Zhonghua Nan Ke Xue. 2024 Oct;30(10):903-909.
ABSTRACT
OBJECTIVE: To investigate the effect of lipopolysaccharide (LPS) on the crawling ability of fibroblasts derived from the scrotal skin under hypoxic conditions, its impact on vascular regeneration during wound healing, and the potential mechanism underlying chronic non-healing scrotal wound caused by gram-negative bacterial infection.
METHODS: We extracted peripheral blood buffy coats from healthy donors after venous blood centrifugation, divided them into four groups (normoxia, normoxia+LPS, hypoxia, and hypoxia+LPS) for pretreatment, and isolated the corresponding cultured supernatant. Then we measured the levels of VEGF-A, FGF and IL-8 secreted using enzyme-linked immunosorbent assay (ELISA), determined the cytotoxicity of different pretreated supernatants by lactate dehydrogenase (LDH) release assay, and detected the cell migration ability and vascular formation in human-umbical vein endothelial cells (HUVEC).
RESULTS: Under hypoxic conditions, the expression of HIF-1α was significantly lower in the fibroblasts from the chronic scrotal wound than in the normal fibroblasts (0.09 ± 0.00 vs 0.48 ± 0.01, P < 0.05), the viability of the cells showed no statistically significant difference from that under normoxic conditions (P > 0.05), and the proliferation and migration abilities of the cells were markedly improved (［83.09 ± 2.56］% vs ［17.84 ± 1.12］%, P < 0.05). The viability of the scrotal fibroblasts was significantly lower in the normoxia+LPS than in the hypoxia+LPS group (［20.15 ± 1.19］% vs ［32.54 ± 1.57］%, P < 0.05). The cells treated with LPS, compared with those in the non-LPS groups, exhibited remarkably decreased endothelial cell tubes formed (5.20 ± 0.15 vs 55.30 ± 1.15, P < 0.05), down-regulated expression of VEGF (OD value: 0.59 ± 0.07 vs 1.25 ± 0.11, P < 0.05) and up-regulated expression of IL-8 (OD value: 2.42 ± 0.07 vs 0.87 ± 0.11, P < 0.05).
CONCLUSION: The supernatant derived from the peripheral blood buffy coats treated with LPS has a stronger cytotoxicity and weaker angiogenesis than that untreated under hypoxic or normoxic conditions, which may be one of the critical factors for difficult healing of scrotal wound.
PMID:40783854
