Skip to main content

Abstract

BACKGROUND:

Subarachnoid hemorrhage (SAH) is characterized by acute and delayed reductions of cerebral blood flow (CBF) caused, among others, by spasms of cerebral arteries and arterioles. Recently, the inactivation of perivascular macrophages (PVM) has been demonstrated to improve neurological outcomes after experimental SAH, but the underlying mechanisms of protection remain unclear. The aim of our exploratory study was, therefore, to investigate the role of PVM in the formation of acute microvasospasms after experimental SAH.

METHODS:

PVMs were depleted in 8- to 10-week-old male C57BL/6 mice (n=8/group) by intracerebroventricular application of clodronate-loaded liposomes and compared with mice with vehicle liposome injections. Seven days later, SAH was induced by filament perforation under continuous monitoring of CBF and intracranial pressure. Results were compared with sham-operated animals and animals who underwent SAH induction but no liposome injection (n=4/group each). Six hours after SAH induction or sham surgery, numbers of microvasospasms per volume of interest and % of affected pial and penetrating arterioles were examined in 9 standardized regions of interest per animal by in vivo 2-photon microscopy. Depletion of PVMs was proven by quantification of PVMs/mm3 identified by immunohistochemical staining for CD206 and Collagen IV. Statistical significance was tested with t tests for parametric data and Mann-Whitney U test for nonparametric data.

RESULTS:

PVMs were located around pial and intraparenchymal arterioles and were effectively depleted by clodronate from 671±28 to 46±14 PVMs/mm3 (P<0.001). After SAH, microvasospasms was observed in pial arteries and penetrating and precapillary arterioles and were accompanied by an increase to 1405±142 PVMs/mm3. PVM depletion significantly reduced the number of microvasospasms from 9 IQR 5 to 3 IQR 3 (P<0.001).

CONCLUSIONS:

Our results suggest that PVMs contribute to the formation of microvasospasms after experimental SAH.

Graphical Abstract

Subarachnoid hemorrhage (SAH) is a severe subtype of stroke with a high mortality and morbidity resulting in a high socioeconomic burden.1–3
In most cases, an intracranial aneurysm ruptures and blood enters the subarachnoid space, resulting in a rapid increase of intracranial pressure and—subsequently—global ischemia, often fatal within minutes. Although interventional and microsurgical techniques are advanced and allow a safe and efficient occlusion of aneurysms to prevent rebleeding, mortality after SAH is high and many patients who survive the initial ictus still suffer from significant morbidity.1
Major features associated with adverse outcome after SAH include early and delayed cerebral ischemia.2 Advanced imaging in patients and after experimental SAH show severe cortical hypoperfusion despite normal cerebral perfusion pressure within hours after aneurysm rupture.4,5 Spasms of pial and penetrating arterioles (microvasospasms) have been demonstrated to be associated with acute cortical hypoperfusion. Options to treat microvasospasms are not available yet, since the mechanisms involved in the formation of microvasospasms are not fully understood.6–8
Recently, Wan et al9 demonstrated that depletion of perivascular macrophages (PVMs) after experimental SAH improves neurological outcome and reduces perivascular inflammation. Since we observed in previous studies that degradation of blood products resulted in the reduction of microvasospasms, we hypothesize that PVMs may be critically involved in the formation of microvasospasms.
In the current study, we therefore used mice depleted of PVMs and visualized pial and penetrating arterioles in vivo by 2-photon microscopy to investigate whether these cells are involved in the formation of microvasospasms after SAH.

METHODS

The authors declare that all supporting data are available within the article and its Supplemental Material.

Animals and Experimental Groups

All procedures on animals, group size calculations, and statistical methods were approved by the Government of Upper Bavaria. The results of the study are reported in accordance with the Animal Research: Reporting of In Vivo Experiments guidelines. Eight- to 10-week-old male C57BL/6 mice (Jackson Laboratory, Bar Harbor) were used for all experiments. We investigated 4 different groups: (1) SAH (n=4) animals without intrathecal injection, (2) Sham (n=4) operated animals, treated with the same protocol except for middle cerebral artery perforation, (3) SAH+Vehicle (n=8), and (4) SAH+clodronate (n=8; Figure S1; Table S1)
All experiments were performed in a strictly randomized and blinded manner, that is, all investigators were unaware of the status of the mice until all experiments were performed and all data was analyzed.

