Objective:
To explore the influence of collagen/fibroin scaffolds containing silver nanoparticles on dermal regeneration of full-thickness skin defect wound in rat.
Methods:
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 (n=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, t test, and Bonferroni correction.
Results:
(1) At PCH 2, 12, and 24, the numbers of human fibroblasts adherent to the scaffolds in the two groups were close (with t values from 1.77 to 2.60, P 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) /mm2] was similar to that in group CS [(78±7) /mm2, t=1.52, P>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) /mm2] were significantly less than those in group CS [(268±16) and (136±13) /mm2, with t values respectively 7.31 and 3.83, P values below 0.01] respectively. (5) Except for the expression in PSW 4 (t=1.23, P>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 t values respectively 13.12 and 4.65, P values below 0.01). Except for the expression in PSW 1 (t=3.08, P<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 t values respectively 2.14 and 0.49, P values above 0.05).
Conclusions
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.