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Wound-Healing Effect of Aqueous Anthocephalus cadamba Leaf Extract [Autosaved].pptx

  1. Wound-Healing Effect of Aqueous Anthocephalus cadamba Leaf Extract in a Diabetic Rat Model. 1 Presented by- Prabhakar Kumar En. no- 67/MPB/SPS/2021 DPSRU, New Delhi Supervisor Dr. Mukesh Nandave Associate Professor Department of Pharmaceutical Biotechnology DPSRU, New Delhi Delhi Pharmaceutical Sciences and Research University, New Delhi-110017 Journal Club Presentation
  2. Title: Accelerative Wound-Healing Effect of Aqueous Anthocephalus Cadamba Leaf Extract in a Diabetic Rat Model Author’s Name: Shoket Ali, Sharmeen Ishteyaque, Foziya Khan, Pragati Singh, Abhishek Soni and Madhav N. Mugale* Journal Name: The International Journal of lower Extremity Wounds Impact factor: 2.057 Publisher: SAGE 2
  3. Introduction Cascade of biomolecular and cellular events like hemostasis, inflammation, cellular migration and remodeling. These processes replace dead debris and cellular layers and restore injured wound tissue to its original state by formation of collagen connective tissue. The pathogenic mechanism of delayed diabetic wound healing includes prolonged inflammatory and oxidative states, deferred proliferation and remodeling phase. 3
  4. Aim & Objective Current study was designed to understand the mechanism involved in wound healing and therapeutic potential of Anthocephalus cadamba in a diabetic rat model. 4
  5. Methodology Schematic Presentation Extract Animal (Disease Induction) Excision wound model Analysis 5 Histopathology Analysis Biochemical Western Blotting
  6. Procurement of Animal The female Sprague Dawley rats of 5 to 7 weeks and body weight 150 to 200 g were used. The total number of animals (n=40) was divided into 4 groups (10 rats/group). Rats were housed in polypropylene cages under relative humidity (RH 55 ±20%) and room temperature (22±3 °C) in a 12 h light/dark cycle. 6
  7. Induction of Diabetes Diabetes was induced by single injection of STZ (Streptozotocin) at the dose of 60 mg/kg intra-peritoneally (dissolved in citrate buffer pH of 4.5; except for the nondiabetic control group (I) rat). After 4 to 5 days of STZ administration, the blood glucose level was measured. Animals with fasting blood glucose levels >250 mg/dL were assumed to be diabetic and used in the study. 7
  8. Excision Diabetic Wound Model All the rats were anesthetized with ketamine (87 mg/kg) and xylazine (13 mg/kg) and their dorsal (last thorax to first sacral vertebra) hair was shaved. Wound field sketching was outlined by tracing a circle on butter paper with a marker. On the dorsal midline, one full-thickness circular excision of 1.77 cm2 area was created (1.5 cm in diameter).  A surgically sterile deep skin wound was created in both diabetic rats and nondiabetic rats. All wounds were cleaned daily with 0.9% sterile normal saline solution. After cleaning, the extract was applied topically at 500 mg/kg evenly to cover the wounds. 8
  9. Experimental Design The animals were randomly divided into 4 groups:- Group I (NDC) (n = 10): normal control without diabetes (served as untreated negative control).14 Group II (DC) (n = 10): diabetic control (wounds allowed to heal naturally without any treatment). Group III (D +KPLE) (n = 10): diabetic rats treated with the topical application of A cadamba aqueous extract (500 mg/kg), once a day for 28 days. Group IV (5% D +PIS) (n = 10): diabetic rats treated with topical application of marketed 5% PIS solution/ marketed standard treatment group on wounds for 28 consecutive days. 9
  10. Statistical analysis was carried out and the results were expressed as mean ± SD. All results were analyzed by one-way analysis of variance at a level of significance P< .05, P < .01, P < .001, and 0.0001 (GraphPad Prism, Version 7.0). 10 Results (D +KPLE) showed a significant increase in the percentage of wound closure (82%) at day 21 as compared to the diabetic control group (42%), nondiabetic control group (I) (49%), and povidone–iodine treatment group (75%) group (IV).
