Follicular unit grafting in chronic ulcers: a valuable technique for integrated management

Abstract Chronic ulcers significantly affect the quality of life of patients and impose a high cost on the healthcare system. The therapeutic management should be comprehensive, taking into consideration the etiological diagnosis of the wound and the characteristics of the wound bed when deciding on a therapeutic proposal appropriate to the healing phase, correcting factors that delay healing. During the epithelialization phase, repair techniques with grafts are recommended to shorten re-epithelialization time, improve the quality of scar tissue, and achieve adequate pain management. Currently, due to the reported benefits of skin appendages, the technique of follicular unit auto-grafting obtained with a scalp punch is among the chosen strategies for wound repair. This is a minimally invasive, outpatient practice, whose technique has advantages over the donor site, patients recovery and well-being.

Tratamento de infecções localizadas em feridas de difícil cicatrização: uma revisão integrativa / Treatment of Localized Infections in Hard-to-Heal Wounds: An Integrative Review / Tratamiento de infecciones localizadas en heridas de difícil cicatrización: una revisión integrativa

Objetivo: Identificar la producción de conocimiento sobre el tratamiento de infecciones localizadas en heridas de difícil cicatrización. Método: Revisión integrativa de la literatura realizada en la Biblioteca Virtual en Salud; Base de datos de enfermería; Scientific Electronic Library; Web of Science; Biblioteca Cochrane; Catálogo de Tesis y Disertaciones de la Coordinación para el Perfeccionamiento del Personal de Educación Superior; y PubMed. Los artículos seleccionados no tienen límite de tiempo. Los estudios fueron exportados a la aplicación Rayyan y sometidos a evaluación doble ciego mediante la lectura del título y el resumen, según los criterios de inclusión y exclusión. La información fue analizada y sintetizada según el nivel de evidencia. Resultados: 19 estudios fueron incluidos para lectura completa. Se encontró como evidencia la higiene de la herida; la limpieza con ácido acético al 1%; la identificación y el tratamiento de biopelículas; el uso de cobertores y soluciones con acción antimicrobiana. Conclusión: La infección localizada de la herida ha sido objeto de varias investigaciones y las prácticas recomendadas se refieren a tratamientos tópicos. (AU)

Objective: To identify the knowledge production on the treatment of localized infections in hard-to-heal wounds. Method: An integrative literature review was conducted in the Virtual Health Library, Nursing Database, Scientific Electronic Library Online, Web of Science, Cochrane Library, Catalog of Theses and Dissertations of the Coordination for the Improvement of Higher Education Personnel, and PubMed. The selected articles had no time limit. The studies were exported to the Rayyan application and subjected to double-blind evaluation through title and abstract reading, based on inclusion and exclusion criteria. The information was analyzed and synthesized according to the level of evidence. Results: A total of 19 publications were fully analyzed. The evidence obtained on the topic includes wound hygiene, cleaning with 1% acetic acid, identification and treatment of biofilms, the use of dressings, and solutions with antimicrobial action. Conclusion: Localized wound infections have been the subject of various research studies, and the recommended practices refer to topical treatments. (AU)

Objetivo: Identificar a produção de conhecimento sobre o tratamento de infecções localizadas em feridas de difícil cicatrização. Método: Revisão integrativa da literatura realizada na Biblioteca Virtual em Saúde, Base de Dados de Enfermagem, Scientific Electronic Library Online, Web of Science, Biblioteca Cochrane, Catálogo de Teses e Dissertações da Coordenação de Aperfeiçoamento de Pessoal de Nível Superior e Public Medline. Os artigos selecionados não possuem limite temporal. Os estudos foram exportados para o aplicativo Rayyan e submetidos à avaliação duplo-cega por meio da leitura do título e do resumo, com base nos critérios de inclusão e exclusão. As informações foram analisadas e sintetizadas de acordo com o nível de evidência. Resultados: Foram analisadas 19 publicações em sua totalidade. Obteve-se como evidências acerca do tema a higienização da ferida, a limpeza com ácido acético 1%, a identificação e o tratamento de biofilmes, o uso de coberturas e as soluções com ação antimicrobiana. Conclusão: A infecção localizada de feridas tem sido objeto de várias pesquisas e as práticas recomendadas referem-se a tratamentos tópicos. (AU)

Effectiveness of leukocyte- and platelet-rich fibrin in the management of medication-related osteonecrosis of the jaws ­ a systematic review / Efectividad de la fibrina rica en leucocitos y plaquetas en el tratamiento de osteonecrosis mandibular relacionada con medicamentos: una revisión sistemática

Background: Medication-related osteonecrosis of the jaw (MRONJ) is a rare, but significant adverse event primarily associated with the intake of antiresorptive and antiangiogenic medications. Although antiresorptive and antiangiogenic the-rapies improve life expectancy, particularly in cancer patients, MRONJ may hamper the patient's quality of life due to pain, discomfort, anxiety, depression, speech impairment, difficulty in swallowing and eating, frequent medical and dental evaluations and treatments, and the possibility of treatment discontinuation. Leukocyte­ and Platelet-rich Fibrin (L-PRF) is an autologous platelet aggregate that promotes wound healing by stimulating re-epithelialization, angiogenesis, and extracellular matrix production. Aim: The present systematic review aimed to compare the results in the published literature on whether L-PRF is an effective and predictable adjuvant to surgical debridement of necrotic bone for improving the healing efficacy in patients with MRONJ. Materials and Methods: The PubMed, Scopus, Cochrane, Science Direct, LILACS, and Web of Science databases were searched using the predetermined MeSH terms and eligibility criteria, and the search yielded a total of five articles. Two studies were retrospective, and three studies were case series. Results: Seventeen participants received a combination of surgical debridement, L-PRF membrane, and antibiotics. Complete wound healing was observed in 70% of the participants, and most of them healed without any complications. Conclusions: L-PRF as an adjuvant to surgical debridement of necrosed bone appears to have a positive association with the healing outcome in patients with MRONJ.

Introducción: La osteonecrosis mandibular relacionada con medicamentos (ONMRM) es un evento adverso raro pero significativo asociado principalmente con la ingesta de medicamentos antirresortivos y antiangiogénicos. Aunque las terapias antirresortivas y antiangiogénicas mejoran la esperanza de vida, particularmente en pacientes con cáncer, la ONMRM puede obstaculizar la calidad de vida del paciente debido a dolor, incomodidad, ansiedad, depresión, discapacidad del habla, dificultad para tragar y comer, evaluaciones y tratamientos médicos y dentales frecuentes, y la posibilidad de interrupción del tratamiento. La fibrina rica en plaquetas y leucocitos (L-PRF) es un agregado de plaquetas autólogo que promueve la curación de heridas al estimular la reepitelización, la angiogénesis y la producción de la matriz extracelular. Objetivo: La presente revisión sistemática tuvo como objetivo comparar los resultados en la literatura publicada sobre si L-PRF es un adyuvante efectivo y predecible al desbridamiento quirúrgico del hueso necrótico para mejorar la eficacia curativa en pacientes con ONMRM. Materiales y Métodos: Las bases de datos de PubMed, Scopus, Cochrane, ScienceDirect, LILACS y Web of Science se registraron utilizando los términos DeCS/MeSH predeterminados y los criterios de elegibilidad, y la búsqueda arrojó un total de cinco artículos. Dos estudios fueron retrospectivos, y tres estudios fueron series de casos. Resultado: Diecisiete participantes recibieron una combinación de desbridamiento quirúrgico, membrana L-PRF y antibióticos. Se observó curación completa de heridas en el 70% de los participantes, y la mayoría de ellos se curaron sin ninguna complicación. Conclusión: L-PRF como adyuvante para el desbridamiento quirúrgico del hueso necrótico parece tener una asociación positiva con el resultado de curación en pacientes con ONMRM.

