Acceleration of Bone Fracture Healing through the Use of Bovine Hydroxyapatite or Calcium Lactate Oral and Implant Bovine Hydroxyapatite-Gelatin on Bone Defect Animal Model.

Bone grafts a commonly used therapeutic technique for the reconstruction and facilitation of bone regeneration due to fractures. BHA-GEL (bovine hydroxyapatite-gelatin) pellet implants have been shown to be able accelerate the process of bone repair by looking at the percentage of new bone, and the contact between the composite and bone. Based on these results, a study was conducted by placing BHA-GEL (9:1) pellet implants in rabbit femoral bone defects, accompanied by 500 mg oral supplement of BHA or calcium lactate to determine the effectiveness of addition supplements. The research model used was a burr hole defect model with a diameter of 4.2 mm in the cortical part of the rabbit femur. On the 7th, 14th and 28th days after treatment, a total of 48 New Zealand rabbits were divided into four groups, namely defect (control), implant, implant + oral BHA, and implant + oral calcium lactate. Animal tests were terminated and evaluated based on X-ray radiology results, Hematoxylin-Eosin staining, vascular endothelial growth Factor (VEGF), osteocalcin, and enzyme-linked immunosorbent assay (ELISA) for bone alkaline phosphatase (BALP) and calcium levels. From this research can be concluded that Oral BHA supplementation with BHA-GEL pellet implants showed faster healing of bone defects compared to oral calcium lactate with BHA-GEL pellet implants.

The Effect of Green Tea with EGCG Active Compound in Enhancing the Expression of M2 Microglia Marker (CD206).

Background: Stroke is a neurological deficit due to vascular disorders. Microglia are the first line of defense against brain injury. Anti-inflammatory cytokines activate M2 microglia, which upregulate CD206. EGCG is abundant in green tea, which has an anti-inflammatory effect. Objective: To know the effect of green tea with its active compound EGCG on CD206 expression. Settings and Design: True experimental trial design. Material and Methods: Rattus Novergicus were divided into six groups: a negative control group (Sham), a positive control group (P0), MCAO mice given 10 mg/kg BW EGCG (P1), 20 mg/kg BW EGCG (P2), 30 mg/kg BW EGCG (P3), and 30 mg/kg BW standardized green tea extract (P4). CD206 expression was measured using immunohistochemistry and scored according to the Allred scoring guidelines. Statistical Analysis Used: Descriptive test, Levine test, Kolmogorov-Smirnoff test, Independent sample t test, Pearson correlation test. Results: We discovered that there is a significant difference in CD206 expression between the Sham and P0 groups (P < 0.05). In addition, there are significant differences in expression between the sham group and the other two groups (P1 and P2) (P < 0.05). Furthermore, when we compared the P0 group with each treatment group, we found that CD206 expression between P0-P2, P0-P3, P0-P4 are significantly different. There is a significant correlation between green tea with its active compound EGCG and CD206 expression enhancement. The correlation is positive. Conclusions: Green tea with EGCG active compound increases CD206 expression as an M2 marker in the Rattus norvegicus with MCAO model.

Attenuation of hyperplasia in lung parenchymal and colonic epithelial cells in DMBA-induced cancer by administering Andrographis paniculata Nees extract using animal model.

OBJECTIVES: This study was designed to evaluate the potential of Andrographis paniculata ethanolic extract to inhibit the increase in proliferation and induction of abnormal cell death. METHODS: The hyperplasia stage as an early stage of cancer development was induced by oral administration of 20 mg/Kg BW DMBA to SD rats twice a week for 5 weeks. There were five groups in this study include negative control, positive control, and treatment groups of DMBA induction followed by administration of A. paniculata ethanolic extract in doses equivalent to 10, 30 or 100 mg/Kg BW andrographolide once per day for 6 consecutive weeks. On the last day, rats were sacrificed, lung and colon tissues were collected. Histological examination by HE staining and immunohistochemistry using p53, telomerase, and caspase-3 antibodies were aimed at observing hyperplasia state in these tissues. RESULTS: DMBA induction to SD rats was able to produce hyperplasia in lung parenchymal and colon epithelial tissue. This can be showed by the increasing number of proliferated cells and as indicated by the number of brown-colored nuclei with sharper intensity. As well telomerase appears to be overexpressed strongly, while p53 and caspase-3 show low intensity. The administration of A. paniculata extract for 6 weeks showed a decrease in the number of cells that actively proliferate, a decrease in telomerase activity, and an increase in caspase-3 levels which indicate cellular death activity. CONCLUSIONS: A. paniculata ethanolic extract can inhibit the development of cancer at the hyperplasia stage by reducing telomerase activity and increasing apoptosis, marked by an increase of caspase-3 expressions.

