| Purpose: To investigate the effect of Hyaluronic acid on the expansion capsular thickness, collagen content, the number of capillaries, the number of myofibroblast and its micro-structure changes, to clarify the impact of Hyaluronic acid on the transformation from fibroblast to myofibroblast, and collagen fiber synthesis during the expansion ,to reveal the mechanism of capsule formation after the intervention of hyaluronic acid, in order to provide a new method to provent reducing the utilization of the Envelope caused by expansion capsula's contracture, to further improve the efficacy of skin expansion techniques.Methods:1 Clinical dataPatients from our Branch during March 2007 and January 2009 were treated with skin soft tissue expansion , male 16 cases, female 12 cases, total 28cases, 18-32 years old, average 22-year-old. Expander implants in head 12 cases , face and neck 8 cases, the trunk 8 cases.2 GroupsThis experiment used oneself contract, dissected lacunar large enough enough in the subcuaneous around the pathological changes, then beried the same capacity soft tissue expander on both sides,and set to one of the side as the experimental group, the other side as the control group.3 Expander implant and injection expansionCoated the hyaluronic acid on the inside flap expansion evenly after the lacuna stripped in place, non-active bleeding, put 1% hyaluronic acid 3ml all around, and then implanted the skin and soft tissue expander in it; control group implanted in the same capacity of soft tissue expansion conventionally,not coated the hyaluronic acid. Started the expansion with water in the 5th day after opertations, with conventional methods of expansion, had each water injection every 3-4 days depending on the situation of expansion, the capacity of every injection of water depended by the pressure response of flap limited to 4-5 second; the same patients had the same each-water-injection in the experimental side and control side. Stop the water rejection after the required capacity (usually 150% -200% of rated capacity) . Started static expansion, it lasted normally 6 weeks.4 BasedDuring the second operations, peeled expansion capsular, tiled up in asepsis gauze, at symmetric parts of the expansion of skin, scissored parts tissue storde in -20°C refrigerator to measure HPr; sutured the remaining part of the corners to gauze, then immersed them in 4% paraformaldehyde fixed, for paraffin-embedded used as HE staining, Masson staining and immunohistochemical staining with CD34 andа-SMA. Then scissored about 0.1cm×0.1cm×0.3cm samples ,blocked them to 4% glutaraldehyde fixed 24h for evacuation TEM examination.5 Observation indicators(1)Expansion capsule thickness : Under light microscope observed the histological changes of experimental group and control group on HE sections, and measured specimens with the application of fiber gauge; (2) the form of collagen: Compared the form of collagen of experimental group with that of control group on the Masson stained sections, contrasted the difference of green between the two group; (3) Immunohistochemical staining of CD34 staining andα-SMA stained sections , Made comparisons between fibrous capsule of CD34 andα-SMA of the experimental group and that of the control group on light microscopy; (4) Messured hydroxyproline envelope (HPr) according to the measurement kit between experimental group and control group; (5) Under transmission electron microscope observed the cell organ and microfilament changes between the experimental group and control group specimens6 StatisticallyApplied SPASS11.5 statistical processing software to make statistical analysis for the experimental results.Results:1 HE staining and the thickness of expansion capsular Optical microscope: there were many fibroblasts (fibroblasts and myofibroblast distinction was not obvious) in the experimental group and control group ,and there was a few of new thin-walled capillaries, scattered in the perspective; the number naive view of the number of fibroblasts in the experimental group was more than that in the control group, while the number mature fibroblasts was reverse. and there were more thick collagen fibers in the control group.The fibrous capsule thickness of experimental group was 451.76±45.32μm, control group, while it of the control group was 838.57±55.49μm, there was significant difference between the two groups (P <0.05).2 Masson staining and collagenObserved under light microscope sections ,on Masson staining, the experimental group: Green staining of collagen fibers was thin and sparse, between which can see mucn red dyed cellulose, fibroblast nucleus were spindle-thin, gray-black, inner layer can be seen in a layer of endothelial cells like lining, occasional red staining of red blood cells can be seen; control group: Green staining of collagen fibers thick, with rules, red dyed cellulose less, many fibroblast nucleus arranged fusiform, scattered.3 Expression of CD34Under a light microscope we can see that capillary endothelial cells were marked with tawny on CD34 stained sections, scattered in the field of vision. The expressions of CD34 are both masculine in experimental group and control group. Image analysis illustrate that the experimental group was lower than the control group, but there was not significant difference between the two groups (P> 0.05).4 Expression ofα-SMAUnder a light microscope we can see that myofibroblast were marked with tawny onα-SMA stained sections, scattered in the field of vision. The expression ofα-SMA was masculine in experimental group, while that was better masculine in control group. Image analysis illustrate that the experimental group was lower than the control group, there was significant difference between the two groups (P <0.05).5 Determination of HPrPut the absorbance values into the formula to calculate the content of hydroxyproline specimens: experimental group 8.20±0.70,control group 10.76±1.00, there was significant difference between the two groups (P <0.05).6 TEM observationExperimental group: the myofibroblast mucleus was ellipse, parts of karyotheca disappearance, endoplasmic reticulum expansion into a pond-like, significantly reducing, and the phenomenon of degranulation Obviously, part of the mitochondria cristae fusion or disappeared, microfilaments with loose, the number of free ribosomes reducing. Control group: Myofibroblast larger, nuclear arranged oval, chromatin-rich, the expansion of intracytoplasmic rough endoplasmic reticulum, rare degranulation phenomenon, well-developed Golgi apparatus, mitochondria cristae basic integrity, microfilament bundles arranged along the with the long axis of the cell membrane, ribosomes no significant reduction in quantity.Conclusion:Hyaluronic acid can inhibit the conversion from fibroblast to myofibroblast, and reduce the synthesis and secretion of collagen, make the expansion of capsular thinning and collagen fiber content decreasing, decrease contractility, thereby reducing the expansion flap contraction and improve surgical outcome. Founded no impact on blood... |
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