Recombinant human epidermal growth factor combined with vacuum sealing drainage for wound healing in Bama pigs

Background:Vacuum sealing drainage(VSD)and epidermal growth factor(EGF)both play an important role in the treatment of wounds.This study aims to explore the effects of the combination of VSD and EGF on wound healing and the optimal concentration and time of EGF.Methods:We tested the proliferation and migration capacity of HaCaT and L929 cells at different EGF concentrations(0,1,5,10,and 100ng/ml)and different EGF action times(2,10,and 30min).A full-thickness skin defect model was established using male,30-week-old Bama pigs.The experiment included groups as follows:routine dressing change after covering with sterile auxiliary material(Control),continuous negative pressure drainage of the wound(VSD),continuous negative pressure drainage of the wound and injection of EGF 10min followed by removal by continuous lavage(V+E 10min),and continuous negative pressure drainage of the wound and injection of EGF 30min followed by removal by continuous lavage(V+E 30min).The wound healing rate,histological repair effect and collagen deposition were compared among the four groups.Results:An EGF concentration of 10ng/ml and an action time of 10min had optimal effects on the proliferation and migration capacities of HaCaT and L929 cells.The drug dispersion effect was better than drug infusion after bolus injection effect,and the contact surface was wider.Compared with other groups,the V+E 10min group promoted wound healing to the greatest extent and obtained the best histological score.Conclusions:A recombinant human epidermal growth factor(rhEGF)concentration of 10 ng/ml can promote the proliferation and migration of epithelial cells and fibroblasts to the greatest extent in vitro.VSD combined with rhEGF kept in place for 10min and then washed,can promote wound healing better than the other treatments in vitro.

Keratinocyte growth factor-2 and autologous serum potentiate the regenerative effect of mesenchymal stem cells in cornea damage in rats

AIM:To investigate the healing process after severe corneal epithelial damage in rats treated with mesenchymal stem cells(MSCs)cultured with or without keratinocyte growth factor(KGF-2)and autologous serum(AS)on amniotic membrane(AM).Many patients are blind and devastated by severe ocular surface diseases due to limbal stem cell deficiency.Bone marrow-derived MSCs are potential sources for cellbased tissue engineering to repair or replace the corneal tissue,having the potential to differentiate to epithelial cells.METHODS:The study included 5 groups each including 10 female'Sprague Dawley'rats in addition to20 male rats used as bone marrow donors.Group I rats received AM+MSCs,Group II rats AM+MSCs cultured with KGF-2,Group III rats AM+MSCs cultured with KGF-2+AS,Group IV rats only AM and Group V rats,none.AS was derived from blood drawn from male rats and bone marrow was obtained from the femur and tibia bones of the same animals.Therapeutic effect was evaluated with clinical,histopathological and immunohistochemical assessment.MSC engraftment was demonstrated via detection of donor genotype(Y+)in the recipient tissue(X)with polymerase chain reaction.RESULTS:Corneal healing was significantly better in Groups I-III rats treated with MSC transplantation compared to Group IV and Group V rats with supportive treatment only.The best results were obtained in Group III rats with 90%transparency,70%lack of neovascularization,and 100%epithelium damage limited to less than 1/4 of cornea.CONCLUSION:We suggest that culture of MSCs with KGF-2 and AS on AM is effective in corneal repair in case of irreversible damage to limbal stem cells.

Growth factor-and cytokine-driven pathways governing liver stemness and differentiation

