高亲水性静电纺丝短纤维海绵促创面愈合的作用

doi:10.3969/j.issn.1674-8115.2023.03.002

上海交通大学学报（医学版） ›› 2023, Vol. 43 ›› Issue (3): 269-277.doi: 10.3969/j.issn.1674-8115.2023.03.002

• 论著 · 基础研究 • 上一篇

高亲水性静电纺丝短纤维海绵促创面愈合的作用

付晓晗¹^,²^,³(), 王娟², 崔文国²(), 王彦¹^,³()

^1.潍坊医学院临床医学院，潍坊 261053
^2.上海交通大学医学院附属瑞金医院上海市伤骨科研究所，上海市中西医结合防治骨与关节病损重点实验室，上海 200025
^3.山东省妇幼保健院整形美容科，济南 250061

收稿日期:2022-10-14 接受日期:2022-02-22 出版日期:2023-03-28 发布日期:2023-03-28
通讯作者: 崔文国,王彦 E-mail:fuxiaohanlisa@163.com;wgcui80@hotmail.com;wangyandr@126.com
作者简介:付晓晗（1992—），女，硕士生；电子信箱：fuxiaohanlisa@163.com。
基金资助:
国家重点研发计划(2020YFA0908200);国家自然科学基金(32000937);山东省高等学校青年创新团队人才引育计划(20190919);济南市科技发展基金(201910)

Effect of high hydrophilic electrospun short fibrous sponge on wound repair

FU Xiaohan¹^,²^,³(), WANG Juan², CUI Wenguo²(), WANG Yan¹^,³()

^1.School of Clinical Medicine, Weifang Medical University, Weifang 261053, China
^2.Shanghai Key Laboratory for Prevention and Treatment of Bone and Joint Diseases, Shanghai Institute of Traumatology and Orthopaedics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
^3.Department of Plastic Surgery, Shandong Provincial Maternal and Child Health Care Hospital, Jinan 250061, China

Received:2022-10-14 Accepted:2022-02-22 Online:2023-03-28 Published:2023-03-28
Contact: CUI Wenguo,WANG Yan E-mail:fuxiaohanlisa@163.com;wgcui80@hotmail.com;wangyandr@126.com
Supported by:
National Key R&D Program of China(2020YFA0908200);National Natural Science Foundation of China(32000937);Youth Innovation Technology Project of Higher School in Shandong Province(20190919);Science and Technology Development Foundation of Jinan(201910)

摘要/Abstract

摘要：

目的·构建一种含有氧化石墨烯（graphene oxide，GO）的高亲水性静电纺丝短纤维海绵（Sponge@GO），探究其对慢性创面的促愈合作用。方法·分别将不含及含有GO的纺丝液经过纺丝成膜、打碎、塑形并用戊二醛交联后，制备出2种短纤维海绵（Sponge和Sponge@GO）。采用扫描电子显微镜（scanning electron microscope，SEM）观察2种海绵的内部结构，通过接触角及吸水率观察两者的亲水性能。经CCK-8法和活/死细胞染色验证海绵的生物相容性。将12只6周龄SD雄性大鼠分为对照组、Sponge组和Sponge@GO组，每组各4只；通过腹腔注射1%的链脲佐菌素溶液建立糖尿病模型，建模后每只大鼠背部制备3块直径1.0 cm的全层皮肤缺损；将材料覆盖于创面，使用医用纱布覆盖创面并固定（对照组仅覆盖无菌纱布包扎）；分别于术后第7、14日测量并计算创面愈合率，同时取创周0.5 cm内组织行苏木精-伊红染色（hematoxylin and eosin staining，H-E染色）、Masson染色观察病理学改变；并在第14日通过α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）免疫荧光染色，观察血管形成情况。结果·SEM观察显示，与Sponge海绵比较，Sponge@GO海绵的纤维直径较细，孔隙率偏大。2种短纤维海绵在10 min内均基本达到最大吸水率，Sponge@GO海绵的吸水率较高。Sponge@GO海绵的接触角小于Sponge海绵，差异有统计学意义（P=0.000）。CCK-8法检测结果显示，术后第3、5日Sponge组与对照组相比，尚呈现出细胞增殖的状态（均P<0.05），而Sponge@GO组与对照组的差异无统计学意义。活/死细胞染色结果显示，3组细胞均呈现良好的增长态势。SEM和荧光染色观察显示，Sponge@GO海绵内部的细胞数量明显较Sponge海绵多。3组大鼠的创面均未见感染。Sponge@GO组和Sponge组术后第7日的创面愈合率均显著高于对照组（均P<0.05）；第14日，Sponge@GO组的创面愈合率仍显著高于对照组（P=0.009），而Sponge组与对照组之间的差异无统计学意义。术后第14日，H-E染色显示Sponge@GO组创面肉芽组织更加成熟，结构更为均匀致密；Masson染色显示Sponge@GO组胶原更加致密，上皮化显著；α-SMA免疫荧光染色显示Sponge@GO组新生血管数量更多，密度更高。结论·Sponge@GO海绵在吸收渗液后可维持创面微湿润环境，有利于促进伤口愈合。

