Bone Marrow Mesenchymal Stem Cell Preconditioning With Alpha Pinene Reverses Ischemia Induced Neuronal Degeneration in Rats

Background: Preconditioned stem cells may offer a promising approach to treat various neurological disorders. Compounds like alpha pinene (AP) possess significant bioactive properties and may enhance the regeneration potential of bone marrow mesenchymal stem cells (BM-MSCs) following cerebral ischemia in rats. Methods: BM-MSCs were cultured, characterized, and treated with the non-cytotoxic concentration of AP and analyzed for the expression of neuronal markers. Rats were randomly divided into four experimental groups; normal control (NC), disease control (DC), and normal MSC (N-MSC) and AP treated MSC (AP-MSC) transplanted groups. Ischemia was induced by ligating the carotid artery. Normal- and AP-MSCs (2 × 10⁶ cells) were transplanted after 24 h of ischemia induction in their respective groups via tail vein. Brain was harvested from each group and assessed at transcriptional (hypoxia and inflammatory mediators), histochemical (TTC staining), and histopathological (H&E staining) levels. Furthermore, in vivo differentiation of MSCs were evaluated by NeuN immunostaining. Results: Normal and AP-MSCs after transplantation showed downregulation of hypoxia and inflammatory markers compared to DC group. Histological analysis of DC indicated neuronal damage, perivascular edema, vacuolation, and distorted tissue architecture in cortex and thalamus, whereas structural integrity was restored in both the transplanted groups. Immunostaining also revealed the expression of NeuN in transplanted MSCs. Conclusion: The study demonstrates the protective role of preconditioned BM-MSCs in reversing the ischemia induced neuronal damage. After subsequent experimental validation, the study findings may offer promising treatment modality against neuronal loss due to ischemic injuries. Lay Summary: Cerebral ischemia also known as ischemic stroke, is the second leading cause of death globally. It induced neuronal injury resulting in complete or partial loss of function of the affected area. Stem cell therapy offers promising option to repair neuronal cells and facilitate recovery of the damaged area. Stem cell treatment with bioactive molecules prior to their transplantation can enhance their regenerative capabilities. The current study treated bone marrow derived mesenchymal stem cells (BM-MSCs) with alpha pinene and explored their regenerative potential in an animal model of cerebral ischemia. The study findings demonstrated that preconditioned BM-MSCs possess the potential to reverse neuronal damage. After conducting subsequent validation through long term functional studies, the current findings may provide a promising therapeutic approach against central nervous system pathologies.