Depletion of PVMs

Intrathecally injected liposomes loaded with clodronate (Liposoma, the Netherlands), a substance which induces apoptosis once taken up by cells, were used to deplete PVMs 7 days before SAH (Figure 1A). Liposomes without clodronate served as vehicle control. For intrathecal injection, animals were anesthetized, fixed in a stereotactic frame (Foehr Medical Instruments, Germany), and the atlanto-occipital membrane covering the cisterna magna was exposed. A custom-made glass capillary and a microsyringe pump (World Precision instruments) were used to inject 10 µL of liposomes into the cisterna magna at a rate of 3 µL/minute.
Figure 1. Experimental setup. Subarachnoid hemorrhage (SAH) and in vivo imaging performed 7 d after intrathecal application of PBS or clodronate-containing liposomes (A). Imaging was done in 9 standardized volumes of interest (VOI) as shown in B. After induction of the SAH, ipsilateral cerebral blood flow (CBF), measured with transcranial laser Doppler probes, was reduced in the SAH groups and the minimal flow was lower after the injection of clodronate (C). Contralateral CBF as indirect sign of intracranial pressure was reduced after SAH induction (C).

Induction of Subarachnoid Hemorrhage

SAH and sham surgery were performed as previously described.7,10 Briefly, mice were anesthetized with a mixture of 0.05 mg/kg fentanyl (Janssen-Cilac, Neuss, Germany), 0.5 mg/kg medetomidine (Pfizer) and 5 mg/kg midazolam (Braun, Germany), intubated, and mechanically ventilated. Core body temperature, pO2, pCO2, mean arterial blood pressure, oxygen saturation, cerebral blood flow, and intracranial pressure were continuously monitored during surgery (Table S2). The Circle of Willis was perforated at the outlet of the left middle cerebral artery with an intravascular filament advanced via the common carotid artery. For sham surgery, the filament was inserted and not advanced far enough to induce perforation. Monitoring was continued for 20 minutes after SAH induction. Anesthesia was then antagonized by subcutaneous injection of 1.2 mg/kg naloxone (Actavis, Ireland), 0.5 mg/kg flumazenil (Inresa, Germany), and 2.5 mg/kg atipamezole (Pfizer).
At the end of the experiment, the amount of subarachnoid blood was quantified according to Sugawara et al11 (Figure S2).

Two-Photon Microscopy

In vivo imaging was performed with a LSM 7 microscope (Zeiss, Germany), equipped with a Li:Ti laser (Chameleon, Coherent), 6 hours after SAH/sham surgery (Figure 1A). Animals were anesthetized as described above and received a femoral artery catheter. A thinned skull window was prepared above the left hemisphere as previously described.7 Nine random 500×500 µm regions of interest were imaged to a depth of 250 µm (Figure 1B) 6 hours after SAH using a 20× objective (Zeiss, Germany). Animals were continuously monitored for end-expiratory pCO2, body temperature, heart rate, and peripheral oxygen saturation during imaging. To visualize the microcirculation, 100 µL fluoresceinisothiocyanate dextran (0.5% in saline, Sigma-Aldrich) was applied by intraarterial injection.

Histological Analysis and Quantification

After in vivo imaging mice were fixed with 4% paraformaldehyde by intracardial perfusion and 50 µm free-floating coronal brain sections were collected. PVMs were labeled with a CD206+ antibody (BIO-RAD, 1:100). The vasculature was stained with Collagen IV (Abcam, 1:100). As secondary antibodies Donkey Anti-Rat IgG (Abcam, 1:200) and Donkey Anti-Rabbit IgG (Jackson, 1:200) were used. Slices were incubated with the primary antibodies for 3 days at 4 °C and with the secondary antibodies for 2 days at 4 °C. Cell nuclei were labeled by DAPI (1 mg/mL, 1:1000, Vector Labs). PVMs were defined by positive staining with CD206 and direct contact to a vessel. Meningeal macrophages were defined as CD206+ meningeal cells without vascular contact. Sections were imaged by confocal microscopy (LSM810, Zeiss, Germany), and 6 standardized ROIs per animal were analyzed in a randomized and blinded fashion.

In Vivo Data Analysis and Quantification

Image analysis was performed with Fiji Image J, Version 2.3. Arteries and arterioles were distinguished from veins by autofluorescence of the vessel wall and blood flow direction. Pial vessels were analyzed in axial planes and penetrating arteries were analyzed in coronal or sagittal planes. Microvasospasms were identified by calculating the constriction grade of the spastic vessel compared with a nonspastic vessel segment as previously described.12 Microvasospasm was defined as a reduction of the vessel diameter ≥15% compared with a nonspastic vessel segment.

Statistical Analysis

Data sets were tested for normal distribution with the D’Agostino & Pearson test and presented as ±SEM (±SEM). Otherwise, medians±percentiles were used. Statistically significant differences between groups were tested with Student t tests for parametric data and with the Mann-Whitney U test for nonparametric data using Prism 8 (Graphpad Software LLC).

RESULTS

After SAH induction, brain perfusion was globally reduced in all mice. There was no difference between animals receiving clodronate (Clo) or phosphate-buffered saline (Figure 1C). Sham-operated animals showed no changes in cerebral blood flow.

