在动物体内具有很好的生物相容性。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

脱细胞天然骨基质与诱导性成骨细胞的体外相容性

背景：前期工作表明TritonX-100处理的脱细胞骨基质已满足组织学和免疫学方面的修复要求。如果细胞能在材料表面很好地生长,将利于进一步进行体内动物实验。 目的：采用细胞培养法在体外评估脱细胞骨基质与诱导后成骨细胞的生物相容性。 方法：第3代骨髓基质干细胞经成骨诱导分化培养液诱导分化为成骨细胞,接种于TritonX-100处理的脱细胞骨基质及羟基磷灰石表面,检测成骨细胞的碱性磷酸酶表达并用扫描电镜观察材料表面的细胞生长情况。 结果与结论：碱性磷酸酶活性分析均表明,TritonX-100处理的脱细胞骨基质在培养48 h之后比羟基磷灰石更利于诱导成骨细胞生长;扫描电镜下可见,成骨细胞在脱细胞骨基质表面呈现立体生长方式,细胞呈球形,并且聚集成簇。体外实验结果显示成骨细胞与脱细胞天然骨基质有较好的生物相容性。     

Autologous satellite cell seeding improves in vivo biocompatibility of homologous muscle acellular matrix implants

Acellular matrix obtained from homologous muscular tissue has been previously used to repair muscular defects. However, the implants, although not rejected, give rise to an intense inflammatory response and are rapidly replaced by fibrous tissue. In this study we examined the possibility that co-culture with autologous satellite cells can improve the efficiency of homologous acellular matrix as skeletal muscle substitute. Satellite cells, isolated from rat dorsal muscle, were cultured in vitro on homologous acellular matrix obtained by detergent-enzymatic treatment of abdominal muscle fragments. Scanning electron microscopy revealed that after 24 h of co-culture satellite cells were attached to the matrix, but still possessed a round shape. After 96 h, seeded cells began to flatten and to differentiate, originating few multinucleated myotubes. Patches of homologous matrix, seeded or not with autologous satellite cells, were implanted in the dorsal muscle of rats. At autopsy, the implants were recovered and processed for light microscopy. Two weeks after surgery, fibrous tissue started to replace the grafts composed only by acellular matrix, which at the 4th week were transformed into a fibrous scar. In contrast, at both times post-surgery the structure of implants containing autologous satellite cells was well preserved. The inflammatory reaction was modest and fibrosis was confined to the periphery of the grafts. It is concluded that the presence of autologous satellite cells is an important factor to preserve the structural integrity and to improve in vivo biocompatibility of homologous muscular acellular matrix implants.     

戊二醛交联优化脱细胞真皮基质的生物相容性

目的 通过体内包埋实验将优化的脱细胞真皮基质(ADM)与成纤维细胞(FB)体外培养,探讨是否具有生物相容性. 方法 用高渗盐-NaOH消蚀法制备得到大鼠ADM,用低浓度戊二醛交联ADM,未交联A组(0min)作为对照,根据交联时间不同分成B组(10min)、C组(15min)和D组(30min).通过大鼠体内包埋实验对比各组ADM 物理性状、包埋后面积、炎性反应、浸润生长的FB和毛细血管数目,选择出最佳交联时间.制备优化的人ADM,用四甲基偶氮唑盐(MTT)比色法检测ADM的细胞毒性;人FB与ADM复合培养检测其细胞相容性,用免疫组织化学和透射电镜观察FB的生长和增殖情况;反转录聚合酶链反应(RT-PCR)检测FBⅠ、Ⅲ型前胶原mRNA的表达. 结果 大鼠体内包埋实验显示,4周取材,未交联组不易触及移植物,植入物和周围的软组织基本融合难以分开;未交联组ADM的面积较包埋前缩小;未交联组ADM内有少量炎性细胞,胶原纤维嗜酸性弱;交联组ADM内未见炎性细胞,浸润生长的FB数目、血管数目与戊二醛交联时间的单因素相关性分析呈负相关;最佳的交联时间为10min.体外培养显示,优化ADM的细胞毒性反应0～1级;人FB能够很好地在优化的ADM表面贴附、生长和增殖;ADM内FBⅠ型、Ⅲ型前胶原mRNA的表达降低. 结论 高渗盐-NaOH消蚀法制备ADM,工艺简单、成本低、生物相容性好,可以提供细胞生长的空间,并可在细胞间转导通讯和信号,下调细胞外基质代谢系统.     