Evidências clínicas do uso de plantas medicinais e fitoterápicos na cicatrização de feridas cutâneas: uma revisão integrativa / Evidencia clínica sobre el uso de plantas medicinales y medicinas a base de hierbas en la curación de heridas en la piel: una revisión integrativa / Clinical evidence on the use of medicinal plants and herbal medicine in healing skin wounds: an integrative review

Introdução: Plantas medicinais e fitoterápicos são tradicionalmente utilizados na cicatrização de diversos tipos de feridas. Muitas plantas contêm compostos bioativos com propriedades anti-inflamatórias, antimicrobianas e cicatrizantes. Objetivo: Reunir evidências clínicas do uso de plantas medicinais e fitoterápicos na cicatrização de feridas cutâneas. Metodologia: Trata-se de uma revisão integrativa da literatura. Os estudos foram coletados das bases de dados MEDLINE/PubMed e LILACS, utilizando como estratégia de busca os termos ((WoundHealing) ou (Inflammation)) e ((MedicinalPlants) ou (Phytotherapy)) com recorte temporal de 2011 a 2021. Resultados: 15 estudos foram incluídos na revisão. A maioria dos estudos foram do tipo ensaio clínico controlado randomizado. Foi investigada a eficácia de 15 espécies medicinais para o tratamento de feridas agudas e crônicas, sendo as mais recorrentes queimaduras de segundo grau, úlceras crônicas e feridas cirúrgicas. As espécies medicinais que apresentaram melhores resultados foram babosa (Aloevera), centela (Centellaasiatica),beldroega (Portulacaoleraceae), Arnebia euchroma, hipérico (Hypericum perforatum) emil-folhas (Achilleamillefolium), melhorando critérios como eritema, edema, tempodere-epitelização, tempo de cicatrização e a aparência geral da ferida. Conclusões: Plantas medicinais e fitoterápicos apresentam eficácia no processo de cicatrização, sendo, portanto, alternativa terapêutica para o tratamento de feridas cutâneas (AU).

Introduction: Medicinal plants and herbal medicines are traditionally used to heal various types of wounds. Many plants contain bioactive compounds with anti-inflammatory, antimicrobial, and wound-healing properties. Objective: Gathering clinical evidence on the use of medicinal plants and herbal medicine in the healing of skin wounds. Methodology:Thisisan integrative review of the literature. The studies were collected from the MEDLINE/PubMed and LILACS data bases, using the terms ((WoundHealing) or (Inflammation)) and ((MedicinalPlants) or (Phytotherapy)) as a search strategy, from 2011 to 2021. Results: Fifteen studies were included in the review. Most of them were of the randomized controlled clinical trial type. The effectiveness of 15 medicinal species was investigated for the treatment of acute and chronic wounds, the most common of which were second-degree burns, chronic ulcers,and surgical wounds. The species with the best results were Aloe vera, Centella asiatica, Portulaca oleraceae, Arnebia euchroma, Hypericum perforatum, and Achillea millefolium, improving criteria such as erythema, edema, re-epithelialization time, healing time, and the general appearance of the wound. Conclusions: Medicinalplants and herbal medicine are effective in the healing process, therefore being a therapeutic alternative for treating skin wounds (AU).

Introducción: Las plantas medicinales y las medicinas a base de hierbas se utilizan tradicionalmente para curar diversos tipos de heridas. Muchas plantas contienen compuestos bioactivos con propiedades antiinflamatorias, antimicrobianas y cicatrizantes. Objetivo: Recopilar evidencia clínica sobre el uso de plantas medicinales y medicinas a base de hierbas en la curación de heridas cutáneas. Metodología: Se trata de una revisión integradora de la literatura. Los estudios fueron obtenidos de las bases de datos MEDLINE/PubMed y LILACS, utilizando como estrategia de búsqueda los términos ((WoundHealing) o (Inflammation)) y ((MedicinalPlants) o (Phytotherapy)) con un marco temporal de 2011 a 2021. Resultados: Se incluyeron 15 estudios en la revisión. La mayoría de los estudios fueron del tipo ensayo clínico controlado aleatorio. Se investigó la eficacia de 15 especies medicinales para el tratamiento de heridas agudas y crónicas, las más comunes fueron quemaduras de segundo grado, úlceras crónicas y heridas quirúrgicas. Las especies medicinales con mejoresr esultados fueron Aloe vera, Centella asiatica, Portulaca oleraceae, Arnebia euchroma, Hypericum perforatum y Achillea millefolium, mejorando criterios como eritema, edema, tiempo de reepitelización, tiempo de cicatrización y aspecto general de la herida. Conclusiones: Plantas medicinales y medicinas a base de hierbas son efectivas en el proceso de cicatrización, siendo portanto un aalternativa terapéutica para el tratamiento de heridas en la piel (AU).

Small Molecules Accelerate Skin Wound Healing: Shikonin Efficacy and Mechanism of Action in Mice / Las Moléculas Pequeñas Aceleran la Curación de Heridas en la Piel: Eficacia de Shikonin y Mecanismo de Acción en Ratones

SUMMARY: The objective of this study was to investigate the therapeutic wound healing potential and molecular mechanisms of shikonin as small molecules in vitro. A mouse burn model was used to explore the potential therapeutic effect of shikonin; we traced proliferating cells in vivo to locate the active area of skin cell proliferation. Through the results of conventional pathological staining, we found that shikonin has a good effect on the treatment of burned skin and promoted the normal distribution of skin keratin at the damaged site. At the same time, shikonin also promoted the proliferation of skin cells at the damaged site; importantly, we found a significant increase in the number of fibroblasts at the damaged site treated with shikonin. Most importantly, shikonin promotes fibroblasts to repair skin wounds by regulating the PI3K/AKT signaling pathway. This study shows that shikonin can effectively promote the proliferation of skin cell, and local injection of fibroblasts in burned skin can play a certain therapeutic role.

