Adipose Mesenchymal Stem Cell Metabolites Oral Gel Enhance Pro-Angiogenic Factors Expression, Angiogenesis, and Clinical Outcome of Oral Ulcer Rat Model.

OBJECTIVE: Enhancing wound healing capacity is one of the main principles in oral ulcer management. Efficient oral ulcer management will accelerate clinical symptom amelioration and prevent complications. Adipose mesenchymal stem cell metabolites (AdMSCM), a novel biological product, contains a plethora of bioactive mediators that can induce a series of processes in wound healing. This study will analyze the clinical outcome, angiogenesis, and expression of FGF-2 and VEGFA in the oral ulcer rat model after AdMSCM oral gel application. MATERIALS AND METHODS: Twenty healthy male Wistar rats (Rattus novergicus) were used to create oral ulcer animal models. AdMSCM oral gel treatment was performed three times daily for 3 and 7 days. Clinical outcome was assessed by measuring the major diameter of the ulcer; the angiogenesis was evaluated through histological assessment; the expression of VEGFA and FGF-2 was assessed using the immunohistochemistry method. STATISTICAL ANALYSIS: This study uses parametric comparative analysis using one-way analysis of variance (ANOVA) and post-hoc Tukey's HSD test RESULTS: The application of AdMSCM oral gel in an oral ulcer rat model significantly enhanced the clinical outcome (p < 0.05). In addition, similar results were shown in the histologic assessment of angiogenesis and supported by the significant increase of VEGFA and FGF-2 expression. CONCLUSIONS: AdMSCM oral gel accelerates oral ulcer healing processes, proven by the enhancement of angiogenesis, pro-angiogenic factors expression, and clinical outcomes.

Modification of the height of a weight drop traumatic brain injury model that causes the formation of glial scar and cognitive impairment in rats.

OBJECTIVE: Traumatic brain injury (TBI) is a chronic, progressive condition associated with permanent disabilities, particularly cognitive impairments. Glial scar formation following TBI is considered a contributing factor to these persistent disabilities. Currently, limited research exists on pharmacological interventions targeting glial scar prevention that require a standard weight drop TBI model for glial scar formation. Since there is no established standard TBI model for glial scar formation, this study aims to validate and modify the height of the weight drop model to identify glial scar formation and cognitive impairments. METHODS: Fifteen male Sprague Dawley rats were randomly divided into sham, WD1, and WD2 groups. The weight drop model with a 10 g load was applied to the right exposed brain of the rats from a height of 5 cm (WD1) and 10 cm (WD2) using a modified Feeney's weight drop device. Cognitive impairments were confirmed using the novel object recognition (NOR) test with ethovision software on day 15. Subsequently, the rats were decapitated on day 16, and GFAP immunohistochemical staining was performed to confirm the presence of glial scarring. RESULTS: The WD1 and WD2 groups exhibited a significant increase in glial scar formation compared to the sham group, with the WD2 group resulting in even more pronounced glial scar formation. Only the WD2 model caused statistically significant cognitive damage. The negative correlation coefficient indicates that an increase in GFAP + cells will decrease the cognitive function. CONCLUSION: Modification of the height of the weight drop model, by dropping a weight of 10 g from a height of 10 cm (WD2 group) onto the right brain exposed of the rat has been proven to induce the formation of a glial scar and cognitive impairment.

RANK-RANKL-OPG expression after gingival mesenchymal stem cell hypoxia preconditioned application in an orthodontic tooth movement animal model.

