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首页> 外文期刊>The Laryngoscope: A Medical Journal for Clinical and Research Contributions in Otolaryngology, Head and Neck Medicine and Surgery, Facial Plastic and Reconstructive Surgery .. >Validating the subcutaneous model of injectable autologous cartilage using a fibrin glue scaffold.
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Validating the subcutaneous model of injectable autologous cartilage using a fibrin glue scaffold.

机译：使用纤维蛋白胶支架验证可注射自体软骨的皮下模型。

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PURPOSE: To create and validate an injectable model for autologous in vivo cartilage engineering with ultimate clinical applicability in human subjects. HYPOTHESIS: Cartilage can be generated subcutaneously using fibrin glue and autologous chondrocyte components. BACKGROUND: To date, cartilage engineering studies have been limited by several factors. Immunocompromised animals and nonautologous chondrocytes have been successfully used to create cartilage, but results using identical designs failed in immunocompetent subjects. Recent studies using more biocompatible tissues and matrices have been performed with both in vitro and in vivo steps. Although successful, several problems are notable. In vitro cartilage displays a poor modulus of elasticity, even after in vivo implantation. Variable deformation and volume loss occurs when in vitro specimens are matured in vivo. These concerns limit the clinical utility of these methods. We therefore set out to create autologous cartilage using a model that was clinically feasible, easy to create, and could be performed with very low patient harvest morbidity. MATERIALS AND METHODS: Eight New Zealand white rabbits underwent a unilateral harvest of ear cartilage. Samples were then digested using standard methods. Cell counts and survival assays were performed before implantation. One sample of fibrin glue (Tisseel) and chondrocytes was injected subcutaneously into each donor rabbit and then left in situ for 3 months. A second sample with both basic fibroblast growth factor (b-FGF) and insulin-like growth factor (IGF)-1 in the injection suspension was also assessed (for a total of 16 samples). After harvest, analysis of overall volume, histology, and chondrocyte drop out counts was performed. RESULTS: Cartilage formation occurred in 8 of 14 (57%) specimens that were obtained at the time of sacrifice. Of note, 6 of 7 (85%) non-growth-factor containing samples yielded positive results. Comparison with the success rate using concomitant growth factors (2/7) showed a negative effect on cartilage yield (P = .015). Chondrocyte survival, based on chondrocyte dropout counts, was not effected. Angiogenesis appeared to correlate with cartilage formation in the central regions of the implant. Alcian blue demonstrated the presence of active matrix deposition, and elastin Verhoff-van Geison (EVG) stains were positive, showing an elastic cartilage phenotype. Very limited osteoid formation was seen in successful implants. Failed implants demonstrated avascular necrosis, giant cell reactions, and inflammatory infiltrates. CONCLUSIONS: This study validates the subcutaneous site as a recipient bed for the engineering of autologous cartilage in vivo. It also represents the first subcutaneous implantation of fibrin glue and chondrocytes in an immunocompetent host as well as the first published report of elastic cartilage generation in vivo. Although the model needs to be further streamlined to increase yields and overall volume, this study clearly demonstrates the feasibility of in vivo chondrogenesis (85% success). The addition of FGF and IGF-1 at the concentrations used negatively influenced cartilage yield. However, extrapolation of these results to other combinations or concentrations can not be done, and this issue deserves further investigation.
机译：目的：创建和验证可注射模型用于自体体内软骨工程，在人类受试者中具有最终的临床适用性。假设：使用纤维蛋白胶和自体软骨细胞成分可在皮下产生软骨。背景：迄今为止，软骨工程研究受到几个因素的限制。免疫功能低下的动物和非自体软骨细胞已成功用于制造软骨，但在免疫功能正常的受试者中使用相同设计的结果却失败了。最近已经在体外和体内步骤中进行了使用更多生物相容性组织和基质的研究。尽管成功，但仍存在一些问题。即使在体内植入后，体外软骨也显示出较差的弹性模量。当体外标本在体内成熟时，会发生各种变形和体积损失。这些问题限制了这些方法的临床实用性。因此，我们着手使用临床上可行，易于创建并且可以在患者收成率非常低的情况下进行的模型创建自体软骨。材料与方法：八只新西兰白兔进行了单侧耳朵软骨收获。然后使用标准方法消化样品。在植入前进行细胞计数和存活测定。将一份纤维蛋白胶（Tisseel）和软骨细胞样品皮下注射到每只供体兔中，然后原位放置3个月。还评估了注射悬浮液中同时具有碱性成纤维细胞生长因子（b-FGF）和胰岛素样生长因子（IGF）-1的第二个样品（总共16个样品）。收获后，进行总体积，组织学和软骨细胞脱落计数的分析。结果：在处死时获得的14个样本中有8个（57％）发生了软骨形成。值得注意的是，在7个非生长因子样本中，有6个（85％）产生了阳性结果。使用伴随的生长因子（2/7）与成功率进行比较显示出对软骨产量的负面影响（P = .015）。基于软骨细胞脱落计数的软骨细胞存活未受影响。血管生成似乎与植入物中心区域的软骨形成相关。阿尔辛蓝表明存在活性基质沉积，弹性蛋白Verhoff-van Geison（EVG）染色呈阳性，显示出弹性软骨表型。在成功的植入物中观察到非常有限的类骨质形成。失败的植入物显示出无血管坏死，巨细胞反应和炎性浸润。结论：本研究证实皮下部位可作为体内自体软骨工程的受体床。它也代表了在具有免疫能力的宿主中首次进行的纤维蛋白胶和软骨细胞的皮下植入，以及体内弹性软骨生成的首次公开报道。尽管需要进一步简化模型以提高产量和总体积，但这项研究清楚地证明了体内软骨形成的可行性（成功率达85％）。以所使用的浓度添加FGF和IGF-1会对软骨产量产生负面影响。但是，无法将这些结果外推到其他组合或浓度，并且这个问题值得进一步研究。

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