Macrophages play a crucial rule in orchestrating immune responses against pathogens and foreign materials. Macrophages have remarkable plasticity in response to environmental cues and are able to acquire a spectrum of activation status, best exemplified by pro-inflammatory (M1) and anti-inflammatory (M2) phenotypes at the two ends of the spectrum. Characterisation of M1 and M2 subsets is usually carried out by quantification of multiple cell surface markers, transcription factors and cytokine profiles. These approaches are time-consuming, require large numbers of cells and are resource intensive. In this study, we used machine learning algorithms to develop a simple and fast imaging-based approach that enables automated identification of different macrophage functional phenotypes using their cell size and morphology. Fluorescent microscopy was used to assess cell morphology of different cell types which were stained for nucleus and actin distribution using DAPI and phalloidin respectively. By only analysing their morphology we were able to identify M1 and M2 phenotypes effectively and could distinguish them from naïve macrophages and monocytes with an average accuracy of 90%. Thus we suggest high-content and automated image analysis can be used for fast phenotyping of functionally diverse cell populations with reasonable accuracy and without the need for using multiple markers.
Macrophages are innate immune cells that have a central role in combating infection and maintaining tissue homeostasis. They exhibit remarkable plasticity in response to environmental cues. At either end of a broad activation spectrum are pro-inflammatory (M1) and anti-inflammatory (M2) macrophages with distinct functional and phenotypical characteristics. Macrophages also play a crucial role in orchestrating immune responses to biomaterials used in the fabrication of implantable devices and drug delivery systems. To assess the impact of different surface chemistries on macrophage polarisation, human monocytes were cultured for 6 days on untreated hydrophobic polystyrene (PS) and hydrophilic O2 plasma-etched polystyrene (O2-PS40) surfaces. Our data clearly show that monocytes cultured on the hydrophilic O2-PS40 surface are polarised towards an M1-like phenotype, as evidenced by significantly higher expression of the pro-inflammatory transcription factors STAT1 and IRF5. By comparison, monocytes cultured on the hydrophobic PS surface exhibited an M2-like phenotype with high expression of mannose receptor (MR) and production of the anti-inflammatory cytokines IL-10 and CCL18. While the molecular basis of such different patterns of cell differentiation is yet to be fully elucidated, we hypothesise that it is due to the adsorption of different biomolecules on these surface chemistries. Indeed our surface characterisation data show quantitative and qualitative differences between the protein layers on the O2-PS40 surface compared to PS surface which could be responsible for the observed differential macrophage polarisation on each surface.
Implantation of medical devices can result in inflammation. A large library of polymers is screened, and a selection found to promote macrophage differentiation towards pro-or anti-inflammatory phenotypes. The bioinstructive properties of these materials are validated within a rodent model. By identifying novel materials with immune-instructive properties, the relationship between material-immune cell interactions could be investigated, and this offers exciting possibilities to design novel bioinstructive materials that can be used for numerous clinical applications including medical implants.
Antigen presenting cells (APCs) such as macrophages and dendritic cells (DCs) play a crucial role in orchestrating immune responses against foreign materials. The activation status of APCs can determine the outcome of an immune response following implantation of synthetic materials, towards either healing or inflammation. A large range of biomaterials are used in the fabrication of implantable devices and drug delivery systems. These materials will be in close contact with APCs and characteristics such as surface chemistry and topography may have a critical role in initiating pro- or anti-inflammatory immune responses. Controlling biomaterial surface attributes provides a powerful tool for modulating the phenotype and function of immune cells with the aim of reducing detrimental pro-inflammatory responses and promoting beneficial healing responses. In this article, we review recent literature on how biomaterial surface topography and chemistry can modulate APC populations towards distinct pro- or anti-inflammatory phenotypes with specific examples of how these properties can be used to control host response in vivo. Topographical and/or chemical design of biomaterial surfaces with respect to the APC responses can pave the way for a new generation of 'cell instructive' materials with immunomodulatory properties with a wide range of clinical applications.