El objetivo de este trabajo fue investigar el potencial terapéutico de cicatrización de heridas y los mecanismos moleculares de la shikonina como moléculas pequeñas in vitro. Se utilizó un modelo de quemaduras en ratones para explorar el posible efecto terapéutico de la shikonina; Rastreamos las células en proliferación in vivo para localizar el área activa de proliferación de células de la piel. A través de los resultados de la tinción para patología convencional, encontramos que la shikonina tiene un buen efecto en el tratamiento de la piel quemada y promueve la distribución normal de la queratina de la piel en el sitio dañado. Al mismo tiempo, la shikonina también promovió la proliferación de células de la piel en el sitio dañado. Es importante destacar que encontramos un aumento significativo en la cantidad de fibroblastos en el sitio dañado tratado con shikonina. Lo más importante es que la shikonina promueve la función reparadora de fibroblastos en las heridas de la piel regulando la vía de señalización PI3K/ AKT. Este estudio muestra que la shikonina puede promover eficazmente la proliferación de células de la piel y que la inyección local de fibroblastos en la piel quemada puede desempeñar un cierto papel terapéutico.

Effect of Curcumin on Promoting Skin Wound Healing in Diabetes Mice / 医药导报

Objective To study the effect of curcumin on wound healing in diabetic mice.Methods The effect of curcumin on fibroblast activity was examined by the MTT assay,and the ROS detection kit was used to detect the effect of curcumin on the hydrogen peroxide-induced scavenging effect of reactive oxygen species(ROS)in fibroblasts.Q-PCR was used to detect the effects of curcumin on the mRNA expression of inflammatory factors CD86,CD206,IL-6 and ARG1 in lipopolysaccharide-induced RAW 264.7macrophage.The wound model of diabetes was established by intraperitoneal injection of streptozotocin.Hematoxylin-eosin(HE)and Masson staining were used to evaluate wound healing and histomorphological changes,and immunofluorescence staining was used to determine skin tissue α-smooth muscle actin,CD86 and CD206 expression.Results Curcumin had no significant effect on fibroblast activity at concentrations less than 20 mol·L-1;curcumin scavenged hydrogen peroxide-induced intracellular ROS in fibroblasts;curcumin decreased the mRNA expression of CD86 and IL-6 while increasing CD206 and ARG1 in lipopolysaccharide-induced RAW 264.7 macrophages.After in vivo administration,compared with the control group,wound healing was significantly faster in the curcumin(15,30 mg·mL-1)group after 7 d and 14 d of wound perforation(P＜0.01).Hematoxylin-eosin(HE)and Masson staining results confirmed a significant increase in granulation tissue and a significant increase in collagen deposition in the curcumin(15,30 mg·mL-1)group.Immunofluorescence assay showed significantly higher expression of CD206(P＜0.01)and significantly reduced expression of CD86(P＜0.01)in the skin wounds of curcumin(15,30 mg·mL-1)for 14 d.In addition,the expression of α-SMA in the wound of the high-dose curcumin(30 mg·mL-1)group was significantly higher than that of the low-dose curcumin group(P＜0.01).Conclusion Curcumin accelerates diabetic wound healing by promoting granulation tissue proliferation and collagen deposition in refractory diabetic wounds in mice through its anti-inflammatory and antioxidant effects.

Effect of oral fish oil on wound healing in patients with diabetic foot ulcers:a randomized,double-blind,placebo-controlled study / 重庆医学

Objective To investigate the effect of oral fish oil on wound healing and related indexes in patients with diabetic foot ulcer(DFU).Methods A randomized,double-blind,placebo-controlled design was used to recruit 68 patients with DFU aged 18-80 years old in the hospital,and the baseline clinical data of the patients were collected.The patients were randomly divided into experimental group(32 cases,fish oil soft capsule,3 g/d)and control group(33 cases,corn oil soft capsule,3 g/d)by random number generated by Ex-cel,and the intervention lasted for 12 weeks.The primary endpoints included the proportion of complete wound healing and healing area≥50%.The secondary endpoints included wound area,healing time,inflamma-tion index,glucose metabolism index,nutrition related index and wound reinfection.Additionally,the influen-cing factors of wound healing were analyzed.Results After intervention,the proportion of complete wound healing and healing area≥50%in the experimental group was significantly higher than that in the control group(P=0.007,0.039).In the subjects with complete wound healing,the mean healing time in the experi-mental group was shorter than that in the control group,but the difference was not statistically significant(P=0.132).The reduction area of wound area in the experimental group was significantly larger than that in the control group(P=0.045).The decrease of interleukin(IL)-6 and IL-8 in the experimental group was significantly higher than that in the control group(P<0.05).There was no significant difference in the reduc-tion of C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),neutrophil-to-lymphocyte ratio(NLR),glycated hemoglobin A1c(HbA1c)and platelet-to-lymphocyte ratio(PLR)between the two groups(P>0.05).The improvement of prealbumin(PA)in the experimental group was higher than that in the control group,but the difference was not statistically significant(P>0.05).Multivariate logistic regression analysis showed that oral fish oil intervention(OR=6.771,95%CI:1.787-25.652),HbA1c(OR=4.149,95%CI:1.026-16.770)and ulcer type(OR=4.319,95%CI:1.026-18.173)were the influencing factors of wound healing(P<0.05).Conclusion Oral fish oil promotes wound healing in patients with DFU,which may be re-lated to improving the level of chronic inflammation in the body.

Huangqi Yanghe Decoction regulates PI3K/AKT/NF-κB signaling pathway to promote wound healing in diabetes foot ulcer rats / 天津医药

Objective To explore the effect of Huangqi Yanghe Decoction on wound healing of diabetic foot ulcer(DFU)rats based on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/nuclear factor-κB(NF-κB)signal pathway.Methods DFU rat model was constructed,and 48 rats successfully modeled were randomly divided into the model group,the Huangqi Yanghe Decoction low(8.5 g/kg)group,the Huangqi Yanghe Decoction high(17 g/kg)dose group and the Huangqi Yanghe Decoction high dose(17 g/kg)+LY294002(PI3K/AKT pathway inhibitor,0.3 mg/kg)group.There were 12 rats in each group.Another 12 rats were selected as the control group.Rats in each group were given corresponding drug intervention for 4 weeks.After the 14th and 28th day-administration,the general state and wound changes of rats were observed,and the wound healing rate was calculated.The fasting blood glucose(FBG)level of rats was measured,and the percutaneous partial pressure of oxygen(TcpO2)of tissue around the wound was detected.Serum levels of vascular endothelial growth factor(VEGF),hypoxia inducible factor-1α(HIF-1α),C-reactive protein(CRP)and interleukin(IL)-6 were determined by enzyme linked immunosorbent assay.Histopathological changes of the wound were observed by hematoxylin-eosin staining.Immunohistochemical staining was used to measure the microvascular density of rat wound tissue.The protein expression levels of PI3K,phosphorylated PI3K(p-PI3K),AKT,phosphorylated AKT(p-AKT),NF-κB p65,phosphorylated NF-κB p65(p-NF-κB p65)and NF-κB inhibitory protein α(IκB-α)in rat wound tissue were determined by Western blot assay.Results Rats in the control group had smooth hair color,normal diet,drinking water and excretion,more active,wound healing fast,less inflammatory reaction in wound tissue,and there were more new blood vessels.Fibroblasts and collagen matrix were abundant in granulation tissue.In the model group,the fur color of rats was dull and matte,and the activity was reduced.The symptoms of polydipsia,polyphagia and polyuria were appeared in the model group,the wound color was dark,and edema and ulcer appeared in the surrounding tissue,a large number of inflammatory cells infiltrated in the wound tissue,accompanied by tissue necrosis and exudation,fewer neovascularization and fibroblasts were observed.Wound healing rate,TcpO2 in wound surrounding tissue,serum VEGF,HIF-1α,microvascular density,p-PI3K,p-AKT and IκB-α protein expression levels in wound tissue were decreased,and FBG,serum CRP,IL-6,p-NF-κB p65 protein expression in wound tissue were increased(P＜0.05).Compared with the model group,the state of rats was gradually improved in the Huangqi Yanghe Decoction low and high dose groups,and the lesion degree of wound tissue was reduced successively,wound healing rate,TcpO2 in wound surrounding tissue,serum VEGF,HIF-1α,microvascular density,p-PI3K,p-AKT and IκB-α protein expression levels in wound tissue were increased in turn(P＜0.05).The FBG,serum CRP,IL-6 and p-NF-κB p65 protein expression in wound tissue were decreased in turn(P＜0.05).LY294002 could partially reverse the therapeutic effect of high-dose Huangqi Yanghe Decoction on DFU rats(P＜0.05).Conclusion Huangqi Yanghe Decoction can regulate PI3K/AKT/NF-κB pathway,inhibit inflammatory response in DFU rats,promote angiogenesis and thus promote wound healing.