Number of Brain Macrophages Before and After Depletion

In naive C57BL/6 mice, we found 101±14 CD206+ cells/mm3 without vessel association (meningeal macrophages). Additionally, we also found 671±28 CD206+ cells/mm3 associated with vessels, that is, PVMs. PVMs have an elongated shape and wrap around microvessels (Figure 2A). Most PVMs were located adjacent to pial arteries (276±24/mm3); PVM density on intraparenchymal vessels was somewhat lower (precapillary arteries: 238±16/mm3 and penetrating arteries; 163±15/ mm3; Figure 2B), but still 3 to 5 times higher than the number of meningeal macrophages.
Figure 2. Cortical CD206+ macrophages are associated with pial membranes and the microvasculature. Immunohistochemical staining of coronal cuts of the superficial (A) and deep (B) cortex of naive 8- to 10-wk-old male C57Bl6/n mice with CD206 (macrophages, red) and Collagen IV (microvessels, green). Meningeal Macrophages are CD206+ cells, adjacent to the pial membrane, not associated with vessels (A). Perivascular macrophages are CD206+ elongated cells embracing pial arteries, penetrating and precapillary arterioles (B). Most CD 206+ cells/ mm3 are vessel associated (C). Scalebar=20 µm.
Application of clodronate-loaded liposomes depleted almost all PVMs within 7 days (Figure 3A). Quantification of the immunohistochemical stainings revealed a reduction in the number of PVMs from around 670 PVMs/mm3 in naïve mice, a number well in line with our previous quantification (Figure 2B), to only 46±14 PVMs/mm3 (P<0.001), that is, a reduction of approximately 93%. Interestingly, we also observed a small but significant increase in the number of PVMs in vehicle-treated mice compared with naive animals (Figure 3B; P<0.001), suggesting that an injection with a thin glass capillary through the atlanto-occipital membrane was sufficient to induce PVM proliferation to a small extent. Since the morphology of the PVMs was the same as in unhandled mice (Figure 3A), a major activation of these cells at the time of SAH could be excluded with a sufficiently high degree of confidence.
Figure 3. Intrathecal liposome injection increases numbers of vessel associated macrophages while clodronate liposome injection leads to a macrophage depletion after 7 d. A, Immunohistochemical staining of coronal cuts of the cortices of naive 8- to 10-wk-old male C57Bl6/n mice (top), 7 d after intrathecal injection of PBS (vehicle) liposomes (middle) and clodronate liposomes (bottom). Shapes of CD206+ cells do not vary between groups. B, Quantification of vessel-associated CD206+ cells shows an increased number after injection of vehicle liposomes and a strong decrease of cell numbers after depletion by clodronate liposomes. Arrow=penetrating arteriole; scale bar=20 µm; ***P<0.001.

Number of Brain Macrophages After SAH

Six hours after SAH, brain macrophages remained unchanged regarding morphology or distribution compared with naive mice (Figure 4A); however, their number increased (Figure 4B and 4C). While the number of meningeal CD206+ cells increased without reaching statistical significance (Figure 4B; P=0.07), the number of PVMs more than doubled after SAH (Figure 4C; P<0.001). These findings suggest that PVMs proliferate particularly rapidly when they come into contact with perivascular blood.
Figure 4. The number of cortical CD206+ macrophages is increased 8 h after subarachnoid hemorrhage (SAH). A, Immunohistochemical staining of coronal cuts of the cortices of 8- to 10-wk-old male C57Bl6/n mice without (top) or with (bottom) SAH induction. CD206+ macrophages are labeled red and Collagen IV+ microvessels are labeled green. Quantification shows a trend toward increasing numbers of meninges associated CD206+ cells without direct vessel contact (B) and significantly more vessel associated CD206+ cells (C) after SAH. Arrow=penetrating arteriole; scale bar=20 µm, ***P<0.001.

Depletion of PVMs Reduces Microvasospasms After SAH

As previously described,7,10,13 SAH induces a high number of microvasospasm (Figure 5A and 5B) in pial and penetrating arterioles (Figure 5C and 5D). In mice depleted of PVMs, the number of microvasospasm was reduced by 66% (Figure 5A and 5B; 9 IQR 5 versus 3 IQR 3) and the proportion of spastic pial and penetrating arterioles was also reduced (Figure 5C and 5D). These findings suggest that PVMs are critically involved in the formation of microvasospasm.
Figure 5. Perivascular macrophage (PVM) depletion reduces microarterial constriction and microvasospasm formation after subarachnoid hemorrhage (SAH). A, Exemplary intravital microscopy images in axial (upper row) and coronal (lower row) projection. High resolution axial and coronal in vivo 2 photon microscopy 6 h after SAH in C57Bl6/n mice with intraarterial labeling 7 d after intrathecal injection of Liposomes containing PBS (left image) or clodronate (right image). Arrows label pearl string shaped spasms in pial arteries and penetrating arterioles labeled with A, veins are labeled with V. Dashed lines show the location of the maximum intensity projection of the coronal image shown below. Scale bar=100 µm. B, Median number±IQR of microvasospasms in the observed VOI. Percentage of spastic/ constricted vessels in the examined volume of interest in pial (C) and (D) penetrating vessels shown as mean±SEM. **P<0.01, ***P<0.001.