Liver is unique in its capacity to regenerate in response to injury or tissue loss. Hepatocytes and other liver cells are able to proliferate and repopulate the liver. However, when this response is impaired, the contribution of hepatic progenitors becomes very relevant. Here, we present an update of recent studies on growth factors and cytokine-driven intracellular pathways that govern liver stem/pro-genitor cell expansion and differentiation, and the rel-evance of these signals in liver development, regeneration and carcinogenesis. Tyrosine kinase receptor signaling, in particular, c-Met, epidermal growth factor receptors or fibroblast growth factor receptors, contribute to prolifera-tion, survival and differentiation of liver stem/progenitor cells. Different evidence suggests a dual role for the trans-forming growth factor (TGF)-β signaling pathway in liver stemness and differentiation. On the one hand, TGF-βmediates progression of differentiation from a progenitor stage, but on the other hand, it contributes to the expan-sion of liver stem cells. Hedgehog family ligands are nec-essary to promote hepatoblast proliferation but need to be shut off to permit subsequent hepatoblast differentiation. In the same line, the Wnt family and β-catenin/T-cell fac-tor pathway is clearly involved in the maintenance of liver stemness phenotype, and its repression is necessary for liver differentiation during development. Collectively, data indicate that liver stem/progenitor cells follow their own rules and regulations. The same signals that are essential for their activation, expansion and differentiation are good candidates to contribute, under adequate conditions, to the paradigm of transformation from a pro-regenerative to a pro-tumorigenic role. From a clinical perspective, this is a fundamental issue for liver stem/progenitor cell-based therapies.

Effect of combination therapy with alginate dressing and mouse epidermal growth factor on epidermal stem cells in patients with refractory wounds

Background The aim of this research was to determine the efficacy of combination therapy using an alginate dressing and mouse epidermal growth factor （mEGF） on proliferation and differentiation of epidermal stem cells （ESCs） in patients with refractory wounds.Methods Eighteen patients （12 males and 6 females,aged from 18 to 61 years （mean 36.4 years）） with various skin wounds,were treated by dressing changing for one month.The wounds were located in the foot （11）,calf （3）,thigh （2）and forearm （2）.The patients were randomly divided into 3 groups：alginate dressing and mEGF （group A; n=6）,mEGF （group B; n=6） and control （group C; n=6）.Wound closure indexes were measured at 7,14,21 and 28 days.Samples were harvested for pathologic examination,at 7 and 14 days following treatment.Cytokeratin 10 （CK10） and cytokeratin 15 （CK15） positive cells were evaluated using the super-sensitivity （SP） immunohistochemical staining technique.Results Wound healing was promoted in groups A and B.In group A,the wound closure index was increased significantly （P 〈0.05）,and in one case the maximum cure area reached 102 cm2.Pathological examination identified a thicker epidermis,active angiogenesis and enhanced granulation in group A compared with groups B and C.Using the SP immunohistochemical staining technique,we showed that ESCs in group A were bigger in size and larger in number than in groups B and C.Overall,there was a significant difference in ESCs proliferation and differentiation between group A and group B （or C）.Conclusions Combination therapy using an alginate dressing and mEGF shows increased proliferation and differentiation of ESCs in patients with refractory wounds compared with those treated with mEGF alone.

Systems biology approach to developing S^2RM-based “systems therapeutics” and naturally induced pluripotent stem cells

The degree to, and the mechanisms through, which stem cells are able to build, maintain, and heal the body have only recently begun to be understood. Much of the stem cell's power resides in the release of a multitude of molecules, called stem cell released molecules(SRM). A fundamentally new type of therapeutic, namely "systems therapeutic", can be realized by reverse engineering the mechanisms of the SRM processes. Recent data demonstrates that the composition of the SRM is different for each type of stem cell, as well as for different states of each cell type. Although systemsbiology has been successfully used to analyze multiple pathways, the approach is often used to develop a small molecule interacting at only one pathway in the system. A new model is emerging in biology where systems biology is used to develop a new technology acting at multiple pathways called "systems therapeutics". A natural set of healing pathways in the human that uses SRM is instructive and of practical use in developing systems therapeutics. Endogenous SRM processes in the human body use a combination of SRM from two or more stem cell types, designated as S2 RM, doing so under various state dependent conditions for each cell type. Here we describe our approach in using statedependent SRM from two or more stem cell types, S2 RM technology, to develop a new class of therapeutics called "systems therapeutics." Given the ubiquitous and powerful nature of innate S2RM-based healing in the human body, this "systems therapeutic" approach using S2 RM technology will be important for the development of anti-cancer therapeutics, antimicrobials, wound care products and procedures, and a number of other therapeutics for many indications.