Hydrogel loaded with platelet-rich plasma promotes wound healing in diabetic rats / 中国组织工程研究

BACKGROUND:Diabetic wounds have complicated conditions such as infection,ischemia,peripheral neuropathy,and vascular disease.Ordinary hydrogel dressings with single structure and function cannot meet the needs of diabetic wound healing. OBJECTIVE:To explore the effect of a hydrogel loaded with platelet-rich plasma on wound healing of full-thickness skin defects in diabetic rats. METHODS:The blood of SD rats was extracted to prepare platelet-rich plasma.Carboxymethyl chitosan/oxychondroitin sulfate hydrogel and carboxymethyl chitosan/oxychondroitin sulfate hydrogel loaded with platelet-rich plasma were prepared separately.Streptozotocin was used to induce diabetes model in adult male SD rats.A round full-thickness skin wound with a diameter of 2 cm was made on the back of diabetic rats.The rats were randomly divided into four groups(n=10 per group).The blank group was applied with gauze on the wound.The hydrogel group,platelet-rich plasma group,and composite hydrogel group were respectively applied with the corresponding hydrogel,platelet-rich plasma and hydrogel loaded with platelet-rich plasma.The wound healing was observed within 20 days after treatment. RESULTS AND CONCLUSION:(1)On day 20 after treatment,the wound healing rate of the hydrogel group,platelet-rich plasma group and composite hydrogel group was significantly higher than that of the blank group(P<0.05).The wound healing rate of the composite hydrogel group was significantly higher than that of the platelet-rich plasma group(P<0.05).(2)The results of hematoxylin-eosin staining on day 5 after treatment showed that compared with the blank group,hydrogel group and platelet-rich plasma group,there were a large number of inflammatory cell infiltration,new granulation tissue and capillary formation in the wound tissue of the composite hydrogel group.(3)On day 5 after treatment,the results of immunohistochemical staining and western blot assay showed that the expression levels of tumor necrosis factor α and interleukin 1β in wound tissue in the composite hydrogel group were significantly lower than those in the blank group,hydrogel group and platelet-rich plasma group(P<0.05).(4)Masson staining results on day 15 after treatment showed that compared with the blank group,hydrogel group and platelet-rich plasma group,there were more collagen fibers in the wound tissue of the composite hydrogel group,which were orderly,evenly distributed and dense.(5)CD31 immunofluorescence staining showed that on day 15 after treatment,the expression of CD31 in wound tissue of the composite hydrogel group was significantly higher than that of the blank group,hydrogel group and platelet-rich plasma group(P<0.05).(6)These results suggest that the hydrogel loaded with platelet-rich plasma can promote the healing of full-thickness skin defect wounds in diabetic rats by promoting granulation tissue,collagen fiber and angiogenesis,and reducing the inflammatory response.

Prussian blue nanoparticles promote wound healing of diabetic skin / 中国组织工程研究

BACKGROUND:Inflammation,oxidative stress and bacterial infection are the main causes of delayed wound healing in diabetes.In recent years,various inorganic nanomaterials have been widely used in the treatment of skin wound healing due to their antibacterial activities,but their effects on anti-oxidation and anti-inflammation are limited. OBJECTIVE:To investigate the effect of Prussian blue nanoparticles on the wound repair of diabetes in terms of antioxidant,anti-inflammatory and photothermal antibacterial activities. METHODS:Prussian blue nanoparticles were prepared and characterized.(1)In vitro:The biocompatibility of Prussian blue nanoparticles with different concentrations was detected by MTT assay.The cytoprotective effect of Prussian blue nanoparticles and the intracellular reactive oxidative species level were examined under the condition of hydrogen peroxide.The ability of Prussian blue nanoparticles to decompose hydrogen peroxide and superoxide anion radicals was tested;the effect of Prussian blue nanoparticles on lipopolysaccharide-induced macrophage inflammation was investigated.The photothermal antibacterial activity of Prussian blue nanoparticles was detected by the plate colony counting method.(2)In vivo:ICR mice were intraperitoneally injected with streptozotocin to establish a diabetes mouse model.After the model was successfully established,a 6 mm wound was created on the back with a hole punch.There were the control group(no treatment),the Prussian blue group and the Prussian blue with light group.The wound healing and histomorphological changes were observed. RESULTS AND CONCLUSION:(1)In vitro:Prussian blue nanoparticles in 25-200 μg/mL were non-toxic to cells.Prussian blue nanoparticles had the extremely strong antioxidant capacity and mitigated the intracellular reactive oxidative species at a high oxidative stress environment,resulting in a pronounced cytoprotective effect.The Prussian blue nanoparticles not only exhibited hydrogen peroxide degradation activity but also showed strong superoxide scavenging ability.Prussian blue nanoparticles also displayed significant anti-inflammatory activity and extremely strong antibacterial ability after light irradiation.(2)In vivo:After 14 days,the wound sizes of the Prussian blue group and Prussian blue with light group were significantly reduced,and the healing speed of Prussian blue with light group was the fastest.Hematoxylin-eosin and Masson staining showed a lot of granulation tissue formation and collagen deposition in the Prussian blue group and the Prussian blue with light group,of which the Prussian blue with light group was the most.Immunofluorescence staining displayed that,compared with the control group,the expressions of α-SMA and CD31 were increased significantly in Prussian blue group and Prussian blue with light group(P<0.05),but F4/80 expression was decreased significantly in Prussian blue group and Prussian blue with light group(P<0.05),indicating more obvious improvement in the Prussian blue with light group.(3)These results showed that Prussian blue nanoparticles could promote the skin wound healing of the diabetes mouse model by exerting anti-inflammatory,antioxidant and antibacterial effects.

