Project Summary: The sequelae of traumatic brain injury (TBI) pathology comprises primary injury where impacting force(es) induce physical damage, including the breakdown of the blood-brain barrier (BBB). Thus the primary injury launches secondary injury comprising chronic inflammation that includes migration of leukocytes, gliosis, and the release of numerous inflammatory mediators. Thus, secondary injury exacerbates the damage incurred during the primary injury and is considered more devastating. Though various drugs are tested to curtail secondary injury, these trials have failed, and the prospect of identifying a TBI drug looks bleak. Hence, the treatment options for TBI survivors are restricted to palliative care. Recently, there have been several publications emphasizing the neuroprotective and anti-inflammatory effects of various diets. Additionally, since cardiovascular diseases (CVDs) are one of the most important comorbid changes in TBI survivors, studies emphasizing the role of CVD in TBI pathology are non-existent. Our proposed research aims to compare the beneficial effects of a low-fat, high-fiber diet (LFHFD) in reducing TBI-associated BBB breakdown, neuroinflammation, and immune response over the standard western diet (WD) that is high-fat and cholesterol, and low in fiber in a mouse model with chronic CVD and TBI. We are employing American Heart Association (AHA) recommended mouse model (LDL receptor-deficient mouse model (LDLr-KO)) for this study. We hypothesize that the uptake of WD further exacerbates the combined effects of CVD and TBI and will cause chronic BBB dysfunction, predisposing individuals to chronic inflammatory changes. To test this hypothesis, we will maintain LDLr-KO mice either on WD or LFHFD from the second month to the sixth month. In the first week of the seventh month, half of the LDLr-KO mice maintained on a WD or LFHFD diet will undergo TBI (Controlled Cortical Impact, CCI) or Sham injury (Craniotomy without CCI). All animals on their respective diets will be maintained for three to six months. Animals will be euthanized at the end of the 9th and 12th month, and brain samples will be extracted and processed for routine paraffin-embedded tissue processing and bright field immunohistochemistry for investing BBB function (aim 1). We will also study the impact of diets on the expression of various neuroinflammatory markers and overall immune responses (aim 2). A second set of animals will be prepared, and their brain samples will be used for protein and gene expression studies of various neuroinflammation markers by employing Western blot and real-time polymerase chain reaction. We will also collect pre-injury and post-injury blood samples from these mice and run a Luminex assay to monitor serum levels of various immune mediators in blood samples. We expect WD to exacerbate BBB dysfunction and inflammatory sequelae compared to LFHFD. Upon completing this study, we will have preliminary data emphasizing the beneficial effects of LFHFD over WD in reducing BBB permeability, neuroinflammation, and immune responses in long-term TBI survivors.

Project Narrative Since most Traumatic Brain Injury (TBI) drug trials have failed, medical intervention options for long-term TBI survivors with chronic TBI and cerebrovascular disease (CVD) are limited. Since a considerable number of research articles highlighting the neuroprotective and anti-inflammatory effects of various diets have been published, we are proposing to study the beneficial effects of a low-fat, high-fiber diet (LFHFD) over high-fat and cholesterol, and low-in-fiber (standard western diet, WD) in an American Heart Association recommended mouse model with chronic CVD and TBI. We will study the impact of these two diets in reinstating BBB integrity and reducing neuroinflammatory and systemic immune responses.