Augmenting Peripheral Nerve Regeneration Using Hair Follicle Stem Cells in Rats.

Introduction: Cell therapy is the most advanced treatment of peripheral nerve injury. This study aimed to determine the effects of transplantation of hair follicle stem cells on the regeneration of the sciatic nerve injury in rats. Methods: The bulge region of the rat whisker were isolated and cultured. Morphological and biological features of the cultured bulge cells were observed by light microscopy and immunocytochemistry methods. Percentages of CD34, K15, and nestin cell markers expression were demonstrated by flow cytometry. Rats were randomly divided into 3 groups of injury, epineurium, and epineurium with cells in which rat Hair Follicular Stem Cells (rHFSCs) were injected into the site of the nerve cut. HFSCs were labeled with Bromodeoxyuridine (BrdU), and double-labeling immunofluorescence was performed to study the survival and differentiation of the grafted cells. After 8 weeks, electrophysiological, histological, and immunocytochemical analysis assessments were performed. Results: Rat hair follicle stem cells are suitable for cell culture, proliferation, and differentiation. The results suggest that transplantation of rat hair follicle stem cells can regenerate sciatic nerve injury; moreover, electrophysiology and histology examinations show that sciatic nerve repair was more effective in the epineurium with cell group than in the other experimental group (P<0.05). Conclusion: The achieved results propose that hair follicle stem cells improve axonal growth and functional recovery after peripheral nerve injury. Highlights: This study showed that rat hair follicle stem cells are suitable for cell culture, proliferation and differentiationThe results suggested that transplantation of rat hair follicle stem cells had the potential capability of regenerating sciatic nerve injuryEvidence of electrophysiology and histology showed Concomitant use of epineurium with hair follicle stem cell was more effective repairment. Plain Language Summary: Although repairing damaged peripheral nerves has always been a medical challenge, but peripheral nerve injury has been successfully repaired using various procedures such as nerve auto-graft or stem cell therapy. The functional reconstruction is the most important after therapy because of that primary nerve repair or use of nerve autograft, are still accepted as golden standard methods for treatment. Considerable recent interest has been focused on adult stem cells for both research and clinical applications. A highly promising source of relatively abundant and accessible, active, multipotent adult stem cells are obtained from hair follicles. In research the hair follicle stem cells implanted into the gap region of a severed sciatic nerve injury greatly enhanced the rate of nerve regeneration and the restoration of nerve function. Time is one of the several aspects require specific attention in the clinical treatment of peripheral nerve injury. Because delay of nerve injury treatment may cause neurobiological alterations in neurons and Schwann cells, impairing nerve functional recovery and affect neuron survival. In this study, concluded that stem cell injection 2 weeks after injury in the damaged nerve epineurium repairs nerve fibers, while electrophysiology of the leg muscles showed that muscle function was significantly improved. It indicates the repair of muscular innervation and nerve repair. The results pave the way for further research on this topic.

Geraniol attenuates oxidative stress, bioaccumulation, serological and histopathological changes during aluminum chloride-hepatopancreatic toxicity in male Wistar rats.

Aluminum chloride (AlCl3) has different industrial applications including manufacturing paint and water treatment. The present study was designed to evaluate the alleviating effect of geraniol against AlCl3-induced hepatopancreatic toxicity. To this end, forty male Wistar rats were divided into control (0.9% NaCl, IP), geraniol (100 mg/kg orally), AlCl3 (70 mg/kg, IP), and AlCl3 (70 mg/kg, IP) plus geraniol (100 mg/kg orally) groups and then were treated daily for 28 days. Based on the results, serum cholesterol, triglyceride, as well as liver and pancreas enzymes increased significantly (P < 0.05) while the level of insulin significantly decreased in AlCl3-treated rats compared to the control group (P < 0.05). The presence of geraniol relieved the toxic effects of AlCl3 as well. On the other hand, the level of malondialdehyde (MDA) increased in the AlCl3-treated group while the activities of glutathione peroxidase and the total antioxidant activity demonstrated a reduction. However, the MDA level decreased while the antioxidant enzymes increased in co-treated with geraniol group. Histopathological examination revealed that simultaneous treatment with geraniol in AlCl3 intoxicated rats ameliorate the liver lesions such as necrosis, inflammatory cell infiltration, vacuolar degeneration, along with hyperemia and the cell density of the Langerhans islands. Finally, the results indicated that geraniol attenuated the side effect of AlCl3-induced hepatopancreatic toxicity.

Protective Effect of Rosa damascena Against Aluminum Chloride-Induced Oxidative Stress.

