RESUMEN
Skin identity is controlled by intrinsic features of the epidermis and dermis and their interactions. Modifying skin identity has clinical potential, such as the conversion of residual limb and stump (nonvolar) skin of amputees to pressure-responsive palmoplantar (volar) skin to enhance prosthesis use and minimize skin breakdown. Greater keratin 9 (KRT9) expression, higher epidermal thickness, keratinocyte cytoplasmic size, collagen length, and elastin are markers of volar skin and likely contribute to volar skin resiliency. Given fibroblasts' capacity to modify keratinocyte differentiation, we hypothesized that volar fibroblasts influence these features. Bioprinted skin constructs confirmed the capacity of volar fibroblasts to induce volar keratinocyte features. A clinical trial of healthy volunteers demonstrated that injecting volar fibroblasts into nonvolar skin increased volar features that lasted up to 5 months, highlighting a potential cellular therapy.
Asunto(s)
Refuerzo Biomédico , Bioimpresión , Dermis , Epidermis , Fibroblastos , Queratinocitos , Adulto , Femenino , Humanos , Masculino , Amputados , Diferenciación Celular , Colágeno/metabolismo , Dermis/citología , Dermis/metabolismo , Elastina/metabolismo , Epidermis/metabolismo , Fibroblastos/citología , Fibroblastos/trasplante , Mano , Queratina-9/metabolismo , Queratinocitos/citología , Queratinocitos/metabolismo , Refuerzo Biomédico/métodosRESUMEN
Marfan syndrome (MFS) is a hereditary condition accompanied by disorders in the structural and regulatory properties of connective tissue, including elastic fibers, due to a mutation in the gene encodes for fibrillin-1 protein (FBN1 gene) and the synthesis of abnormal fibrillin-1 glycoprotein. Despite the high potential of mast cells (MCs) to remodel the extracellular matrix (ECM), their pathogenetic significance in MFS has not been considered yet. The group of patients with Marfan syndrome included two mothers and five children (three girls aged 4, 11, and 11 and two boys aged 12 and 13). Normal skin was examined in two children aged 11 and 12. Histochemical, monoplex, and multiplex immunohistochemical techniques; combined protocols of simultaneous histochemical and immunohistochemical staining (the results of staining were assessed using light, epifluorescence, and confocal microscopy); and bioinformatics algorithms for the quantitative analysis of detected targets were used to evaluate mast cells and their relationship with other cells from extracellular structures in the skin dermis. Analysis of the skin MC population in children with Marfan syndrome revealed a considerably increased number of intra-organic populations with the preservation of the specific Tryptase+Chymase+CPA3+ protease profile typical of the skin. The features of the MC histotopography phenotype in MFS consisted of closer colocalization with elastic fibers, smooth muscle cells, and fibroblasts. MCs formed many intradermal clusters that synchronized the activity of cell functions in the stromal landscape of the tissue microenvironment with the help of spatial architectonics, including the formation of cell chains and the creation of fibrous niches. In MCs, the expression of specific proteases, TGF-ß, and heparin increased, with targeted secretion of biologically active substances relative to the dermal elastic fibers, which had specific structural features in MFS, including abnormal variability in thickness along their entire length, alternating thickened and thinned areas, and uneven surface topography. This paper discusses the potential role of MCs in strain analysis (tensometry) of the tissue microenvironment in MFS. Thus, the quantitative and qualitative rearrangements of the skin MC population in MFS are aimed at altering the stromal landscape of the connective tissue. The results obtained should be taken into account when managing clinical signs of MFS manifested in other pathogenetically critical structures of internal organs, including the aorta, tendons, cartilage, and parenchymal organs.