DISCUSSION

SAH leads to microvascular constriction and microvasospasm formation in the subacute and acute phase, which decreases cortical perfusion thereby causing tissue ischemia and subsequent early brain injury.6–8,13 PVMs were shown to improve outcome after SAH9; however, it remains unclear how this protective effect was mediated in regard to microvasospasm formation. In the present study, we reproduced previous results from our laboratory that pial and penetrating arterioles constrict acutely after SAH in vivo7,10,13 performed the first detailed characterizations of the location and morphology of macrophages in relation to the cortical microcirculation and confirmed that intrathecal injection of clodronate-loaded liposomes depleted >90% of PVMs. The main finding of the current study is that depletion of PVMs reduces early microvasospasm formation to a large degree (−70%). These results suggest that PVMs play an important and so far unrecognized role in the formation of microvasospasm after SAH.
Resident brain macrophages originate from yolk sac progenitors, populate the brain in early development, and have been identified to play a critical role in the maintenance of brain homeostasis.14 Depending on their anatomic location, macrophages are categorized as PVM, associated with pial arteries or penetrating arterioles, or meningeal (MM). PVMs have been shown to be a major source of reactive oxygen species, to mediate neurovascular dysfunction,15 and to be involved in the pathophysiology of diverse brain diseases such as Alzheimer disease, multiples sclerosis, CNS infections, and arterial hypertension.14,15 Intrathecal injection of clodronate liposomes, which are taken up by brain macrophages and induce apoptosis, allows to specifically diminish this distinct cell population from the brain and to study their role in various disease models.16
All experiments were performed and analyzed in a fully randomized and blinded manner using a clinically relevant SAH model. Since humans also have PVMs and a very similar microvascular anatomy compared with mice,17 we have sufficient evidence to believe that our results are robust and mirror the events which also occur in the human brain after SAH.
The endovascular filament perforation model employed in our study is the most frequently used preclinical SAH model since it can replicate important pathophysiological features of SAH in a satisfactory manner, for example, vessel rupture and endothelial damage.18 Other mouse models are not able to reproduce these features, which underlines the relevance and great translational potential of the SAH model used in our study, indicating that our results can be considered robust, while minimizing methodological bias.
Despite the apparent involvement of PVMs in microvasospasm formation, the underlying molecular mechanisms need further investigation. The starting point of a cascade of events finally leading to microvasospasm seems to be found in blood degradation products,19 which already have been demonstrated to negatively impact the cerebral microcirculation after SAH.10 Within hours after bleeding onset, erythrocytes are phagocytosed and degraded by macrophages as already shown after intracerebral hemorrhage.20,21 Alternatively, erythrocytes may also decompose by autolysis.22 These processes result in the release of blood degradation products, for example, free iron and hemoglobin, which then accumulate at high concentrations in the narrow perivascular space.19 Free hemoglobin is a very potent nitric oxide (NO) scavenger and may therefore cause local depletion of NO und subsequent vasoconstriction.23 A reason to think that this mechanism occurs after SAH is supported by experiments demonstrating that application of NO to cerebral microvessels resolves microvasospasm and improves outcome after experimental SAH.8,24 Further, hemoglobin degradation products, for example, propentdyopents or bilirubin oxidation end products, may directly induce constriction of cerebral microvessels.25,26 Another possible mechanism by which PVMs could trigger microvasospasm (that may be used for therapeutic purposes) is the activation by blood or blood degradation products and the subsequent release of inflammatory cytokines and free radical species.17 Free radicals are potent NO scavengers and may, in the end, cause local vasoconstriction. The fact that free radicals do not have a long half-live in living tissues and cannot readily pass cell membranes may explain why microvasospasm have a pearl string like morphology, that is, spasms occur only at the site where free radicals are produced by PVMs. Further experiments measuring free radical species in spastic microvessels, however, are needed to clarify this process in the future.
Apart from PVMs, neutrophil-triggered inflammation seems to be a relevant factor in early microvascular changes following SAH.27 In the endovascular filament model, neutrophils have been shown to mediate early cerebral cortical hypoperfusion and oxidative stress after SAH, suggesting that targeting neutrophil function and neutrophil-induced oxidative stress may serve a novel approach to mitigate cerebral hypoperfusion early after SAH.28 Analogous to our results where PVM depletion leads to decreased microvasospasm formation and potentially improves cortical perfusion and tissue ischemia, reduction of neutrophil activity seems to decrease early microvascular injury after SAH and implies that neutrophil-targeted interventions may ameliorate early brain injury or at least limit microvascular injury after SAH.27 Moreover, neutrophil extracellular traps (NETs) that promote neuroinflammation after SAH may be potential therapeutic targets since pharmacological inhibition of NETs demonstrated anti-inflammatory effects by decreasing the levels of proinflammatory factors caused by SAH.29 Similarly, inhibiting NETs may reduce microthrombosis and therefore attenuate early brain injury after SAH.30
Despite its apparent advantages, the current study has also some limitations. Clodronate had to be applied by intrathecal injection; however, the application of vehicle alone already increased the number of CD206+ cells compared with naive mice. Although these cells had the same morphology as perivascular and meningeal macrophages, we cannot completely rule out that these cells were not blood macrophages, which infiltrated the subarachnoid space after injection rather than proliferated PVMs. To take the potential influence by blood borne macrophages on our results into consideration, we always used naive mice as controls for all depletion experiments (where all macrophages were depleted by clodronate treatment).16
Another potential limitation is that we performed in vivo microscopy through a relatively small cranial window, which allowed us to analyze only a certain fraction of the cerebral cortex. Thus, we cannot generalize our findings to the entire brain.
In conclusion, our results suggest that PVMs mediate the formation of microvasospasm early after SAH. We think that the underlying mechanisms are either the release of free radicals by PVMs and subsequent NO scavenging or the degradation of red blood cells by PVMs, which result in increase of perivascular blood degradation and subsequent NO depletion. Since distinguishing between these 2 mechanistic pathways may have therapeutic consequences, further studies are needed to fully understand the molecular mechanisms of microvasospasm formation after SAH.

