Clinic and Policlinic for Dermatology and Venereology, University Medical Center Rostock, Rostock, Germany;
BerlinAnalytix GmbH, Berlin, Germany.
In Vivo. 2022 Sep-Oct;36(5):2042-2051. doi: 10.21873/invivo.12930.
BACKGROUND/AIM: Macrophages and biomaterial-induced multinucleated giant cells (BMGCs) are central elements in the tissue reaction cascade towards bone substitute materials (BSM). The enzymatic detection of the lytic enzyme tartrate-resistant acid phosphatase (TRAP) has manifoldly been used to examine the so-called "bioactivity" of BSM. The present study aimed to compare the detection validity and expression pattern of the TRAP enzyme using enzymatic and immunohistochemical detection methods in the context of biocompatibility analyses of BSM.
Biopsies from 8 patients were analyzed after sinus augmentation with a xenogeneic bone substitute. Analysis of both macrophage and BMGC polarization were performed by histochemical TRAP detection and immunohistochemical detection of TRAP5a. Histomorphometrical analysis was used for comparison of the TRAP detection of BMGCs.
The enzymatic TRAP detection method revealed that in 7 out of 8 biopsies only single cells were TRAP-positive, whereas most of the cells and especially the BMGCs were TRAP-negative. The immunohistochemical detection of TRAP5a showed moderate numbers of stained mononuclear cells, while the majority of the BMGCs showed signs of TRAP5a-expression. The enzymatic TRAP detection was comparable to the results obtained via immunohistochemistry only in one case. The histomorphometrical analysis showed that significantly more mononuclear and multinucleated TRAP-positive cells were found using immunohistochemical TRAP5a-staining compared to the enzymatic TRAP detection method. Also, significantly more TRAP-negative BMGCs were found using the enzymatic TRAP detection.
The immunohistochemical detection of TRAP is more accurate for examination of the bioactivity and cellular degradability of BSM.
背景/目的：巨噬细胞和生物材料诱导的多核巨细胞（BMGCs）是骨替代材料（BSM）组织反应级联中的核心元素。抗酒石酸酸性磷酸酶（TRAP）的酶检测已被广泛用于检查 BSM 的所谓“生物活性”。本研究旨在比较酶法和免疫组织化学检测方法在 BSM 生物相容性分析中检测 TRAP 酶的检测有效性和表达模式。
对 8 例接受异种骨替代物鼻窦增大的患者进行活检。通过组织化学 TRAP 检测和 TRAP5a 的免疫组织化学检测分析巨噬细胞和 BMGC 极化。组织形态计量学分析用于比较 BMGCs 的 TRAP 检测。
酶法 TRAP 检测显示，在 8 例活检中有 7 例仅单个细胞呈 TRAP 阳性，而大多数细胞，特别是 BMGCs 呈 TRAP 阴性。TRAP5a 的免疫组织化学检测显示中等数量的染色单核细胞，而大多数 BMGCs 显示出 TRAP5a 表达的迹象。仅在一种情况下，酶法 TRAP 检测与免疫组织化学检测结果相当。组织形态计量学分析显示，与酶法 TRAP 检测相比，免疫组织化学 TRAP5a 染色检测到的单核和多核 TRAP 阳性细胞明显更多。此外，用酶法 TRAP 检测还发现了更多的 TRAP 阴性 BMGCs。
免疫组织化学检测 TRAP 更准确地检查 BSM 的生物活性和细胞降解性。