Asunto(s)
Dermis , Tejido Elástico , Síndrome de Marfan , Mastocitos , Humanos , Síndrome de Marfan/metabolismo , Síndrome de Marfan/patología , Síndrome de Marfan/genética , Mastocitos/metabolismo , Mastocitos/patología , Niño , Masculino , Femenino , Tejido Elástico/metabolismo , Tejido Elástico/patología , Preescolar , Dermis/patología , Dermis/metabolismo , Adolescente , Fibrilina-1/metabolismo , Fibrilina-1/genética , Piel/metabolismo , Piel/patología , Matriz Extracelular/metabolismo , AdipoquinasRESUMEN
The pemphigoid disease epidermolysis bullosa acquisita (EBA) is an autoimmune blistering skin disease characterized by autoantibodies against type VII collagen (COL7), immune cell infiltrates at the dermal-epidermal junction and subepidermal blistering. Proteases, particularly granzyme B (GzmB), have been established as therapeutic targets for the treatment of EBA and other pemphigoid diseases. We investigated the impact of the novel GzmB inhibitor SNT-6935 on anti-COL7 IgG-induced subepidermal blistering in a well-established EBA ex vivo model. Our findings demonstrate that pharmacological targeting of GzmB with its selective inhibitor SNT-6935 significantly reduced autoantibody-induced dermal-epidermal separation in human skin cryosections. Interestingly, treatment of skin cryosections with recombinant human GzmB alone did not cause dermal-epidermal separation, suggesting that additional mechanisms alongside GzmB are required for tissue damage in EBA. In conclusion, our study highlights the significant contribution of GzmB to the pathogenesis of EBA and supports the notion of GzmB as a therapeutic target in EBA and other pemphigoid diseases.
Asunto(s)
Autoanticuerpos , Colágeno Tipo VII , Epidermis , Epidermólisis Ampollosa Adquirida , Granzimas , Epidermólisis Ampollosa Adquirida/tratamiento farmacológico , Epidermólisis Ampollosa Adquirida/inmunología , Humanos , Granzimas/metabolismo , Granzimas/antagonistas & inhibidores , Colágeno Tipo VII/inmunología , Epidermis/patología , Dermis/patología , Piel/patologíaRESUMEN
The hair follicle is the basis of hair regeneration, and the dermal papilla is one of the most important structures in hair regeneration. New intervention and reversal strategies for hair loss may arise due to the prevention of oxidative stress. GC/MS analysis was used to determine the compounds contained in NSO. Then, NSO was applied to DPC for cell proliferation and oxidative stress experiments. RNA-seq was performed in cells treated with NSO and minoxidil. The quantitative real-time polymerase chain reaction (qRT-PCR) was applied to verify the gene expression. The effects of NSO on hair length, weight, the number and depth of hair follicles, and the dermal thickness were also studied. GC/MS analysis showed that the main components of NSO were eicosapentaenoic acid, palmitic acid, and linoleic acid. NSO promotes DPC proliferation and reduces H2O2-mediated oxidative damage. NSO can also activate hair growth-related pathways and upregulate antioxidant-related genes analyzed by gene profiling. The topical application of NSO significantly promotes hair growth and increases hair length and weight in mice. NSO extract promotes hair growth and effectively inhibits oxidative stress, which is beneficial for the prevention and treatment of hair loss.
Asunto(s)
Proliferación Celular , Folículo Piloso , Cabello , Estrés Oxidativo , Proliferación Celular/efectos de los fármacos , Animales , Humanos , Folículo Piloso/efectos de los fármacos , Folículo Piloso/metabolismo , Folículo Piloso/crecimiento & desarrollo , Folículo Piloso/citología , Ratones , Estrés Oxidativo/efectos de los fármacos , Cabello/efectos de los fármacos , Cabello/crecimiento & desarrollo , Antioxidantes/farmacología , Dermis/metabolismo , Dermis/citología , Dermis/efectos de los fármacosRESUMEN
Electronic skin (e-skin) is considered as a highly promising interface for human-computer interaction systems and wearable electronic devices. Through elaborate design and assembly of various materials, it possesses multiple characteristics similar to human skin, including remarkable flexibility, stretchability, sensitivity to temperature and humidity, biocompatibility, and efficient interfacial ion/electron transport capabilities. Here, we innovatively integrate multifunctional carbon quantum dots (CQDs), which exhibit conductivity, antibacterial properties, ultraviolet absorption, and fluorescence emission, with poly(acrylic acid) and glycerin (Gly) into a three-dimensional network structure of natural goatskin collagen fibers. Through a top-down design strategy enhanced by hydrogen bond reconstruction, we successfully fabricated a novel transparent e-skin (PAC-eSkin). This e-skin exhibited significant tensile properties (4.94 MPa of tensile strength and 263.42% of a maximum breaking elongation), while also possessing Young's modulus similar to human skin (2.32 MPa). It is noteworthy that the functionalized CQDs used was derived from discarded goat hair, and the addition of Gly gave PAC-eSkin excellent antifreezing and moisturizing properties. Due to the presence of ultrasmall CQDs, which creates efficient ion/electron transport channels within PAC-eSkin, it could rapidly sense human motion and physiological signals (with a gauge factor (GF) of 1.88). Furthermore, PAC-eSkin had the potential to replace traditional electrode patches for real-time monitoring of electrocardiogram, electromyogram, and electrooculogram signals, with a higher SNR (signal-to-noise ratio) of 25.1 dB. Additionally, the customizable size and shape of PAC-eSkin offer vast possibilities for the construction of single-electrode triboelectric nanogenerator systems. We have reason to believe that the design and development of this transparent e-skin based on CQDs-functionalized dermal collagen matrices can pave a new way for innovations in human-computer interaction interfaces and their sensing application in diverse scenarios.
