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Original Research

Evaluation of wound healing properties of bioactive aqueous fraction from Moringa oleifera Lam on experimentally induced diabetic animal model

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Pages 1715-1730 | Published online: 24 May 2016

Figures & data

Table 1 Animal grouping

Table 2 Antibacterial activity of the aqueous fraction of Moringa oleifera against bacterial isolates

Table 3 Minimum inhibitory concentration (MIC) of aqueous fraction of Moringa oleifera

Figure 1 A graph comparing the concentration of blood glucose level of normal rats with streptozotocin-induced diabetic rats at day 0, 3, 7, 14, and 21.

Notes: The increased blood glucose in diabetic rats is highly significant compared to normal rats (*P<0.05 diabetic vs control groups). Values are mean ± standard deviation of three independent readings.
Figure 1 A graph comparing the concentration of blood glucose level of normal rats with streptozotocin-induced diabetic rats at day 0, 3, 7, 14, and 21.

Figure 2 A graph comparing the concentration of insulin level of normal rats with streptozotocin-induced diabetic rats at 72 hours and 21 days after induction of diabetes.

Notes: The high level of insulin in normal rats was more significant compared to diabetic rats. *P<0.05 normal control group versus diabetic groups. Values are mean ± standard deviation of three independent readings.
Figure 2 A graph comparing the concentration of insulin level of normal rats with streptozotocin-induced diabetic rats at 72 hours and 21 days after induction of diabetes.

Figure 3 Histological features of hematoxylin–eosin stained sections of pancreatic tissue 21 days after induction of diabetes using a combination of streptozotocin and nicotinamide.

Notes: n= A high intensity normal colored structure indicating presence of normal arrangement of islet of Langerhans in normal control rats. p= A low intensity and partially destroyed colored structure indicating a partially destroyed islet of Langerhans (×20 magnification).
Figure 3 Histological features of hematoxylin–eosin stained sections of pancreatic tissue 21 days after induction of diabetes using a combination of streptozotocin and nicotinamide.

Figure 4 Digital photographs of excision wounds induced on normal control animals without treatment on day 0, 3, 7, 14, and 21 postwounding.

Notes: The progress of healing indicated by the arrows showed healing was achieved by day 14 postwounding and progressed to complete wound closure after 21 days in the aqueous fraction treated groups compared to diabetic control that remained unhealed after 21 days treatment period.
Figure 4 Digital photographs of excision wounds induced on normal control animals without treatment on day 0, 3, 7, 14, and 21 postwounding.

Figure 5 A graph of wound area analysis showing decrease in wound in diabetic-treated rats in a dose-dependent manner and the decrease was significant compared to untreated diabetic control (*P<0.05).

Note: Data are mean ± standard deviation of triplicate values. *Indicates greater significance.
Figure 5 A graph of wound area analysis showing decrease in wound in diabetic-treated rats in a dose-dependent manner and the decrease was significant compared to untreated diabetic control (*P<0.05).

Figure 6 A graph of epithelialization period in control and diabetic-treated rats.

Notes: The decrease in epithelialization period in diabetic treated groups was significant compared to untreated diabetic control animals. *P<0.05. Data are mean ± standard deviation of triplicate values.
Figure 6 A graph of epithelialization period in control and diabetic-treated rats.

Figure 7 Hematoxylin–eosin histological sections of excisional wound site obtained from a diabetic wound treated with silver sulfadiazine (SSD).

Notes: Wistar rats on day 21 after wounding. (h): healed area with multiple layers of fibrous connective tissue; (br): boundary between unhealed and healed tissue; (ie): immature epidermis; (ig): immature granulation tissue; (bv): blood vessels; (id): interdigitation; (e): epithelium. Magnifications (×10).
Figure 7 Hematoxylin–eosin histological sections of excisional wound site obtained from a diabetic wound treated with silver sulfadiazine (SSD).

Table 4 Semiquantitative evaluation of histological parameters for the assessment of wound healing

Figure 8 A graph of cytokine regulation showing downregulation of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) in wound tissue treated 21 days after treatment.

Notes: The downregulation of cytokines level was highly significant in diabetic-treated rats compared to untreated diabetic control rats (P<0.05). Data are mean ± standard deviation of triplicate readings. *Indicates greater significance.
Figure 8 A graph of cytokine regulation showing downregulation of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) in wound tissue treated 21 days after treatment.

Figure 9 Effect of aqueous fraction on expression of cytokines showing upregulation and downregulation of various cytokines in tissues 21 days after treatment detected by Western blotting.

Abbreviations: SSD, silver sulfadiazine; VEGF, vascular endothelial growth factor; iNOS, inducible nitric oxide synthase.
Figure 9 Effect of aqueous fraction on expression of cytokines showing upregulation and downregulation of various cytokines in tissues 21 days after treatment detected by Western blotting.

Figure 10 A graph showing analysis of expression of cytokines based on intensity of their bands obtained by image Lab software (Bio-Rad Laboratories Inc.).

Notes: *P<0.05 versus untreated diabetic control. Data are mean ± standard deviation of triplicate readings.
Abbreviations: VEGF, vascular endothelial growth factor; iNOS, inducible nitric oxide synthase.
Figure 10 A graph showing analysis of expression of cytokines based on intensity of their bands obtained by image Lab software (Bio-Rad Laboratories Inc.).

Figure 11 Effect of aqueous fraction of Moringa oleifera on vascular endothelial growth factor expression showing relative brown stain indicated by the black arrows in wound tissue of treated diabetic rats in skin wound tissues 21 days postwounding.

Notes: (i): Normal control. (ii): Untreated diabetic control. (iii, iv, and v): 0.5%, 1%, and 2% aqueous fraction treated groups, respectively. (vi): Positive control treated with standard silver sulfadiazine (magnification ×20).
Figure 11 Effect of aqueous fraction of Moringa oleifera on vascular endothelial growth factor expression showing relative brown stain indicated by the black arrows in wound tissue of treated diabetic rats in skin wound tissues 21 days postwounding.

Figure 12 Possible mechanisms by which aqueous fraction enhances wound healing in diabetic condition.

Figure 12 Possible mechanisms by which aqueous fraction enhances wound healing in diabetic condition.

Figure S1 A graph of wound contraction rate in control and diabetic treated animals.

Notes: This improved contraction rate in diabetic-treated rats was significant compared to the untreated diabetic control (*P<0.05). Data are mean ± standard deviation of triplicate values. *Indicates greatest significance.
Figure S1 A graph of wound contraction rate in control and diabetic treated animals.

Figure S2 A graph showing estimates of amount of granulation tissue formed in control and diabetic rats.

Notes: The amount of granulation tissue formed in diabetic-treated rats was significantly higher compared to untreated diabetic control rats. *P<0.05. Data are mean ± standard deviation of triplicate readings.
Figure S2 A graph showing estimates of amount of granulation tissue formed in control and diabetic rats.