中药浸浴联合重组表皮细胞在压疮患者中的应用效果

目的探究中药浸浴联合重组表皮细胞在压疮患者中的应用效果。方法选取2022年1月至2023年6月贵溪市中医院收治的60例压疮患者作为研究对象,采用随机数字表法将其分为对照组与观察组,每组各30例。对照组采用重组表皮细胞生长因子方法,观察组在对照组基础上使用中药浸浴,分析两组干预后的创面愈合有效率、炎症因子水平、症状缓解时间、转化生长因子(TGF-β1)、血管内皮生长因子(VEGF)表达水平以及不良反应发生率。结果治疗前两组的TGF-β1、VEGF表达水平、炎症因子水平比较,差异无统计学意义(P>0.05)。观察组治疗后TGF-β1、VEGF表达水平以及炎症因子水平均低于对照组,差异有统计学意义(P<0.05)。观察组症状缓解时间、不良反应发生率低于对照组,差异有统计学意义(P<0.05)。观察组创面愈合总有效率高于对照组,差异有统计学意义(P<0.05)。结论在临床上对压疮创面修复中采用重组表皮细胞生长因子联合中药浸浴效果更好,患者创面愈合有效率提高,症状缓解时间缩短,不良反应发生率少,安全性高且效果显著,临床应用价值高。

Transfection of the glial cell line-derived neurotrophic factor gene promotes neuronal differentiation

Glial cell line-derived neurotrophic factor recombinant adenovirus vector-transfected bone marrow mesenchymal stem cells were induced to differentiate into neuron-like cells using inductive medium containing retinoic acid and epidermal growth factor. Cell viability, micro- tubule-associated protein 2-positive cell ratio, and the expression levels of glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein-43 protein in the su- pernatant were significantly higher in glial cell line-derived neurotrophic factor/bone marrow mesenchymal stem cells compared with empty virus plasmid-transfected bone marrow mes- enchymal stem cells. Furthermore, microtubule-associated protein 2, glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein743 mRNA levels in cell pellets were statistically higher in glial cell line-derived neurotrophic factor/bone marrow mesen- chymal stem cells compared with empty virus plasmid-transfected bone marrow mesenchymal stem cells. These results suggest that glial cell line-derived neurotrophic factor/bone marrow mesenchymal stem cells have a higher rate of induction into neuron-like cells, and this enhanced differentiation into neuron-like cells may be associated with up-regulated expression of glial cell line-derived neurotrophic factor, nerve growth factor and growth-associated protein-43.

Novel nervous and multi-system regenerative therapeutic strategies for diabetes mellitus with mTOR

Throughout the globe,diabetes mellitus（DM） is increasing in incidence with limited therapies presently available to prevent or resolve the significant complications of this disorder.DM impacts multiple organs and affects all components of the central and peripheral nervous systems that can range from dementia to diabetic neuropathy.The mechanistic target of rapamycin（m TOR） is a promising agent for the development of novel regenerative strategies for the treatment of DM.m TOR and its related signaling pathways impact multiple metabolic parameters that include cellular metabolic homeostasis,insulin resistance,insulin secretion,stem cell proliferation and differentiation,pancreatic β-cell function,and programmed cell death with apoptosis and autophagy.m TOR is central element for the protein complexes m TOR Complex 1（m TORC1） and m TOR Complex 2（m TORC2） and is a critical component for a number of signaling pathways that involve phosphoinositide 3-kinase（PI 3-K）,protein kinase B（Akt）,AMP activated protein kinase（AMPK）,silent mating type information regulation 2 homolog 1（Saccharomyces cerevisiae）（SIRT1）,Wnt1 inducible signaling pathway protein 1（WISP1）,and growth factors.As a result,m TOR represents an exciting target to offer new clinical avenues for the treatment of DM and the complications of this disease.Future studies directed to elucidate the delicate balance m TOR holds over cellular metabolism and the impact of its broad signaling pathways should foster the translation of these targets into effective clinical regimens for DM.