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Carbono , Puntos Cuánticos , Dispositivos Electrónicos Vestibles , Puntos Cuánticos/química , Humanos , Carbono/química , Animales , Resinas Acrílicas/química , Glicerol/química , Cabras , Dermis , Resistencia a la Tracción , Colágeno/química , Conductividad EléctricaRESUMEN
Crucial for skin homeostasis, synthesis and degradation of extracellular matrix components are orchestrated by dermal fibroblasts. During aging, alterations of component expression, such as collagens and enzymes, lead to reduction of the mechanical cutaneous tension and defects of skin wound healing. The aim of this study was to better understand the molecular alterations underwent by fibroblasts during aging by comparing secretomic and proteomic signatures of fibroblasts from young (<35years) and aged (>55years) skin donors, in quiescence or TGF-stimulated conditions, using HLPC/MS. The comparison of the secretome from young and aged fibroblasts revealed that 16 proteins in resting condition, and 11 proteins after a 24h-lasting TGF-ß1-treatment, were expressed in significant different ways between the two cell groups (fold change>2, p-value <0.05), with a 77% decrease in the number of secreted proteins in aged cells. Proteome comparison between young and aged fibroblasts identified a significant change of 63 proteins in resting condition, and 73 proteins in TGF-ß1-stimulated condition, with a 67% increase in the number of proteins in aged fibroblasts. The majority of the differentially-expressed molecules belongs to the cytoskeleton-associated proteins and aging was characterized by an increase in Coronin 1C (CORO1C), and Filamin B (FLNB) expression in fibroblasts together with a decrease in Cofilin (CFL1), and Actin alpha cardiac muscle 1 (ACTC1) detection in aged cells, these proteins being involved in actin-filament polymerization and sharing co-activity in cell motility. Our present data reinforce knowledge about an age-related alteration in the synthesis of major proteins linked to the migratory and contractile functions of dermal human fibroblasts.
