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Cranial Osteopathic Manipulation Alters Alzheimer’s Disease Phenotype in Wild Type and Transgenic Rats

Journal: Journal of Osteopathic Medicine Date: 2023/12, 123(12):Pages: A17-A19. doi: Subito , type of study: animal experiment

Full text    (https://www.degruyter.com/document/doi/10.1515/jom-2023-2000/html)

Keywords:

animal experiment [36]
cranio-sacral osteopathy [158]

Abstract:

Statement of Significance: Reduced fluid circulation in the brain can lead to a build-up of macromolecules like amyloid beta (Aβ), which cause neuroinflammation and has been linked to Alzheimer’s [1,2]. Manipulating the CNS lymphatic vessels with Cranial Osteopathic Manipulation (COM) to increase fluid circulation has reduced Aβ, activated astrocytes, and improved excitatory neurotransmission for aged rats in our pilot study. To study the effect of Cranial Osteopathic Manipulative Medicine on biochemical and cognitive function in transgenic and naturally aged rat models of Alzheimer’s disease. Research Methods: This part of the study was split into two phases, Phase 4 & Phase 5. Phase 4 had 11 three-month-old wild-type rats while Phase 5 had 12 Fischer 344 transgenic rats. Male and female subjects were used for both phases. Phase 4 serves as a negative control group for Phase 5 and future studies, since subjects will not have a build-up of Aβ at a younger age. Rats that had open wounds or reproductive tumors were excluded from participating. All remaining rats that met this criterion were randomly split, using simple randomization, into COM (treated) and untreated (UT) groups, with the same number of males and females in each group before exclusion. To minimize treatment errors an osteopathic doctor performed the COM treatment. To quantify pressure applied to the occipital squama, FingerTPS nanosensor gloves were used. The gloves fit individually over each finger and have a sensor that can be calibrated per user. To assess changes in learning and memory the Morris Water Maze assay was used for both phases, and the Novel Object Recognition Assay (NORA) was used for rats in Phase 5. MWM assessed this by analyzing the path trajectory and quickness of a subject as they attempt to reach a platform located in one of four quadrants in a circular tub filled with water. MWM assay was performed over 8 days with days 1-4 being training days and 5-8 being experimental days. The platform was submerged on days 5-7 and missing on day 8. NORA was performed to assess the ability to recognize a novel object, under the principle that rodents prefer novelty. Handlers were blinded for both behavioral assays. To assess molecular changes, a proteome analysis was performed. The proteome analysis identified genes that were differentially expressed between COM and UT groups on the protein level. These experiments were conducted to evaluate the effect of COM on cognitive function. Data Analysis: The average duration of COM treatment was 200-250 seconds for 6 rats from Phase 4. Average force varied with the male rats receiving less force on average than the female rats. For behavioral assays, the Holm-Šídák multiple comparisons T-Test for days 5-8 was used for analysis. Phase 4 had 15 measures that were statistically significant with p-values less than 0.05: (1) Shortest Visit to the NW Zone, (2) Maximum Distance from the NW Zone, (3) Maximum Distance from the Platform Zone, (4) Time Moving away from the Platform Zone, (5) Distance Traveled in the Platform zone, (6) Average Speed in the Platform Zone, (7) Average Absolute Heading Error to the Platform Zone, (8) Number of Entries to the NW Zone, (9) Number of Exits from the NW Zone, (10) Number of Line Crossings in the NW Zone, (11) Number of Entries to the Platform Zone, (12) Number of Exits from the Platform Zone, (13) Time in the Platform Zone, (14) Time Moving towards the Platform Zone, and (15) Number of Line Crossings in the Platform Zone. While Phase 5 had 10 measures: (1) Average speed, (2) Time Moving towards the NW Zone, (3) Signed Initial Heading Error, (4) Shortest Visit to the NW Zone, (5) Signed Initial Heading Error to the Platform Zone, (6) Maximum Distance to the Platform Zone, (7) Average Distance to the Platform Zone, (8) Path efficiency, (9) Clockwise Rotations of the Animal’s Body, and (10) Latency to Last Entry to the Platform Zone. For NORA the Phase 4 COM-treated rats spent numerically more time (COM, 31.4±13.7 vs 26.6±16.51s; UT, 15.5±3.25 vs 24.0 ±12.0s) exploring the novel object. For the proteomic analysis, the abundance ratio between COM and UT was used to identify p-values less than 0.05 for medium confidence and 0.01 for high confidence. The collected hippocampal tissue from the Phase 5 transgenic rats identified that COM significantly increased the expression of serine-threonine kinase P21-activated kinase (PAK3), and fifty other proteins. Conclusion: These results indicate that COM induced improvements in spatial learning and memory behavior parameters for transgenic and wild-type rats. In addition, the proteome analysis highlights increased expression of proteins associated with Alzheimer’s Disease and other neurological disorders. Further analysis on two cohorts with larger sample sizes, Phase 6 and Phase 7, are underway to corroborate current findings. The outcomes of this study would help justify the clinical use of COM as a minimally invasive non-pharmacological treatment strategy for Alzheimer’s disease.


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