ARTICLE INFORMATION

Supplemental Material

ARRIVE Checklist
Tables S1–S2
Figures S1–S2

Acknowledgments

We would like to thank Uta Mamrak for excellent technical and organizational support.

Footnote

Nonstandard Abbreviations and Acronyms

NO
nitric oxide
PVM
perivascular macrophage
SAH
subarachnoid hemorrhage

Supplemental Material

File (str_stroke-2022-042290_supp1.pdf)
File (str_stroke-2022-042290_supp2.pdf)

REFERENCES

1.
Hoogmoed J, de Oliveira Manoel AL, Coert BA, Marotta TR, Macdonald RL, Vandertop WP, Verbaan D, Germans MR. Why do patients with poor-grade subarachnoid hemorrhage die? World Neurosurg. 2019;131:e508–e513. doi: 10.1016/j.wneu.2019.07.221
2.
Macdonald RL, Schweizer TA. Spontaneous subarachnoid haemorrhage. Lancet. 2017;389:655–666. doi: 10.1016/S0140-6736(16)30668-7
3.
Neifert SN, Chapman EK, Martini ML, Shuman WH, Schupper AJ, Oermann EK, Mocco J, Macdonald RL. Aneurysmal subarachnoid hemorrhage: the last decade. Transl Stroke Res. 2021;12:428–446. doi: 10.1007/s12975-020-00867-0
4.
Schubert GA, Seiz M, Hegewald AA, Manville J, Thome C. Acute hypoperfusion immediately after subarachnoid hemorrhage: a xenon contrast-enhanced ct study. J Neurotrauma. 2009;26:2225–2231. doi: 10.1089/neu.2009.0924
5.
Schubert GA, Seiz M, Hegewald AA, Manville J, Thome C. Hypoperfusion in the acute phase of subarachnoid hemorrhage. Acta Neurochir Suppl. 2011;110:35–38. doi: 10.1007/978-3-7091-0353-1_6
6.
Liu H, Dienel A, Scholler K, Schwarzmaier SM, Nehrkorn K, Plesnila N, Terpolilli NA. Microvasospasms after experimental subarachnoid hemorrhage do not depend on endothelin a receptors. Stroke. 2018;49:693–699. doi: 10.1161/STROKEAHA.117.020028
7.
Schwarting J, Nehrkorn K, Liu H, Plesnila N, Terpolilli NA. Role of pial microvasospasms and leukocyte plugging for parenchymal perfusion after subarachnoid hemorrhage assessed by in vivo multi-photon microscopy. Int J Mol Sci. 2021;22:8444. doi: 10.3390/ijms22168444
8.
Terpolilli NA, Brem C, Buhler D, Plesnila N. Are we barking up the wrong vessels? Cerebral microcirculation after subarachnoid hemorrhage. Stroke. 2015;46:3014–3019. doi: 10.1161/STROKEAHA.115.006353
9.
Wan H, Brathwaite S, Ai J, Hynynen K, Macdonald RL. Role of perivascular and meningeal macrophages in outcome following experimental subarachnoid hemorrhage. J Cereb Blood Flow Metab. 2021;41:1842–1857. doi: 10.1177/0271678X20980296
10.
Liu H, Schwarting J, Terpolilli NA, Nehrkorn K, Plesnila N. Scavenging free iron reduces arteriolar microvasospasms after experimental subarachnoid hemorrhage. Stroke. 2021;52:4033–4042. doi: 10.1161/STROKEAHA.120.033472
11.
Sugawara T, Ayer R, Jadhav V, Zhang JH. A new grading system evaluating bleeding scale in filament perforation subarachnoid hemorrhage rat model. J Neurosci Methods. 2008;167:327–334. doi: 10.1016/j.jneumeth.2007.08.004
12.
Lenz IJ, Plesnila N, Terpolilli NA. Role of endothelial nitric oxide synthase for early brain injury after subarachnoid hemorrhage in mice. J Cereb Blood Flow Metab. 2021;41:1669–1681. doi: 10.1177/0271678X20973787
13.
Friedrich B, Muller F, Feiler S, Scholler K, Plesnila N. Experimental subarachnoid hemorrhage causes early and long-lasting microarterial constriction and microthrombosis: an in-vivo microscopy study. J Cereb Blood Flow Metab. 2012;32:447–455. doi: 10.1038/jcbfm.2011.154
14.
Faraco G, Park L, Anrather J, Iadecola C. Brain perivascular macrophages: characterization and functional roles in health and disease. J Mol Med (Berl). 2017;95:1143–1152. doi: 10.1007/s00109-017-1573-x
15.
Santisteban MM, Iadecola C. Hypertension, dietary salt and cognitive impairment. J Cereb Blood Flow Metab. 2018;38:2112–2128. doi: 10.1177/0271678X18803374
16.
