You Z G, Zhang L P, Wang X G, Zhou H L, Guo S X, Wu P, Han C M
Department of Burns, the Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou 310009, China.
Zhonghua Shao Shang Za Zhi. 2017 Feb 20;33(2):103-110. doi: 10.3760/cma.j.issn.1009-2587.2017.02.011.
To explore the influence of collagen/fibroin scaffolds containing silver nanoparticles on dermal regeneration of full-thickness skin defect wound in rat. Eighty-one collagen/fibroin scaffolds containing silver nanoparticles (with the mass concentration of silver nanoparticles as 10 mg/L) and 81 collagen/fibroin scaffolds without silver nanoparticles were produced respectively with freeze-drying method and enrolled as silver nanoparticles scaffold group (SNS) and control scaffold group (CS). Nine scaffolds in each group were cultured with human fibroblasts. At post culture hour (PCH) 2, 12, and 24, the human fibroblasts adherent to the scaffolds (=3) in two groups were counted. Four full-thickness skin defect wounds were reproduced on the back of each one of the 36 SD rats. The rats were divided into groups SNS (wounds were transplanted with collagen/fibroin scaffolds containing silver nanoparticles) and CS (wounds were transplanted with collagen/fibroin scaffolds without silver nanoparticles) according to the random number table, with 18 rats in each group. In post surgery week (PSW) 1, 2, and 4, 6 rats in each group were sacrificed respectively for general observation, observation of histological structure, inflammatory cell infiltration, and collagen deposition with HE staining, count of CD68 positive cells with immunohistochemical staining, and mRNA expressions of interleukin-6 (IL-6) and IL-10 with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, test, and Bonferroni correction. (1) At PCH 2, 12, and 24, the numbers of human fibroblasts adherent to the scaffolds in the two groups were close (with values from 1.77 to 2.60, values above 0.05). (2) In PSW 1, no obvious symptom of infection was observed in wound or wound edge of rats in group SNS with obvious vascularization of scaffolds, while obvious symptoms of infection were observed in wounds of rats in group CS with some scaffolds exfoliated. In PSW 2, the scaffolds were firmly attached to the wounds of rats in group SNS, while obvious contracture was observed in the wounds of rats in group CS with a lot of scaffolds exfoliated. In PSW 4, the scaffolds covered the wounds of rats in group SNS with obvious epithelization on the surface of the scaffolds, while all the scaffolds exfoliated, leaving obvious contracture of residual wounds of rats in group CS. (3) In PSW 1 and 2, compared with those in group CS, more collagen secretion and tissue regeneration and less inflammatory cell infiltration in the scaffolds were observed in the wounds of rats in group SNS. In PSW 4, obvious epithelization was observed in the wounds of rats in group SNS, while inflammatory cell infiltration was observed without obvious epithelization in the wounds of rats in group CS. (4) In PSW 1, the number of CD68 positive cells in the wounds of rats in group SNS [(54±10) /mm(2)] was similar to that in group CS [(78±7) /mm(2,) =1.52, >0.05]. In PSW 2 and 4, the numbers of CD68 positive cells in the wounds of rats in group SNS [(154±10) and (77±7) /mm(2)] were significantly less than those in group CS [(268±16) and (136±13) /mm(2,) with values respectively 7.31 and 3.83, values below 0.01] respectively. (5) Except for the expression in PSW 4 (=1.23, >0.05), the mRNA expressions of IL-6 in the wounds of rats in group SNS in PSW 1 and 2 were significantly lower than those in group CS (with values respectively 13.12 and 4.65, values below 0.01). Except for the expression in PSW 1 (=3.08, <0.05), the mRNA expressions of IL-10 in PSW 2 and 4 in the wounds of rats in the two groups were similar (with values respectively 2.14 and 0.49, values above 0.05). Besides good biocompatibility, collagen/fibroin scaffolds containing silver nanoparticles have obvious effect in modulating inflammation, thus they can accelerate dermal regeneration induced by collagen/fibroin scaffolds for wound repair.
探讨含银纳米颗粒的胶原/丝素蛋白支架对大鼠全层皮肤缺损创面真皮再生的影响。采用冷冻干燥法分别制备81个含银纳米颗粒的胶原/丝素蛋白支架(银纳米颗粒质量浓度为10 mg/L)和81个不含银纳米颗粒的胶原/丝素蛋白支架,分别作为银纳米颗粒支架组(SNS)和对照支架组(CS)。每组取9个支架与人成纤维细胞共培养。在培养后2、12和24小时,计数两组中附着于支架上的人成纤维细胞数量(每组 =3)。在36只SD大鼠的背部各制备4个全层皮肤缺损创面。根据随机数字表将大鼠分为SNS组(创面移植含银纳米颗粒的胶原/丝素蛋白支架)和CS组(创面移植不含银纳米颗粒的胶原/丝素蛋白支架),每组18只。在术后第1、2和4周,每组分别处死6只大鼠,进行大体观察、组织学结构观察、HE染色观察炎症细胞浸润和胶原沉积情况、免疫组化染色计数CD68阳性细胞以及实时荧光定量逆转录聚合酶链反应检测白细胞介素-6(IL-6)和IL-10的mRNA表达。数据采用析因设计方差分析、t检验和Bonferroni校正进行处理。(1)在培养后2、12和24小时,两组中附着于支架上的人成纤维细胞数量相近(t值为1.77至2.60,P值均大于0.05)。(2)在术后第1周,SNS组大鼠创面及创缘无明显感染症状,支架血管化明显,而CS组大鼠创面有明显感染症状,部分支架脱落。在术后第2周,SNS组大鼠创面支架与创面紧密附着,而CS组大鼠创面出现明显挛缩,大量支架脱落。在术后第4周,SNS组大鼠创面被支架覆盖,支架表面有明显上皮化,而CS组所有支架均脱落,大鼠残余创面挛缩明显。(3)在术后第1和2周,与CS组相比,SNS组大鼠创面支架内胶原分泌、组织再生更多,炎症细胞浸润更少。在术后第4周,SNS组大鼠创面有明显上皮化,而CS组大鼠创面有炎症细胞浸润但无明显上皮化。(4)在术后第1周,SNS组大鼠创面CD68阳性细胞数量为(54±10)/mm²,与CS组的(78±7)/mm²相似(t =1.52,P>0.05)。在术后第2和4周,SNS组大鼠创面CD68阳性细胞数量分别为(154±10)和(77±7)/mm²,明显少于CS组的(268±16)和(