Wound Healing Histopathological Analysis

On postoperative days 5, 7, 14, 21 and 35, the wound and surrounding tissue (0.5 cm) were carefully excised, rinsed in PBS, and then fixed in 4% Paraformaldehyde (PFA). Samples were dehydrated in a graded ethanol series (70–100%) and embedded in paraffin. Five-micrometer sections were prepared. According to the standard procedures, samples were stained with either Hematoxylin and Eosin (HE) or Masson trichrome, and immunohistochemistry including α-SMA (1:100, Boster), K1 (1:100, Abcam), K6 (1:500, Covance), CD86 and CD163 (1:200, Bioss)38 (link).
Image-Pro Plus was used to analyse the average optical density values for K1, K6, α-SMA expression. Five randomly selected fields of view were examined for each group at each time point and used to assess the average optical density value per unit area. Statistics regarding the number of positively stained cells for CD86 and CD163 were obtained using five randomly selected fields of view for each group at each time point.

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Chen S., Shi J., Zhang M., Chen Y., Wang X., Zhang L., Tian Z., Yan Y., Li Q., Zhong W., Xing M., Zhang L, & Zhang L. (2015). Mesenchymal stem cell-laden anti-inflammatory hydrogel enhances diabetic wound healing. Scientific Reports, 5, 18104.

Publication 2015

CD163

Cd163 Dehydrated ethanol Embedded paraffin Eosin Immunohistochemistry Optical Paraformaldehyde Tissue Wound

Corresponding Organization :

Other organizations : Southern Medical University, Children's Hospital Research Institute of Manitoba, University of Manitoba

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Variable analysis

independent variables

Postoperative days (5, 7, 14, 21, 35)

dependent variables

Wound and surrounding tissue (0.5 cm) histological analysis
Average optical density values for K1, K6, α-SMA expression
Number of positively stained cells for CD86 and CD163

control variables

Wound and surrounding tissue samples were carefully excised, rinsed in PBS, and then fixed in 4% Paraformaldehyde (PFA)
Samples were dehydrated in a graded ethanol series (70–100%) and embedded in paraffin
Five-micrometer sections were prepared
Samples were stained with either Hematoxylin and Eosin (HE) or Masson trichrome, and immunohistochemistry including α-SMA (1:100, Boster), K1 (1:100, Abcam), K6 (1:500, Covance), CD86 and CD163 (1:200, Bioss)

controls

No specific positive or negative controls were mentioned in the input protocol.

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