Therapeutic effect of platelet-rich plasma and chondroitinase abc combination on rat spinal cord injuries

Traumatic spinal cord injury is an important health problem with high mortality-morbidity rates and devastating social, physiological and psychological effects worldwide. Despite all the advances in treatment methods, the survival and well-being rate in patients with spinal cord injuries is very low. Despite all experimental and clinical research, neuroprotection and regeneration of the injured spinal cord have not been achieved in human spinal cord injuries. Neuroprotection studies have been carried out for a long time to stop the secondary spinal cord injury cascade, which includes molecular and cellular changes that begin within minutes following primary spinal cord injury and can continue for days or even weeks. Targets for the prevention of secondary damage in spinal cord injury are: glutamatergic, cholinergic and catholinergic neurotransmission systems, free radical production, lipid peroxidation, calcium and other ion channels, growth factors, neurotrophic factors, inflammation19 process, endogenous opioid receptors, enzymes, apoptotic cell death and regeneration mechanisms. Platelet-rich plasma (PRP) is used after centrifugation of fresh blood with an anticoagulant. It is prepared by collecting the platelet-rich part. It has been reported that intrathecal (i.t.) PRP application induces microglia and astrocyte activation and PDGF-B, ICAM-1, GAP43 expression. A soluble enzyme chondroitinase ABC (ChABC) forms the carbohydrate branch of CSPG molecules. CSPGs are extracellular cell molecules that are upregulated around sites of damage in the central nerve, inhibiting the neural growth process in vitro and in vivo. ChABC allows further growth and increases central axon regeneration. When we scanned the literature, we did not find any publications containing the effects of intrathecal (i.t.) PRP and intrathecal (i.t.) ChABC combined application as spinal cord injury treatment. In this research project, we planned to show that the combined application of intrathecal (i.t.) PRP and i.t. ChABC will induce GAP43 expression, and also significantly increase axonal regeneration and significantly support spinal cord recovery. Method: Our study was carried out with the approval decision of Balıkesir University Faculty of Medicine Animal Experiments Ethics Committee dated 24/03/2022 and numbered 2022/2-5. The experimental part of the study was carried out in the Experimental Animals Laboratory of Balıkesir University Faculty of Medicine, and the histopathological studies were carried out in the Pathology Laboratory of Balıkesir University. 31 female rats, 2.5 months old and weighing 250-300 g, were used in our experimental groups.20 One rat was separated to prepare PRP. Randomly selected rats were divided into 5 groups, with 6 rats in each group: Group1 (Control Group): Only T9-T10 laminectomy was performed. Group 2 (Physiological Saline Group): T9-T10 laminectomy was performed and spinal cord damage was done. 1st hour after cord injury, 1st,3rd, 5th. and 6. A total of 5 times per day. Physiological Saline was applied. Group 3 (PRP Group): T9-T10 laminectomy was performed and spinal cord damage was done. 1st hour after cord injury, 1st,3rd, 5th. and 6. A total of 5 times per day. PRP was applied. Group 4 (Chondroitinase ABC Group): The rats in this group underwent T9-T10 laminectomy and spinal cord damage. 1st hour after cord injury, 1st,3rd, 5th. and 6. A total of 5 times per day. Chondroitinase ABC was applied. Group 5 (Chondroitinase ABC +PRP Group): The rats in this group underwent T9-T10 laminectomy and spinal cord damage. 1st hour after cord injury, 1st,3rd, 5th. and 6. A total of 5 times per day. Chondroitinase ABC and PRP were applied. In the 2nd week, the experimental animals were sacrificed and the spinal cord samples were taken and delivered in tubes containing formaldehyde to Pathology Department Laboratory of Balıkesir University. Sections were stained with hematoxylin and eosin for histopathological examination. Histomorphological changes between groups were evaluated. Additionally, GAP43 was used as the primary antibody for immunohistochemical staining.21 Statistical Analysis IBM SPSS V23 was used for all statistical comparisons. Fisher Freeman Halton test was used to compare categorical data, and multiple comparisons were examined with the Bonferroni Corrected Z test. Results: There is a statistically significant connection between groups and axonal regeneration (p<0.001). There were differences between the control and saline groups without axonal regeneration and other groups. There were differences between ChABC and Control, PRP+ChABC and SF groups in 1+ regeneration. In 2+ regeneration, PRP+ChABC and ChABC, control and SF groups showed differences. In the ChABC group, 20% had +1 staining, 80% had +2 staining, 0% had +3 staining ,100% of the control group had +1 staining, 0% had +2 staining, 0% had +3 staining.+3 staining 20% of the PRP group had +1 staining, 80% had +2 staining, 0% had +3 staining,In the PRP+ChABC group, 0% had +1 staining, 0% had +2 staining, 100% had +3 staining, 100% of the SF group had +1 staining, 0% had +2 staining, and 0% had +3 staining. There is a statistically significant connection between the groups and GAP43 staining rate (p<0.001). Control and SF and PRP+ChABC groups showed differences in +1 staining. Other groups showed differences in +3 staining with PRP+ChABC. Conclusion: In our study, the combined İt administration of PRP + ChABC has been shown to have a significant histopathological benefit compared to separate PRP and ChABC applications, and these results are statistically significant.22 The results of our study is stimulating a new researches in this field. Further cellular-based and clinical studies on this subject may be promising for patients and their relatives in the future.