Analysis of the immune response induced by a single xenoantigen in vivo

Transgenic mice expressing human major histocompatibility complex (MHC) class II molecules would provide a valuable model system for studying murine anti-human MHC immune response. We have previously shown that skin from HLA-DR1 transgenic mice was rejected by control littermates and spleen cells from rejecting mice were able to proliferate to donor cells. The aim of this paper is to analyze the mechanism of recognition of this xenoantigen and the possible involvement of antibody response in anti-HLA-DR1 immune response. Control littermates were immunized with spleen cells from HLA-DR1 transgenic (TG) mice; at indicated times, xenoantigen-specific proliferation and IFNgamma production was assessed using APC obtained from HLA-DR1 TG mice. Mixed direct-indirect pathway of xenoantigen recognition was suggested by the following findings: i)T cell response to HLA-DR1 was inhibited adding in culture monoclonal antibodies directed either to donor (HLA-DR) or to recipient MHC (I-A); ii) APC from control mice pulsed with purified DR1 molecules were able to induce proliferation by FVB/N mice immunized with transgenic spleen cells. HLA-DR1 recognition permits DR peptide-specific T cell response by lymphocytes of control littermates immunized with the xenoantigen. In addition, we detected xenoreactive IgM and IgG2 antibodies. Our data suggest that HLA-DR1 xenoantigen may be recognized through direct or indirect pathway and provide additional information on mouse anti-human HLA immune response.     

异体脱细胞真皮基质的研究进展

异体脱细胞真皮基质是天然真皮的脱细胞基质,经过冷冻干燥形成的三维支架结构。作为一种新兴的生物工程支架,它通过空间诱导和组织替代作用修复组织缺损。由于其源于人体正常组织,经临床应用证实,异体脱细胞真皮基质拥有优秀的组织相容性,现已广泛地应用于临床各科的创伤修复。     

异体脱细胞真皮基质的研究进展

异体脱细胞真皮基质是天然真皮的脱细胞基质，经过冷冻干燥形成的三维支架结构。作为一种新兴的生物工程支架，它通过空间诱导和组织替代作用修复组织缺损。由于其源于人体正常组织，经临床应用证实，异体脱细胞真皮基质拥有优秀的组织相容性，现已广泛地应用于临床各科的创伤修复。     

去细胞真皮基质的黏弹性研究

目的通过对比分析皮肤的物理参数值评价去细胞真皮基质的黏弹性性质。方法将清洁级日本大耳白兔背部皮肤进行3种不同的创面处理,分别为薄自体皮加上去细胞真皮基质移植到创面组(PADM组)、薄自体皮原位回植组(TS组)、正常皮肤真皮组(NS组)。对该3组皮片进行应力-应变关系的实验。结果从PADM、TS、NS 3组皮肤的应力松弛曲线、蠕变曲线和应力-应变关系曲线可知,在一定应力下,PADM组的应变最小,NS组的应变最大,TS组居于两者之间。结论皮肤的黏弹性力学模型是4参量黏弹性模型。用去细胞真皮基质作为皮肤替代物移植到皮肤创面后,该植皮的弹性较差,变形后的恢复能力也较差,与临床结果相符合。     

异种脱细胞真皮基质在难愈性创面修复中的应用研究

目的探讨异种脱细胞真皮基质（ADM）在难愈性创面修复中的应用价值。方法选取新疆维吾尔自治区人民医院烧伤、创面修复外科2005年6月至2011年6月收住的各种难愈性创面患者84例,按随机数字表法分为试验组41例和对照组43例。两组常规清创后,试验组创面使用ADM覆盖,对照组常规应用生理盐水、凡士林纱布覆盖,每例患者选取4cm×4cm大小创面作为观察区域,比较两组治疗后1d、3d、7d、12d肉芽组织生长情况及临床治疗时间。结果异种脱细胞真皮基质移植术后第5天两组创面肉芽组织生长开始有区别,治疗后7d,试验组肉芽创面（24.87±0.24）%与B组（18.66±0.45）%相比差异有统计学意义（P〈0.01）;试验组创面愈合时间为（16.17±1.87）d,较B组（32.71±3.95）d明显缩短,差异有高度统计学意义（P〈0.01）。结论异种脱细胞真皮基质作为创面覆盖物,能有效地保护创面,促进创面血管化形成,缩短临床治疗周期。     

Host response to human acellular dermal matrix transplantation in a primate model of abdominal wall repair

