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人成骨细胞对不同形式脱蛋白牛骨的细胞反应。

Cellular Response of Human Osteoblasts to Different Presentations of Deproteinized Bovine Bone.

作者信息

de Azambuja Carvalho Pedro Henrique, Al-Maawi Sarah, Dohle Eva, Sader Robert Alexander, Pereira-Filho Valfrido Antonio, Ghanaati Shahram

机构信息

Department of Diagnosis and Surgery, School of Dentistry, Sao Paulo State University (Unesp), Araraquara 14801-060, Brazil.

FORM-Lab, Frankfurt Oral Regenerative Medicine, Department for Oral, Cranio-Maxillofacial and Facial Plastic Surgery, Medical Center of the Goethe University, 60323 Frankfurt, Germany.

出版信息

Materials (Basel). 2022 Jan 27;15(3):999. doi: 10.3390/ma15030999.

Abstract

: This study evaluated the cellular response of primary osteoblasts exposed to two different presentations of a low-temperature non-sintered deproteinized bovine bone matrix (DBBM). : Six different baths of a commercially available DBBM block (Bonefill Porous Block) and one of DBBM granule (Bonefill Porous) were evaluated to identify the mineral structure and organic or cellular remnants. Samples of the same baths were processed in TRIZOL for RNA extraction and quantification. For the immunologic cell reaction assay, primary human osteoblasts (pOB) were exposed to DBMM block (pOB + B) or granules (pOB + G), or none (control) for 1, 3, or 7 days of cell cultivation. Expression of proinflammatory cytokines by pOB was evaluated by crosslinked ELISA assay. In addition, total DNA amount, as well as cell viability via LDH evaluation, was assessed. : Organic remnants were present in DBBM blocks; 45.55% (±7.12) of osteocytes lacunae presented cellular remnants in blocks compared to 17.31% (±1.31) in granules. In three of five batches of blocks, it was possible to isolate bovine RNA. The highest concentration of TGF-β1 was found in supernatants of pOB + G on day 7 (218.85 ± 234.62 pg/mL) ( < 0.05), whereas pOB + B presented the lowest amount of TGF-β1 secretion at the end of evaluation (30.22 ± 14.94 pg/mL, < 0.05). For IL-6 and OPG, there was no statistical difference between groups, while pOB + G induced more IL-8 secretion than the control (3.03 ± 3.38 ng/mL, < 0.05). Considering the kinetics of cytokine release during the study period, all groups presented a similar pattern of cytokines, estimated as an increasing concentration for IL-6, IL-8, and OPG during cultivation. Adherent cells were observed on both material surfaces on day 7, according to H&E and OPN staining. : Neither tested material induced a pronounced inflammatory response upon osteoblast cultivation. However, further studies are needed to elucidate the potential influence of organic remnants in bone substitute materials on the regeneration process.

摘要

本研究评估了原代成骨细胞暴露于两种不同形式的低温非烧结脱蛋白牛骨基质(DBBM)后的细胞反应。评估了市售DBBM块(骨填充多孔块)的六种不同批次以及一种DBBM颗粒(骨填充多孔),以确定矿物质结构以及有机或细胞残余物。将相同批次的样品用TRIZOL处理以进行RNA提取和定量。对于免疫细胞反应测定,将原代人成骨细胞(pOB)暴露于DBMM块(pOB + B)或颗粒(pOB + G),或不进行处理(对照),培养1、3或7天。通过交联ELISA测定评估pOB中促炎细胞因子的表达。此外,评估了总DNA量以及通过LDH评估的细胞活力。DBBM块中存在有机残余物;与颗粒中17.31%(±1.31)相比,骨陷窝中有45.55%(±7.12)在块中呈现细胞残余物。在五批块中的三批中,可以分离出牛RNA。在第7天,pOB + G的上清液中发现TGF-β1的浓度最高(218.85±234.62 pg/mL)(P<0.05),而在评估结束时,pOB + B的TGF-β1分泌量最低(30.22±14.94 pg/mL,P<0.05)。对于IL-6和OPG,各组之间无统计学差异,而pOB + G诱导的IL-8分泌比对照更多(3.03±3.38 ng/mL,P<0.05)。考虑到研究期间细胞因子释放的动力学,所有组呈现出相似的细胞因子模式,估计在培养过程中IL-6、IL-8和OPG的浓度会增加。根据苏木精-伊红(H&E)和骨桥蛋白(OPN)染色,在第7天在两种材料表面均观察到贴壁细胞。两种测试材料在成骨细胞培养时均未诱导明显的炎症反应。然而,需要进一步研究以阐明骨替代材料中有机残余物对再生过程的潜在影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/136a/8839050/12fea1e13d04/materials-15-00999-g001.jpg

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