Asunto(s)
Envejecimiento , Citoesqueleto , Fibroblastos , Proteómica , Humanos , Fibroblastos/metabolismo , Citoesqueleto/metabolismo , Adulto , Persona de Mediana Edad , Envejecimiento/metabolismo , Factor de Crecimiento Transformador beta1/metabolismo , Anciano , Piel/metabolismo , Piel/citología , Proteoma/metabolismo , Células Cultivadas , Masculino , Secretoma/metabolismo , Femenino , Dermis/citología , Dermis/metabolismoRESUMEN
OBJECTIVE: To explore the perioperative nursing methods of autologous dermal transplantation for penile girth enhancement combined with penile lengthening surgery. METHODS: Summarize the perioperative nursing data of 5 patients with small penis who underwent autologous groin dermal transplantation for penile girth enhancement combined with penile lengthening surgery. RESULTS: After comprehensive perioperative nursing, all 5 patients recovered well after the surgery. The preoperative APPSSI scores of the patients were 4.60±0.48, which were all less than 6 points. The postoperative APPSSI scores at 2 months, 6 months, and 12 months were 9ï¼12 (10.6±1.02), 10ï¼12 (11.2±0.98), and 10ï¼12 (11.2±0.98) respectively, showing satisfaction with the surgical outcomes. There was a statistically significant difference compared to the preoperative APPSSI scores (ï¼°<0.05). The preoperative SAS scores were 45ï¼58 (52.2±4.35), and the SAS scores at 2 months, 6 months, and 12 months postoperatively were 31ï¼40 (34.2±3.31), 30-41 (35.8±3.65), and 33ï¼40 (35.6±2.33) respectively, indicating a reduction in anxiety levels after the surgery, with a statistically significant difference compared to the preoperative SAS scores (P<0.05). The preoperative IIEF-5 scores were 7ï¼15 (10.4±2.87), and the IIEF-5 scores at 2 months, 6 months, and 1 year postoperatively were 16ï¼24 (19.8±2.71), 18ï¼25 (21.2±2.48), and 18ï¼24 (20.8±2.39) respectively, showing a significant improvement postoperatively, with statistical significance (P<0.05). The preoperative NPTR examination showed a sustained erection time of 18ï¼25 (21.2±2.59) minutes, and the NPTR examination at 2 months, 6 months, and 1 year postoperatively showed sustained erection times of 18ï¼24 (21.8±2.28), 20-25 (23.4±2.30), and 24ï¼27 (25.4±1.14) minutes respectively. There was no statistically significant difference in the sustained erection time at 2 months and 6 months postoperatively compared to preoperative NPTR examination, but there was a statistically significant difference at 12 months postoperatively (P<0.01). CONCLUSION: Comprehensive perioperative nursing is an important factor in achieving high satisfaction with the surgery, promoting postoperative recovery, and improving the quality of sexual life for patients undergoing autologous groin dermal transplantation for penile girth enhancement combined with penile lengthening surgery.
Asunto(s)
Pene , Trasplante de Piel , Trasplante Autólogo , Humanos , Masculino , Pene/cirugía , Trasplante de Piel/métodos , Procedimientos de Cirugía Plástica/métodos , Dermis/trasplante , Resultado del Tratamiento , Adulto , Atención PerioperativaRESUMEN
There is a need in clinical practice for new wound healing techniques to address full thickness skin injuries, particularly in individuals with diabetes. Herein we investigated whether dermal derived matrix hydrogel (DMH) loaded with curcumin (Cur) could promote healing in diabetic rats. Sixty diabetic rats were randomly assigned into the non-treated group, DMH group, Cur group, and DMH+Cur group. According to the phases of wound healing, sampling was done on days 7, 14, and 21 for further assessments. Our results indicated that the wound contraction rate, new epidermal length and thickness, number of fibroblasts and vascular length, collagen deposition, and strength properties of the healed wounds were meaningfully increased in the treatment groups than in the non-treated group, and these changes were more obvious in the DMH+Cur ones. In addition, the expression of VEGF and IL-10 genes were meaningfully upregulated in all treatment groups compared to the non-treated group and were greater in the DMH+Cur group. This is while the number of neutrophils and expression levels of TNF-α and IL-1ß genes decreased more significantly in the DMH+Cur group compared to the other groups. In conclusion, it was found that using both DMH and curcumin has a greater impact on diabetic wound healing.
