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Evaluation of the Effectiveness of Nano-Hydroxyapatite Particles in Wound Healing in an Animal Study

Sahar Sadat Lalehzar1*, Rokhsareh Meamar2, Ardeshir Talebi3, Mehrafarin Fesharaki2

Background: The main activity of the skin is to create a protective barrier against damage. Loss of the skin due to injury or disease and failure to regenerate the affected area may result in disability, infection, or even death. We conducted the animal study to evaluation of the effectiveness of nano-hydroxyapatite particles in wound healing.

Method: This animal study performed in Isfahan university of medical science animal lab. Experiments were performed on 30 wistar in 5 groups. Biopsies 5 mm × 5 mm were obtain of abdominal, and were transferred to the cell culture laboratory into the Phosphate Buffer Saline (PBS). The cell proliferation was determined using the colorimetric MTS assay. The type and approach of this animal study is to create a deep skin wound and try to treat the wound with drug (nano hydroxyapatite 10%, nano hydroxyapatite 40%, combination of nickle ion with nanohydroxyapatite 10%, and 40%) intervention on an animal model of rat. Macroscopic evaluation and pathological examination were done. For pathological and histological examination of the wound, sampling was done on the seventh and fourteenth days after ulcer induction. All continuous and categorical data are presented as mean ± Standard Deviation (SD) and frequency (percentage), respectively. Paired sample T-test and repeated measure Analysis of Variance (ANOVA), chi-squared test was used.

Results: During this study, MTS assay was carried out to evaluate the proliferation of mice fibroblast on the gelatin without hydroxyapatite, and with 10%, 40% hydroxyapatite after 1 day, 2 days and 3 days of culture. Significant enhancement of cell proliferation was observed in nano HA 10%, 40% and nano HA 10% with nickel in comparison when the cells seeded on gelatin and HA 10%. The best result was shown in 24 hours after seeding the cells in gelatin in comparison with 48 hours and 72 hours. Indeed, after 48 hours and 72 hours, the cell proliferation on gelatin decreased. In evaluation of wound area with image j soft ward, the wound area between 3rd day, 7th day and 14th day of treatment after wound induction there were no significant difference between groups. In microscopic study and analysis for evaluation and comparing wound length with the michrome camera and mosaic soft ward, there were no significant relation in time (p1=0.77). There is a difference is close to significant between the groups (p2=0.065). There was no significant difference between time and group (p3=0.323). In day 14 the wound length between groups had significant difference (p4=0.049).

Conclusion: In conclusion, hydroxyapatites and its combination with nickle ion have significant effect on wound healing and cell proliferation.