The Study Of Influence Of L-Arginine Mixed Fibronectin On The Wound Healing Of Partial Thickness Burn Rats

Objective: By making the production of rat skin of partial thickness burn rats model, force-fed rats given L-arginine ( L-Arg), and burn local joint application fibronectin ( FN), intervent wound healing process and thereby observe the L-arginine and fibronectin combination of partial thickness burn wound healing speed and the quality of healing, and by immunohistochemistry revealed burn healing process of nitric oxide synthase (NOS) the expression of the laws of light microscopy observation during different stages of healing fibroblast cells and collagen fiber content changes detected by biochemical methods granulation tissue in the hydroxyproline content before and after the experiment by contrast, observation wound healing situation. Research on L-arginine and fibronectin deepâ…¡application to the promotion of healing of burn wounds, burns to promote the exploration of an effective method of wound healing, and further deepen the repair of burn wound healing clinical study provide an objective experimental basis.Method: 1, Laboratory animal: healthy male Wistar rats 60, weighing a number. 2, Experimental animal groups: The rats were divided into four groups: the control group (0.9% saline), the experimental group A (L-Arg 400mg/kg/d Joint 0.5 mg / ml FN0.05ml), B (L-Arg 400mg/kg/d), C (0.5mg/ml the FN0.05ml). 15 each. 3, Partial thickness burn rats model Preparation Methods: Rats with 3% intraperitoneal injection of pentobarbital anesthesia (35mg/kg), back cuts, and 8% sodium sulfide shed, wash temperature, Shian. Keeping with the same 24 hours after intraperitoneal injection of pentobarbital anesthesia successful, it will be fixed in the prone position in the operating table. On the back side on both sides of a spinal 1 cm, cranio-caudal ends of the spine parallel to the edge marked a 1.5 cm long, with an area of 2.25 square cm2 pretreatment marker. With the same area of kerosene gauze review in the back of the marker, with wet gauze to protect the surrounding skin, the light of kerosene combustion gauze and 10 seconds after saline gauze covered with anti-formed four deep squareâ…¡degree burn (pathology confirmed). From 0.5 mg / ml FN0.05ml injection in the experimental group A and C each wound and wound subcutaneous fascia, the control group and experimental group B to 0.9% saline bandaging supplies. Question rats regained consciousness, according to 400 mg / kg / d calculated and the experimental group were given A and B L-Arg gavage. The dressing was changed every other day one (the day the animals were killed not dressing). Experimental group A and C each day wound injection of 0.5 mg / ml FN0.05ml until the animals were killed. 4, drawing: each animal divided into five groups, each with three, respectively 3,5,7,14,21 days after the original burned cut within the framework of full-thickness skin, random organizations will be fixed in a 4% POM, conventional embedded in paraffin, sliced, respectively conventional HE staining, Masson staining and immunohistochemical staining. The remaining tissues were frozen at -20℃preserved tissue chemical detection. 5, observed: (1) general observation. (2) fibroblast density (numerical density on area, NA): 400 times the light microscope observation HE staining biopsy, in the granulation tissue central shallow, the central deep, on both sides of the randomly selected 10 of the rectangular field of vision (0.0052 mm2 /VF), visual count and calculate slice of units within the area of fibroblast cells, the results are from a few. (3) surface density of collagen fibers (area density on area AA): Masson staining tissue, in the granulation tissue central shallow, the central deep, on both sides of the Department of randomly selected five horizons, using computer-assisted image analysis system pathology, Green staining of collagen fibers of the surface density, the results are from a few. (4) Determination of hydroxyproline in accordance with the kit brochures operation of granulation tissue hydroxyproline content. (5) immunohistochemical staining biopsy, light microscopic observation NOS2 location and distribution of deep coloring degree. Digital medical image analysis system, the ratio test positive cells. 6, statistically: SPSS12.0 for windows statistical software, a single-factor analysis of variance methods of statistical analysis, data±s, with P <0.05 for the difference between statistical significance. Results: 1, morphological observation: in the control group post-traumatic 16 to 17 can basically healing; experimental group A good granulation tissue growth, healing time compared with the control group three to four days in advance. Experimental Group B, C healing well, compared with the control group 1 to 2 days in advance. 2, light microscopic observation: the experimental group A in the three days after burn, seven days than the control group B, C and the experimental group were more capillary formation, 14 days after burn, 21 Tiancheng fibroblast neat experimental group B , C after three days in the burn, seven days but also many capillary formation, but less than the experimental group A, and the inflammatory response in the experimental group A serious. 3, fibroblast density (NA): the experimental group A, B, C, 3,5,7 days in the burn NA control group (P <0.05), and in 14, 21 days after burns are low in the control group (P <0.05), while the experimental group A than group B and C more significantly (P <0.01). 4, collagen fiber areal density (AA): four groups showed continuous increase in the experimental group A at all time points were higher than the controls (P <0.01) in group B and C at all time points above the control group (P <0.05). 5, hydroxyproline content: the control group hydroxyproline content from trauma after 3 days to 14 days for sexual increase in the experimental group A after burn each time point hydroxyproline content of the control group (P <0.01), the experimental group B, group C at different time points after burn hydroxyproline content of the control group (P <0.05). 6, NOS₂ positive cells ratio: the control group after three days of burns positive staining intensity increased and reached a peak 3 to 7 days in the NOS₂ cells in the cell relative expression level remained stable, 14 days after the burn cells, the positive expression once again reached a peak, then gradually reduced expression. And the experimental group A, B and C group in three days after the start of positive staining increased to section 14 Tianshida to a climax, with the most positive expression of group A (group A, P <0.01, B, C, P <0.05). and subsequently expression gradually weakened.Conclusion: The burn wound healing process in the use of L-arginine alone or fibronectin can promote the healing of burn wounds, and locally with fibronectin at the same time the application of L-arginine has a significant promotion burn wound healing the role of rats can significantly accelerate the back skin deepâ…¡degree burn healing time.