Effects of advanced platelet-rich fibrin/chitosan thermosensitive hydrogel on full-thickness skin defect wound healing in diabetic rats

Xun Haoyi; Su Xiaowei; Hu Fangchao; Liu Xiangyu; Wu Yushou; Liu Tian; Sun Ran; Duan Hongjie; Chi Yunfei; Chai Jiake

doi:10.3760/cma.j.cn501225-20231020-00127

Volume 40 Issue 5

May 2024

Turn off MathJax

Article Contents

Abstract

References

Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > 40(5): 451-460

Yan ZZ,Wang YX,Zhang TL,et al.Properties of gelatin-polyethylene glycol hydrogel loaded with silver nanoparticle Chlorella and its effects on healing of infected full-thickness skin defect wounds in mice[J].Chin J Burns Wounds,2024,40(1):33-42.DOI: 10.3760/cma.j.cn501225-20231020-00126.

Citation:

Xun HY,Su XW,Hu FC,et al.Effects of advanced platelet-rich fibrin/chitosan thermosensitive hydrogel on full-thickness skin defect wound healing in diabetic rats[J].Chin J Burns Wounds,2024,40(5):451-460.DOI: 10.3760/cma.j.cn501225-20231020-00127.

Citation:

PDF( 9179 KB)

Effects of advanced platelet-rich fibrin/chitosan thermosensitive hydrogel on full-thickness skin defect wound healing in diabetic rats

doi: 10.3760/cma.j.cn501225-20231020-00127

1.
Graduate School, Chinese PLA General Hospital, Beijing 100853, China
2.
Burn Institute, the Fourth Medical Center, Chinese PLA General Hospital, Beijing 100048, China
3.
Hebei North University, Zhangjiakou 075132, China

Funds:

General Program of National Natural Science Foundation of China 81772067

General Program of Beijing Natural Science Foundation of China 7222179

More Information

Corresponding author: Chai Jiake, Email: cjk304@163.com
Received Date: 2023-10-20