Commercially available human acellular dermal matrix (HADM), AlloDerm((R)), was implanted as an interpositional graft in the abdominal wall of adult vervet monkeys. Host response to implanted HADM was evaluated and compared with a human cellular dermal matrix (HCDM) and a primate acellular dermal matrix (PADM). Clinical acceptance of the acellular grafts (HADM and PADM) and graft remodeling were evidenced by fibroblast repopulation and neoangiogenesis. A mild inflammatory response marked predominantly by macrophages and T-cells was present in both HADM and PADM during the first month but was absent by 3 months. Similarly, antibody and complement deposition into the grafts as well as in the serum was evident only at the early time points. Interleukin-6 (IL-6) or IL-10 was induced in some acellular graft-implanted monkeys at the early time points, but tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), or IL-2 was not detected over the study period. In contrast, significant inflammation was observed in HCDM-implanted animals, as evidenced by immune cell infiltration (p 相似文献   

脱细胞基质软骨支架的制备

背景：脱细胞基质材料去除了天然材料中的细胞成分,保留了基质成分,有效降低了天然材料的免疫原性,同时能够保持材料的机械强度。 目的：拟利用洗脱方法去除兔肋软骨中的细胞基质,制备天然生物支架材料。 方法：取新西兰大白兔肋软骨,清除周围组织后随机分组处理,以未经处理的肋软骨作为正常对照组;48 h处理组以去污剂-酶化学消化48 h;96 h处理组以去污剂-酶化学消化96 h,3组均通过苏木精-伊红染色及电镜观察脱细胞效果。同时收集诱导第7天的兔骨髓间充质干细胞3×109 L-1,与同种异体肋软骨脱细胞基质体外复合培养,于第3,7天取复合物行电镜观察细胞在脱细胞基质表面的黏附生长情况。 结果与结论：新鲜肋软骨标本每个软骨陷窝内均有排列紧密的二三个软骨细胞,去污剂-酶化学消化后软骨陷窝内的细胞逐渐脱失,至消化处理96 h后,软骨陷窝内的细胞完全脱失。共培养第3天时,脱细胞基质表面有大量骨髓间充质干细胞分布,细胞为多角形,有伪足伸出,锚定在基质表面,部分区域可见细胞在基质表面增殖分裂;第7天时,脱细胞基质表面大部分均为细胞覆盖,细胞呈扁平状,有多个足突充分伸展,细胞之间互相连接,分泌大量细胞外基质沉积在基质表面,呈冰霜样改变,表明制备的脱细胞基质具有良好的细胞相容性。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

兔肋软骨脱细胞基质的制备

背景：研究表明新西兰兔软骨组织可作为组织工程支架材料,其中关节软骨及耳软骨的脱细胞基质的研究较多,但采用肋软骨作为组织工程软骨支架的研究较少。 目的：制备新西兰兔肋软骨脱细胞基质,探讨天然软骨支架作为组织工程支架的可行性。 方法：用联合去垢剂-酶法获得软骨支架,根据脱细胞过程中Triton X-100第2次处理时间0,24,48,96 h分为4组。脱细胞完毕后各组支架固定行扫描电镜采集图像观察计算支架孔隙率、孔径长度,并对支架进行苏木精-伊红染色、甲苯胺蓝及Ⅱ型胶原免疫组织化学染色,并将脱细胞支架植入异体新西兰兔皮下观察其相容性。 结果与结论：兔肋软骨脱细胞基质呈乳白色,大小均一,染色示支架结构完整,仍保存大量酸性黏多糖及Ⅱ型胶原成分,扫描电镜观察经一定时间的脱细胞处理后可得到结构完整,孔隙均匀的天然软骨支架,其孔隙率为(61.31±8.45) %;孔径长度为(32.80±5.15) μm,符合正态性分布,各组脱细胞支架植入异体新西兰兔皮下7 d后生物相容性良好,周围软组织无明显充血、化脓等炎症排斥反应出现。结果显示,兔肋软骨脱细胞支架具有良好的基质组成,有较完整、均匀的孔隙结构及孔径分布,可作为组织工程支架材料。     

异种脱细胞真皮基质在开放性乳突病变切除术中的应用

目的探讨开放性乳突病变切除术后加速乳突术腔上皮化及缩小乳突术腔的有效办法。方法63例行开放性乳突病变切除术的患者按照复查治疗方法分为两组，异种脱细胞真皮组36例，应用异种脱细胞真皮基质覆盖术腔，碘仿纱条组27例，以碘仿纱条填塞术腔。术后随访两组患者，对比观察术腔愈合情况和上皮化时间。结果异种脱细胞真皮组术腔上皮化时间为2～4周，平均2．2周；碘仿纱条组术腔上皮化时间为9～35周，平均13．7周，异种脱细胞真皮组的上皮化时间短于碘仿纱条组（P〈0．01）。结论异种脱细胞真皮基质可促进上皮组织再生，减少术后感染及肉芽发生，加速乳突腔上皮化，提高开放性乳突病变切除术的疗效。     

