白芨胶载高分子神经生长因子促进伤口的愈合

背景:生长因子开创了伤口愈合研究与应用的新纪元,祖国医学与生物医学相结合对伤口愈合的影响尚在探索中。目的:用中药白芨胶作为外源性高分子神经生长因子的载体,观察促进大鼠伤口愈合的作用。设计:高分子神经生长因子活性鉴定采用单一样本观察;观察白芨胶载高分子神经生长因子对伤口愈合的作用采用随机对照动物实验。单位:深圳市布吉人民医院骨科,华中科技大学同济医院骨科。材料:90～120dSD大鼠40只,雌雄不限,体质量220～280g;牛精液高分子神经生长因子1mg/支,冻干制剂。方法:实验于2001-09/2004-12在深圳市布吉人民医院骨科及华中科技大学同济医院骨科实验室完成。实验分两部分进行:①将生理盐水、白芨胶、高分子神经生长因子及高分子神经生长因子 白芨胶分别加入无血清培养基中,观察其对鸡胚背根神经节的影响。②选用SD大鼠40只,分为4组,每组大鼠均为10只。空白对照组,伤口不用药;白芨胶组、高分子神经生长因子组及高分子神经生长因子 白芨胶组,在各大鼠背部切取2cm的切口,分别给予外用白芨胶、高分子神经生长因子及高分子神经生长因子 白芨胶,用药1次/d。在用药后第3天及第10天测量伤口面积,并用图像分析仪计算伤口面积,用光学显微镜观察细胞渗出类型和数量,并观察伤口愈合时间。主要观察指标:①高分子神经生长因子活性鉴定。②伤口愈合速率的观察。③伤口愈合时间观察。④光镜组织学观察。结果:①高分子神经生长因子活性鉴定:鸡胚背根神经节培养24h后,高分子神经生长因子与高分子神经生长因子 白芨胶的稀液倍数以1∶106～1∶108神经节周围有突起生长反应最明显。而生理盐水及白芨胶,均未见神经节有突起生长。②伤口愈合速率的观察:高分子神经生长因子 白芨胶组伤口愈合速率显著高于空白对照组、白芨胶组、高分子神经生长因子组。③伤口愈合时间观察:空白对照组、白芨胶组、高分子神经生长因子组及高分子神经生长因子 白芨胶组伤口愈合时间依次为(19.5±0.7),(17.3±0.6),(16.6±0.7)及(14.9±0.4)d。④光镜组织学观察:用药后3d组织切片观察:白芨胶组、高分子神经生长因子组和高分子神经生长因子 白芨胶组渗出的多形核白细胞、单核细胞及成纤维细胞数量明显多于空白对照组,且高分子神经生长因子组及高分子神经生长因子 白芨胶组可见毛细血管。用药后第10天组织切片观察:高分子神经生长因子组、高分子神经生长因子 白芨胶组与空白对照、白芨胶组比较,肉芽组织较致密的,含有较多毛细血管。结论:高分子神经生长因子 白芨胶在伤口愈合早期及后期均有显著促进伤口愈合作用,且作用优于单用白芨胶或高分子神经生长因子。

High-molecular-weight nerve growth factor with bletilla colloid as a carrier in the facilitation of wound healing

BACKGROUND:. Growth factor starts a new epoch of the research and application of wound healing, and the effect of the combination of Chinese tradition medicine and biomedicine in wound healing is still in the discussion.OBJECTIVE: To observe the effect in promoting wound healing of rats with bletilla colloid, a Chinese herbal medicine, as a carrier of exogenous high molecular weight nerve growth factor (HMW-NGF).DESIGN: The single sample observation was adopted in the identification of activities of HMW-NGF; The randomized control experiment with animal as subjects was adopted in the observation of the effect of the HMW-NGF mixed with the bletilla colloid as a carrier in promoting wound healing.SETTING: Department of Orthopaedics, Shenzhen Buji People's Hospital,and Department of Orthopaedics, Tongji Hospital Affiliated to Huazhong University of Science and Technology.MATERIALS: Forty SD rats aged 90-120 days of either gender were employed, with the body mass of 220-280 g; The freeze dried preparation of cattle seminal fluid with HMW-NGF with 1mg each.METHODS: From September 2001 to December 2004, the experiment was carried out in the Department of Orthopaedics in Shenzhen Buji People's Hospital and Central Laboratory of Department of Orthopaedics in Tongji Hospital Affiliated to Huazhong University of Science and Technolcolloid, HMW-NGF, HMW-NGF+ bletilla colloid were added into the serum-free medium (SFM), and the influences of them on the dorsal root lected and divided into 4 groups with 10 rats in each group. The rats of normal control group had no medicine on the wound. In the bletilla colloid group, the HMW-NGF group and the HMW-NGF+bletilla colloid group, incision size 2 cm was made at the back of each rat, and bletilla colloid,HMW-NGF, HMW-NGF+ bletilla colloid were applied respectively on the wound, once per day. The wound area was measured at day 3 and 10 after treatment, and the wound area was calculated by the image analysator. The category and the quantity of the emigrated cells were observed with light microscope, and the healing time of the wound was observed.tion under light microscope.the DRG of chick embryo was cultured, the reaction of evection growth around the ganglion in the HMW-NGF, HMW-NGF+ bletilla colloid with the dilution desity of 1:106-1: 108 was most significant, which was not seen in healing with HMW-NGF + bletilla colloid was significantly increased as compared with that in normal control group, the bletilla colloid group and the The healing time of the wound of the normal control group, the bletilla colloid group and the HMW-NGF group and the HMW-NGF+ bletilla colloid group were (19.5±0.7), (17.3±0.6), ( 16.6±0.7 )and ( 14.9±0.4 ) days respecvation of cut sheets 3 days after treatment: The quantity of polymorphonuclear leucocytes, histoleueocyteand fibroblast in the bletilla colloid group,the HMW-NGF group and the HMW-NGF+ bletilla colloid group were significantly larger than that in the normal control group, and the micrangium could be seen in the HMW-NGF group and the HMW-NGF+ bletilla colloid group. The observation of cut sheets 10 days after treatment: The granulation tissue in the HMW-NGF group and the HMW-NGF+ bletilla colloid group was compacter and had more micrangium as compared with the normal control group and the bletilla colloid group.CONCLUSION: HMW-NGF+ bletilla colloid has significant effect on promoting the healing of the wound at the earlier and later period, and the effect is better than that of bletilla colloid or HMW-NGF.