Abstract

Abstract

Objective To prepare advanced platelet-rich fibrin (A-PRF)/chitosan thermosensitive hydrogel (hereinafter referred to as composite hydrogel) and explore the effects of composite hydrogel on full-thickness skin defect wound healing in diabetic rats. Methods This study was an experimental study. The composite hydrogel with porous mesh structure and thermosensitive characteristics was successfully prepared, containing A-PRF with mass concentrations of 10, 15, 20, 50, and 100 g/L. Diabetic model was successfully established in male Sprague-Dawley rats aged 6-8 weeks by intraperitoneal injection of streptozotocin, and 4 full-thickness skin defect wounds were established on the back of each rat (finally the model was successfully established in 36 rats). Three wounds of each rat were divided into blank group (no drug intervention), positive control group (dropping recombinant human granulocyte-macrophage stimulating factor gel), and chitosan hydrogel group (dropping chitosan hydrogel solution). Thirty rats were collected, and the remaining one wound of each rat (totally 30 wounds) was divided into 10, 15, 20, 50, and 100 g/L composite hydrogel groups, with 6 wounds in each group, which were dropped with composite hydrogel solution containing 10, 15, 20, 50, and 100 g/L A-PRF, respectively. Taking the remaining six rats, the remaining one wound from each rat was dropped with composite hydrogel solution containing 100 g/L A-PRF. On 14 d after injury, 6 rats with one wound dropped with composite hydrogel containing 100 g/L A-PRF were selected for hematoxylin-eosin (HE) staining to observe the inflammation, hemorrhage, or necrosis of the heart, liver, spleen, lung, and kidney. On 10 d after injury, 6 rats with one wound dropped with composite hydrogel containing 15 g/L A-PRF were selected to observe the blood perfusion of wounds in the four groups (with sample size of 6). On 7 and 14 d after injury, the wound healing rates in the eight groups were calculated. On 14 d after injury, the wound tissue in the eight groups was taken for HE and Masson staining to observe the formation of new epithelium and collagen formation, respectively; the positive expressions of CD31 and vascular endothelial growth factor A (VEGFA) were detected by immunohistochemistry, and the percentages of positive areas were calculated; the protein expressions of CD31 and VEGFA were detected by Western blotting; the mRNA expressions of CD31 and VEGFA were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction method (with all sample sizes of 4). Results On 14 d after injury, no obvious inflammation, hemorrhage, or necrosis was observed in the heart, liver, spleen, lung, and kidney in the 6 rats. On 10 d after injury, the blood perfusion volume of wound in 15 g/L composite hydrogel group was significantly more than that in blank group, positive control group, and chitosan hydrogel group, respectively (with P values all <0.05). On 7 and 14 d after injury, the wound healing rates of blank group were (26.0±8.9)% and (75.0±1.8)%, which were significantly lower than those of positive control group, chitosan hydrogel group, and 10, 15, 20, 50, and 100 g/L composite hydrogel groups, respectively ((45.8±3.2)%, (49.8±3.7)%, (51.2±2.9)%, (68.5±2.4)%, (68.8±1.5)%, (72.7±2.1)%, (75.0±3.7)% and (79.1±1.9)%, (77.2±1.7)%, (82.3±1.3)%, (89.6±1.9)%, (89.8±1.3)%, (87.3±1.1)%, (87.9±1.3)%), P<0.05; the wound healing rates of positive control group, chitosan hydrogel group, and 10 g/L composite hydrogel group were significantly lower than those of 15, 20, 50, and 100 g/L composite hydrogel groups (P<0.05). On 14 d after injury, the wound epithelialization degrees of 15, 20, 50, and 100 g/L composite hydrogel groups were higher than those of the other 4 groups, the new microvascular situation was better, and the collagen was more abundant and arranged more neatly. On 14 d after injury, the percentages of CD31 and VEGFA positive areas in wounds in positive control group and the percentage of VEGFA positive area in wounds in chitosan hydrogel group were significantly higher than those in blank group (P<0.05), the percentage of VEGFA positive area in wounds in 10 g/L composite hydrogel group was significantly higher than that in blank group, chitosan hydrogel group, and positive control group (with P values all <0.05), and the percentages of CD31 and VEGFA positive areas in wounds in 15, 20, 50, and 100 g/L composite hydrogel groups were significantly higher than those in blank group, positive control group, chitosan hydrogel group, and 10 g/L composite hydrogel group (P<0.05). On 14 d after injury, the protein and mRNA expressions of CD31 and VEGFA in wound tissue in chitosan hydrogel group, positive control group, and 10 g/L composite hydrogel group were significantly higher than those in blank group (P<0.05); the protein expression of VEGFA in wound tissue in 10 g/L composite hydrogel group was significantly higher than that in positive control group (P<0.05), and the mRNA expressions of CD31 and VEGFA in wound tissue in 10 g/L composite hydrogel group were significantly higher than those in positive control group and chitosan hydrogel group (P<0.05); the protein and mRNA expressions of CD31 and VEGFA in wound tissue in 15, 20, 50, and 100 g/L composite hydrogel groups were significantly higher than those in blank group, positive control group, chitosan hydrogel group, and 10 g/L composite hydrogel group (P<0.05); the mRNA expressions of CD31 and VEGFA in wound tissue in chitosan hydrogel group were significantly lower than those in positive control group (P<0.05). Conclusions The composite hydrogel has high biological safety, can improve wound blood perfusion, effectively promote the formation of blood vessels and collagen in wound tissue, thus promoting the wound healing of full-thickness skin defects in diabetic rats. 15 g/L is the optimal mass concentration of A-PRF in composite hydrogel.
- Biocompatible materials,
- Hydrogel,
- Diabetes mellitus,
- Wound repair,
- Advanced platelet-rich fibrin

FullText(HTML)

References(30)