Effects of allogeneic PRP combined with ADSC on wound healing and serum HA and VEGF levels in mice

Objective:Elet-rich plasma(PRP)combined with adipose-derived mesenchymal stem cells(ADSC)on skin wound healing and serum hyaluronic acid(HA)and vascular endothelial growth factor(VEGF)levels in mice.Methods:Forty-five mice were randomly divided into three groups(n=15),which were control,ADSC,and ADSC+PRP.The wound injury model was established in all three groups.The control group was injected subcutaneously with normal saline,the ADSC group was injected subcutaneously with allogeneic PRP,and the ADSC+PRP was injected subcutaneously with ADSC and PRP.Wound healing was observed at 3 d,5 d,and 7 d after modeling,and histomorphological features were observed by HE staining.CollagenⅠand CollagenⅢlevels were detected using Western blot.The levels of serum HA and VEGF in each group were detected and compared.Results:The wound healing rate of ADSC group and ADSC+PRP group was significantly higher than that of control group at 3 d,5 d and 7 d after modeling(P<0.01).The wound healing rate of the ADSC+PRP group was significantly higher than that of the ADSC group(P<0.01).The inflammatory response of the control group and the ADSC group was significant at 3 and 5 d after modeling,and there was a small amount of granulation tissue in the ADSC group.The ADSC+PRP group had a milder inflammatory response and more granulation tissue;After 7 d,the ADSC+PRP group had more neovascularization and higher re-epithelialization rate.The control group and the ADSC group had poorer epithelialization and fewer new blood vessels.The expression levels of CollagenⅠand CollagenⅢin ADSC group and ADSC+PRP group were significantly higher than those in control group at 3 d,5 d and 7 d after modeling(P<0.01).CollagenⅠand CollagenⅢin the ADSC+PRP group were significantly higher than those in the ADSC group(P<0.01).Serum HA and VEGF in ADSC group and ADSC+PRP group were significantly higher than those in control group at 3 d,5 d and 7 d after modeling(P<0.01).Serum HA and VEGF were significantly higher in the ADSC+PRP group than in the ADSC group(P<0.01).Conclusion:Allogeneic PRP combined with ADSC can promote the regeneration and healing of skin wounds in mice.The mechanism may be related to the up-regulation of the expression of vascular endothelial growth factor and HA and the promotion of the formation of blood vessels and granulation tissue.

Epithelial stem cell islands in the regenerated epidermis

Objective: The effects of growth factors on wound healing have been studied extensively, however, their molecular and genetic mechanisms that regulate epidermal regeneration are not fully understood. In this study, we explore the cell reversion characteristics and epithelial stem cell distribution in human regenerated epidermis treated with recombinant human epidermal growth factor (rhEGF). Methods:Tissue biospies from 8 regenerated skins treated with rhEGF were used to evaluate the cell reversion.

手术切除联合自体断层皮片移植治疗面部鳞状细胞癌

目的:分析手术切除联合自体断层皮片移植治疗面部鳞状细胞癌(Squamous cell carcinoma,SCC)的临床疗效。方法:选择2019年1月-2021年1月笔者医院收治的53例面部SCC患者,按入院顺序依随机数字表法分组,对照组(n=26)给予手术切除联合常规外科修复,研究组(n=27)给予手术切除联合自体断层皮片修复。比较两组临床疗效、血管内皮生长因子(Vascular endothelial growth factor,VEGF)与转化生长因子β1(Transforming growth factor-β1,TGF-β1)水平、美学效果、创面愈合时间及肿瘤复发情况。结果:术后14 d,研究组总有效率高于对照组(P<0.05);术后7 d,研究组VEGF与TGF-β1水平均高于对照组(P<0.05);出院时,研究组美学效果优于对照组(P<0.05);研究组创面愈合时间短于对照组(P<0.05),术后1年内复发率低于对照组(P<0.05)。结论:手术切除联合自体断层皮片移植治疗面部SCC创面愈合时间短,美学效果好,复发率低,整体疗效肯定。

