| Objective:To explore the influence of recombinant humangranulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel ondeep partial thickness burn and the mechanism of action on promoting woundhealing by establishing the model of deep partial burn in rats, coated withrecombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF) hydrogel and bandaged with iodophor gauze. Methods:1.Modeling:40SD rats were randomly divided into experimental and controlgroups, each group of20, average body weight (250±25) g, male or female.0.3%sodium pentobarbital (1ml/100g) used for anesthesia by intraperitonealinjection, and then with6%sodium sulfide on the back of rat to remove the hair.A size of22cm×11cm wooden pre-production, middle to make a5cm x6cmrectangular hole.Twenty four hours later, rats were taken supine limbs fixed inthe wood, make it back hair removal skin of exposed wood hole, and then woodplaced in heated hot water bath of80℃, mild pressing the wood to the backhair removal skin of SD rats immersed into hot water for15s, then the model ofdeep partial burn in rats were established.The depth was histologicallyconfirmed. Wipe dry the injured area with dry gauze, and recovery the rat withintraperitoneal injection of5ml saline. The experimental group, recombinanthuman granulocyte-macrophage colony stimulating factor hydrogel evenly spread in the wound, the thickness of about1mm, bandaged with a layer ofVaseline gauze and3-4layers of sterile gauze. The control group, bandagedwith gauze soaked in iodine solution of0.5‰and3-4layers of sterilegauze.Each wound respectively treated with rhGM-CSF hydrogel and iodinesolution of0.5‰,once a day for13d. Select5rats from each group randomlyto execute4d,7d,10d and13d after administration respectively.2. The generalobservation of wound: Observing and recording the experimental group’s andcontrol group’s speed of scabbing and scabbing off, the situation ofsecretions, and a margin of reaction on4:00pm every day.3. The calculations ofwound healing rate (WHR): Select5rats from each group randomly to execute4d,7d,10d and13d after administration respectively.And then calculate the areaof each wound with transparent graph paper.The wound healing rate (woundhealing rate WHR)=the wound area before treatment minus the area of thewound after treatment/the area of the wound before treatment.4.The count offibroblasts (FB):Select5rats from each group randomly to execute4d,7d,10dand13d after administration respectively. The wound skin in the wound centerwas drawed as sample,and then place it in4%formaldehyde solution.Samplewas progressively dehydrated in ethanol (75%'85%'95%'100%), andthen be cleared twice with xylene.The specimen was embedded in Paraffin, HEstained to observe and count the fibroblasts at400magnification in fivehorizons taken randomly.5.Wound microvascular density (MVD): Specimenswere immunohistochemistry stainned with antibodies to anti-angiogenic factor Ⅷ. Observing and counting the microvessels at400magnification in fiveconsecutive rather than overlapping horizons.6. The area of the collagen fibersin wound: Specimens were trichrome masson stainned to observe and count thecollagen fibers area in wound center by Image-Pro-Plus image analysissoftware at400magnification in five horizons taken randomly.Results:1. Thegeneral observation of wound: Handled by two different treatment methods, thetwo groups of the burn wound area were gradually reduced increased with timeprogress.Both the speed of scabbing and scabbing off of the experimental groupwere faster than the control group. Wound secretion is relatively large and someof which are purulent secretions in the control group.The duration of marginalreaction in the control group is longer than it in the experimental group.2. Thecalculations of wound healing rate (WHR):4d of the wound healing rate in theexperimental group and control group:5.39±0.45and4.92±0.69, thedifference being not statistically significant (P>0.05). The wound healing ratein the experimental group was17.54±1.49,27.67±1.00,52.40±4.84respectivelyon7d,10d,13d after treatment,which were obviously higher than that in thecontrol group(11.48±0.80、15.59±0.50、31.91±2.67, respectively,P<0.05).3. Thecount of fibroblasts (FB): The count of fibroblasts in the experimental groupwas obviously higher than that in the control group on4d,7d,10d after treatment(P<0.05).Nevertheless, The count of fibroblasts in the control group wasobviously higher than that in the experimental group on13d after treatment(P<0.05).4. Wound microvascular density (MVD): The number of microvessels in the experimental group was1.623±0.272、2.358±0.135、5.557±0.479、7.807±0.242respectively on4d,7d,10d,13d after treatment,which wereobviously higher than that in the control group(0.856±0.109、1.497±0.282、3.446±0.286、6.002±0.313, respectively,P<0.05).5. The area of the collagenfibers in wound: The area of the collagen fibers in the experimental group wasobviously higher than that in the control group on4d,7d,10d,13d after treatment(P<0.05). The collagen fibers in the experimental group was arraged tidily andorderly, but which in the control group was relativelydisorder.Conclusion:Recombinant human granulocyte-macrophage colonystimulating factor (rhGM-CSF) hydrogel can promote the formation andshedding of the burn wound scab. To increase wound healing and improve therepaired quality by promoting microvascular growth, promoting fibroblastformation and wound collagen formation. Compared to conventional treatmentmethods, rhGM-CSF hydrogel can significantly accelerate wound healing ondeep partial burn. |
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