Assessment of IL-6 in the Oral Mucosal Healing with PerioKIN Hyaluronic 1% ® and OzOHEAL-M ® in Rabbits

,

viscoelastic nature make it unique to be used in several applications such as facilitating scar-less healing and regeneration of incision wounds. (3,4) Ozone therapy is a safe bio-oxidative therapy in which a mixture of oxygen and ozone is administered to obtain therapeutic effects such as; wound healing, postsurgical pain, and many other uses in dental practice. (5).
A biomarkers are an indicators of a particular physiological state, pathogenic processes, or pharmacological responses to any therapeutic intervention.Biomarkers can be measured objectively, and be accurately reproduced.Many researchers have identified several cellular events and mediators involved in wound healing that can function as biomarkers. (6,7) Immunohistochemistry (IHC) is a method of identifying antigens in tissue sections using antibodies.The main advantage of IHC is the ability to recognize the location of an antigen in a tissue or cell for future study of cell function and characterization of lesions.The basic feature of the IHC process is the detection of a protein, more precisely an epitope that is a part of the protein known as the antigenic determinant, which can be recognized specifically by antibodies. (8).

MATERIALS AND METHODS
Twelve (12) male New Zealand rabbits, three to four months in age, and 1.2∼1.4Kg in weight were included.They were kept in clean cages and housed in a standard environment (a temperature of 18∼24°C).
They were fed equal amounts of standard food (grass and fresh vegetables) and water.unclear histology were excluded from the analysis.The scoring system was used for IL-6 staining intensity evaluation (9)(10)(11)(12) as shown in ( Ozoheal group showed that at day three perioKIN group showed higher expression of the marker but at day seven there was high scores for Ozoheal group and finally at day fourteen there is no expression of IL-6 marker in Ozoheal group, so there was a statistically significant difference at all periods for Ozoheal group as shown in (Figure 1 and Table 2).For more details (Figures 2 and 3

Days (P-value)
On the third day Group I day3 vs Group II day3 p< 0.001 (0.0001) Highly-significant On the seventh day Group I day7 vs Group II day7 p< 0.01 (0.005) Highly-significant On the fourteenth day Group I day14 vs Group II day14 p< 0.001 (0.0001) Highly-significant      We agree with the proposal recently done (13)   Although IL-6 is an important proinflammatory cytokine closely implicated in the progression of the inflammatory process, (14,15) high concentrations of pro-inflammatory cytokines inhibit the migration of epithelial cells and gingival fibroblasts, and enhance the expression of TNF-a and IL-1b at the wound site, thereby reducing cell survival and delaying wound closure. (16)The proinflammatory cytokines proved to be where IL-6 and TNF-α also be seen in the chronic granulation tissue of the mouse after a wound age of up to 28 days in macrophages (17)   .
The distribution pattern of proinflammatory mediators in normal human skin is described as nonuniform.Nevertheless, there is accordance in the basics in the expression pattern of IL-1β, IL-6, and TNF-α in normal skin (18) which we found and those of most literature information, predominantly a slight (up to negative) preexisting, frequently granular or intercellular formed expression of mediators.

CONCLUSION
Ozoheal drug is considered to be better than perioKIN gel in the treatment of wound healing also to accelerate the healing process as it is regarded as a -DOI: 10.33899/RDENJ.2021.131056.1127, © 2023, College of Dentistry, University of Mosul.This is an open access article under the CC BY 4.0 license (http://creativecommons.org/licenses/by/4.0/)INTRODUCTION A wound is defined as a break in the continuity of the covering skin, and/or lining mucosa.The wound healing process is a complex mechanism involving a cascade of coordinated inflammatory and proliferative steps.Different biomaterials might be required to improve clinical outcomes, and have a potential function in wound healing and regeneration, thereby playing an important role in tissue repair mechanisms in many oral surgical procedures. (1, 2).Hyaluronic acid (HA) is a polysaccharide found in the extracellular matrix of connective tissue.The biological action of HA comes from its potential relationship with growth factors, and serves as a lubricant for various body tissues.thus maintaining the structural integrity of tissues.The biocompatible properties of hyaluronic acid being biocompatible, with ) showed photomicrograph of oral mucosa for day 3 of the perioKIN group with moderate positive reaction (++), And the Ozoheal group showed mild positive reaction (+) of IL-6 expression to immunohistochemistry in the submucosa at the site of the wound, (Figure 4 and 5) showed day 7 with mild positive reaction (+) for perioKIN group and shows moderate positive reaction (++) for another group, (Figures 6 and 7) showed day 14 with moderate positive reaction (++) of IL-6 expression for perioKIN group and negative reaction (-) for the ozoheal group.

Figure 2 :
Figure 2: photomicrograph of oral mucosa of group I (3 days) shows moderate positive reaction (+2) of IL-6 expression to immunohistochemistry in the submucosa (arrow) at the site of wound.IHC for IL-6, 400X.

Figure 3 :
Figure 3: photomicrograph of oral mucosa of group II (3 days) shows mild positive reaction (+1) of IL-6 expression to immunohistochemistry in the submucosa (arrow) at the site of wound.IHC for IL-6, 400X.

Figure 4 :
Figure 4: photomicrograph of oral mucosa of group I (7 days) shows mild positive reaction (+1) of IL-6 expression to immunohistochemistry in the submucosa (arrow) at the site of wound.IHC for IL-6, 400X.

Figure 5 :
Figure 5: photomicrograph of oral mucosa of M2 group (7 days)shows moderate positive reaction (+2) of IL-6 expression to immunohistochemistry in the submucosa (arrow) at the site of wound.IHC for IL-6, 400X.

Figure 6 :
Figure 6: photomicrograph of oral mucosa of M1 group (14 days)shows moderate positive reaction (2+) of IL-6 expression to immunohistochemistry in the submucosa (arrow) at the site of wound.IHC for IL-6, 400X.

Figure 7 :
Figure 7: photomicrograph of oral mucosa of M2 group (14 days)shows negative reaction (0) of IL-6 expression to immunohistochemistry in the submucosa at the site of wound.IHC for IL-6, 400X.

Table 1
).The median and interquartile range were used to present the data, and a non-parametric test (Mann-Whitney U test) was used for statistical analysis