糖尿病合并慢性难愈合创面患者血清VEGF、bFGF和创面组织EGFR的表达及其临床意义

目的探讨糖尿病合并慢性难愈合创面患者血清血管内皮细胞生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)和创面组织表皮生长因子受体(EGFR)的表达及其临床意义。方法回顾性选取2020年1月至2022年1月联勤保障部队第九〇九医院(厦门大学附属东南医院)收治的糖尿病合并慢性难愈合创面患者76例纳入难愈组,另选取同期接受治疗的糖尿病创面良好愈合的76例患者纳入康复组。对比两组患者治疗前血清VEGF、bFGF及创面组织EGFR的表达水平,分析3项指标表达水平与患者创面难愈的关系;对难愈组患者开展专项治疗,统计临床疗效及创面闭合指数,分析3项指标与难愈组患者临床疗效、闭合指数的相关性。结果治疗前,难愈组患者血清VEGF、bFGF及创面组织EGFR的表达水平分别为(184.69±35.83)pg/mL、(300.24±33.62)pg/mL、(5.67±1.01)分,均低于康复组[(206.01±42.03)pg/mL、(327.91±45.74)pg/mL、(8.20±1.11)分],差异均有统计学意义(P<0.05)。Logistic分析显示,血清VEGF、bFGF及创面组织EGFR均为糖尿病合并慢性难愈合创面的保护因素(P<0.05),且血清VEGF、bFGF及创面组织EGFR联合检测可提高诊断价值(P<0.05)。难愈组治疗1个疗程后临床治愈35.53%,显效40.79%,闭合指数为(81.95±8.48)%;血清VEGF、bFGF及创面组织EGFR与糖尿病合并慢性难愈合创面患者临床疗效间具有高度负相关性(r=-0.938、-0.938、-0.938,P<0.05),与创面闭合指数间均具有高度正相关性(r=0.984、0.983、0.987,P<0.05)。结论血清VEGF、bFGF及创面组织EGFR的表达水平为糖尿病合并慢性难愈合创面的保护因素,较高的血清VEGF、bFGF及创面组织EGFR表达水平有利于降低糖尿病创伤患者并发慢性难愈合创面,且联合检测三项指标对糖尿病合并慢性难愈合创面具有较高的诊断价值。同时糖尿病合并慢性难愈合创面患者3项指标表达水平越高则临床疗效越理想,创面闭合指数亦随着3项指标水平的升高而升高。

纳米脂肪在皮肤修复中的应用与机制

纳米脂肪是将抽取出的颗粒脂肪机械乳化提纯后留下的微小颗粒组织。一方面,机械乳化过程去除了移植后难以存活的含有大油滴的成熟脂肪细胞,可减轻移植后的炎症反应与吸收;另一方面,随着针对纳米脂肪成分及作用机制研究的不断深入,人们发现纳米脂肪中含有大量脂肪来源干细胞、血管内皮细胞、免疫细胞,以及细胞外基质与生长因子等,这些成分具有促进组织再生的可能。尤其是在皮肤修复领域,纳米脂肪不单能起充填作用,还能促进创面愈合,并可抑制修复后瘢痕,改善皮肤质地与色泽,甚至对毛发等皮肤附属器具有促进修复作用。本文就纳米脂肪在皮肤修复中的综合作用及可能机制展开综述。

