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一种无菌和非无菌低成本和商业网片的成纤维细胞生物相容性的体外研究。

An in vitro study on the biocompatibility of fibroblasts in sterile and non-sterile low-cost and commercial meshes.

机构信息

Department of General an Visceral Surgery, Bodden-Kliniken Ribnitz-Damgarten, Ribnitz-Damgarten, Germany.

Heart und Vascular Center, Albertinen Hospital, Hamburg, Germany.

出版信息

Hernia. 2019 Dec;23(6):1163-1174. doi: 10.1007/s10029-019-01932-w. Epub 2019 Apr 4.

Abstract

INTRODUCTION

Despite several successful studies with low-cost meshes (LCM) for the treatment of inguinal hernias in India and Africa, a nationwide application has not been possible for a variety of reasons. One problem is the special preparation and sterilization of these meshes-naturally, they should comply with international standards and demands, which is often difficult to achieve in Africa. Our primary approach was to determine whether there are differences in the biocompatibility of fibroblasts between non-sterile and sterile LCMs and commercial meshes (CM).

MATERIALS AND METHODS

Two polyester CMs with different pore size and a polyester LCM were examined as both sterile and non-sterile. LCM was plasma sterilized at 60 °C and steam sterilized at 134 °C. Sterile and non-sterile meshes were soaked with an antibiotic (penicillin/streptomycin) and antimycotic solution (amphotericin B). Human fibroblasts from healthy subcutaneous tissue were used. Various tests for evaluating the growth behavior and cell morphology of human fibroblasts were conducted. Semiquantitative (light microscopy) and qualitative (scanning electron microscopy) analyses were performed after 1 week and again after 12 weeks. The metabolism of fibroblasts was checked by pH measurements and glucose analyses. Biocompatibility of fibroblasts on sterile and non-sterile meshes was carried out by luminescence methods (cell viability and apoptosis) as well as calorimetric methods for proliferation determination (BrDU assay) and cytotoxicity (LDH assay).

RESULTS

Light and electron microscopy revealed a moderate growth of fibroblasts on all investigated mesh types. The results of glycolysis and the pH value were within the normal range for all sterile and non-sterile meshes. In biocompatibility studies, no elevated level of apoptosis was detected. The viability measurement of mitochondrial activity of fibroblasts showed a 50% inhibition of mitochondria in all nets, with the exception of non-sterile CM, whereas mitochondrial activity was increased in the non-sterile CM. A proliferation measurement (BrdU test) revealed different growth inhibition in the sterile and non-sterile meshes. This growth inhibition was significantly stronger, particularly for non-sterile CM light meshes, than it was for the non-sterile LCM.

CONCLUSION

Again, our studies show no significant differences in biocompatibility of fibroblasts between expensive and low-cost meshes. In addition, we detected fibroblast growth even in sterile meshes, independent of the mesh group. To our knowledge, the present study is the first of its kind in terms of qualitative equivalence of sterile and non-sterile in vitro mesh samples. We do not wish to create future patient studies with non-sterilized meshes saturated with antibiotics/antimycotics. However, perhaps we can prove in future studies that under semi-sterile conditions with certain LCMs, wound infection rates can be acceptable.

摘要

简介

尽管在印度和非洲有几项使用低成本网片(LCM)治疗腹股沟疝的成功研究,但由于多种原因,尚未在全国范围内推广应用。其中一个问题是这些网片的特殊准备和消毒——它们自然应该符合国际标准和要求,但这在非洲往往难以实现。我们的主要方法是确定非无菌和无菌 LCM 与商业网(CM)之间成纤维细胞的生物相容性是否存在差异。

材料和方法

检查了两种不同孔径的聚酯 CM 和一种聚酯 LCM,既作为无菌也作为非无菌。LCM 在 60°C 下进行等离子体消毒,在 134°C 下进行蒸汽消毒。无菌和非无菌网片用抗生素(青霉素/链霉素)和抗真菌溶液(两性霉素 B)浸泡。使用来自健康皮下组织的人成纤维细胞。进行了各种测试以评估人成纤维细胞的生长行为和细胞形态。在第 1 周和第 12 周后进行半定量(相差显微镜)和定性(扫描电子显微镜)分析。通过 pH 值测量和葡萄糖分析检查成纤维细胞的代谢。通过发光法(细胞活力和细胞凋亡)以及细胞增殖测定的量热法(BrDU 试验)和细胞毒性测定(LDH 试验)来评估成纤维细胞在无菌和非无菌网片上的生物相容性。

结果

光镜和电镜显示所有研究的网片类型上成纤维细胞的生长均适中。所有无菌和非无菌网的糖酵解和 pH 值结果均在正常范围内。在生物相容性研究中,未检测到凋亡水平升高。成纤维细胞线粒体活性的活力测量显示,所有网片中的线粒体有 50%受到抑制,非无菌 CM 除外,而非无菌 CM 中的线粒体活性增加。增殖测量(BrdU 试验)显示在无菌和非无菌网片中存在不同的生长抑制。这种生长抑制在无菌和非无菌 CM 光网片中更为明显,尤其是在非无菌 CM 光网片中,比在无菌 LCM 中更为明显。

结论

再次,我们的研究表明,昂贵和低成本网之间的成纤维细胞生物相容性没有显著差异。此外,我们甚至在无菌网片中检测到成纤维细胞的生长,而与网片类型无关。据我们所知,目前的研究是首次对无菌和非无菌体外网片样本进行定性等效性研究。我们不希望在未来的患者研究中使用非无菌且用抗生素/抗真菌药物饱和的网片。然而,也许我们可以在未来的研究中证明,在某些 LCM 的半无菌条件下,伤口感染率是可以接受的。

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