3D-printed multifunctional wound dressing for combined radiation and wound injury / 中国组织工程研究

BACKGROUND:Combined radiation and wound injury appeared mainly in patients with tumor radiotherapy and nuclear radiation accidents.The radiation destroys the repair mechanism,resulting in delayed or prolonged wound healing.It still lacks an effective therapeutic strategy currently. OBJECTIVE:To prepare multifunctional wound dressings based on the multiple clinical symptoms of combined radiation and wound injury,which are designed to be antibacteria,promoted healing and analgesics. METHODS:Using levofloxacin,fibroin and lidocaine hydrochloride as raw materials,3D bioprinting technology was applied to prepare the multifunctional wound dressing.(1)The multifunctional dressing was placed on a fixed culture plate coated with Staphylococcus aureus,Escherichia coli and Pseudomonas aeruginosa,and incubated at 37 ℃ overnight to detect the diameter of the antibacterial zone.(2)40 Kunming mice were randomly divided into trauma group,radiation and trauma model group,treatment group and positive drug group,with 10 mice in each group.Mice in the radiation and trauma model group,treatment group and positive drug group were irradiated by 60Co gamma rays.After 1 hour of radiation,a full-layer skin defect wound with a diameter of 1 cm was made on the back of each mouse in the four groups.Normal saline was applied to the wounds of the trauma group and the radiation and trauma model group.Trethanolamine cream was applied to the wounds of the positive drug group.Multifunctional dressing was applied to the wounds of the treatment group.The dressing was changed every 2 days,and the treatment was continued for 14 days.Wound healing rate and serum interleukin-6 level were measured at 3,7 and 14 days after wound modeling.14 days after the wound modeling,the skin tissue of the wound was obtained and received hematoxylin-eosin staining,Masson staining and cytokeratin-14 immunohistochemical staining. RESULTS AND CONCLUSION:(1)3D-printed multifunctional wound dressing had good antibacterial activity.The antibacterial zone diameters against Staphylococcus aureus,Escherichia coli and Pseudomonas aeruginosa were(4.15±0.09),(4.18±0.23)and(4.35±0.13)cm,respectively.(2)With the extension of modeling time,the wound healed gradually.The wound healing rate of the treatment group and the positive drug group was higher than that of the radiation and trauma model group at 3,7 and 14 days after modeling(P<0.01,P<0.001).The wound healing rate of the treatment group was higher than that of the positive drug group.With the extension of modeling time,the serum interleukin level of mice increased first and then decreased.The serum interleukin level in the treatment group at 3,7 and 14 days after modeling was lower than that in the radiation and trauma model group.Hematoxylin-eosin staining and Masson staining exhibited that inflammatory cells infiltrated the granuloma tissue in the trauma group,and the dermal collagen fibers were densely arranged.The normal structure of epidermis and dermis was destroyed and inflammatory cells were infiltrated in the radiation and trauma model group.In the treatment group,normal skin mucosal tissue was observed,the epidermis was arranged closely,and the sweat glands,hair follicles and dermal collagen fibers were arranged regularly.In the positive drug group,the arrangement of epidermal layer was tight,and the arrangement of sweat glands,hair follicles and dermal collagen fibers was regular.Cytokeratin-14 immunohistochemical staining displayed that the epidermal tissue thickness in the treatment group was lower than that in the other three groups(P<0.01,P<0.001).(3)The results confirm that the 3D-printed multifunctional dressing has multiple functions of local anesthesia,anti-infection and promoting healing.

Placenta-derived mesenchymal stem cells in promoting acute skin wound healing in rats / 中国组织工程研究

BACKGROUND:Currently,a variety of mesenchymal stem cells have been confirmed to have the effect of promoting wound repair,but there is still a lack of relevant research on whether placenta-derived mesenchymal stem cells can promote acute skin wound healing. OBJECTIVE:To investigate the effect of placenta-derived mesenchymal stem cell transplantation on the healing of acute skin wound in rats. METHODS:Twenty SD rats were divided into PBS group and stem cell group by the random number table method,with 10 rats in each group.All rats were selected to establish a full-thickness skin defect model.In the PBS group and stem cell group,PBS buffer and placenta-derived mesenchymal stem cells were immediately injected on the wound surface and wound margin immediately and on day 8 after modeling.The wound healing was observed immediately and on days 2,4,6,8,10,12,and 14 after modeling.The skin tissue of the wound surface was taken on day 14 and treated with hematoxylin-eosin staining,Masson staining,immunohistochemical staining and immunofluorescence staining. RESULTS AND CONCLUSION:(1)The wound surface of the rats in each group decreased with the prolongation of treatment time.The wound healing rate and wound epithelization rate of the stem cell group at 14 days were higher than those of the PBS group(P<0.01),and the wound contracture rate was lower than that of the PBS group(P<0.01).(2)The results of hematoxylin-eosin staining showed that the skin wound healing of the stem cell group was better than that of the PBS group;the degree of wound epithelization was higher,and the morphology of collagen fibers was close to that of normal skin.(3)Masson staining results showed that compared with the PBS group,collagen fibers in the skin wound tissue of the stem cell group were significantly increased and thicker,and the content of collagen fibers in the new tissue was significantly higher than that of the PBS group(P<0.01).(4)Immunohistochemical staining showed that the number of new capillaries in the stem cell group was higher than that in the PBS group(P<0.01),while the expressions of tumor necrosis factor-α and interleukin-6 were lower than those in the PBS group(P<0.01).(5)Immunofluorescence staining showed that the number of M2 macrophages in the new wounds of the stem cell group was higher than that of the PBS group(P<0.01),while the number of M1 macrophages was less than that in the PBS group(P<0.01).These findings indicate that placenta-derived mesenchymal stem cells can accelerate skin wound healing,promote wound epithelization,and reduce wound contracture,which may be related to the promotion of capillary angiogenesis,regulation of collagen fiber production,inhibition of inflammation,and regulation of macrophage polarization to M2 type.