EGFR shRNA联合雷帕霉素对Colo-16细胞增殖、迁移及侵袭的影响

目的:明确表皮生长因子受体(EGFR)shRNA联合雷帕霉素(RAPA)对皮肤鳞癌Colo-16细胞增殖、迁移和侵袭的影响,并对其可能机制进行初步探究。方法:Colo-16细胞分为5组:正常细胞组(Colo-16细胞+磷酸盐缓冲液PBS)、阴性对照组(转染shRNA-NC质粒的Colo-16细胞+PBS)、EGFR shRNA组(转染EGFR shRNA质粒的Colo-16细胞+PBS)、RAPA组(Colo-16细胞+RAPA)、联合组(转染EGFR shRNA质粒的Colo-16细胞+RAPA)。采用细胞计数试剂盒(CCK-8)和克隆形成实验测定细胞增殖能力;划痕实验和Transwell法检测细胞迁移和侵袭情况,采用Western blot、RT-PCR检测增殖、侵袭相关基因Ki-67、MMP-2、MMP-9蛋白和mRNA水平表达。多组间比较采用单因素方差分析,组间两两多重比较采用SNK-q检验。结果:EGFR shRNA组、RAPA组及联合组Colo-16细胞吸光度A值、细胞克隆形成率、划痕愈合率、侵袭细胞数目、Ki-67、MMP-2、MM-P9等基因蛋白和mRNA表达显著低于正常细胞组与阴性对照组,差异有统计学意义(均P<0.05),且联合的效果更加明显。正常细胞组与阴性对照组差异均无统计学意义(均P>0.05)。结论:EGFR shRNA联合雷帕霉素在抑制Colo-16细胞增殖、迁移及侵袭方面有协同增效的作用,其机制可能与EGFR/PI3K/AKT/mTOR信号通路的双重抑制作用相关。

双特异性CAR-T细胞对EGFRvⅢ^(+)/CD133^(+)胶质瘤干细胞的靶向杀伤

目的:制备双特异性CAR-T(bs CAR-T)细胞,观察其对表达表皮生长因子Ⅲ型突变阳性(EGFRvⅢ^(+),简称vⅢ^(+))和CD133^(+)胶质瘤干细胞的靶向杀伤作用。方法:基于前期研制的vⅢ/CD133双特异性微抗体和二代CAR构建的双特异性CAR(bs CAR),制备慢病毒载体转染人外周血T细胞,FCM和WB法检测bs CAR转染效率和表达水平。bs CAR-T细胞和vⅢ^(+)/CD133^(+)U87胶质瘤干细胞共培养,乳酸脱氢酶(LDH)释放实验、IFN-γ分泌实验检测其特异性杀伤作用和对IFN-γ分泌的促进作用。制备裸鼠vⅢ^(+)/CD133^(+)U87干细胞移植瘤模型检测bs CAR-T细胞对移植瘤生长的抑制作用。结果:vⅢsc Fv和CD133sc Fv通过重叠PCR无缝连接入二代CAR表达框(S-vⅢscFv/CD133scFv-Hinge-TM-CD137-CD3ζ)中,然后克隆入p CDH-MSCV-MCS-EF1-copGFP载体的Eco RⅠ和Bam HⅠ位点(pbs CAR)。3种质粒(p VSV-G、p CMV-d R8.9和pbs CAR)共转染HEK293T细胞制备慢病毒载体,转染外周血T细胞,FCM检测bs CAR表达率为71.1%,WB法结果显示bs CAR表达正确。bs CAR-T细胞和vⅢ^(+)/CD133^(+)U87干细胞共培养检测结果显示,bs CAR-T细胞对胶质瘤干细胞具有特异性杀伤作用,与效靶比呈正比;IFN-γ分泌量为(2350.6±92)pg·m L^(-1),明显高于对照组(P<0.01)。裸鼠移植瘤动物模型显示,bs CAR-T细胞在体内具有明显的移植瘤抑制作用(P<0.01)。结论:bs CAR-T细胞能够特异性靶向杀伤vⅢ^(+)/CD133^(+)胶质瘤干细胞,实验结果为促进实体瘤的细胞免疫治疗提供了实验依据。

