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氨基乙酰丙酸光动力疗法增强了用于牙周病学的猪脱细胞真皮基质膜上培养细胞的细胞活性。

Photodynamic Therapy with Aminolevulinic Acid Enhances the Cellular Activity of Cells Cultured on Porcine Acellular Dermal Matrix Membranes Used in Periodontology.

作者信息

Petrini Morena, D'Amico Emira, Pierfelice Tania Vanessa, Aceto Gitana Maria, Karaban Maryia, Felice Pietro, Piattelli Adriano, Barone Antonio, Iezzi Giovanna

机构信息

Department of Medical, Oral and Biotechnological Sciences, University G. d'Annunzio of Chieti-Pescara, 66100 Chieti, Italy.

Department of Biomedical and Neuromotor Sciences, University of Bologna, 40126 Bologna, Italy.

出版信息

Gels. 2023 Jul 20;9(7):584. doi: 10.3390/gels9070584.

Abstract

This study aims to test a photodynamic protocol based on a gel containing aminolevulinic acid followed by red-LED (ALAD-PDT) irradiation on human gingival fibroblasts (hGFs) and osteoblasts (hOBs) cultured on a porcine acellular dermal matrix membrane (PADMM). In the previous literature, ALAD-PDT showed solid antibacterial activity and proliferative induction on HGFs cultured on plates and HOBs cultured on a cortical lamina. PADMMs are used in dentistry and periodontology to treat gingival recessions and to increase the tissue thickness in the case of a thin biotype without the risks or postoperative discomfort associated with connective tissue grafts. However, one of the possible complications in this type of surgery is represented by bacterial invasion and membrane exposition during the healing period. We hypothesized that the addition of ALAD-PDT to PADMMs could enhance more rapid healing and decrease the risks connected with bacterial invasion. In periodontal surgery, PADMMs are inserted after a full-thickness flap elevation between the bone and the flap. Consequently, all procedures were performed in parallel on hOBs and hGFs obtained by dental patients. The group control (CTRL) was represented by the unexposed cells cultured on the membranes, group LED (PDT) were the cells subjected to 7 min of red LED irradiation, and ALAD-PDT were the cells subjected to 45 min of ALAD incubation and then to 7 min of red LED irradiation. After treatments, all groups were analyzed for MTT assay and subjected to histological examination at 3 and 7 days and to the SEM observations at 3, 7, and 14 days. Different bone mineralization assays were performed to quantify the effects of ALAD-PDT on hOBs: ALP activity, ALP gene expression, osteocalcin, and alizarin red. The effects of ALAD-PDT on hGFs were evaluated by quantifying collagen 1, fibronectin, and MMP-8. Results showed that ALAD-PDT promoted cellular induction, forming a dense cellular network on hOBs and hGFs, and the assays performed showed statistically significantly higher values for ALAD-PDT with respect to LED alone and CTRLs. In conclusion, ALAD-PDT could represent a promising aid for enhancing the healing of gingival tissues after PADMM applications.

摘要

本研究旨在测试一种光动力方案,该方案基于含氨基乙酰丙酸的凝胶,随后对培养在猪脱细胞真皮基质膜(PADMM)上的人牙龈成纤维细胞(hGFs)和成骨细胞(hOBs)进行红光发光二极管(LED)照射(ALAD-PDT)。在先前的文献中,ALAD-PDT对培养在平板上的hGFs和培养在皮质板上的hOBs显示出可靠的抗菌活性和增殖诱导作用。PADMMs用于牙科和牙周病学领域,以治疗牙龈退缩,并在生物型较薄的情况下增加组织厚度,且不存在与结缔组织移植相关的风险或术后不适。然而,这类手术中一种可能的并发症是愈合期细菌入侵和膜暴露。我们假设在PADMMs中添加ALAD-PDT可以促进更快的愈合,并降低与细菌入侵相关的风险。在牙周手术中,PADMMs在骨和皮瓣之间进行全厚皮瓣翻起后植入。因此,对牙科患者获取的hOBs和hGFs并行进行所有操作。对照组(CTRL)由培养在膜上未暴露的细胞组成,LED组(PDT)是接受7分钟红光LED照射的细胞,ALAD-PDT组是先接受45分钟ALAD孵育然后再接受7分钟红光LED照射的细胞。处理后,对所有组进行MTT分析,并在第3天和第7天进行组织学检查,在第3天、第7天和第14天进行扫描电子显微镜(SEM)观察。进行不同的骨矿化分析以量化ALAD-PDT对hOBs的影响:碱性磷酸酶(ALP)活性、ALP基因表达、骨钙素和茜素红。通过量化I型胶原、纤连蛋白和基质金属蛋白酶-8(MMP-8)来评估ALAD-PDT对hGFs的影响。结果表明,ALAD-PDT促进细胞诱导,在hOBs和hGFs上形成密集的细胞网络,并且所进行的分析显示,与单独的LED组和对照组相比,ALAD-PDT组的值在统计学上显著更高。总之,ALAD-PDT可能是一种有前景的辅助手段,有助于增强PADMM应用后牙龈组织的愈合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/16a1/10379034/39b9061c3373/gels-09-00584-g001.jpg

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