Asunto(s)
Curcumina , Diabetes Mellitus Experimental , Hidrogeles , Cicatrización de Heridas , Animales , Curcumina/farmacología , Cicatrización de Heridas/efectos de los fármacos , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/patología , Hidrogeles/química , Ratas , Masculino , Ratas Sprague-Dawley , Dermis/metabolismo , Dermis/patología , Dermis/efectos de los fármacos , Dermis/lesionesRESUMEN
BACKGROUND: Our goal was to investigate linkages between skin color parameters and skin hydration. Since most prior studies focused on stratum corneum hydration, we focused on epidermal and dermal hydration in relation to skin color parameters in both sexes. MATERIALS AND METHODS: Thirty adults (16 female) with an age ± SD of 24.3 ± 0.6 years participated. Three sites on both volar forearms were evaluated for melanin index (MI), erythema index (EI), Individual Typology Angle (ITA), tissue dielectric constant (TDC) values to depths of 0.5 mm (TDC0.5) and 2.5 mm (TDC2.5), and Fitzpatrick skin type (FST). RESULTS: MI and EI were highly correlated (r = 0.800, p < 0.001) with maximum differences in MI and ITA along the arm of 3% and 6.3% with no difference between arms. Male MI was greater than females (p < 0.01). Male TDC2.5 was 36.1 ± 5.4 and correlated with EI (r = 0.231, p = 0.035). Contrastingly, female TDC25 was 28.5 ± 3.6 with no correlation with EI but was correlated with MI (r = -0.301, p = 0.003). These differential patterns held true for TDC0.5. For both sexes, FST and ITA were highly correlated (r = -0.756, p < 0.001). CONCLUSIONS: The findings revealed several correlations between skin color parameters and hydration that differed between males in females in some cases. The observed correlations may indicate that melanin may differentially impact water-holding capacity between sexes and provides a future research target. Further, these initial findings also may hold significance for dermatological assessments and the customization of skincare treatments tailored to individual skin types and demographics.
Asunto(s)
Epidermis , Melaninas , Pigmentación de la Piel , Humanos , Femenino , Masculino , Pigmentación de la Piel/fisiología , Adulto , Epidermis/metabolismo , Adulto Joven , Melaninas/metabolismo , Agua Corporal/metabolismo , Eritema/patología , Eritema/fisiopatología , Piel , Agua/metabolismo , DermisRESUMEN
Human mesenchymal stromal cells (MSCs) have gained significant interest as cell-based therapeutics for organ restoration in the field of regenerative medicine. More recently, substantial attention has been directed toward cell-free therapy, achieved through the utilization of soluble factors possessing trophic and immunomodulatory properties present in the MSC secretome. This collection of soluble factors can be found either freely in the secretome or packed within its vesicular fraction, known as extracellular vesicles (EVs). MSCs can be derived from various tissue sources, each involving different extraction methods and yielding varying cell amounts. In this study, we describe a nonenzymatic procedure for a straightforward isolation of MSCs from the fetal dermis and the adult dermis. The results demonstrate the isolation of a cell population with a uniform MSC immunophenotype from the earliest passages (approximately 90% positive for the classical MSC markers CD90, CD105, and CD73, while negative for the hematopoietic markers CD34 and CD45, as well as HLA-DR). Additionally, we describe the procedures for cell expansion, banking, and secretome collection.
Asunto(s)
Separación Celular , Dermis , Células Madre Mesenquimatosas , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Dermis/citología , Dermis/metabolismo , Separación Celular/métodos , Inmunofenotipificación , Técnicas de Cultivo de Célula/métodos , Biomarcadores , Células Cultivadas , Vesículas Extracelulares/metabolismo , Secretoma/metabolismoRESUMEN
Prostaglandin E2 (PGE2) is known to be effective in regenerating tissues, and bimatoprost, an analog of PGF2α, has been approved by the FDA as an eyelash growth promoter and has been proven effective in human hair follicles. Thus, to enhance PGE2 levels while improving hair loss, we found dihydroisoquinolinone piperidinylcarboxy pyrazolopyridine (DPP), an inhibitor of 15-hydroxyprostaglandin dehydrogenase (15-PGDH), using DeepZema®, an AI-based drug development program. Here, we investigated whether DPP improved hair loss in human follicle dermal papilla cells (HFDPCs) damaged by dihydrotestosterone (DHT), which causes hair loss. We found that DPP enhanced wound healing and the expression level of alkaline phosphatase in DHT-damaged HFDPCs. We observed that DPP significantly down-regulated the generation of reactive oxygen species caused by DHT. DPP recovered the mitochondrial membrane potential in DHT-damaged HFDPCs. We demonstrated that DPP significantly increased the phosphorylation levels of the AKT/ERK and activated Wnt signaling pathways in DHT-damaged HFDPCs. We also revealed that DPP significantly enhanced the size of the three-dimensional spheroid in DHT-damaged HFDPCs and increased hair growth in ex vivo human hair follicle organ culture. These data suggest that DPP exhibits beneficial effects on DHT-damaged HFDPCs and can be utilized as a promising agent for improving hair loss.