糖尿病皮肤慢性创面无细胞治疗的进展与问题

背景:糖尿病皮肤溃疡是糖尿病的难治并发症之一,发生后迁延不愈。以往研究发现,间充质干细胞具有组织修复和炎症免疫调节能力,并且相关研究表明间充质干细胞可能主要是通过旁分泌效应分泌各种细胞因子和外泌体等,发挥组织与创面修复作用。因而可使用生长因子、间充质干细胞条件培养基及外泌体等治疗糖尿病皮肤慢性创面,以直接或间接机制对伤口愈合产生积极影响。目的:总结生长因子、条件培养基和外泌体等促进糖尿病皮肤慢性创面修复的相关机制、应用效果,以及无细胞治疗在当前研究的局限性。方法:由第一作者检索PubMed及Web of Science数据库中收录的相关文献,检索词为“Diabetes,Diabetic foot,Diabetic foot ulcers,Chronic wound,Mesenchymal stem cells,Cell-free,Exosomes,Growth factors,Conditioned mediums,Wound healing,Tissue repair,Angiogenesis,Regeneration,Biomaterial”,文献检索时限为2005年1月至2022年2月,经过阅读并筛选整理,选择与综述内容相符的文献,最终确定整理出75篇文献进行综述。结果与结论:①间充质干细胞条件培养基、外泌体和生长因子可通过不同机制改善局部微环境,促进慢性创面的愈合;②通过条件培养基、外泌体和生长因子与新兴的组织工程生物学材料(如细胞支架、水凝胶等)相结合,可使其发挥更强的促进慢性创面愈合的作用;③基于间充质干细胞条件培养基、外泌体、生长因子的无细胞疗法是促进慢性创面修复的极有前景的治疗策略,要实现由临床前研究转化到临床研究,确保其用于患者治疗的安全性,还需要进行更多的研究,以查明相应的治疗作用机制。

Effect of Optimized Concentrations of Basic Fibroblast Growth Factor and Epidermal Growth Factor on Proliferation of Fibroblasts and Expression of Collagen： Related to Pelvic Floor Tissue Regeneration

Background： Fibroblasts were the main seed cells in the studies of tissue engineering of the pelvic floor ligament. Basic fibroblast growth factor （bFGF） and epidermal growth factor （EGF） were widely studied but at various concentrations. This study aimed to optimize the concentrations of combined bFGF and EGF by evaluating their effects on proliferation and collagen secretion of fibroblasts. Methods： Fibroblasts were differentiated from rat adipose mesenchymal stem cells （ADSCs）. Flow cytometry and immunohistochemistry were used for cell identification. The growth factors were applied at concentrations of 0, 1, 10, and 100 ng/ml as three groups： （1） bFGF alone, （2） EGF alone, and （3） bFGF mixed with EGF. Cell proliferation was evaluated by Cell Counting Kit-8 assays. Expression of Type I and III collagen （Col-I and Col-1II） mRNAs was evaluated by real-time quantitative reverse transcription-polymerase chain reaction. Statistical analysis was performed with SPSS software and GraphPad Prism using one-way analysis of variance and multiple t-test. Results： ADSCs were successfully isolated from rat adipose tissue as identified by expression of typical surface markers CD29, CD44, CD90, and CD45 in flow cytometry. Fibroblasts induced from ADSC, compared with ADSCs, were with higher mRNA expression levels of Col I and Col III （F = 1.29, P = 0.0390）. bFGF, EGF, and the mixture ofbFGF with EGF can enhanced fibroblasts proliferation, and the concentration of 10 ng/ml of the mixture ofbFGF with EGF displayed most effectively （all P 〈 0.05）. The expression levels of Col-I and Col-Ill mRNAs in fibroblasts displayed significant increases in the 10 ng/ml bFGF combined with EGF group （all P 〈 0.05）. Conclusions： The optimal concentration of both bFGF and EGF to promote cell proliferation and collagen expression in fibroblasts was 10 ng/ml at which fibroblasts grew faster and secreted more Type I and III collagens into the extracellular matrix, which might contribute to the stability of the pelvic floor microenvironment.