References

[1]	HealdAH, StedmanM, DaviesM, et al. Estimating life years lost to diabetes: outcomes from analysis of National Diabetes Audit and Office of National Statistics data[J]. Cardiovasc Endocrinol Metab, 2020,9(4):183-185. DOI: 10.1097/XCE.0000000000000210.
[2]	RøikjerJ,WerkmanNCC,EjskjaerN,et al.Incidence, hospitalization and mortality and their changes over time in people with a first ever diabetic foot ulcer[J].Diabet Med,2022,39(4):e14725.DOI: 10.1111/dme.14725.
[3]	ApelqvistJ, BakkerK, van HoutumWH, et al. International consensus and practical guidelines on the management and the prevention of the diabetic foot. International Working Group on the Diabetic Foot[J]. Diabetes Metab Res Rev, 2000,16 Suppl 1:S84-92.DOI: 10.1002/1520-7560(200009/10)16:1+<::aid-dmrr113>3.0.co;2-s.
[4]	HangaardS,RasmussenA,AlmdalT,et al.Standard complication screening information can be used for risk assessment for first time foot ulcer among patients with type 1 and type 2 diabetes[J].Diabetes Res Clin Pract,2019,151:177-186.DOI: 10.1016/j.diabres.2019.04.021.
[5]	MaZ,DingJ,WangY,et al.Study of platelet-rich fibrin promoting endothelial cell differentiation and angiogenesis induced by transplantation of adipose-derived stem cells[J].Acta Histochem,2023,125(6):152059.DOI: 10.1016/j.acthis.2023.152059.
[6]	HeidariM,SadeghifardL,YaghobiR,et al.An investigation of the association between vascular endothelial growth factor +405 G/C polymorphism and acute liver transplant rejection in Iranian liver transplant recipients[J].Exp Clin Transplant,2022,20(6):564-568.DOI: 10.6002/ect.2020.0515.
[7]	GhanaatiS,BoomsP,OrlowskaA,et al.Advanced platelet-rich fibrin: a new concept for cell-based tissue engineering by means of inflammatory cells[J].J Oral Implantol,2014,40(6):679-689.DOI: 10.1563/aaid-joi-D-14-00138.
[8]	BrizuelaC,HuangGT,DiogenesA,et al.The four pillars for successful regenerative therapy in endodontics: stem cells, biomaterials, growth factors, and their synergistic interactions[J].Stem Cells Int,2022,2022:1580842.DOI: 10.1155/2022/1580842.
[9]	SmoczerC,YuthKR,AskarMA,et al.Growth factors released from advanced platelet-rich fibrin in the presence of calcium-based silicate materials and their impact on the viability and migration of stem cells of apical papilla[J].Dent J (Basel),2023,11(9):220.DOI: 10.3390/dj11090220.
[10]	WarinR,VongchanP,SuriyasathapornW,et al.In vitro assessment of lyophilized advanced platelet-rich fibrin from dogs in promotion of growth factor release and wound healing[J].Vet Sci,2022,9(10):566.DOI: 10.3390/vetsci9100566.
[11]	MoradiL, WitekL, Vivekanand NayakV, et al. Injectable hydrogel for sustained delivery of progranulin derivative Atsttrin in treating diabetic fracture healing[J]. Biomaterials, 2023,301:122289. DOI: 10.1016/j.biomaterials.2023.122289.
[12]	HerronC,HastingsCL,Herron-RiceC,et al.A thermoresponsive chitosan/β-glycerophosphate hydrogel for minimally invasive treatment of critical limb ischaemia[J].Polymers (Basel),2021,13(20):3568.DOI: 10.3390/polym13203568.
[13]	LinCJ,LinHL,YouWC,et al.Composite hydrogels of ultrasound-assisted-digested formic acid-decellularized extracellular matrix and sacchachitin nanofibers incorporated with platelet-rich plasma for diabetic wound treatment[J].J Funct Biomater,2023,14(8):423. DOI: 10.3390/jfb14080423.