While many patients infected by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) eventually produce neutralising antibodies, the degree of susceptibility of previously infected individuals to reinfection by SARS-CoV-2 is currently unknown. To better understand the impact of the immunoglobulin (IgG) level on reinfection in recovered coronavirus disease 2019 (COVID-19) patients, anti-nucleocapsid IgG levels against SARS-CoV-2 were measured in 829 patients with previously confirmed infection just after their recovery. Notably, 87 of these patients had no detectable IgG concentration. While there was just one case of asymptomatic reinfection 4.5 months after the initial recovery amongst patients with detectable anti-nucleocapsid IgG levels, 25 of the 87 patients negative for anti-nucleocapsid IgG were reinfected within one to three months after their first infection. Therefore, patients who recover from COVID-19 with no detectable anti-nucleocapsid IgG concentration appear to remain more susceptible to reinfection by SARS-CoV-2, with no apparent immunity. Also, although our results suggest the chance is lower, the possibility for recovered patients with positive anti-nucleocapsid IgG findings to be reinfected similarly exists.
ABSTRACT:Macrophages are master regulators of immune responses towards implanted biomaterials. The activation state adopted by macrophages in response to biomaterials determines their own phenotype and function as well as those of other resident and infiltrating immune and non-immune cells in the area. A wide spectrum of macrophage activation states exists, with M1 (pro-inflammatory) and M2 (anti-inflammatory) representing either ends of the spectrum. In biomaterials research, cellinstructive surfaces that favour or induce M2 macrophages have been considered as beneficial due to the anti-inflammatory and pro-regenerative properties of these cells. In this study, we used a gelatin methacryloyl (GelMA) hydrogel platform to determine whether micropatterned surfaces can modulate the phenotype and function of human macrophages. The effect of microgrooves/ridges and micropillars on macrophage phenotype, function, and gene expression profile were assessed using conventional methods (morphology, cytokine profile, surface marker expression, phagocytosis) and gene microarrays.Our results demonstrated that micropatterns did induce distinct gene expression profiles in human macrophages cultured on microgrooves/ridges and micropillars. Significant changes were observed in genes related to primary metabolic processes such as transcription, translation, protein trafficking, DNA repair and cell survival. However, interestingly conventional phenotyping methods, relying on surface marker expression and cytokine profile, were not able to distinguish between the different conditions, and indicated no clear shift in cell activation towards an M1 or M2 phenotypes. This highlights the limitations of studying the effect of different physicochemical conditions on macrophages by solely relying on conventional markers that are primarily developed to differentiate between cytokine polarised M1 and M2 macrophages. We therefore, propose the adoption of unbiased screening methods in determining macrophage responses to biomaterials. Our data clearly shows that the exclusive use of conventional markers and methods for determining macrophage activation status could lead to missed opportunities for understanding and exploiting macrophage responses to biomaterials.
Background: Abnormal inflammation coagulation biomarker levels of troponin, C-reactive protein (CRP), and D-dimer levels in serum have been demonstrated to be associated and involved in the disease progression of coronavirus disease 2019 .Methods: First: the study aimed to investigate the correlation of troponin, CRP, D-dimer, white blood cell (WBC) and polymerase chain reaction-cycle threshold (PCR-Ct) within COVID-19 survivors (143 patients; 79 males, 64 females) and in deceased (30 patients; 12 males, 18 females) group. Also, assessing any differences between both groups in studied parameters. Second: a correlation study of studied parameters' level has been conducted within families (41 patients; 23 males [seven deaths] and 18 females [eight deaths]) that lost more than one member due to the severity of the disease. Also, differences between these family and control group (132 patients; 69 males and 63 females) group in studied parameters have been assessed. Results: In the first week of hospitalization, there were significant differences in D-dimer, CRP and troponin level between survived and deceased patient groups. In the second week of the admission, both groups had significant differences in the level of all studied parameters; troponin I, D-dimer, CRP, and WBCs. WBC levels positively correlated to CRP in male survivors (r = 0.75, p < 0.0001), and to troponin in deceased male patients (r = 0.74, p = 0.007). The second week of patient admission was critical in the group of families who lost more than one person, when troponin was correlated positively with D-dimer, CRP, and WBCs. Conclusion:Troponin, D-dimer, CRP, and WBCs level were significantly higher in COVID-19 patients who died than in COVID-19 survivors. High troponin and WBC levels, were considerably associated with families that lost more than one member, when compared with the unrelated COVID-19 patient control.
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