纳米纤维素蛋白和脱细胞基质修复口腔黏膜

 BACKGROUND: The sensitivity and mucus secretion of the oral mucosa make oral soft tissues difficult to repair, so patients cannot achieve satisfactory outcomes after treatment. Nano-cellulose protein mainly composed of glycine, alanine and serine has good histocompatibility. However, there is a lack of comparative study about the effect of nano-cellulose protein and acellular matrix in oral mucosa repair, and the clinical effects of the two materials are still under discussion.     

Regeneration of abdominal wall musculofascial defects by a human acellular collagen matrix

This work studied the reconstruction of an abdominal wall defect by a human acellular collagen matrix. The abdominal wall defect was cured in 40 rats by implanting (i) polypropylene (Pro), (ii) polyester (Mers) meshes, and (iii) human acellular collagen matrix with two orientations: fibres in parallel (fascia lata longitudinal [FLL]) or perpendicular (fascia lata transversal [FLT]) to native rats' abdominal walls. Hernia recurrence, adhesions, and histology (for inflammation and remodelling) were assessed at 4 and 8 weeks after implantation. Two large abdominal eventrations were cured by a human acellular matrix in human patients. A higher hernia recurrence rate was observed for rats transplanted with FLL than with FLT/Pro/Mers at 4 and 8 weeks after implantation. A lower adhesion rate was achieved for FLL/FLT than for Pro/Mers meshes (p<0.05). A decrease in immunologic cell infiltrations in FLL/FLT was observed between day 30 and day 60 (p<0.05). Collagen, elastin, and muscular tissues were found only in FLL/FLT matrix; a weaker muscular cell infiltration for FLL occurred at 8 weeks. Human abdominal eventrations were totally cured by using FLT as confirmed by computed tomography scanning at 12 and 16 months after implantation. In conclusion, human acellular collagen matrix, placed in an FLT position, can induce an abdominal wall reconstitution without adhesions and hernia recurrence.     

无细胞胶原基质的研究进展

天然胶原类组织,经过脱细胞处理后,降低免疫原性,可作为组织工程的支架材料,本文就脱细胞的方法以及无细胞胶原基质的研究进展作一综述。     

In vitro and in vivo autoimmune response to enamel matrix proteins.

Enamel matrix proteins taken from neonatal C57Bl/6 mice were shown to be able to elicit in vitro proliferative responses in C57Bl/6 splenic lymphocytes taken from animals which had not been exposed to exogenous enamel proteins. Animals which had been injected with enamel matrix proteins in Freund's complete adjuvant made IgG antibodies against enamel proteins which were detected by indirect immunofluorescence. Spleen cells from the immune animals gave an augmented in vitro proliferative response to enamel proteins, while spleen cells from C57Bl/6 nu/nu mice or anti-Thy 1 and complement-treated normal C57Bl/6 spleen cells were incapable of responding to enamel proteins in vitro. Thus, enamel matrix proteins appear to be T-cell dependent autoantigens. The natural history of enamel matrix proteins is reviewed, and it is suggested, based on the anatomic details of enamel synthesis, secretion, and maturation, that enamel matrix proteins are autoantigenic because they are anatomically sequestered.     

组织工程血管基质的制备与改性

目的：完全脱除猪胸主动脉细胞，对脱细胞血管基质进行改性，增强基质的力学强度，制备组织工程血管支架材料。方法：取家猪的新鲜去除外膜胸主动脉20根，随机分成4组，分别采用胰蛋白酶、TritonX-100及十二烷基硫酸钠（SDS）作为脱细胞试剂对猪胸主动脉进行脱细胞处理，并对脱细胞后的血管基质进行改性，采用HE染色、弹力纤维染色、力学性能测试，以评价脱细胞效果以及材料的力学性能。结果：采用1％TritonX-100单独作用84h既能完全脱除细胞，同时又可完整保留血管基质的三维结构，对胶原纤维、弹力纤维的损伤小，是一种较理想的脱细胞方法。对脱细胞后的基质进行冷冻干燥及真空热交联处理，能有效提高材料的机械强度，冷冻干燥24h后真空120℃下热交联处理12h所获得的材料的机械强度最好，断裂强度可达到1．70MPa。结论：以脱细胞血管基质经冷冻干燥和真空热交联处理后，可以作为血管组织工程的支架材料。