[14]	顾雅男,徐翔昊,王彦平,等.氧化铈纳米酶-甲基丙烯酸酐化明胶水凝胶在小鼠全层皮肤缺损感染创面修复中的作用[J].中华烧伤与创面修复杂志,2024,40(2):131-140.DOI: 10.3760/cma.j.cn501225-20231120-00201.
[15]	CaoX,CaiX,ChenR,et al.A thermosensitive chitosan-based hydrogel for sealing and lubricating purposes in dental implant system[J].Clin Implant Dent Relat Res,2019,21(2):324-335.DOI: 10.1111/cid.12738.
[16]	SunH,SaeediP,KarurangaS,et al.IDF diabetes atlas: global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045[J].Diabetes Res Clin Pract,2022,183:109119.DOI: 10.1016/j.diabres.2021.109119.
[17]	黄玉林,马坤岭.糖尿病肾病遗传调控机制研究进展[J].解放军医学杂志,2023,48(7):856-862.DOI: 10.11855/j.issn.0577-7402.2952.2022.1124.
[18]	VijayakumarV,SamalSK,MohantyS,et al.Recent advancements in biopolymer and metal nanoparticle-based materials in diabetic wound healing management[J].Int J Biol Macromol,2019,122:137-148.DOI: 10.1016/j.ijbiomac.2018.10.120.
[19]	SawayaAP, StoneRC, BrooksSR, et al. Deregulated immune cell recruitment orchestrated by FOXM1 impairs human diabetic wound healing[J]. Nat Commun, 2020,11(1):4678. DOI: 10.1038/s41467-020-18276-0.
[20]	DasariN,JiangA,SkochdopoleA,et al.Updates in diabetic wound healing, inflammation, and scarring[J].Semin Plast Surg,2021,35(3):153-158.DOI: 10.1055/s-0041-1731460.
[21]	LiuL,HuJ,WangY,et al.The role and research progress of the balance and interaction between regulatory T cells and other immune cells in obesity with insulin resistance[J].Adipocyte,2021,10(1):66-79.DOI: 10.1080/21623945.2021.1876375.
[22]	GenesiBP,de Melo BarbosaR,SeverinoP,et al.Aloe vera and copaiba oleoresin-loaded chitosan films for wound dressings: microbial permeation, cytotoxicity, and in vivo proof of concept[J].Int J Pharm,2023,634:122648.DOI: 10.1016/j.ijpharm.2023.122648.
[23]	CraciunAM,Mititelu TartauL,PintealaM,et al.Nitrosalicyl-imine-chitosan hydrogels based drug delivery systems for long term sustained release in local therapy[J].J Colloid Interface Sci,2019,536:196-207.DOI: 10.1016/j.jcis.2018.10.048.
[24]	LiuS,LiuZ,WuM,et al.NIR as a "trigger switch" for rapid phase change, on-demand release, and photothermal synergistic antibacterial treatment with chitosan-based temperature-sensitive hydrogel[J].Int J Biol Macromol,2021,191:344-358.DOI: 10.1016/j.ijbiomac.2021.09.093.
[25]	ValachovaK,SvikK,BiroC,et al.Impact of ergothioneine, hercynine, and histidine on oxidative degradation of hyaluronan and wound healing[J].Polymers (Basel),2020,13(1):95.DOI: 10.3390/polym13010095.
[26]	Dohan EhrenfestDM, RasmussonL, AlbrektssonT.Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF)[J]. Trends Biotechnol, 2009,27(3):158-167. DOI: 10.1016/j.tibtech.2008.11.009.
[27]	ClarkD, RajendranY, PaydarS, et al. Advanced platelet-rich fibrin and freeze-dried bone allograft for ridge preservation: a randomized controlled clinical trial[J]. J Periodontol, 2018,89(4):379-387. DOI: 10.1002/JPER.17-0466.
[28]	MirhajM,SalehiS,TavakoliM,et al.Comparison of physical, mechanical and biological effects of leucocyte-PRF and advanced-PRF on polyacrylamide nanofiber wound dressings: in vitro and in vivo evaluations[J].Biomater Adv,2022,141:213082.DOI: 10.1016/j.bioadv.2022.213082.
[29]	张清荣,陈长友,徐娜,等.载P311微球的温敏壳聚糖水凝胶对大鼠全层皮肤缺损创面愈合的影响[J].中华烧伤与创面修复杂志,2022,38(10):914-922.DOI: 10.3760/cma.j.cn501225-20220414-00135.
[30]	郑力铭,刘钟元,颜鸿宇,等.小檗碱对糖尿病小鼠全层皮肤缺损创面愈合的影响及其机制[J].中华烧伤与创面修复杂志,2023,39(11):1072-1082.DOI: 10.3760/cma.j.cn501225-20230411-00120.