Aluminum is considered an essential element endowed with toxicity potentials in human and animal. Thus, intoxication with aluminum can lead to oxidative stress, which is associated with oxidative damage to various macromolecules. Moreover, antioxidants from natural sources can play an important role in human health. Accordingly, the purpose of this study was to investigate the protective effect of Rosa damascena extract against aluminum-induced oxidative stress. In this study, 60 male rats were randomly divided into six groups and then they were given daily aluminum chloride and Rosa damascena extract. After 8 weeks of treatment, the levels of total antioxidant and malondialdehyde, as well as antioxidant enzymes including catalase, glutathione S-transferase, and myeloperoxidase, were measured in all experimental groups in this study. A significant increase was found in the total antioxidant level in the rats treated with aluminum, Rosa damascena extract, and aluminum plus Rosa damascena extract compared with those in the control group. Also, malondialdehyde levels were not significantly different in all the studied groups. Glutathione S-transferase activity levels in rats receiving the Rosa damascena extract as well as rats taking aluminum with Rosa damascena extract increased significantly compared with the ones in the control group. Catalase activity in the aluminum-treated group also increased significantly compared with the rates in the control group (31.34 ± 4.50 U/gHb vs. 14.04 ± 6.17 U/gHb, p = 0.014). Furthermore, myeloperoxidase activity in the aluminum-treated group increased significantly compared with the control group (49.47 ± 5.12 U/L vs. 25.28 ± 2.18 U/L, p < 0.001). The Rosa damascena extract could improve antioxidant capacity and reduce oxidative conditions in rats receiving aluminum chloride as evidenced by assays of the ferric reducing ability of plasma and activity of antioxidant enzymes. According to the findings of this study, it can be concluded that the Rosa damascena extract with its high antioxidant content is able to exert a protective effect against aluminum-induced oxidative stress.

The role of biodegradable engineered nanofiber scaffolds seeded with hair follicle stem cells for tissue engineering.

BACKGROUND: The aim of this study was to fabricate the poly caprolactone (PCL) aligned nanofiber scaffold and to evaluate the survival, adhesion, proliferation, and differentiation of rat hair follicle stem cells (HFSC) in the graft material using electrospun PCL nanofiber scaffold for tissue engineering applications. METHODS: The bulge region of rat whisker was isolated and cultured in DMEM: nutrient mixture F-12 supplemented with epidermal growth factor. The morphological and biological features of cultured bulge cells were observed by light microscopy using immunocytochemistry methods. Electrospinning was used for production of PCL nanofiber scaffolds. Scanning electron microscopy (SEM), 3-(4, 5-di-methylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and histology analysis were used to investigate the cell morphology, viability, attachment and infiltration of the HFSC on the PCL nanofiber scaffolds. RESULTS: The results of the MTT assay showed cell viability and cell proliferation of the HFSC on PCL nanofiber scaffolds. SEM microscopy images indicated that HFSC are attached, proliferated and spread on PCL nanofiber scaffolds. Also, immunocytochemical analysis showed cell infiltration and cell differentiation on the scaffolds. CONCLUSION: The results of this study reveal that PCL nanofiber scaffolds are suitable for cell culture, proliferation, differentiation and attachment. Furthermore, HFSC are attached and proliferated on PCL nanofiber scaffolds.

BD PuraMatrix peptide hydrogel as a culture system for human fetal Schwann cells in spinal cord regeneration.

BD PuraMatrix peptide hydrogel, a three-dimensional cell culture model of nanofiber scaffold derived from the self-assembling peptide RADA16, has been applied to regenerative tissue repair in order to develop novel nanomedicine systems. In this study with PuraMatrix, self-assembling nanofiber scaffold (SAPNS) and Schwann cells (SCs) were isolated from human fetal sciatic nerves, cultured within SAPNS, and then transplanted into the spinal cord after injury (SCI) in rats. First, the peptide nanofiber scaffold was evaluated via scanning electron microscopy and atomic force microscopy. With phase-contrast microscopy, the appearance of representative human fetal SCs encapsulated in PuraMatrix on days 3, 5, and 7 in 12-well plates was revealed. The Schwann cells in PuraMatrix were cultured for 2 days, and the SCs had active proliferative potential. Spinal cord injury was induced by placing a 35-g weight on the dura of T9-T10 segments for 15 min, followed by in vivo treatment with SAPNS and human fetal SCs (100,000 cells/10 µl/injection) grafted into spinal cord 7 days after SCI. After treatment, the recovery of motor function was assessed periodically using the Basso, Beattie, and Bresnahan scoring system. Eight weeks after grafting, animals were perfusion fixed, and the survival of implanted cells was analyzed with antibody recognizing SCs. Immunohistochemical analysis of grafted lumber segments at 8 weeks after grafting revealed reduced asterogliosis and considerably increased infiltration of endogenous S100(+) cells into the injury site, suggesting that PuraMatrix may play an important role in the repair observed after SAPNS and human fetal SC transplantation.