Influence of adipose tissue and its derivative on wound repair and vascularization / 中国组织工程研究

BACKGROUND:Vascularization is essential for wound healing and functional recovery during soft tissue repair.Adipose tissue is believed to be the body's largest source of stem cells,and a number of different fat complexes have been developed for research and treatment.Its ability to promote angiogenesis and soft tissue repair has been extensively studied. OBJECTIVE:To review the progress of vascularization in soft tissue repair,and to summarize the preparation methods of adipose tissue and its derivative and their applications in vascularization and soft tissue repair.It is proven that adipose tissue and its derivative have excellent research value and clinical application prospects in vascular and soft tissue engineering. METHODS:PubMed,Web of Science and CNKI databases were used to search the related articles published from January 2010 to February 2023.The search terms were"soft tissue repair,wound healing,vascularization,angiogenesis,adipose tissue,stromal vascular fraction,adipose tissue-derived microvascular fragment,nanofat,adipose extracellular matrix/stromal vascular fraction gel"in Chinese and English.A small number of old classic literature was also included.An initial screening was performed by reading the titles and abstracts to exclude literature that was not relevant to the topic of the article,and 69 papers were finally included for the analysis of the result. RESULTS AND CONCLUSION:(1)Wound healing is an important physiological process,which mainly occurs when tissue is damaged,such as injury,surgery,burn,tumor,infection and vascular disease caused by tissue damage and defects.(2)Adequate vascularization of the wound site is essential for tissue repair,reconstruction of local homeostasis and functional recovery.(3)Adipose tissue is believed to be the body's largest pool of stem cells and a number of different fat components have been used for research and treatment.(4)Due to its inherent composition and preparation advantages,adipose tissue will continue to play an important role in tissue engineering research and therapy.

Moxibustion and reduced graphene oxide/cerium dioxide nanocomposites for repairing infectious wounds / 中国组织工程研究

BACKGROUND:The repair process of skin trauma is complex and susceptible to infection,easy to lead to poor healing,is the current difficulty and hot spot in wound repair research,and has received extensive attention in the fields of traditional Chinese medicine and tissue engineering. OBJECTIVE:To investigate the effect of moxibustion and reduced graphene oxide/cerium oxide nanocomposite on promoting the healing of infectious wounds. METHODS:(1)Reduced graphene oxide/cerium dioxide nanocomposites with mass ratios of 2:1,1:1 and 1:2 were synthesized by hydrothermal method.The resulting composites were recorded as G2C1,G1C1 and G1C2,respectively.The photothermal properties,cytotoxicity and antibacterial properties of the three kinds of materials were tested.After taking moxa sticks,three kinds of moxibustion distances were set(3.0-3.5 cm,recorded as moxibustion 1;2.5-3.0 cm,recorded as moxibustion 2;2.0-2.5 cm,recorded as moxibustion 3).Moxibustion was applied to the surface of human skin for 10 minutes to detect the photothermal properties.The antibacterial properties of moxibustion were tested at three different distance intervals.Simultaneously,the back body surface infrared imaging of rats with different mass concentrations of G1C1 material,moxibustion(three kinds of moxibustion distances)and moxibustion 2+G1C1 material was detected.(2)Sixty male Sprague-Dawley rats were selected to model the wound of Staphylococcus aureus infection.48 hours later,they were randomly divided into 10 groups with 6 rats in each group:control group(did not receive any treatment),mupirocin group,moxibustion 2+G1C1 group,moxibustion 1 group,moxibustion 2 group,moxibustion 3 group and 60,80,100,and 120 μg/mL G1C1 groups(The G1C1 group was given 808 nm near-infrared laser irradiation for 10 min/time,and the G1C1 suspension was loaded on the wound surface before each treatment.Each group of moxibustion underwent in-situ suspension moxibustion,and the intervention time was 10 min/time.Moxibustion 2+G1C1 group was loaded with G1C1 suspension on the wound surface before each treatment,and moxibustion was suspended in situ with moxa strips,and the intervention time was 10 min/time).The frequency of treatment was 2 days once.Wound healing,wound colony count and repair were detected after 7 days of intervention. RESULTS AND CONCLUSION:(1)The three kinds of reduced graphene oxide/cerium dioxide nanocomposites had good photothermal properties,and the higher the mass concentration of the composites,the better the photothermal properties.The temperature of the moxibustion 2 group reached 47.6 ℃for 10 minutes without causing thermal damage,which was more suitable for animal experiments.The results of co-culture with NIH-3T3 cells exhibited that 60,80,and 100 μg/mL G1C1 had good biocompatibility.The results of a co-culture experiment with Staphylococcus aureus suspension displayed that G2C1,G1C1 and G1C2 had good antibacterial activity,among which G1C1 group demonstrated excellent antibacterial performance,and the antibacterial rate reached 100％when its mass concentration was 80 μg/mL.60-120 μg/mL G1C1 could effectively remove Staphylococcus aureus biofilm,and the higher the material mass concentration,the better the removal effect.Moxibustion could also effectively remove Staphylococcus aureus biofilm,and the closer the moxibustion was,the better the removal effect.(2)Compared with the control group,the wound area of the mupirocin group,moxibustion 2 group,moxibustion 2+G1C1 group and 80,100 μg/mL G1C1 groups was significantly reduced on day 7 of treatment,and the quality of wound repair was better.Mupirocin,G1C1,moxibustion and moxibustion 2+G1C1 could effectively remove the residual bacteria on the wound surface,and the higher the mass concentration of G1C1,the lower the residual bacteria.Among them,the wound repair efficiency and bacterial residue of 80 μg/mL G1C1 group and moxibustion 2 group were very similar,and the wound repair efficiency of both was better than that of mupirocin group.In addition,it was also observed that the combination of materials and moxibustion had a better ability to clear wound bacteria than that used alone.(3)The results confirm that moxibustion,reduced graphene oxide/cerium dioxide nanocomposites and their combination have good anti-infection and wound healing effects.

Platelet-camouflaged silver nanoparticle hydrogel accelerates wound healing in type 1 diabetic rats / 中国组织工程研究

BACKGROUND:Bacterial infections and impaired angiogenesis have been obstacles to diabetic wound healing,and the problem of multidrug resistance cannot be ignored,so there is an urgent need to find a new therapeutic strategy. OBJECTIVE:To prepare platelet-camouflaged silver nanoparticle hydrogel and observe its therapeutic effect on diabetic wounds. METHODS:(1)In vitro test:The ultra-small silver nanoparticles(usAgNPs)were obtained by reflux heating.usAgNPs-PL/CMC hydrogel was prepared by coupling it with platelets(PL)and adding it to the carboxymethyl cellulose(CMC)hydrogel.The microstructures,swelling properties,and mechanical properties of the hydrogel were characterized.The antibacterial activity of the hydrogel was tested by 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)5[(phenylamino)carbonyl]-2H-tetrazolium hydroxide method,reactive oxygen species method,bacterial biofilm method,and bacterial activity.(2)In vivo test:Forty SD rats were randomly divided into sham operation group,model group,CMC hydrogel group,usAgNPs/CMC hydrogel group,and usAgNPs-PL/CMC hydrogel group,with eight rats in each group.The type 1 diabetes model was established in the other four groups except the sham operation group.After successful model establishment,a skin wound(20 mm in diameter)deep to the fascia layer was made on the back of rats of the five groups.Rats in the sham operation group and model group were injected with normal saline.The rats in the CMC hydrogel group,usAgNPs/CMC hydrogel group,and usAgNPs-PL/CMC hydrogel group were implanted with the corresponding hydrogel,separately.The wound healing condition and healing quality were observed after 14 days of treatment. RESULTS AND CONCLUSION:(1)In vitro test:The usAgNPs-PL/CMC hydrogel had a uniform three-dimensional network structure,good mechanical properties,and strong water absorption ability,which could remarkably inhibit the growth of Escherichia coli and Staphylococcus aureus,and had a good inhibition and clearance effect on the formation of biofilms of the two bacteria,and had a good antibacterial ability.(2)In vivo test:After 14 days of treatment,the wounds of the usAgNPs-PL/CMC hydrogel group were basically closed.The wound healing rate was the highest,and the number of wound colonies was significantly lower than that of the model group.After 14 days of treatment,hematoxylin-eosin and Masson staining showed that the tissue structure of regenerated dermis appeared on the wounds of the usAgNPs-PL/CMC hydrogel group,usAgNPs/CMC hydrogel group,and CMC hydrogel group,and the epidermis of the new granulation tissue was complete and thick.Parallel mature collagen fibers were observed in the usAgNPs-PL/CMC hydrogel group.After 7 days of treatment,CD31 immunohistochemistry and Ki67 immunofluorescence staining showed that the usAgNPs-PL/CMC hydrogel group had the highest number of microvessels and fibroblasts.(3)The results showed that usAgNPs-PL/CMC hydrogel could accelerate wound healing in type 1 diabetic rats by rapidly inhibiting bacteria and promoting angiogenesis.

