Carlos Garaicoa-Pazmino1, Tobias Fretwurst1,2, Cristiane H Squarize1,3, Tord Berglundh4, William V Giannobile1,5, Lena Larsson1,4, Rogerio M Castilho1,3. 1. Department of Periodontics and Oral Medicine, University of Michigan School of Dentistry, Ann Arbor, Michigan, USA. 2. Department of Oral and Craniomaxillofacial Surgery, Center for Dental Medicine, University Medical Center Freiburg, Freiburg, Germany. 3. Laboratory of Epithelial Biology, University of Michigan School of Dentistry, Ann Arbor, Michigan, USA. 4. Department of Periodontology Institute of Odontology, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden. 5. Department of Biomedical Engineering, College of Engineering and Biointerfaces Institute, University of Michigan, Ann Arbor, Michigan, USA.
Abstract
AIM: To explore the M1/M2 status of macrophage polarization from healthy, gingivitis, and periodontitis patient samples. MATERIALS AND METHODS: Gingival biopsies were collected from 42 individuals (14 gingivitis, 18 periodontitis, and 10 healthy samples) receiving periodontal therapy. Histomorphology analysis was performed with haematoxylin and eosin staining. Immunofluorescence was performed using a combination of CD68 (macrophages), iNOS (M1), and CD206 (M2) in order to acquire changes in macrophage polarization at a single-cell resolution. Macrophages were quantified under microscopy using narrow wavelength filters to detect Alexa 488, Alexa 568, Alexa 633 fluorophores, and Hoechst 33342 to identify cellular DNA content. RESULTS: Gingivitis and periodontitis samples showed higher levels of macrophages compared with healthy samples. Unexpectedly, periodontitis samples displayed lower levels of macrophages dispersed in the stromal tissues compared with gingivitis samples; however, it remained higher than healthy tissues. The polarization of macrophages appears to be reduced in periodontitis and showed similar levels to those observed in healthy tissues. CONCLUSIONS: Our study found that gingivitis and periodontitis differ from each other by the levels of macrophage infiltrate, but not by changes in macrophage polarization.
AIM: To explore the M1/M2 status of macrophage polarization from healthy, gingivitis, and periodontitispatient samples. MATERIALS AND METHODS: Gingival biopsies were collected from 42 individuals (14 gingivitis, 18 periodontitis, and 10 healthy samples) receiving periodontal therapy. Histomorphology analysis was performed with haematoxylin and eosin staining. Immunofluorescence was performed using a combination of CD68 (macrophages), iNOS (M1), and CD206 (M2) in order to acquire changes in macrophage polarization at a single-cell resolution. Macrophages were quantified under microscopy using narrow wavelength filters to detect Alexa 488, Alexa 568, Alexa 633 fluorophores, and Hoechst 33342 to identify cellular DNA content. RESULTS:Gingivitis and periodontitis samples showed higher levels of macrophages compared with healthy samples. Unexpectedly, periodontitis samples displayed lower levels of macrophages dispersed in the stromal tissues compared with gingivitis samples; however, it remained higher than healthy tissues. The polarization of macrophages appears to be reduced in periodontitis and showed similar levels to those observed in healthy tissues. CONCLUSIONS: Our study found that gingivitis and periodontitis differ from each other by the levels of macrophage infiltrate, but not by changes in macrophage polarization.