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Vet World
2023 May 01;165:1084-1091. doi: 10.14202/vetworld.2023.1084-1091.
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Hydroxyapatite-chitosan composites derived from sea cucumbers and shrimp shells ameliorate femoral bone defects in an albino rat model.
Safira A
,
Rani CAM
,
Fikri F
,
Purnomo A
,
Khairani S
,
Chhetri S
,
Maslamama ST
,
Purnama MTE
.
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BACKGROUND AND AIM: A bone defect is defined as a critically sized autologous bone and a bone gap. Bone grafting is one of the most commonly used surgical methods to enhance bone regeneration in orthopedic procedures. A composite of collagen, hydroxyapatite (HA), and chitosan (Ch) is suitable as a bone matrix and stimulates ossification. This study aimed to evaluate the use of natural HA-Ch composites derived from sea cucumbers and shrimp shells and quantify the levels of cytokines, polymorphonuclear neutrophils (PMNs), serum liver enzymes, calcium, phosphate, and procollagen type 1 N-terminal propeptide (PINP) in albino rats with femoral bone defects.
MATERIALS AND METHODS: A total of 48 albino rats with femoral bone defects were divided into 4 groups (n = 12 each): (C-) placebo, (C+) polyethylene glycol, (T1) HA, and (T2) HA-Ch groups. Each group was divided into two subgroups (n = 6 each), with euthanization on 7- and 42-day post-treatment, respectively. Procollagen Type 1 N-terminal propeptide and the cytokines interleukin (IL)-4, IL-6, IL-10, and tumor necrosis factor-alpha were quantified using enzyme-linked immunosorbent assay. Flow cytometry was performed to evaluate PMNs. A clinical chemistry analyzer was used to measure the serum levels of liver enzymes, calcium, and phosphate.
RESULTS: There was a significant decrease in the level of IL-6 on 7 days and in the level of IL-10 on 42 days in the HA-Ch group. The level of PMNs also decreased significantly on 7 and 42 days in the HA-Ch group. Regarding serum liver enzymes, alkaline phosphatase (ALP) levels in the HA-Ch group increased significantly on 42 days. Calcium and phosphate levels increased significantly on 7 and 42 days in the HA and HA-Ch groups, and PINP levels increased significantly on 7 and 42 days in the HA-Ch group.
CONCLUSION: The HA-Ch composite derived from sea cucumbers and shrimp shells ameliorated femoral bone defects in albino rats. The HA-Ch composite modulated the levels of IL-6, IL-10, PMNs, ALP, calcium, phosphate, and PINP on 7- and 42-day post-treatment.
Figure-1. Cytokines evaluation of interleukin (IL)-4, IL-6, IL-10, and tumor necrosis factor-α on 7- and 42-day post-treatment. Values are expressed in mean ± standard error (SE) (n = 6 animals in respective four groups). Values are represented statistically when a,b,c, in comparison with C-group; *p < 0.05, **p < 0.01, ***p < 0.001, in comparison with C+ group.
Figure-2. polymorphonuclear neutrophils percentage on 7- and 42-day post-treatment. Values are expressed in mean ± Standard error (n = 6 animals in respective four groups). Values are represented statistically when a,b,c,d, in comparison with C-group; *p < 0.05, **p < 0.01, ***p < 0.001, in comparison with C+ group.
Figure-3. Enzyme serum evaluation of alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyl transferase on 7- and 42-day post-treatment. Values are expressed in mean ± SE (n = 6 animals in respective four groups). Values are represented statistically when a,b, in comparison with C-group; *p < 0.05, **p < 0.01, ***p < 0.001, in comparison with C+ group.
Figure-4. Evaluation of calcium and phosphate levels on 7- and 42-day post-treatment. Values are expressed in mean ± Standard error (n = 6 animals in respective four groups). Values are represented statistically when a,b, in comparison with C-group; *p < 0.05, **p < 0.01, ***p < 0.001, in comparison with C+ group.
Figure-5. Propeptide of type I collagen level on 7- and 42-day post-treatment. Values are expressed in mean ± Standard error (n = 6 animals in respective four groups). Values are represented statistically when a,b,c, in comparison with C-group; *p < 0.05, **p < 0.01, ***p < 0.001, in comparison with C+ group.
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