Functional characteristics and clinical applications of MXene nanoparticles in wound healing / 中国组织工程研究

BACKGROUND:MXene nanoparticles have considerable application prospects as effective functional components of skin wound dressings due to their unique properties of conductivity,hydrophilicity,antibacterial activity,and biocompatibility. OBJECTIVE:To review the synthesis methods,functional properties,and application of MXene nanoparticles in skin injury repair. METHODS:"MXene,nanoparticles,nanomaterials,bioactive nanoparticles","wound dressing,wound dressing,wound repair materials","wound repair,wound healing,wound surface"were used as Chinese search terms to search Wanfang and CNKI databases."MXene,nanoparticles,nano-materials,bioactive nanoparticles","wound dressing,wound healing material","wound healing,wound repair,wound"were used as English search terms to search the PubMed database.Finally,88 articles were included for review analysis. RESULTS AND CONCLUSION:(1)MXene synthesis can be divided into two methods:bottom-up synthesis and top-down synthesis.The synthesized MXene can be further modified to enhance biocompatibility for better application in biomedicine.(2)MXene has a series of excellent properties such as hydrophilicity,antibacterial,photothermal properties,electrical conductivity,and good biocompatibility,all of which make it the basis for excellent skin repair materials.(3)At present,many scholars have developed new composite materials for wound dressings.Currently,these composite materials are based on MXene nanomaterials and make full use of its excellent characteristics as mentioned above,which play a role in local skin wound sterilization,drug delivery,and sustained release,active regulation of cytokines,and can integrate the advantages of other biologically active agents.It plays a better role in wound healing,especially in the treatment of complex chronic wounds.(4)Various composite materials such as MXene@PVA hydrogel and MXene@CH sponge developed based on various properties of MXene have shown good effects on improving tissue repair performance and repairing skin damage as drug carriers.It shows that MXenes nanoparticles are in the initial stage of development and have great prospects in the field of promoting skin repair.The characteristics and surface modification of MXenes nanoparticles have been well studied,but the molecular mechanism of dose-dependent biotoxicity is relatively incomplete.

Exosomes and skin wound healing / 中国组织工程研究

BACKGROUND:Exosomes play a role in all stages of wound repair,and there is currently a large body of research on exosomes in skin wound repair,which has been shown to have great potential for clinical applications. OBJECTIVE:To summarize and discuss the main mechanisms and clinical applications of exosomes in the treatment of skin wounds,in order to promote the clinical translation of exosomes. METHODS:PubMed,clinicaltrials.gov,China National Knowledge Infrastructure,Food and Drug Administration database,and Chinese Clinical Trial Register were searched from inception to March 2023.The English search terms were"exosomes,wound healing,stem cells,chronic wound,immunoregulation,inflammation,skin,therapeutic use,isolation,characterization,infections".The Chinese search terms were"exosomes,wound healing,stem cells,immunomodulation,clinical applications".A total of 79 articles were included for the summary. RESULTS AND CONCLUSION:(1)Exosomes can improve and accelerate wound healing through inflammation regulation,immune protection,angiogenesis,cell proliferation and migration,and collagen remodeling.(2)Exosomes derived from stem cells have mature preparation techniques and related mechanism research,which is currently the mainstream research direction.Non-stem cell-derived exosomes have the advantages of convenience,economy,and easy production,and can be used as a supplement for clinical applications.(3)The clinical application of exosomes is still in its infancy,but has great potential for application.Various exosome modification techniques have laid the foundation for the future development of clinically personalized services and require further research.(4)The clinical translation of exosomes faces many challenges,such as low yield,high heterogeneity,lack of unified standards for isolation,purification,and quality control,and difficulties in storage.

Liposome gel loaded oleic acid promotes the repair of chronic burn wounds / 中国组织工程研究

BACKGROUND:Oleic acid can regulate inflammation and immune responses,and has the potential to repair skin wounds.Oleic acid has a short retention time at the lesion.It is prone to self oxidation and deterioration in the air,and suitable drug carriers are needed to fully exert the therapeutic effect of oleic acid. OBJECTIVE:To investigate the efficacy of oleic acid-liposome gel in the treatment of chronic burn wounds. METHODS:Oleic acid liposome solution was prepared by thin film dispersion method,and then dissolved in Poloxamer gel matrix to prepare oleic acid-liposome gel.(1)In vitro experiment:Oleic acid-liposome gel solution was prepared by adding different volumes of oleic acid-liposome gel into cell medium(volume ratio:1:3,1:9,1:27,respectively).Alma-blue reagent was used to detect the effects of different concentrations of oleic acid-liposome gel on the proliferation of human keratinocytes and human fibroblasts.Crystal violet staining was used to observe cell morphology.(2)In vivo experiment:The animal model of chronic burn wounds was established by using full-thickness burn of SD rat back skin combined with local subcutaneous injection of epirubicin.The 30 successfully modeled rats were randomly divided into five groups with six rats in each group.The wounds of oleic acid liposome gel group,oleic acid group,liposome gel group,positive control group and negative control group were applied with gauze of oleic acid liposome gel,oleic acid,liposome gel,recombinant human epidermal growth factor gel and normal saline.The dressing was changed once every other day.A total of 16 doses were administered.The wound healing was observed. RESULTS AND CONCLUSION:(1)In vitro experiments:Alma-blue reagent detection and crystal violet staining showed that oleic acid liposome gel solution with volume ratio of 1:9 could promote the proliferation of human keratinocytes and human fibroblasts.(2)In vivo experiment:The wound healing time of the oleic acid liposome gel group was shorter than that of the other four groups(P＜0.01),and the wound healing rate at 4,8,12,16,and 20 days was higher than that of the other four groups(P＜0.01).After administration,hematoxylin-eosin staining showed epithelialization and healing of wounds in all five groups,and the epidermal thickness of oleic acid liposome gel group was the closest to normal skin and better than the other four groups.Immunohistochemical staining showed that the expressions of cytokeratin 10,tumor protein 63,α-smooth muscle actin,collagen I,tumor necrosis factor α,interleukin 6,malonaldehyde,and superoxide dismutase in oleic acid liposome gel group were closest to those in normal skin,and superior to those in other four groups.On days 12 and 32 of administration,the expressions of tumor necrosis factor α,interleukin 6,malondialdehyde,and superoxide dismutase in wound homogenate supernatant in oleic acid liposome gel group were closest to those in normal skin,and superior to those in other four groups.(3)The results showed that oleic acid liposome gel could promote the proliferation of keratinocytes and fibroblasts,reduce inflammation and oxidative stress injury,and promote the healing of chronic burn wounds.