同种异体间充质干细胞在慢性创面修复中的疗效研究

目的探究同种异体间充质干细胞在慢性创面修复中的疗效研究。方法选取2021年9月—2022年9月在广东省韶关市粤北人民医院进行慢性创面治疗的60例患者作为研究对象,根据随机数表法分为对照组和观察组,每组30例,对照组采取常规治疗,观察组在常规治疗操作的基础上实施间充质干细胞治疗干预,比较两组临床治疗有效率、创面愈合时间,检测创面组织血管内皮细胞因子(vascular endothelial growth factor,VEGF)表达情况、记录不良事件发生率。结果观察组总有效率为93.33%,显著高于对照组,差异有统计学意义(χ^(2)=6.093,P<0.05)。观察组创面愈合时间为(12.43±2.18)d,短于对照组的(18.69±2.08)d,差异有统计学意义(t=13.029,P<0.001)。干预后,观察组VEGF表达水平为(240.29±19.39)pg/mg,显著高于对照组,差异有统计学意义(t=11.279,P<0.001)。治疗期间,密切关注两组患者每日状态,记录伤口愈合情况,未发现有不良事件发生。结论对进行慢性创面修复的患者实施间充质干细胞治疗干预,能够有效提高患者临床治疗有效率,缩短修复时间,患者治疗的安全性得到保障。

Stem cell therapy for chronic skin wounds in the era of personalized medicine:From bench to bedside

With the significant financial burden of chronic cutaneous wounds on the healthcare system,not to the personal burden mention on those individuals afflicted,it has become increasingly essential to improve our clinical treatments.This requires the translation of the most recent benchtop approaches to clinical wound repair as our current treatment modalities have proven insufficient.The most promising potential treatment options rely on stem cellbased therapies.Stem cell proliferation and signaling play crucial roles in every phase of the wound healing process and chronic wounds are often associated with impaired stem cell function.Clinical approaches involving stem cells could thus be utilized in some cases to improve a body’s inhibited healing capacity.We aim to present the laboratory research behind the mechanisms and effects of this technology as well as current clinical trials which showcase their therapeutic potential.Given the current problems and complications presented by chronic wounds,we hope to show that developing the clinical applications of stem cell therapies is the rational next step in improving wound care.

Adipose-derived stem cells improve grafted burn wound healing by promoting wound bed blood flow

Background:Researchers have explored the use of adipose-derived stem cells(ASCs)as a cellbased therapy to cover wounds in burn patients;however,underlying mechanistic aspects are not completely understood.We hypothesized that ASCs would improve post-burn wound healing after eschar excision and grafting by increasing wound blood flow via induction of angiogenesis-related pathways.Methods:To test the hypothesis,we used an ovine burn model.A 5 cm^(2) full thickness burn wound was induced on each side of the dorsum.After 24 hours,the burned skin was excised and a 2 cm^(2) patch of autologous donor skin was grafted.The wound sites were randomly allocated to either topical application of 7 million allogeneic ASCs or placebo treatment(phosphate-buffered saline[PBS]).Effects of ASCs culture media was also compared to those of PBS.Wound healing was assessed at one and two weeks following the application of ASCs.Allogeneic ASCs were isolated,cultured and characterized from non-injured healthy sheep.The identity of the ASCs was confirmed by flow cytometry analysis,differentiation into multiple lineages and gene expression via real-time polymerase chain reaction.Wound blood flow,epithelialization,graft size and take and the expression of vascular endothelial growth factor(VEGF)were determined via enzyme-linked immunosorbent assay and Western blot.Results:Treatment with ASCs accelerated the patch graft growth compared to the control(p<0.05).Topical application of ASCs significantly increased wound blood flow(p<0.05).Expression of VEGF was significantly higher in the wounds treated with ASCs compared to control(p<0.05).Conclusions:ASCs accelerated grafted skin growth possibly by increasing the blood flow via angiogenesis induced by a VEGF-dependent pathway.