Polfliet MM, Goede PH, van Kesteren-Hendrikx EM, van Rooijen N, Dijkstra CD, van den Berg TK. A method for the selective depletion of perivascular and meningeal macrophages in the central nervous system. J Neuroimmunol. 2001;116:188–195. doi: 10.1016/s0165-5728(01)00282-x
17.
Lapenna A, De Palma M, Lewis CE. Perivascular macrophages in health and disease. Nat Rev Immunol. 2018;18:689–702. doi: 10.1038/s41577-018-0056-9
18.
Muroi C, Fujioka M, Marbacher S, Fandino J, Keller E, Iwasaki K, Mishima K. Mouse model of subarachnoid hemorrhage: technical note on the filament perforation model. Acta Neurochir Suppl. 2015;120:315–320. doi: 10.1007/978-3-319-04981-6_54
19.
Plesnila N. Are we looking into an iron age for subarachnoid hemorrhage? Stroke. 2022;53:1643–1644. doi: 10.1161/STROKEAHA.121.037670
20.
Zhao X, Grotta J, Gonzales N, Aronowski J. Hematoma resolution as a therapeutic target: the role of microglia/macrophages. Stroke. 2009;40:S92–S94. doi: 10.1161/STROKEAHA.108.533158
21.
Chang CF, Massey J, Osherov A, Angenendt da Costa LH, Sansing LH. Bexarotene enhances macrophage erythrophagocytosis and hematoma clearance in experimental intracerebral hemorrhage. Stroke. 2020;51:612–618. doi: 10.1161/STROKEAHA.119.027037
22.
Peterson JW, Roussos L, Kwun BD, Hackett JD, Owen CJ, Zervas NT. Evidence of the role of hemolysis in experimental cerebral vasospasm. J Neurosurg. 1990;72:775–781. doi: 10.3171/jns.1990.72.5.0775
23.
Hugelshofer M, Buzzi RM, Schaer CA, Richter H, Akeret K, Anagnostakou V, Mahmoudi L, Vaccani R, Vallelian F, Deuel J, et al. Haptoglobin administration into the subarachnoid space prevents hemoglobin-induced cerebral vasospasm. J Clin Invest. 2019;129:5219–5235. doi: 10.1172/JCI130630
24.
Terpolilli NA, Feiler S, Dienel A, Muller F, Heumos N, Friedrich B, Stover J, Thal S, Scholler K, Plenila N. Nitric oxide inhalation reduces brain damage, prevents mortality, and improves neurological outcome after subarachnoid hemorrhage by resolving early pial microvasospasms. J Cereb Blood Flow Metab. 2016;36:2096–2107. doi: 10.1177/0271678X15605848
25.
Joerk A, Ritter M, Langguth N, Seidel RA, Freitag D, Herrmann KH, Schaefgen A, Ritter M, Guenther M, Sommer C, et al. Propentdyopents as heme degradation intermediates constrict mouse cerebral arterioles and are present in the cerebrospinal fluid of patients with subarachnoid hemorrhage. Circ Res. 2019;124:e101–e114. doi: 10.1161/CIRCRESAHA.118.314160
26.
Fumoto T, Naraoka M, Katagai T, Li Y, Shimamura N, Ohkuma H. The role of oxidative stress in microvascular disturbances after experimental subarachnoid hemorrhage. Transl Stroke Res. 2019;10:684–694. doi: 10.1007/s12975-018-0685-0
27.
Friedrich V, Flores R, Muller A, Bi W, Peerschke EI, Sehba FA. Reduction of neutrophil activity decreases early microvascular injury after subarachnoid haemorrhage. J Neuroinflammation. 2011;8:103. doi: 10.1186/1742-2094-8-103
28.
Neulen A, Pantel T, Kosterhon M, Kramer A, Kunath S, Petermeyer M, Moosmann B, Lotz J, Kantelhardt SR, Ringel F, et al. Neutrophils mediate early cerebral cortical hypoperfusion in a murine model of subarachnoid haemorrhage. Sci Rep. 2019;9:8460. doi: 10.1038/s41598-019-44906-9
29.
Zeng H, Fu X, Cai J, Sun C, Yu M, Peng Y, Zhuang J, Chen J, Chen H, Yu Q, et al. Neutrophil extracellular traps may be a potential target for treating early brain injury in subarachnoid hemorrhage. Transl Stroke Res. 2022;13:112–131. doi: 10.1007/s12975-021-00909-1
30.
Hao X, Zeng Z, Liang L, Feng Z, Li W, Xiong B, Guo P, Zhang Q, Chen Y, Feng H, et al. The role of neutrophil extracellular traps in early microthrombosis and brain injury after subarachnoid hemorrhage in mice. Transl Stroke Res. 2022;1–14. doi: 10.1007/s12975-022-01074-9

