Mechanism of Co-transplantation of Platelet Rich Plasma and Human Hair Follicle Stem Cells on Deep Second-degree Burn Wound Healing in Rats

XU Linshengzi; QIU Weiming; WANG Yu; LIU Yueping; DIAO Bo

doi:10.13730/j.issn.2097-2148.2025.05.003

Abstract：Objective To investigate the effects and mechanism of co-transplantation of platelet rich plasma(PRP)and human hair follicle stem cells(HFSCs)on deep second-degree burn wounds in rats.Methods Cell counting kit 8(CCK-8)cell proliferation experiment and cell scratch experiment were used to explore the effect of PRP on the prolifera-tion and migration of human HFSCs under in vitro culture conditions.Twenty-eight SD rats were used to establish deep second-degree burn wound model,the burn wound tissues and normal skin tissues of 4 rats after modeling were randomly selected to observe and identify whether the model was successfully established by hematoxylin and eosin(HE)staining.Twenty-four rats with successful modeling were randomly divided into control group[injection of 500 μl phosphate buffered saline(PBS)under burn wound tis-sues at multiple point injections],HFSCs group(the hu-man HFSCs suspension mixed with 500 μl PBS was injected at multiple points under the burn wound tissues),PRP group(500 μl PRP was injected at multiple points under the burn wound tissues)and PRP+HFSCs group(the HFSCs suspen-sion mixed with PRP was injected at multiple points under the burn wound tissues)by random number table method,with 6 rats in each.Injections were administered twice weekly around the burn area.Burn wound healing rates were calculated on the 7th,14th and 20th days of the interventions;on the 20th day of the interventions,burn wound tissues was harves-ted for further analysis,immunohistochemical staining was conducted to quantify number of CD31 positive vessels,and Masson staining was performed to observe collagen fiber regeneration in burn wound tissues;reverse transcription quanti-tative polymerase chain reaction(RT-PCR)was used to assess the mRNA relative expression levels of interleukin 1 beta(IL-1 β),superoxide dismutase 1(SOD1),B-cell lymphoma 2(Bcl-2),transforming growth factor beta(TGF-β)and ma-trix metalloproteinase 9(MMP-9)in burn wound tissues;Western blot was conducted to evaluate the protein relative ex-pression levels of IL-1β,SOD1 and Bcl-2 in burn wound tissues.Results The 10％PRP in the medium enhanced the pro-liferation and migration of HFSCs.Compared with the control group,HFSCs group and PRP group,the PRP+HFSCs group demonstrated the highest wound healing rates on the 7th,14th and 20th days of the interventions(all P＜0.05);on the 20th day of the interventions,the PRP+HFSCs group exhibited the highest number of CD31 positive vessels(P＜0.05),the mRNA and protein expression levels of pro-inflammatory factor IL-1β,TGF-β and MMP-9 were the lowest,and the mRNA expression levels of Bcl-2 and SOD1 were the highest(all P＜0.05),the protein expression level of IL-1βwas the lowest(all P＜0.05).Furthermore,compared with the control,HFSCs and PRP groups,the burn wound tissues of rats in PRP+HFSCs group exhibited the most compact and organized collagen fibers without excessive collagen deposi-tion.Conclusion PRP can promote the proliferation and migration of human HFSCs in vitro.The co-transplantation of PRP and human HFSCs can control the inflammatory response and oxidative stress at the burn site,promote the forma-tion of new blood vessels,thereby accelerating the wound healing.