Background: The expression of receptor activator of Nuclear Factor Kappa Beta (RANK) and its ligand (RANKL), as well as osteoprotegrin (OPG), in the alveolar bone (AB), may improve bone remodeling during orthodontic tooth movement (OTM). It is hypothesized that hypoxia-preconditioned gingival mesenchymal stem cells (GMSC) may be more effective than normoxia-preconditioned GMSC in this regard. This study aims to investigate the expression of RANK, RANKL, and OPG in the compression and tension sides of AB after allogeneic administration of GMSC that were normoxia or hypoxia-preconditioned in rabbits (Oryctolagus cuniculus) undergoing OTM. Methods: Twenty-four healthy young male rabbits were divided into two groups: T1, which underwent OTM and received normoxia-preconditioned GMSC, and T2, which underwent OTM and received hypoxia-preconditioned GMSC. A ligature wire was attached to the mandibular first molar and connected to a 50 g/mm2 closed coil spring, exerting force on the central incisor and left mandibular molar of the experimental animals. After 24 h of OTM, either normoxia- or hypoxia-preconditioned GMSC were injected into the gingiva of the samples in a single dose of 20 µl of phosphate-buffered saline (PBS). All samples were sacrificed on days 7, 14, and 28, and immunohistochemistry was performed to analyze the expression of RANK, RANKL, and OPG on the tension and compression sides. Results: The expressions of RANK-RANKL-OPG in the alveolar bone of the compression and tension sides were significantly different during the 14-day period of OTM following allogeneic administration of GMSC that were normoxia or hypoxia-preconditioned (p < 0.05). Conclusion: The expression of RANK-RANKL was significantly increased on the compression side of the alveolar bone during OTM after the administration of hypoxia-preconditioned allogeneic GMSC but not on the tension side. Conversely, RANKL-OPG expression was enhanced on the tension side but not on the compression side, as observed through immunohistochemical analysis in vivo.

Gingiva Mesenchymal Stem Cells Normoxic or Hypoxic Preconditioned Application Under Orthodontic Mechanical Force on Osterix, Osteopontin, and ALP Expression.

OBJECTIVES: The aim of this article was to investigate Osterix, ALP, and osteopontin expression in the compression and tension sides of alveolar bone after the application of normoxic/hypoxic-preconditioned GMSCs in rabbits (Oryctolagus cuniculus) induced with OMF. MATERIALS AND METHODS: Forty-eight healthy, young male rabbits were divided into four groups: [-] OMF; [+] OMF; OMF with GMSCs normoxic-preconditioned; and OMF and GMSCs hypoxic-preconditioned. The central incisor and left mandibular molar in the experimental animals were moved, the mandibular first molar was moved mesially using nickel titanium (NiTi) and stainless steel ligature wire connected to a 50 g/mm2 light force closed coil spring. Allogeneic application of normoxic or hypoxic-preconditioned GMSCs was used in as many as 106 cells in a 20 µL phosphate buffered saline single dose and injected into experimental animals' gingiva after 1 day of OTM. On days 7, 14, and 28, all experimental animals were euthanized. Osterix, ALP, and osteopontin expressions were examined by immunohistochemistry. RESULTS: Osterix, ALP, and osteopontin expressions were significantly different after allogeneic application of hypoxic-preconditioned GMSCs than normoxic-preconditioned GMSCs in the tension and compression of the alveolar bone side during OMF (p < 0.05). CONCLUSION: Osterix, ALP, and osteopontin expressions were significantly more enhanced post-transplantation of GMSCs with hypoxic-preconditioning than after transplantation of normoxic-preconditioned GMSCs in rabbits (O. cuniculus) induced with OMF.

Effect of the olive leaf extract in chronic spinal cord injury model: an experimental research.

Posttraumatic myelopathy is defined as a spinal cord injury (SCI) that results in varying degrees of motor and sensory deficits. The degree of 'secondary damage,' which is caused by a variety of cellular, molecular, and biochemical cascades is linked to the outcome of SCI. According to research, the beneficial effects of oleuropein and its derivatives have been linked to radical scavenging/antioxidant actions and anti-inflammatory effects. Materials and Methods: This study was divided into six groups: control negative (sham-operated) group, control positive 1 and 2 (early chronic and chronic), treatment groups 1, 2, and 3 (prophylactic, concomitant, and late). Olive leaf extract (OLE) given dose was 350 mg/kg body weight. Blood was taken from the left corotic artery before the animals were terminated, seromarker assessment, enzyme-linked immunosorbent assay of IL-6, TNF-α, brain-derived neurotrophic factor (BDNF), and assessment of functional motoric outcome before the animal was terminated. Results: Chronic spinal cord compression increased serum levels of IL-6, TNF-α, and decreased serum level of BDNF. OLE 350 mg/kg body weight decreased serum levels of IL-6, TNF-α and increased functional motoric outcome, especially in prophylactic and concomitant therapy. Discussion: These findings indicate that OLE may be effective in protecting chronic SCI model. Conclusion: Oleuropein has a potential effect to reduce the IL-6 and TNF-α in rabbit model of SCI, and the BDNF value risen after the administration of Oleuropein.

Gingival Mesenchymal Stem Cells Metabolite Decreasing TRAP, NFATc1, and Sclerostin Expression in LPS-Associated Inflammatory Osteolysis In Vivo.