eLetters(0)

eLetters should relate to an article recently published in the journal and are not a forum for providing unpublished data. Comments are reviewed for appropriate use of tone and language. Comments are not peer-reviewed. Acceptable comments are posted to the journal website only. Comments are not published in an issue and are not indexed in PubMed. Comments should be no longer than 500 words and will only be posted online. References are limited to 10. Authors of the article cited in the comment will be invited to reply, as appropriate.

Comments and feedback on AHA/ASA Scientific Statements and Guidelines should be directed to the AHA/ASA Manuscript Oversight Committee via its Correspondence page.

Information & Authors

Information

Published In

Go to Stroke
Go to Stroke
Stroke
Pages: 2126 - 2134
PubMed: 37325921

Versions

You are viewing the most recent version of this article.

History

Received: 20 December 2022
Revision received: 8 May 2023
Accepted: 16 May 2023
Published online: 16 June 2023
Published in print: August 2023

Permissions

Request permissions for this article.

Keywords

  1. arteriole
  2. morbidity
  3. perfusion
  4. spasms
  5. subarachnoid hemorrhage

Subjects

Authors

Affiliations

Xiangjiang Lin, MD
Institute for Stroke and Dementia Research (ISD) (X.L., I.K., B.N.H., N.A.T., J.S., N.P.), Munich University Hospital, Germany.
Munich Cluster for Systems Neurology (SyNergy), Germany (X.L., I.K., B.N.H., N.A.T., J.S., N.P.).
Igor Khalin, MD
Institute for Stroke and Dementia Research (ISD) (X.L., I.K., B.N.H., N.A.T., J.S., N.P.), Munich University Hospital, Germany.
Munich Cluster for Systems Neurology (SyNergy), Germany (X.L., I.K., B.N.H., N.A.T., J.S., N.P.).
Biyan Nathanael Harapan, MD https://orcid.org/0000-0003-4358-3405
Institute for Stroke and Dementia Research (ISD) (X.L., I.K., B.N.H., N.A.T., J.S., N.P.), Munich University Hospital, Germany.
Department of Neurosurgery, Munich University Hospital, Germany (B.N.H., N.A.T., J.S.).
Munich Cluster for Systems Neurology (SyNergy), Germany (X.L., I.K., B.N.H., N.A.T., J.S., N.P.).
Nicole Angela Terpolilli, MD https://orcid.org/0000-0001-7070-3113
Institute for Stroke and Dementia Research (ISD) (X.L., I.K., B.N.H., N.A.T., J.S., N.P.), Munich University Hospital, Germany.
Department of Neurosurgery, Munich University Hospital, Germany (B.N.H., N.A.T., J.S.).
Munich Cluster for Systems Neurology (SyNergy), Germany (X.L., I.K., B.N.H., N.A.T., J.S., N.P.).
Julian Schwarting, MD, BSc* https://orcid.org/0000-0003-0616-5706
Institute for Stroke and Dementia Research (ISD) (X.L., I.K., B.N.H., N.A.T., J.S., N.P.), Munich University Hospital, Germany.
Department of Neurosurgery, Munich University Hospital, Germany (B.N.H., N.A.T., J.S.).
Department of Diagnostic and Interventional Neuroradiology, Klinikum rechts der Isar, Technische University Munich, Germany (J.S.).
Munich Cluster for Systems Neurology (SyNergy), Germany (X.L., I.K., B.N.H., N.A.T., J.S., N.P.).
Institute for Stroke and Dementia Research (ISD) (X.L., I.K., B.N.H., N.A.T., J.S., N.P.), Munich University Hospital, Germany.
Munich Cluster for Systems Neurology (SyNergy), Germany (X.L., I.K., B.N.H., N.A.T., J.S., N.P.).