关键词: 氧化石墨烯, 短纤维, 静电纺丝, 创面修复

Abstract:

Objective ·To construct an electrospun short fibrous sponge (Sponge@GO) laden with graphene oxide (GO) for chronic wound healing. Methods ·Two types of short fibrous sponges (Sponge and Sponge@GO) without and with GO were prepared by means of electrospinning, homogenizing, shaping and crosslinking with glutaraldehyde, respectively. The internal structures of the two sponges were observed with a scanning electron microscope (SEM), and their hydrophilic properties were observed via contact angle and water absorption rate. The biocompatibility of the sponge was verified by CCK-8 and live/dead staining. Twelve 6-week-old SD male rats were divided into control group, Sponge group and Sponge@GO group, with 4 rats in each group. The diabetes models were established by intraperitoneal injection of 1% streptozotocin solution, and three full-layer skin defects with a diameter of 1.0 cm were prepared on the back of each rat after modelling. Covering on the wound, the material was fixed with medical gauze. The control group was only covered with sterile gauze dressing. The wound healing rate was measured and calculated on Day 7 and 14, respectively, while hematoxylin-eosin (H-E) staining and Masson staining were performed on tissues within 0.5 cm around the wound to observe pathological changes. The angiogenesis was observed by α-smooth muscle actin (α-SMA) immunofluorescence staining on Day 14. Results ·SEM observation showed that the fiber diameter of Sponge@GO was significantly thinner and the porosity increased. The two types of short fiber scaffolds basically reached the maximum water uptake within 10 min, but the Sponge@GO scaffold showed better water absorption performance. The water contact angle of Sponge@GO scaffold was significantly smaller than that of Sponge, and the difference was statistically significant (P=0.000). The results of CCK-8 method showed that on Day 3 and 5, the Sponge group had better cell proliferation compared with the control group (both P<0.05), while there was no statistical significance between Sponge@GO group and control group. The results of live/dead staining showed that all the three groups of cells showed good cell growth trend. SEM and fluorescence staining showed that there were more cells in the Sponge@GO scaffold. In vivo experiment, no infection was found on the wound surface of the three groups of rats. The wound healing rate of Sponge@GO and Sponge groups was significantly higher than that of control group on Day 7 (both P<0.05). On Day 14, the wound healing rate of the Sponge@GO group was still significantly higher than that of the control group (P=0.009), while the difference between the Sponge group and the control group was not statistically significant. On Day 14, H-E staining showed more mature granulation tissue and more uniform and dense structure in the Sponge@GO group; Masson staining showed more dense collagen and significant epithelialization in the Sponge@GO group; α-SMA immunofluorescence staining showed more neovascularization and higher density in the Sponge@GO group. Conclusion ·Sponge@GO sponge can ensure micro-moist environment on the wound surface after absorbing exudate and has shown promising results in promoting wound healing.

Key words: graphene oxide (GO), short fiber, electrospinning, wound repair

中图分类号:

R641

付晓晗, 王娟, 崔文国, 王彦. 高亲水性静电纺丝短纤维海绵促创面愈合的作用[J]. 上海交通大学学报（医学版）, 2023, 43(3): 269-277.

FU Xiaohan, WANG Juan, CUI Wenguo, WANG Yan. Effect of high hydrophilic electrospun short fibrous sponge on wound repair[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2023, 43(3): 269-277.

图/表 6

图1 Sponge及Sponge@GO的形貌及吸湿性能Note： A. Appearance of Sponge. B. SEM image of Sponge. C. Appearance of Sponge@GO. D. SEM image of Sponge@GO. E. Water absorbtion. F. Contact angle.

