明胶/白芨胶-微米三七海绵材料对创面愈合影响的实验研究

The preparation of gelatine/Bletilla striata gelatin-micron Panax notoginseng porous biomaterial and its effects on wound healing

目的将微米三七赋以新型复合材料载体,制作出一种新型止血材料,并通过理化试验初步评估其性能及生物相容性。研究该材料对大鼠背部创伤模型愈合的影响,探讨其促进创面愈合的机制。方法应用高能球磨法制备超微米级的三七粉,与明胶、白芨胶按一定比例通过冷冻干燥法制备出可降解的复合性海绵。经过常规理化分析及溶血率测定,证实具有良好的生物相容性。选取SPF级健康SD雄性大鼠60只,将其随机分为空白对照组(A组)、明胶海绵组(B组)、明胶/白芨胶海绵材料组(C组)和明胶/白芨胶-微米三七海绵材料组(D组),各15只。先制备全层皮肤缺损的大鼠动物模型,再分别用无菌纱布、明胶海绵、明胶/白芨胶海绵材料(不载有微米三七)、明胶/白芨胶-微米三七海绵材料覆盖创面,并分别于术后第3、7、14天测量伤口面积并取材(每组每次各取5只),计算各组的创面愈合速率,并用免疫组织化学染色(SABC法)检测各组干预措施对创面组织VEGF(血管内皮生长因子)的表达情况。结果明胶/白芨胶-微米三七海绵材料的理化性能符合创面敷料基本要求,具有组织相容性;各组在术后第3、7天的创面愈合速率差异均有统计学意义(P<0.05),在第14天,各组间比较差异无统计学意义(P>0.05);各组各时间点VEGF的阳性表达差异均有统计学意义(P<0.05),其中材料治疗组创面肉芽组织中VEGF阳性表达显著性升高。结论明胶/白芨胶-微米三七海绵可作为一种新型创面止血敷料,它具有良好的组织相容性、制备工艺简便、成本低等特点。通过动物模型实验初步证实该材料可促进创面的愈合,其作用机理可能与促进创面VEGF的高表达相关。

Objective To make a novel wound dressing-a composite carrier for micron-sized particles of Panax notoginseng, to evaluate the property and biocompatibility of the newly composited material by a series of physical and chemical testing, and to evaluate the effects of the novel wound-dressing on wound healing in rats by the preparation of the traumatic model. Methods High-energy ball milling was used to prepare micron-sized particles of Panax notoginseng, and it was compounded with scaffolds. Then the mixture was lyophilized to make a spongy and porous material. Sixty healthy SPF level male SD rats were randomly divided into blank control group （group A）, gelatin sponge group （group B）, gelatine/Bletilla striata gelatin sponge group （group C） and gelatine/Bletilla striata gelatin-micron Panax notoginseng sponge group （group D）, with fifteen SD rates in each group. And then the circular skin loss （Φ=1.8 cm） were prepared on dorsal skin. The wounds were covered by sterile gauze, absorbable gelatin sponge, porous gelatine/Bletilla striata gelatin （without micron-sized particles of Panax notoginseng） and this kind of porous biomaterial carrying micron-sized particles of Panax notoginseng respectively. The areas of the wound were measured 3, 7 and 14 days after the surgery to calculate the wound healing rate, and the full-thickness of the skin which was 2 cm inside the wounds was taken from five rats of each group each time respectively to detect the expression of VEGF by immunohistochemistry detection. Results Physical and chemical tests showed that gelatine/Bletilla striata gelatin-micron Panax notoginseng porous biomaterial would meet the standards of an eligible wound dressing. Significant difference are found among the four groups in wound healing rates 3 and 7 days after the surgery respectively （P〈0.05）, and no significant difference are found among the four groups 14 days after the surgery （P〉0.05）. Increased expression of VEGF is found in each group at the three time points, and the difference among these four groups are all significant （P〈0.05）. Furthermore, group D has the highest VEGF expression. Conclusion As a novel wound dressing, gelatine/Bletilla striata gelatin-micron Panax notoginseng porous biomaterial is safety for wound healing dressing. The preparation technology of this gelatine/Bletilla striata gelatin-micron Panax notoginseng porous biomaterial is approaching perfection. Through the animal model of rats, we have proved that this material can shorten would healing time and accelerate wound healing. The mechanics of it may be that this kind of material can promote the expression of VEGF at the surface of the wound.