OBJECTIVE: Bone is a dynamic tissue that undergoes remodeling. During bone remodeling, there are transcription factors such as nuclear factor-activated T cells-1 (NFATc1), sclerostin, and tartrate-resistant acid phosphatase (TRAP) that are released for bone resorption. Metabolite from gingival mesenchymal stem cells (GMSCs) has the ability to activate proliferation, migration, immunomodulation, and tissue regeneration of bone cells and tissues. Furthermore, the aim of this study is to investigate the metabolite of GMSCs' effect on expression of NFATc1, TRAP, and sclerostin in calvaria bone resorption of Wistar rats. MATERIALS AND METHODS: Twenty male healthy Wistar rats (Rattus norvegicus), 1 to 2 months old, 250 to 300 g body were divided into four groups, namely group 1 (G1): 100 µg phosphate-buffered saline day 1 to 7; group 2 (G2): 100 µg lipopolysaccharide (LPS) day 1 to 7; group 3 (G3): 100 µg LPS + 100 µg GMSCs metabolite day 1 to 7; and group 4 (G4): 100 µg GMSCs metabolite day 1 to 7. Escherichia coli LPS was used to induce inflammatory osteolysis on the calvaria with subcutaneous injection. GMSCs metabolite was collected after passage 4 to 5, then injected subcutaneously on the calvaria. All samples were sacrificed on the day 8 through cervical dislocation. The expression of TRAP, NFATc1, and sclerostin of osteoclast in the calvaria was observed with 1,000× magnification. STATISTICAL ANALYSIS: One-way analysis of variance and Tukey honest significant different were conducted to analyze differences between groups (p < 0.05). RESULTS: The administration of GMSCs metabolite can significantly decrease TRAP, NFATc1, and sclerostin expression (p < 0.05) in LPS-associated inflammatory osteolysis calvaria in Wistar rats (R. norvegicus). There were significantly different TRAP, NFATc1, and sclerostin expressions between groups (p < 0.05). CONCLUSION: GMSCs metabolite decrease TRAP, NFATc1, and sclerostin expression in LPS-associated osteolysis calvaria in Wistar rats (R. norvegicus) as documented immunohistochemically.

Kombucha drink enriched with sea grapes (Caulerpa racemosa) as potential functional beverage to contrast obesity: An in vivo and in vitro approach.

BACKGROUND AND AIMS: Obesity is currently a global issue and is a major cause of the metabolic disorder, including dyslipidemia. However, currently approved treatments have various limitations including serious side effects, numerous contraindications, and lack of acceptance. Caulerpa racemosa, also referred as Sea grapes, is a seaweed known for its various benefits. C. racemosa extract has the potential to improve lipid profile and role as an anti-obese agent. In order to maximize its health benefits, C. racemosa was made using kombucha drink as a carrier medium. This study aims to assess the effect of Sea grapes kombucha drink on lipase activity in vitro and lipid profile in vivo. METHODS: A lipase inhibition test was carried out by incubating Sea grapes kombucha drink compared with orlistat as the control in porcine pancreatic lipase and p-nitrophenyl butyrate in reaction buffer. A total of four groups were made, each containing 10 male swiss webster albino mice; group A received standard dry pellet diet as control, group B received cholesterol and fat-enriched diets (CFED), group C and D received CFED and 150 and 300 mg/kgBW of kombucha drink from Sea grapes respectively for 4 weeks. RESULTS: Sea grapes kombucha drink improved lipid profiles in the way of reducing total cholesterol, triglyceride, LDL, and increasing HDL levels compared to CFED and normal groups. The effect was more robust following the incrementing dose of the Sea grapes excluding total cholesterol. The lipase inhibitory activity of Sea grapes kombucha drink was similar to orlistat at a dose of 250 µg/mL, otherwise, orlistat was superior in the lower doses. CONCLUSIONS: Sea grapes kombucha drink treatment also induced weight loss and increased level of liver SOD. Kombucha drink from C. racemosa has good potential as a functional beverage with anti-obese and lipid improving activity.

Study of Alveolar Bone Remodeling Using Deciduous Tooth Stem Cells and Hydroxyapatite by Vascular Endothelial Growth Factor Enhancement and Inhibition of Matrix Metalloproteinase-8 Expression in vivo.