The effect of Camellia sinensis (green tea) with its active compound EGCG on neuronal cell necroptosis in Rattus norvegicus middle cerebral artery occlusion (MCAO) model.

OBJECTIVES: To determine the inhibition effect of epigallocatechin gallate (EGCG) and green tea extract on neuronal necroptosis based on necroptosis morphology. METHODS: In vivo study was performed on male Rattus norvegicus middle cerebral artery occlusion (MCAO) model divided into five groups, MCAO-control groups, EGCG 10 mg/kg BW/day, EGCG 20 mg/kg BW/day, EGCG 30 mg/kg BW/day, and green tea extract 30 mg/kg BW/day for 7 days treatment. MCAO model was made by modification method using Bulldog clamp. After 7 days of treatment, all R. norvegicus were sacrificed. After that, examination using Hematoxylin-Eosin stain was conducted to look at necroptosis morphology in each group. RESULTS: We found that there are significant differences between control group and the other three groups (EGCG 20 mg/kg BW/day, EGCG 30 mg/kg BW/day, and green tea extract (p<0.05). There is a significant correlation between the number of neuron cell necroptosis and both EGCG and green tea extract (p<0.05). The correlation is negative, which means both EGCG and green tea extract will decrease the number of neuron cell necroptosis. EGCG will decrease neuron cell necroptosis starting from the dose of 20 mg/kg BW/day. EGCG 30 mg/kg BW/day produces the best result compared to other doses. CONCLUSIONS: Camellia sinensis (green tea) with its active compound EGCG decreases neuronal necroptosis morphology in MCAO models.

Green tea and its active compound epigallocathechin-3-gallate (EGCG) inhibit neuronal apoptosis in a middle cerebral artery occlusion (MCAO) model.

OBJECTIVES: To determine the effect of green tea with the active ingredient epigallocathechin-3-gallate (EGCG) on the inhibition of apoptosis in the middle cerebral artery occlusion (MCAO) model. METHODS: Four month old male Rattus norvegicus rats with a body weight of 200-275 g was used for the MCAO model and divided into five groups, and the treatment was carried out for 7 days. Before being sacrificed, the subject had 1 cc of blood drawn for high mobility group box 1 (HMGB-1) examination using enzyme-linked immunosorbent assay (ELISA), and after being sacrificed, the brain tissue specimen was taken to examine caspase-3 and B-cell lymphoma 3 (BCL-3) using immunohistochemistry methods. RESULTS: There was no significant difference in HMGB-1 results for the treatment group compared to the control group (P1: 384.20 ± 231.72 [p = 0.553]; P2: 379.11 ± 268.4 [p = 0.526]; P3: 284, 87 ± 276.19 [p = 0.140]; P4: 435.32 ± 279.95 [p = 0.912]). There is a significant increase in BCL-2 expression between the treatment group compared to the control group (P1: 2.58 ± 0.51 [p = 0.04]; P2: 3.36 ± 0.50 [p<0.001]; P3: 4.00 ± 0.42 [p<0.001]; P4: 3.60 ± 0.52 [p<0.001]). There was a significant difference in caspase-3 expression compared to the control group in the P3 group (P1: 4.33 ± 0.49 [p = 0.652]; P2: 4.09 ± 0.30 [p = 0.136]; P3: 3.58 ± 0.51 [p = 0.01]; P4: 3.89 ± 0.42 [p = 0.063]). There is no correlation between HMGB-1 and caspase-3 (r = -0.063; p = 0.613) or BCL-2 (r = -0.106; p = 0.396). There is significant negative correlation between caspase-3 and BCL-2 (r = -0.459; p = 0.000). CONCLUSIONS: Green tea with the active ingredient EGCG can inhibit neuronal cell death through the apoptotic pathway and not through the activation of HMGB-1.

