Table 3 Summarizing the data of all preclinical studies in AD specifying the design, objectives, methodology, results, and conclusion

35

Animal study (5xFAD mice model)

To assess the effect of rTMS on glial activation and Aβ accumulation

High-frequency wide-field rTMS applied to 5xFAD mice

Downregulation of microglia (Iba +) and astrocyte (GFAP +/VIM +) activation. Reduction of Aβ deposits in mPFC, dentate gyrus, and CA3

rTMS suppresses plaque accumulation by regulating glial activation and reducing Aβ deposits

36

Animal study (STZ-induced AD model)

To examine rTMS effects on glial activation and neuroinflammation

Intermittent theta-burst stimulation (iTBS) in AD mice model

Significant downregulation of microglia and astrocyte activation in hippocampus and periventricular area

rTMS reduces glial activation and neuroinflammation in AD models, improving brain function

37

Animal study (5xFAD mice model)

To investigate rTMS effects on cytokine release and the PI3 K/Akt/NF-κB pathway

20-Hz rTMS applied to 5xFAD mice. Measurement of cytokine levels and PI3 K/Akt/NF-κB signaling

Decreased TNF-α levels and regulation of the PI3 K/Akt/NF-κB pathway

rTMS reduces pro-inflammatory cytokines through the modulation of immune signaling pathways

38

In-vitro and animal study (mouse organotypic brain cultures)

To assess the role of microglia and cytokine release in rTMS-induced plasticity

10-Hz rTMS applied to brain tissue cultures and in-vivo microglial depletion

rTMS induced synaptic plasticity via cytokine release; microglial depletion abolished the changes in neurotransmission

Microglia and cytokine release are key targets for rTMS-induced changes in neural excitability and plasticity

39

Animal study (APP/PS1 dE9 mice model)

To assess the effect of FUS combined with microbubbles (FUS-BBBD) on microglia and plaque suppression

Bilateral FUS-BBBD with 07/2a mAb injection in APP/PS1 dE9 mice

Increased monocyte infiltration and recruitment to plaques, enhanced plaque suppression, and improved cognitive function

FUS-BBBD combined with monoclonal antibody treatment enhances plaque suppression and cognitive function

40

Animal study (rTg4510 tau pathology model)

To investigate the effect of FUS-BBBD on tau pathology and immune response

Unilateral FUS-BBBD treatment in tau pathology model mice

Reduction in phosphorylated tau levels, correlated with microglial activation

FUS-BBBD treatment reduces tau pathology and activates immune responses, potentially aiding disease modification

41

Animal study (APP/PS1 dE9 mice model)

To investigate the effects of FUS-BBBD combined with mAb treatment on immune responses and plaque pathology

FUS-BBBD treatment combined with 07/2a mAb injection

Significant immune responses, upregulation of IgG2a mAb levels, increased phagocytosis, and plaque suppression

FUS-BBBD enhances immune response and plaque suppression, improving cognitive outcomes

42

Animal study (Sprague Dawley rats)

To assess the long-term effects of SIR induced by pFUS-MB

pFUS-MB treatment in rats with MRI analysis of brain changes

Persistent BBB disruption, cortical atrophy, and astrogliosis in treated rats

pFUS-MB treatment induces long-term changes in the brain, including cortical atrophy and gliosis

43

Animal study (TgCRND8 mice)

To assess the effects of MRIgFUS and scyllo-inositol on amyloid plaques and astrocyte activation

MRIgFUS followed by scyllo-inositol treatment in TgCRND8 mice

Decreased amyloid concentrations and reduced astrocyte activation in hippocampus and cortex

MRIgFUS combined with scyllo-inositol treatment decreases amyloid burden and astrocyte activation

44

Animal study (TgCRND8 mice model)

To investigate the effect of FUS-BBBD on leukocyte infiltration and β-amyloid pathology

FUS-BBBD and 07/2a mAb treatment in TgCRND8 mice

Increased leukocyte infiltration and reduced β-amyloid pathology

FUS-BBBD enhances leukocyte infiltration, helping to reduce amyloid pathology