Background: Periodontitis progression is characterized by alveolar bone loss, and its prevention is a major clinical problem in periodontal disease management. Matrix metalloproteinase-8 (MMP-8) has been shown to adequately monitor the treatment of chronic periodontitis patients as gingival crevicular fluid MMP-8s were positively associated with the severity of periodontal disease. Moreover, modulating the vascular endothelial growth factor (VEGF) levels in bones could be a good way to improve bone regeneration and cure periodontitis as VEGF promotes endothelial cell proliferation, proteolytic enzyme release, chemotaxis, and migration; all of which are required for angiogenesis. Purpose: The aim of this study was to determine the effect of hydroxyapatite incorporated with stem cells from exfoliated deciduous teeth (SHED) in Wistar rats' initial alveolar bone remodeling based on the findings of MMP-8 and VEGF expressions. Methods: A hydroxyapatite scaffold (HAS) in conjunction with SHED was transplanted into animal models with alveolar mandibular defects. A total of 10 Wistar rats (Rattus norvegicus) were divided into two groups: HAS and HAS + SHED. Immunohistochemistry staining was performed after 7 days to facilitate the examination of MMP-8 and VEGF expressions. Results: The independent t-test found significant downregulation of MMP-8 and upregulation VEGF expressions in groups transplanted with HAS in conjunction with SHED compared with the HAS group (p < 0.05). Conclusion: The combination of SHED with HAS on alveolar bone defects may contribute to initial alveolar bone remodeling as evident through the assessments of MMP-8 and VEGF expressions.

Comparative performance of ELISA and dot blot assay for TSH-receptor antibody detection in Graves' disease.

BACKGROUND: Graves' disease (GD) is an autoimmune disease, and it accounts for major cases of hyperthyroidism. Antibody against thyroid-stimulating hormone receptor/TSHR (TRAb) is responsible for hyperthyroidism and is considered as a diagnostic marker for GD. Therefore, we developed a recombinant protein of human TSHR-169 (hTSHR-169), which was specifically recognized TRAb in the serum of GD patients and then compare the diagnostic performance between ELISA and dot blot of TRAb tests for their ability to diagnose GD. METHODS: 20 GD patients and 20 healthy individuals from the Indonesian population were enrolled. TRAb concentration and density were quantified. Comparative analysis was performed using receiver-operating curve (ROC) analysis. RESULTS: For dot blot assay, the minimum concentration to detect TRAb requiring 100 ng of antigen with antiserum diluted at 1:60. For diagnosing GD, the ELISA yielded a higher AUC compared with the dot blot assay (0.95 and 0.85, respectively). Using the recommended cutoff values, the efficiency of both assays was examined by comparing the specificity and sensitivity of the assays to the clinical diagnosis. The ELISA showed 80% and 95%, while the dot blot assay showed 70% and 95% sensitivity and specificity, respectively. CONCLUSION: Although the dot blot assay exhibited lower performance than the ELISA method, the dot blot assay is a simple and rapid diagnostic assay that is suitable for diagnosing GD in rural areas, in which healthcare facilities sometimes are not accessible.

Potential benefit of olive leaf extract in cervical spondylotic myelopathy model.

INTRODUCTION: Cervical spondylosis is the most common cause of myelopathy in the cervical due to chronic compression of the spinal cord in patients aged 55 years or older. Recent studies suggest that olive extracts suppress inflammation and reduce stress oxidative injury. The purpose of this study was to determine the potential neuroprotective effects of olive leaf extract (OLE) in an experimental cervical spondylotic myelopathy model. METHODS: This study was divided into 6 groups; Control Negative (Sham-Operated) Group, Control Positive 1 & 2 (early chronic and chronic), Treatment Groups 1, 2 & 3 (prophylactic, concomitant & late). Olive leaf extract (OLE) give 350 mg/kg BW and spinal cord sample was taken at the compression level C5. Histopathological assessment and immunohistochemistry of Amyloid-ß, p-Tau, TDP-43 dan CD-68 dan evaluation of functional motoric outcome was done before animals were terminated. RESULTS: Chronic spinal cord compression increased the expression of Amyloid-ß, p-Tau, TDP-43 dan CD-68. OLE 350 mg/kg BW decreased the expression of these biomarkers and increased functional motoric outcome, especially as prophylactic dan concomitant treatment. DISCUSSION: These findings indicate that OLE may be effective in protecting cervical spondylotic myelopathy.