Fig 1 Morphology and hygroscopic properties of Sponge and Sponge@GO

图2 Sponge及Sponge@GO的材料相容性Note： A. CCK-8. B. Live/dead staining. ①P=0.014, ②P=0.000, compared with the control group.

Fig 2 Biocompatibility of Sponge and Sponge@GO

图3 材料的细胞黏附性Note： A. SEM images of HUVECs after 7 d of incubation on Sponge. B. Fluorescence staining of HUVECs after 7 d of incubation on Sponge. C. SEM images of HUVECs after 7 d of incubation on Sponge@GO. D. Fluorescence staining of HUVECs after 7 d of incubation on Sponge@GO.

Fig 3 Cell adhesion of the materials

图4 糖尿病创面愈合情况观察(n=4)Note： A. Wound healing at Day 0, 7 and 14. B. Wound healing rate. ①P=0.024, ②P=0.000, ③P=0.009, compared with the control group at the same point.

Fig 4 Observation of diabetic wound healing (n=4)

图5 创面组织病理学观察Note： A. H-E staining. B. Masson staining.

Fig 5 Histopathological observation of wound

图6 术后第14日α-SMA免疫荧光染色观察创面血管形成情况

Fig 6 Observation of angiogenesis by α-SMA immunofluorescence staining at Day 14

参考文献 32

1	NAKAMURA Y, ISHIKAWA H, KAWAI K, et al. Enhanced wound healing by topical administration of mesenchymal stem cells transfected with stromal cell-derived factor-1[J]. Biomaterials, 2013, 34(37): 9393-9400.
2	NUNAN R, HARDING K G, MARTIN P. Clinical challenges of chronic wounds: searching for an optimal animal model to recapitulate their complexity[J]. Dis Model Mech, 2014, 7(11): 1205-1213.
3	CHEN T Y, WEN T K, DAI N T, et al. Cryogel/hydrogel biomaterials and acupuncture combined to promote diabetic skin wound healing through immunomodulation[J]. Biomaterials, 2021, 269: 120608.
4	陈军, 张楠. 负压封闭引流联合邻近皮瓣转位修复压疮的临床效果[J]. 临床骨科杂志, 2022, 25(4): 515-517.
	CHEN J, ZHANG N. Clinical effect of vacuum sealing drainage combined with adjacent flap transposition in repairing pressure ulcers[J]. J Clin Orthop, 2022, 25(4): 515-517.
5	ZHENG Y, JI S, WU H, et al. Topical administration of cryopreserved living micronized amnion accelerates wound healing in diabetic mice by modulating local microenvironment[J]. Biomaterials, 2017, 113: 56-67.
6	CHO N H, SHAW J E, KARURANGA S, et al. IDF Diabetes Atlas: global estimates of diabetes prevalence for 2017 and projections for 2045[J]. Diabetes Res Clin Pract, 2018, 138: 271-281.
7	SHIEKH P A, SINGH A, KUMAR A. Exosome laden oxygen releasing antioxidant and antibacterial cryogel wound dressing OxOBand alleviate diabetic and infectious wound healing[J]. Biomaterials, 2020, 249: 120020.
8	SCHÖNFELDER U, ABEL M, WIEGAND C, et al. Influence of selected wound dressings on PMN elastase in chronic wound fluid and their antioxidative potential in vitro[J]. Biomaterials, 2005, 26(33): 6664-6673.
9	SPEAR M. Wound exudate: the good, the bad, and the ugly[J]. Plast Surg Nurs, 2012, 32(2): 77-79.
10	STRECKER-MCGRAW M K, JONES T R, BAER D G. Soft tissue wounds and principles of healing[J]. Emerg Med Clin North Am, 2007, 25(1): 1-22.
11	DHIVYA S, PADMA V V, SANTHINI E. Wound dressings-a review[J]. Biomedicine (Taipei), 2015, 5(4): 22.
12	PURNA S K, BABU M. Collagen based dressings: a review[J]. Burns, 2000, 26(1): 54-62.
13	BOATENG J, CATANZANO O. Advanced therapeutic dressings for effective wound healing: a review[J]. J Pharm Sci, 2015, 104(11): 3653-3680.
14	JAIN R K, AU P, TAM J, et al. Engineering vascularized tissue[J]. Nat Biotechnol, 2005, 23(7): 821-823.
15	KONG X, FU J, SHAO K, et al. Biomimetic hydrogel for rapid and scar-free healing of skin wounds inspired by the healing process of oral mucosa[J]. Acta Biomater, 2019, 100: 255-269.