Mechanism by which lycium barbarum polysaccharides inhibit keratinocyte apoptosis in burn wounds via autophagy / 中国组织工程研究

BACKGROUND:Lycium barbarum polysaccharide has many biological activities and has been found to have potential effects on promoting wound healing. OBJECTIVE:To investigate the mechanism of lycium barbarum polysaccharide in tumor necrosis factor-α-mediated keratinocyte apoptosis and its effect on the healing of burn wounds. METHODS:(1)In vitro experiment:Keratinocytes were divided into four groups:cells were cultured in the α-MEM medium(complete medium)containing 15%fetal bovine serum and 1%glutamine in normal group,cultured in the complete medium containing lycium barbarum polysaccharide in positive control group,cultured in the complete medium containing tumor necrosis factor-α in model group,and cultured in the complete medium containing lycium barbarum polysaccharide and tumor necrosis factor-α in experimental group.After 24 hours of culture,cell proliferation was detected using cell counting kit-8 assay;Cleaved caspase-8,TNF R1,FADD,and LC3 were detected using western blot.Then an autophagy inhibitor group(the complete medium containing 3-methyladenine)and an autophagy inhibitor+lycium barbarum polysaccharide group(the complete medium containing lycium barbarum polysaccharide,tumor necrosis factor-α,and 3-methyladenine)were set up.After 24 hours of culture,keratinocyte apoptosis was detected by flow cytometry.(2)In vivo experiment:Six Sprague-Dawley rats were randomly divided into a control group and an experimental group,with three rats in each group.Four deep Ⅱ degree burn wounds with a diameter of 2 cm were made on the back of each rat.At 24 hours after modeling,mice in the control and experimental groups were coated with normal saline and lycium barbarum polysaccharide solution,respectively,once a day.Wound healing was observed regularly after treatment.Samples were taken 28 days after treatment and the pathologic pattern of the wound was observed. RESULTS AND CONCLUSION:(1)In vitro experiment:Addition of lycium barbarum polysaccharide alone did not affect cell proliferation and apoptosis and the expression of apoptotic and autophagic proteins in keratinocytes.After the addition of tumor necrosis factor α,the proliferation of keratinocytes was inhibited,the apoptotic rate increased,and the expression of apoptotic and autophagic proteins was elevated,while lycium barbarum polysaccharide could antagonize the above effects of tumor necrosis factor-α.Lycium barbarum polysaccharide combined with autophagy inhibitors further reduced the apoptotic rate of keratinocytes.(2)In vivo experiment:The wound healing rate of rats in the experimental group was higher than that of the control group at 12,16,20,24,and 28 days after treatment(P＜0.05,P＜0.01).Hematoxylin-eosin staining results at 28 days after treatment showed an intact and well-defined epidermis of the wound in the experimental group compared with the control group.To conclude,lycium barbarum polysaccharide protects the integrity of skin epidermal tissue and promotes wound healing by inhibiting autophagy and apoptosis of keratinocytes.

Oxymatrine hydrogel promotes wound healing by activating Nrf2/HO-1 pathway in keratinocytes / 中国组织工程研究

BACKGROUND:Inflammation and oxidative stress contribute to the barriers of regeneration in chronic wound.Oxymatrine has various biological activities,such as anti-oxidation,anti-inflammation and so on,which may have the potential effect of promoting wound healing. OBJECTIVE:To investigate the effect of oxymatrine on wound healing and the protective effect on H2O2-induced oxidative stress injury in human keratinoid cell line HaCaT cells. METHODS:(1)In vivo experiment:Hyaluronic acid methacryloyl hydrogels containing 0,0.05,0.1,0.2 g/L oxymatrine were prepared.A full-layer skin defect model with a diameter of 12 mm was made in the back of 75 diabetic mice and randomly divided into five groups for intervention,with 15 mice in each group.The wounds of the model group were bandaged and fixed.The wounds of the hydrogel group were covered with hyaluronic acid methacryloyl hydrogel.The wounds of the low-dose,moderate-dose and high-dose oxymatrine groups were covered with hyaluronic acid methacryloyl hydrogel containing 0.05,0.1,and 0.2 g/L oxymatrine,respectively,and then bandaged and fixed after light curing.Relevant indicators were detected within 14 days.(2)In vitro experiment:Human keratinocyte line HaCaT was divided into five groups.The normal group was cultured conventionally.H2O2 group and low-,moderate-and high-concentration oxymatrine groups were treated with H2O2 for 4 hours,and then the medium was replaced with medium containing 0,0.05,0.1,and 0.2 g/L oxymatrine,respectively,and the relevant indexes were detected after 24 hours of culture. RESULTS AND CONCLUSION:(1)In vivo experiment:Compared with the model group,the wound healing rate of mice in the hydrogel group had no significant change.The wound healing rate of mice in the low-,moderate-and high-dose oxymatrine group was increased at 7 and 14 days after treatment(P<0.05).Pathological observation of wound section 14 days after treatment showed that compared with the model group,the thickness of regenerated epidermal layer,the number of microvessels,and collagen deposition in the moderate-and high-dose oxymatrine groups were increased(P<0.05).Western blot assay analysis of wound samples 7 days after surgery showed that compared with the model group,the protein expressions of tumor necrosis factor α and interleukin 6 in the moderate-and high-dose oxymatrine groups were decreased(P<0.05).(2)In vitro experiment:CCK8 assay,EdU and Ki67 staining showed that compared with the H2O2 group,the cell proliferation ability of the moderate-and high-concentration oxymatrine groups was significantly increased(P<0.05).Compared with the H2O2 group,mitochondrial membrane potential was increased(P<0.05)and reactive oxygen species content was decreased(P<0.05)in the moderate-and high-concentration oxymatrine groups.Western blot assay results showed that compared with the H2O2 group,the expression levels of Nrf2 nuclear protein,Nrf2 total protein,HO-1 protein,and superoxide dismutase 1 protein were increased in the high-concentration oxymatrine group(P<0.05).(3)These findings confirm that oxymatrine can alleviate oxidative stress damage in HaCat cells and accelerate wound healing by upregulating the levels of Nrf2 and HO-1 protein.