In vitro and in vivo study: Ethanolic extract leaves of Azadirachta indica Juss. variant of Indonesia and Philippines suppresses tumor growth of hepatocellular carcinoma by inhibiting IL-6/STAT3 signaling.

Background: Azadirachta indica Juss. has been shown to suppress cancer progression through a variety of mechanisms. In order to treat cancer progression, cancer immunotherapy is used to stimulate the immune system where immunosuppression is present in tumor microenvironments. Many cancer cells produce a lot of interleukin-6 (IL-6) and signal transducer activator of transcription 3 (STAT3). STAT3 plays a key role in suppressing the expression of critical immune activation regulators. IL-6-mediated STAT3 activation is common in the tumor microenvironment. Inhibiting the IL-6/STAT3 signaling pathway has become a therapeutic option for cancer progression. As vimentin is also expressed in hepatic stellate cells boosting cancer survival. We focused on the precise effect of extract from leaves of Azadirachta indica Juss, on inhibiting the IL-6/STAT3 signaling cascade on hepatocellular carcinoma by in vitro and in vivo study. Methods: In the in vitro study, the effect of Azadirachta indica Juss. variant Indonesia and Philippines against the expression of IL-6 and STAT3 was examined in liver cancer cell line. In the in vivo study, 24 male rats ( Rattus norvegicus) strain Wistar were induced by diethylnitrosamine and carbon tetrachloride (CCl 4). Based on the therapy given, the groups were divided into negative control, positive control, Indonesia extract, and Philippine extract. Expression of IL-6, STAT3, and vimentin were tested using immunohistochemistry staining. The data were analyzed using analysis of variance, which was then followed by the Tukey test. Results: Statistically significant difference in IL-6 and STAT3 was observed between the treatment groups and positive control group by in vitro study and in vivo study. Generally, there is no significant difference between treatment using Indonesian and Philippine leaves. Conclusion: Both therapy doses of Azadirachta indica variant in Indonesia and Philippines were able to reduce IL-6, STAT3 and vimentin expression of hepatocellular carcinoma cell by in vitro and in vivo experiment.

Progressive spinal cord compression technique in experimental rabbit animal model for cervical spondylotic myelopathy.

INTRODUCTION: Cervical spondylotic myelopathy (CSM) presently estimated at 54% population, commonly cause of myelopathy due to chronic compression of the spinal cord in older people. Physiological injuries caused by static and dynamic forces including compressed, pinched, and pulled out inducing secondary injuries at the molecular level. METHODS: We examined the rabbit model approach with the clinical case of spondylotic myelopathy, in which the disk and facet maintained the cervical spine mobility, and compression was given 0.5 mm per week three times in this model. In this study, a group of 14 days was made (early into the chronic phase) and the 21 day group had a chronic process for 1 week, that period can be categorized as a chronic process and CSM is a chronic process. By examining motor scores, histological examination and immunohistochemistry of the spinal cord, this model efficiently produces myelopathy. The distribution of microglia expressing GFAP, S100-ß, and Neurofilaments were observed by immunohistochemical techniques. RESULTS: There was a significant difference in the number of cells expressing GFAP between the control group and the 21-day compression group (p = 0.001). There is a decrease in S100-ß expression of spinal cord tissue after receiving compression exposure. There was a significant difference in the number of cells expressing NF between the control group, the 14-day compression group (p = 0.04) and 21-day compression group (p = 0.04). DISCUSSION: Neurons have the intrinsic ability to regenerate after injury, although not spontaneously. Cervical spondylotic myelopathy causes permanent neurological disorders, partly due to glial scar formation consisting of astrocytes and microglia. The difference between our study and previous research methods is that we perform compression of the spinal cord in stages (0.5 mm, 1.0 mm & 1.5 mm) so that it is more like the natural occurrence of chronic spinal cord compression. CONCLUSION: An increasing of GFAP value in this study indicates the presence of astrocyte activity which can be associated with chronic spinal cord injury. There is a decrease in S100-ß expression of spinal cord tissue neuron cells after receiving compression exposure. The expression of NF decreased indicating degenerative axons.

Expression of AIF and Caspase-3 in New Zealand rabbit with Cervical Spondylosis Myelopathy model.