Apoptosis-Inducing Factor, Protein Expression, and Apoptosis Changes with Glutamine in Podocytes Cells Exposed with Cisplatin.

Cisplatin is a well-known chemotherapeutic drug. It is one of the most effective anticancer agents and is widely used for the treatment of several types of tumors. However, side effects in normal tissues and organs, such as nephrotoxicity that induces apoptosis in epithelial cells in the kidney, limit the use of cisplatin. Glutamine is a substrate for the synthesis of glutathione as an antioxidant and promotes HSP70 release, protecting cells from apoptosis induced by different stimuli. In the present study, we investigated the protective effect of glutamine on cisplatin nephrotoxicity in the kidney. Mice were divided into three groups such as a group of control (P0), a group of intraperitoneal injection of a single dose cisplatin 20 mg/kg BW at 7th day (P1), and a group of intravenous glutamine injection 100 mg/kg BW at days 1-7 and given an intraperitoneal injection of single dose cisplatin 20 mg/kg BW at 7th day (P2). Measurement of AIF expression and apoptotic cells was carried out by immunohistochemical methods. The number of AIF expressions and apoptotic cells is expressed in the Allred score. AIF expression result is as follows: P0: 3.29 ± 0.79, P1: 5.32 ± 0.68, and P2: 4.49 ± 0.47. Apoptosis result is as follows: P0: 3.04 ± 0.70, P1: 5.26 ± 0.53, and P2: 4.44 ± 0.41. There is a decreased expression of AIF on intravenous glutamine administration, followed by a decrease in apoptosis in the podocyte. In conclusion, glutamine administration might represent the treatment of nephrotoxic-induced cisplatin.

Effect of Intravenous Glutamine on Caspase-12 Expression in the Apoptosis of the Glomerular Epithelial Cells of Male Rats Exposed to Cisplatin.

OBJECTIVE: Cisplatin is potent chemotherapy for broad-spectrum malignancies treatment, but its use is limited by organ toxicity effects, including nephrotoxicity. Glutamine prevents cisplatin nephrotoxicity by inhibiting the oxidative stress in kidney cell apoptosis. METHODS: This research examined the nephroprotective effects of intravenous glutamine on the glomerular epithelium of male rats (Rattus norvegicus). 30 male rats were randomly divided into (1) P0 as the control group; (2) P1 that was administered with single dose cisplatin (20 mg/kg BW) intraperitoneal injection; and (3) P2 that was administered with intravenous injection of glutamine (100 mg/kg BW) and single-dose cisplatin (20 mg/kg BW) intraperitoneal injection. The measurement of caspase-12 expression and apoptotic cells was performed using immunohistochemical methods. RESULTS: The caspase-12 expression are as follows: P0 = 0.5 ± 0.15; P1 = 4.1 ± 0.86; P2 = 2.54 ± 0.72. The apoptotic cells are as follows: P0 = 14.5 ± 5.23 cells/field of view; P1 = 52.7 ± 17.06 cells/field of view; P2 = 31.5 ± 6.73 cells/field of view. There is a decrease in the caspase-12 expression and apoptotic cells after intravenous glutamine administration in male white rats' glomerular epithelial cells exposed to cisplatin. The decrease of caspase-12 expression is followed by a decrease in glomerular epithelium apoptosis after intravenous glutamine administration. CONCLUSION: Immunohistochemical examination can be used as a marker of the nephrotoxic effect of cisplatin on the renal glomerular epithelium. Glutamine has been observed to give nephroprotective effect to cisplatin nephrotoxic effects.
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