  11. 11 Groups Day 1 Day 7 Day 14 Day 21 Day 28 NDC Gropus 1.77±0 1.41±0.13 0.88±0.3 0.58±0.4 Scar DC group II 1.77±0 1.37±0.08 0.95±0.16 0.64±0.28 0.2±0.4 D+KPLE group IIII 1.77±0 1.21±0.05 0.36±0.09*** 0.071 ±0.08*** Scar 5% D+PIS group IV 1.77±0 1.33±0.08 0.28±0.1*** 0.15±0.05** Scar DC, diabetic control; NDC, nondiabetic control; D+KPLE, diabetic+Kadam plant leaf extract; D+PIS, diabetic+povidone–iodine sol.
  12. Results:- Body Weight and Hydroxyproline 12 Figure 1. Change in body weight of different groups. *P < .05, **P < .01, ***P < .001 (significant when compared to the nondiabetic control group). Figure 2. Hydroxyproline content in the wound tissue. *P < .05, **P < .01, ***P < .001 (significant when compared to the nondiabetic control group).
  13. Results:- Feed Intake and Blood glucose 13 Figure 4. Blood glucose levels of different groups. *P < .05, **P < .01, ***P < .001 (significant when compared to the nondiabetic control group). Figure 3. Feed intake (gram) in different treatment groups (no statistical differences were found between groups).
  14. Results:- Vascular endothelial growth factor 14 Figure 5. Quantification of vascular endothelial growth factor (VEGF) in different groups evaluated by normalizing against β-actin. *P < .05, **P < .01, ***P < .001 (significant when compared to the nondiabetic control group).
  15. Results:- Histopathology 15 Figure 6. Rat: wound area histopathology at 21 and 28 post wounding days in different groups (scale bar=10×, hematoxylin & eosin [H&E] staining).
  16. Results:- 16 Figure 7. Rat: wound area Masson’s trichrome staining in different groups (scale bar=10×, hematoxylin & eosin [H&E] staining).
  17. 17 Conclusion The present study demonstrates that the topical application of aqueous leaf extract:  Kadam plant indicates novel healing properties of wounds in diabetic rats.  The healing effects seemed to be due to an increased amount of hydroxyproline, fibroblasts, granulation tissue, total protein, and collagen deposition in the wound.  Therefore, the findings of the current study showed the extract promotes healing of diabetic wound in rats and provides a scientific perspective for the use of A cadamba aqueous leaf extract as an alternative to synthetic drugs with expensive costs and adverse effects.  However, to the best of our knowledge, this is probably the first study that evaluates the wound-healing effect of A cadamba leaf extract in diabetic wounds.
  18. 18 References 1. Hashemnia M, Nikousefat Z, Mohammadalipour A, Zangeneh MM, Zangeneh A. Wound healing activity of Pimpinella anisum methanolic extract in streptozotocin-induced diabetic rats. J Wound Care. 2019;28(Sup10):S26-S36. 2. Ponrasu T, Suguna L. Efficacy of Annona squamosa L in the synthesis of glycosaminoglycans and collagen during wound repair in streptozotocin induced diabetic rats. Biomed Res Int. 2014;2014:1-10. doi:10.1155/2014/124352 3. TanWS, Arulselvan P, Ng SF, Taib CN, SarianMN, Fakurazi S. Improvement of diabetic wound healing by topical application of vicenin-2 hydrocolloid film on Sprague Dawley rats. BMC Compl Alternative Med. 2019;19(1):1-6. 4. Harsha L, Brundha MP. Role of collagen in wound healing, Drug Invent Today. 2020;13(1):55-57. 5. Barrett EJ, Liu Z, Khamais M, et al. Diabetic microvascular disease: an endocrine society scientific statement. J Clin Endocrinol Metab. 2017;102(12):4343-4410.
  19. Thank You 19

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  1. There was significant decrease in body weight in the Kadam extract group as compared to the nondiabetic group (I) (Figure 2). Polyphagia/increase in feed intake was observed in the diabetic control group (II) rats as compared to the nondiabetic group (I) whereas in the Kadam extract group (III) and povidone group (IV) it was almost same as in the nondiabetic group (I) (Figure 3).