Asunto(s)
Folículo Piloso , Hidroxiprostaglandina Deshidrogenasas , Humanos , Folículo Piloso/efectos de los fármacos , Folículo Piloso/metabolismo , Hidroxiprostaglandina Deshidrogenasas/metabolismo , Hidroxiprostaglandina Deshidrogenasas/antagonistas & inhibidores , Dihidrotestosterona/farmacología , Dihidrotestosterona/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Dermis/metabolismo , Dermis/citología , Dermis/efectos de los fármacos , Células Cultivadas , Vía de Señalización Wnt/efectos de los fármacos , Alopecia/tratamiento farmacológico , Alopecia/metabolismo , Cicatrización de Heridas/efectos de los fármacos , Cabello/efectos de los fármacos , Cabello/crecimiento & desarrollo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Inhibidores Enzimáticos/farmacologíaRESUMEN
Wool is generated by hair follicles (HFs), which are crucial in defining the length, diameter, and morphology of wool fibers. However, the regulatory mechanism of HF growth and development remains largely unknown. Dermal papilla cells (DPCs) are a specialized cell type within HFs that play a crucial role in governing the growth and development of HFs. This study aims to investigate the proliferation and induction ability of ovine DPCs to enhance our understanding of the potential regulatory mechanisms underlying ovine HF growth and development. Previous research has demonstrated that microRNA-181a (miR-181a) was differentially expressed in skin tissues with different wool phenotypes, which indicated that miR-181a might play a crucial role in wool morphogenesis. In this study, we revealed that miR-181a inhibited the proliferation and induction ability of ovine DPCs by quantitative Real-time PCR (qRT-PCR), cell counting Kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, and alkaline phosphatase staining. Then, we also confirmed G protein subunit alpha i2 (GNAI2) is a target gene of miR-181a by dual luciferase reporter assay, qRT-PCR, and Western blot, and that it could promote the proliferation and induction ability of ovine DPCs. In addition, GNAI2 could also activate the Wnt/ß-Catenin signaling pathway in ovine DPCs. This study showed that miR-181a can inhibit the proliferation and induction ability of ovine DPCs by targeting GNAI2 through the Wnt/ß-Catenin signaling pathway.
Asunto(s)
Proliferación Celular , Folículo Piloso , MicroARNs , Vía de Señalización Wnt , MicroARNs/genética , MicroARNs/metabolismo , Animales , Ovinos , Folículo Piloso/metabolismo , Folículo Piloso/citología , Subunidades alfa de la Proteína de Unión al GTP Gi-Go/metabolismo , Subunidades alfa de la Proteína de Unión al GTP Gi-Go/genética , Dermis/citología , Dermis/metabolismo , Células Cultivadas , Lana/metabolismo , beta Catenina/metabolismo , beta Catenina/genéticaRESUMEN
Dermal fibroblasts deposit type I collagen, the dominant extracellular matrix molecule found in skin, during early postnatal development. Coincident with this biosynthetic program, fibroblasts proteolytically remodel pericellular collagen fibrils by mobilizing the membrane-anchored matrix metalloproteinase, Mmp14. Unexpectedly, dermal fibroblasts in Mmp14-/- mice commit to a large-scale apoptotic program that leaves skin tissues replete with dying cells. A requirement for Mmp14 in dermal fibroblast survival is recapitulated in vitro when cells are embedded within, but not cultured atop, three-dimensional hydrogels of crosslinked type I collagen. In the absence of Mmp14-dependent pericellular proteolysis, dermal fibroblasts fail to trigger ß1 integrin activation and instead actuate a TGF-ß1/phospho-JNK stress response that leads to apoptotic cell death in vitro as well as in vivo. Taken together, these studies identify Mmp14 as a requisite cell survival factor that maintains dermal fibroblast viability in postnatal dermal tissues.