INTRODUCTION: Cervical spondylosis myelopathy (CSM) is a clinical syndrome of motoric or sensoric, caused by degenerative process chronically causing narrowing of cervical canal and compressing the spinal cord. The narrowing of canalis spinalis causing chronic compression and disrupting vascular patency in spinal cord. This is worsened on repetitive trauma on flexion, extension and rotation. CSM has an incidence of 4.04 in 100.000 cases per year and the total patients undergoing treatment operative or nonoperatively in year increasing for 7 times. Apoptosis plays a critical role in important biological processes such as morphogenesis, tissue homeostasis, and immunity; furthermore, its aberrant activation or impairment may contribute to a number of diseases. The understanding of CSM pathophysiology from apoptotic pathway is an essential topic to discussed and the treatment of this case in future. METHOD: This study uses experimental study with Post test Only Control Group, using New Zealand rabbits. The rabbits given the compression on cervical as high as C5 to induce CSM. The tissue was taken from spinal cord on compression area and histopathology examination was done to calculate apoptotic factor expression such as AIF and caspase-3. RESULT: Chronic compression on spinal cord causing myelopathy clinically on animal study, resulted in weakness of all extremities. Based on this study, the expression of AIF and caspase-3 is increasing in compression group in day 14 to day 21. CONCLUSION: Chronic compression in spinal cord causing increase in AIF and caspase-3 in day 14 and day 21, and this may be caused by increasing of apoptotic expression on animal study.

New insight into the role of a combination of zinc oxide and turmeric rhizome liquid extract in osteogenic marker expression.

Aim: This research was aimed to determine the potential for treating osteogenesis with a combination of zinc oxide and turmeric (ZOT) rhizome liquid extract. Setting and Design: In vivo, post test-control group design. Material and Methods: The mandibular incisors of Wistar rats were extracted and left untreated or received an application of zinc oxideeugenol (ZOE) 10% or ZOT rhizome liquid extract at various concentrations (10%, 20%, and 40%). The mandible was then subjected to immunohistochemical analysis to detect RUNX2 and alkaline phosphatase (ALP) activity. Statistical Analysis Used: One-way ANOVA and Tukey HSD using SPSS software. Results: All groups demonstrated increasing RUNX2 and ALP activity. ZOT 40% showed the highest activity in all groups on day 3 and day 7, although there were no significant differences with ZOE 10%. Conclusion: A combination of ZOT rhizome liquid extract can induce the osteogenic process in postextraction sockets. The results highlight the need for further investigation of the potential osteogenesis of curcumin in humans.

Wound healing properties of Gliricidia sepium leaves from Indonesia and the Philippines in rats (Rattus norvegicus).

BACKGROUND AND AIM: Gliricidia sepium is a medium-sized leguminous plant found widely in tropical to subtropical areas. It has been used as a medicinal ingredient and in rodenticides by local communities in both Indonesia and the Philippines. This study aimed to investigate the wound healing effects of an ointment containing G. sepium leaves on inflammatory cells using a rat model. We also determined its effect on the expression of interleukin (IL) 6 and IL-1ß. MATERIALS AND METHODS: We used 16 Wistar male rats aged approximately 2 months and weighing 150-200 g. They were divided into four treatment groups (T1, positive control; T2, negative control; T3, wounds treated with G. sepium from Indonesia; and T4, wounds treated with G. sepium from the Philippines), and the ointment therapies were applied to wounds for 3 days. Hematoxylin and eosin staining was performed to examine the inflammatory cells microscopically. IL-1ß and IL-6 expression were observed immunohistochemically. RESULTS: G. sepium leaves significantly (p<0.05) decreased the number of inflammatory cells, and the expression of IL-1ß and IL-6 in the group treated with Indonesian G. sepium leaves was higher than that in the group treated with G. sepium leaves from the Philippines. The leaves contain flavonoids, saponins, and tannins, which act as anti-inflammatory agents to enhance the wound healing process. CONCLUSION: Our findings suggest that G. sepium leaves from both the Philippines and Indonesia possess wound healing properties.

Correlation of signal to noise ratio (SNR) value on distortion product otoacoustic emission (DPOAE) and expression of nuclear factor erythroid 2-related factor 2 (NRF2) in cochlear organ of Corti in rat exposed to noise.