16	LI Y, WANG J, QIAN D, et al. Electrospun fibrous sponge via short fiber for mimicking 3D ECM[J]. J Nanobiotechnology, 2021, 19(1): 131.
17	FU X H, WANG J, QIAN D J, et al. Living electrospun short fibrous sponge via engineered nanofat for wound healing[J]. Adv Fiber Mater, 2022. DOI : 10.1007/s42765-022-00229-5.
18	SENTHIL R, BERLY R, BHARGAVI RAM T, et al. Electrospun poly(vinyl) alcohol/collagen nanofibrous scaffold hybridized by graphene oxide for accelerated wound healing[J]. Int J Artif Organs, 2018, 41(8): 467-473.
19	EROL O, UYAN I, HATIP M, et al. Recent advances in bioactive 1D and 2D carbon nanomaterials for biomedical applications[J]. Nanomedicine, 2018, 14(7): 2433-2454.
20	QIAN Y, SONG J, ZHAO X, et al. 3D fabrication with integration molding of a graphene oxide/polycaprolactone nanoscaffold for neurite regeneration and angiogenesis[J]. Adv Sci (Weinh), 2018, 5(4): 1700499.
21	LÓPEZ-DOLADO E, GONZÁLEZ-MAYORGA A, GUTIÉRREZ M C, et al. Immunomodulatory and angiogenic responses induced by graphene oxide scaffolds in chronic spinal hemisected rats[J]. Biomaterials, 2016, 99: 72-81.
22	GOUGH J E, SCOTCHFORD C A, DOWNES S. Cytotoxicity of glutaraldehyde crosslinked collagen/poly(vinyl alcohol) films is by the mechanism of apoptosis[J]. J Biomed Mater Res, 2002, 61(1): 121-130.
23	LIU G S, YAN X, YAN F F, et al. In situ electrospinning iodine-based fibrous meshes for antibacterial wound dressing[J]. Nanoscale Res Lett, 2018, 13(1): 309.
24	BLAKENEY B A, TAMBRALLI A, ANDERSON J M, et al. Cell infiltration and growth in a low density, uncompressed three-dimensional electrospun nanofibrous scaffold[J]. Biomaterials, 2011, 32(6): 1583-1590.
25	WANG J, CHENG Y, WANG H, et al. Biomimetic and hierarchical nerve conduits from multifunctional nanofibers for guided peripheral nerve regeneration[J]. Acta Biomater, 2020, 117: 180-191.
26	WANG J, LIN J, CHEN L, et al. Endogenous electric-field-coupled electrospun short fiber via collecting wound exudation[J]. Adv Mater, 2022, 34(9): e2108325.
27	SINHA RAY S. Polylactide-based bionanocomposites: a promising class of hybrid materials[J]. Acc Chem Res, 2012, 45(10): 1710-1720.
28	INKINEN S, HAKKARAINEN M, ALBERTSSON A C, et al. From lactic acid to poly(lactic acid) (PLA): characterization and analysis of PLA and its precursors[J]. Biomacromolecules, 2011, 12(3): 523-532.
29	SENTHIL R, BERLY R, BHARGAVI RAM T, et al. Electrospun poly(vinyl) alcohol/collagen nanofibrous scaffold hybridized by graphene oxide for accelerated wound healing[J]. Int J Artif Organs, 2018, 41(8): 467-473.
30	FU C, BAI H, ZHU J, et al. Enhanced cell proliferation and osteogenic differentiation in electrospun PLGA/hydroxyapatite nanofibre scaffolds incorporated with graphene oxide[J]. PLoS One, 2017, 12(11): e0188352.
31	YANG Y, HUANG K, WANG M, et al. Ubiquitination flow repressors: enhancing wound healing of infectious diabetic ulcers through stabilization of polyubiquitinated hypoxia-inducible factor-1α by theranostic nitric oxide nanogenerators[J]. Adv Mater, 2021, 33(45): e2103593.
32	HINZ B. The role of myofibroblasts in wound healing[J]. Curr Res Transl Med, 2016, 64(4): 171-177.

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高亲水性静电纺丝短纤维海绵促创面愈合的作用

Effect of high hydrophilic electrospun short fibrous sponge on wound repair

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