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bFGF/胶原蛋白复合海绵的研制及体内外药理学研究

Preparation and in vitro /in vivo pharmacological studies of bFGF/collagen composite sponge

  • 摘要: 目的: 研制碱性成纤维细胞生长因子(bFGF)/胶原蛋白复合海绵并对其体外活性和体内过敏性进行研究。 方法: 以新鲜猪皮和bFGF为原料,制成bFGF/胶原蛋白复合海绵,并检测其理化性质,采用MTT法测定其浸提液对小鼠胚胎成纤维细胞3T3小鼠细胞的体外促增殖活性,采用迟发型超敏反应封闭贴敷试验研究其体内过敏作用。 结果: 高低剂量bFGF/胶原蛋白复合海绵的表观密度等理化性质指标与空白海绵比较均没有显著性差异;电泳检查结果表明复合海绵在约18 kD处具有明显的条带;体外活性检测表明高低剂量bFGF/胶原蛋白复合海绵生理盐水浸提液均对3T3细胞具有明显的促细胞增殖作用,与PBS对照组比较具有显著性差异(P<0.01,P<0.05)。在过敏试验中,高低剂量bFGF/胶原蛋白复合海绵组以及生理盐水浸提液组动物在诱导和激发期间均未见过敏症状,病理检查结果表明,动物皮肤未见炎症病变;而阳性对照2,4-二硝基氯苯组动物过敏反应发生率为100%,且具有明显的炎症病变。 结论: bFGF/胶原蛋白复合海绵理化性质较好,具有明显的促细胞增殖活性,对皮肤无过敏反应,有望开发成为新型外用治疗创伤的材料。

     

    Abstract: Aim: To investigate a new type wound dressing,basic fibroblast growth factor(bFGF)/collagen composite sponge,and conduct its pharmacological studies in vitro and in vivo. Methods: bFGF/collagen composite sponge was prepared using fresh pig skin and bFGF.The sponge′s physicochemical properties were studied.MTT assay was used to detect the proliferation effect of the sponge extract on 3T3 cells.Delayed allergy of the sponge was tested for the assurance of its biosafety. Results: Results showed that the physicochemical properties of bFGF/collagen composite sponge with high and low doses of bFGF have no significant difference from those of blank collagen sponge.SDS-page analysis indicated that the composite sponge has apparent strip in 18 kD.It was also found that bFGF/collagen composite sponge was responsible for significant effects on 3T3 cell proliferation in comparison to saline treatement (P<0.01 ,P<0.05).In the allergy study,during the periods of the induction and stimulation,no allergic reaction was found in bFGF/collagen composite sponge groups with high and low doses of bFGF,while severe reactions and inflammation occurred in positive group (2,4-dinitrochlorobenzene). Furthermore,pathological examination indicated the intact dermal structure and no sign of inflammation. Conclusion: The developed sponge has good physicochemical propertis and noticed cellular proliferation without dermal irritation.There is much potential to develop bFGF/collagen composite sponge into a new kind of wound dressing material for clinical use.

     

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