Aim To investigate the changes in the value of the signal to noise ratio (SNR) and to assess changes in the expression of nuclear factor erythroid 2-related factor 2 (NRF2) in the organ of Corti of rat exposed to noise. Methods This study used a randomized post test only control group laboratory experimental design with 27 male Wistar strain Rattus norvegicus. The study group was divided into 3 groups (n = 9): group I (control), group 2 (2 hours of 100 dB noise exposure) and group 3 (2 hours of 110 dB noise exposure). Results There was no significant difference in the SNR in the group 1 on day 0, 2 and 4 (p>0.05). However, there was a significant difference in the SNR in the group 2 and the group 3 on day 0, 2 and 4 (p<0.05). There was a significant difference in the mean levels of NRF2 expression in the cochlear organ of Rattus norvegicus in all groups (p<0.05). There was no correlation between the SNR and the NRF2 expression in group 2 (p>0.05), but there was a correlation between the SNR and the NRF2 expression in the group 3 (p<0.05). Conclusion There was found a correlation between the SNR value on distortion product otoacoustic emission (DPOAE) examination and NRF2 expression in the cochlear organ of Corti of Rattus norvegicus exposed to 110 dB noise.

Immunohistochemical analysis of stem cells from human exfoliated deciduous teeth seeded in carbonate apatite scaffold for the alveolar bone defect in Wistar rats ( Rattus novergicus).

Background: Stem cells from human exfoliated deciduous teeth (SHED) seeded in carbonate apatite scaffold (CAS) may have multiple functions that could be used to regenerate the alveolar bone defects. The purpose of this study is to examine the ability of SHED and CAS in alveolar bone defects using an immunohistochemical analysis. Methods: ten three-month-old healthy male Wistar rats (R. novergicus) that weighed between 150-250 grams (g) were used as animal models. A simple blind random sampling method was used to select the sample that was assigned to the study group for CAS and SHED seeded in CAS (n=5). The animal study model of the alveolar bone was established by extracting the anterior mandible teeth. Rodent anesthesia was applied to relieve the pain during the procedure for all test animals. Immunohistochemistry was performed after seven days to facilitate the examination of the receptor activator of NF-κß ligand (RANKL), osteoprotegrin (OPG), transforming growth factor-ß (TGF-ß), vascular endothelial growth factor (VEGF), runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osteocalcin, and osteopontin expression. The data was analyzed using the unpaired t-test (p<0.01) and Pearson's correlation test (p<0.05). Results: The OPG, RUNX2, TGF-ß, VEGF, ALP, osteocalcin, and ostepontin expressions were higher in SHED seeded in CAS than CAS only with a significant difference between the groups (p<0.01). Furthermore, the RANKL expression was lower in SHED seeded in CAS compared to CAS only. There was a strong reverse significant correlation between OPG and RANKL expression (p<0.05). Conclusions: The number of osteogenic marker expressing cells, such as OPG, RUNX2, TGF-ß, VEGF, ALP, osteocalcin, and ostepontin, increased. However, RANKL expression in the alveolar bone defects that were implanted with SHED seeded in CAS did not increase after seven days.

Effectivity of zinc oxide-turmeric extract dressing in stimulating the reepithelization phase of wound healing.

BACKGROUND AND AIM: Reepithelialization can be described as the resurfacing of a wound with new epithelium in the process of healing, with the overlapping step from keratinocyte migration and proliferation to the tissue contraction. Zinc oxide-turmeric extract dressing has been proven to have anti-inflammatory properties, but its effectivity in the reepithelialization process is still unknown. This study aimed to determine the effect of a wound dressing consisting of zinc oxide and turmeric extract on wound reepithelialization by assessing the expression of cytokeratin 14 (CK14), epidermal growth factor receptor (EGFR), and epithelial cadherin (E-cadherin). MATERIALS AND METHODS: A total of 40 Wistar rats were randomized into four control and four treatment groups (n=5 per group). On day 1, a square-shaped full-thickness skin excision measuring 6×6 mm in size was created in the dorsal thoracic area of the rats, and the wounds were either dressed with a combination of zinc oxide and turmeric extract in the treatment groups or left undressed in the control groups. Then, the rats were sequentially sacrificed on days 3, 5, 7, and 14 to obtain subepithelial excision samples, which were subsequently subjected to immunohistochemistry analysis for the expression of CK14, EGFR, and E-cadherin to ascertain wound reepithelization. The data were tabulated and analyzed using a one-way analysis of variance and least significant difference test. RESULTS: The highest expression levels of CK14, EGFR, and E-cadherin were observed on days 7 and 14 in the treatment and control groups, respectively. While the expression levels of these markers on day 7 were found to be significantly higher in the treatment than the control groups, no significant difference in the expression levels on day 14 was detected between the control and treatment groups (p<0.05). CONCLUSION: A wound dressing consisting of zinc oxide and turmeric extract can help accelerate reepithelization in the wound healing process.

