Dipartimento di Scienze biotecnologiche di base, cliniche intensivologiche e perioperatorie, Sezione di Biochimica, Università Cattolica del Sacro Cuore, Largo F. Vito, 1, 00168, Rome, Italy.
Fondazione Policlinico Universitario A. Gemelli, IRCCS, Largo A. Gemelli 1, 00168, Rome, Italy.
Odontology. 2022 Oct;110(4):710-718. doi: 10.1007/s10266-022-00705-9. Epub 2022 Mar 30.
Membrane exposure is a widely reported and relatively common complication in Guided Bone Regeneration (GBR) procedures. The introduction of micro-porous dPTFE barriers, which are impervious to bacterial cells, could reduce the technique sensitivity to membrane exposure, even if there are no studies investigating the potential passage of bacterial metabolites through the barrier. Aim of this study was the in vitro evaluation of the permeability of three different GBR membranes (dPTFE, native and cross-linked collagen membranes) to Porphyromonas gingivalis; in those cases, where bacterial penetration could not be observed, another purpose was the analysis of the viability and differentiation capability of an osteosarcoma (U2OS) cell line in presence of bacteria eluate obtained through membrane percolation. A system leading to the percolation of P. gingivalis broth culture through the experimental membranes was arranged to assess the permeability to bacteria after 24 and 72 h of incubation. The obtained solution was then added to U2OS cell cultures which underwent, after 10 days of incubation, MTT and red alizarin essays. The dPTFE membrane showed resistance to bacterial penetration, while both types of collagen membranes were crossed by P. gingivalis after 24 h. The bacteria eluate filtered through dPTFE membrane didn't show any toxicity on U2OS cells. Results of this study demonstrate that dPTFE membranes can contrast the penetration of both P. gingivalis and its metabolites toxic for osteoblast-like cells. The toxicity analysis was not possible for the collagen membranes, since permeability to bacterial cells was observed within the first period of incubation.
膜暴露是引导骨再生（GBR）过程中广泛报道的相对常见的并发症。微孔 dPTFE 屏障的引入，其对细菌细胞是不可渗透的，即使没有研究调查细菌代谢产物是否可以通过屏障，也可以降低该技术对膜暴露的敏感性。本研究的目的是体外评估三种不同 GBR 膜（dPTFE、天然和交联胶原膜）对牙龈卟啉单胞菌的通透性；在无法观察到细菌穿透的情况下，另一个目的是分析在膜渗滤获得的细菌洗脱液存在的情况下，骨肉瘤（U2OS）细胞系的活力和分化能力。建立了一个使牙龈卟啉单胞菌培养液通过实验膜渗滤的系统，以评估孵育 24 和 72 小时后细菌的通透性。然后将获得的溶液添加到 U2OS 细胞培养物中，孵育 10 天后进行 MTT 和红色茜素实验。dPTFE 膜显示出对细菌穿透的抵抗力，而两种类型的胶原膜在 24 小时后都被牙龈卟啉单胞菌穿透。dPTFE 膜过滤的细菌洗脱液对 U2OS 细胞没有任何毒性。本研究的结果表明，dPTFE 膜可以阻止牙龈卟啉单胞菌及其对成骨样细胞有毒的代谢物的穿透。由于在孵育的第一阶段观察到细菌细胞的通透性，因此无法对胶原膜进行毒性分析。