Notes

For Sources of Funding and Disclosures, see page 2133.
*
J. Schwarting and N. Plesnila contributed equally.
Supplemental Material is available at Supplemental Material.
Correspondence to: Nikolaus Plesnila, MD, Institute for Stroke and Dementia Research, University Hospital, LMU Munich, Feodor-Lynen Strasse 17, 81377 Munich, Germany. Email [email protected]

Disclosures

Disclosures None.

Sources of Funding

This project was supported by the “Fakultätsförderprogramm für Forschung und Lehre” (FöFoLe; project No. 1075) to Dr Schwarting by the Medical Faculty of the University of Munich and by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy within the framework of the Munich Cluster for Systems Neurology (EXC 2145 SyNergy – ID 390857198). Dr Lin was funded by the China Scholarship Council (CSC).

Metrics & Citations

Metrics

Citations

Download Citations

If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Select your manager software from the list below and click Download.

  1. Emerging Role of Neutrophil Extracellular Traps in Subarachnoid Hemorrhage, Stroke, 55, 12, (2882-2884), (2024).https://doi.org/10.1161/STROKEAHA.124.049321
    Crossref
  2. Perivascular Neutrophil Extracellular Traps Exacerbate Microvasospasm After Experimental Subarachnoid Hemorrhage, Stroke, 55, 12, (2872-2881), (2024)./doi/10.1161/STROKEAHA.124.047574
    Abstract
  3. CT perfusion-guided administration of IV milrinone is associated with a reduction in delayed cerebral infarction after subarachnoid hemorrhage, Scientific Reports, 14, 1, (2024).https://doi.org/10.1038/s41598-024-65706-w
    Crossref
  4. A cell-autonomous role for border-associated macrophages in ApoE4 neurovascular dysfunction and susceptibility to white matter injury, Nature Neuroscience, 27, 11, (2138-2151), (2024).https://doi.org/10.1038/s41593-024-01757-6
    Crossref
  5. Perivascular macrophages in cerebrovascular diseases, Experimental Neurology, 374, (114680), (2024).https://doi.org/10.1016/j.expneurol.2024.114680
    Crossref
  6. Single-Cell Transcriptomics Revealed White Matter Repair Following Subarachnoid Hemorrhage, Translational Stroke Research, (2024).https://doi.org/10.1007/s12975-024-01265-6
    Crossref
  7. All Three Supersystems—Nervous, Vascular, and Immune—Contribute to the Cortical Infarcts After Subarachnoid Hemorrhage, Translational Stroke Research, (2024).https://doi.org/10.1007/s12975-024-01242-z
    Crossref
  8. Endothelial cells and macrophages as allies in the healthy and diseased brain, Acta Neuropathologica, 147, 1, (2024).https://doi.org/10.1007/s00401-024-02695-0
    Crossref
  9. Depletion of perivascular macrophages delays ALS disease progression by ameliorating blood-spinal cord barrier impairment in SOD1G93A mice, Frontiers in Cellular Neuroscience, 17, (2023).https://doi.org/10.3389/fncel.2023.1291673
    Crossref
Loading...

View Options

View options

PDF and All Supplements

Download PDF and All Supplements

PDF/EPUB

View PDF/EPUB
Login options

Check if you have access through your login credentials or your institution to get full access on this article.

Personal login Institutional Login
Purchase Options

Purchase this article to access the full text.

Purchase access to this article for 24 hours

Perivascular Macrophages Mediate Microvasospasms After Experimental Subarachnoid Hemorrhage
Stroke
  • Vol. 54
  • No. 8

Purchase access to this journal for 24 hours

Stroke
  • Vol. 54
  • No. 8
Restore your content access

Enter your email address to restore your content access:

Note: This functionality works only for purchases done as a guest. If you already have an account, log in to access the content to which you are entitled.

Media

Figures

Other

Tables

Share

Share

Share article link

Share

Comment Response