Exfoliated Human Deciduous Tooth Stem Cells Incorporating Carbonate Apatite Scaffold Enhance BMP-2, BMP-7 and Attenuate MMP-8 Expression During Initial Alveolar Bone Remodeling in Wistar Rats (Rattus norvegicus).

BACKGROUND: Post-tooth extraction socket preservation is necessary due to alveolar bone resorptive patterns through regenerative dentistry approaches that involve the use of stem cells, scaffold and growth factor. Stem cells derived from human exfoliated deciduous teeth (SHED) are known to potentially possess the osteogenic ability. Meanwhile, carbonate apatite scaffold (CAS) can act as a biocompatible scaffold capable of supporting mesenchymal stem cells (MSCs) to proliferate and differentiate optimally. The aim of this study is to investigate the expression of bone morphogenic protein-2 and 7 (BMP2, BMP7) and Matrix Metalloproteinase-8 (MMP-8) after the transplantation of SHED-incorporated CAS during in vivo bone remodeling. MATERIAL AND METHODS: A total of 14 healthy, male, Wistar rats, whose mandible anterior teeth were extracted by means of sterile needle holder clamps, constituted the subjects of this study of alveolar bone defects. Two research groups were created: a control group (CAS) as group I and an experimental group (CAS + SHED) as group II. SHED with a density of 106 cells were incorporated into CAS before being transplanted into the experimental group. After 7 days, all the animals were sacrificed and their mandible anterior region extracted. The BMP2, BMP7 and MMP-8 expression were subsequently analyzed by means of immunostaining. An unpaired t-test was conducted to analyze the treatment and control group (p<0.01) data. RESULTS: The expression of BMP-2 and BMP-7 was higher in group II compared to group I. Meanwhile, the level of MMP-8 was lower in group II than group I. There was greater significant increased expression of BMP-2 and BMP-7 expression in Group II compared to Group I. There was significant decreased expression of MMP-8 between group II than group I (p<0.01). CONCLUSION: SHED-incorporated CAS can enhance BMP-2 and BMP-7 expression while attenuating MMP-8 expression during in vivo alveolar bone remodeling.

Protective Effects of Propolis Extract in a Rat Model of Traumatic Brain Injury via Hsp70 Induction.

BACKGROUND: Traumatic brain injury (TBI) is one of the major global health problems. Secondary brain injury is a complex inflammation cascades process that causes brain cell apoptosis. Propolis is a natural product that has neuroprotective property. AIM: This study aimed to investigate the effect of propolis toward Hsp70 expression with apoptosis marker in brain tissue after TBI. METHODS: Thirty-three Sprague Dawley rats were randomised into three treatments group, i.e. sham-operated controls, closed head injury (CHI), and CHI with propolis extract (treatment group). In the treatment group, propolis was given 200 mg/kg per oral for 7 days then harvested brain tissues after sacrificed by cervical dislocation at day 8. We investigated Hsp70, Caspase 3, apoptosis-inducing factor (AIF), and TUNEL assay expression using immunohistochemistry staining. Statistical test using one-way ANOVA test and Tukey HSD as post hoc test. RESULTS: Mean of positive Hsp70 stained cells in group 1 was 6.82 ± 2.14, group 2 was 3.91 ± 2.26, and group 3 was 9.64 ± 3.53 with a significant difference of Hsp70 expression distribution within groups (p = 0.0001). Mean of positive caspase 3 stained cells in group 1 was 5.45 ± 2.30, group 2 was 13.82 ± 2.44, and group 3 was 7.03 ± 1.54 with a significant difference of caspase3 expression distribution within groups (p=0.0001). Mean of positive AIF stained cells in group 1 was 5.36 ± 2.11, group 2 was 12.82 ± 1.40, and group 3 was 8.09 ± 1.81 with a significant difference of AIF expression distribution within groups (p = 0.0001). Mean of positive TUNEL assay stained cells in group 1 was 4.82 ± 2.04, group 2 was 11.55 ± 1.51, and group 3 was 7.64 ± 1.96 with a significant difference of TUNEL test expression distribution within groups (p = 0.0001). CONCLUSION: Propolis may protect brain cell from apoptosis after injury by maintaining Hsp70 expression in addition to antioxidant and anti-inflammatory.