Asunto(s)
Apoptosis , Supervivencia Celular , Fibroblastos , Metaloproteinasa 14 de la Matriz , Animales , Metaloproteinasa 14 de la Matriz/metabolismo , Metaloproteinasa 14 de la Matriz/genética , Fibroblastos/metabolismo , Ratones , Ratones Noqueados , Colágeno Tipo I/metabolismo , Colágeno Tipo I/genética , Integrina beta1/metabolismo , Integrina beta1/genética , Factor de Crecimiento Transformador beta1/metabolismo , Dermis/metabolismo , Dermis/citología , Células Cultivadas , Matriz Extracelular/metabolismo , Ratones Endogámicos C57BL , Piel/metabolismoRESUMEN
Tendons and ligaments (T/L) are strong hierarchically organized structures uniting the musculoskeletal system. These tissues have a strictly arranged collagen type I-rich extracellular matrix (ECM) and T/L-lineage cells mainly positioned in parallel rows. After injury, T/L require a long time for rehabilitation with high failure risk and often unsatisfactory repair outcomes. Despite recent advancements in T/L biology research, one of the remaining challenges is that the T/L field still lacks a standardized differentiation protocol that is able to recapitulate T/L formation process in vitro. For example, bone and fat differentiation of mesenchymal precursor cells require just standard two-dimensional (2D) cell culture and the addition of specific stimulation media. For differentiation to cartilage, three-dimensional (3D) pellet culture and supplementation of TGFß is necessary. However, cell differentiation to tendon needs a very orderly 3D culture model, which ideally should also be subjectable to dynamic mechanical stimulation. We have established a 3-step (expansion, stimulation, and maturation) organoid model to form a 3D rod-like structure out of a self-assembled cell sheet, which delivers a natural microenvironment with its own ECM, autocrine, and paracrine factors. These rod-like organoids have a multi-layered cellular architecture within rich ECM and can be handled quite easily for exposure to static mechanical strain. Here, we demonstrated the 3-step protocol by using commercially available dermal fibroblasts. We could show that this cell type forms robust and ECM-abundant organoids. The described procedure can be further optimized in terms of culture media and optimized toward dynamic axial mechanical stimulation. In the same way, alternative cell sources can be tested for their potential to form T/L organoids and thus undergo T/L differentiation. In sum, the established 3D T/L organoid approach can be used as a model for tendon basic research and even for scaffold-free T/L engineering.
Asunto(s)
Técnicas de Cultivo de Célula , Fibroblastos , Ligamentos , Organoides , Tendones , Humanos , Tendones/citología , Fibroblastos/citología , Organoides/citología , Ligamentos/citología , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/fisiología , Dermis/citologíaRESUMEN
The aging process is linked to numerous cellular changes, among which are modifications in the functionality of dermal fibroblasts. These fibroblasts play a crucial role in sustaining the healing of skin wounds. Reduced cell proliferation is a hallmark feature of aged dermal fibroblasts. Long intergenic non-coding RNA (lincRNAs), such as LincRNA-EPS (Erythroid ProSurvival), has been implicated in various cellular processes. However, its role in aged dermal fibroblasts and its impact on the cell cycle and its regulator, Cyclin D1 (CCND1), remains unclear. Primary dermal fibroblasts were isolated from the skin of 17-week-old (young) and 88-week-old (aged) mice. Overexpression of LincRNA-EPS was achieved through plasmid transfection. Cell proliferation was detected using the MTT assay. Real-time PCR was used to quantify relative gene expressions. Our findings indicate a noteworthy decline in the expression of LincRNA-EPS in aged dermal fibroblasts, accompanied by reduced levels of CCND1 and diminished cell proliferation in these aging cells. Significantly, the overexpression of LincRNA-EPS in aged dermal fibroblasts resulted in an upregulation of CCND1 expression and a substantial increase in cell proliferation. Mechanistically, LincRNA-EPS induces CCND1 expression by sequestering miR-34a, which was dysregulated in aged dermal fibroblasts, and directly targeting CCND1. These outcomes underscore the crucial role of LincRNA-EPS in regulating CCND1 and promoting cell proliferation in aged dermal fibroblasts. Our study provides novel insights into the molecular mechanisms underlying age-related changes in dermal fibroblasts and their implications for skin wound healing. The significant reduction in LincRNA-EPS expression in aged dermal fibroblasts and its ability to induce CCND1 expression and enhance cell proliferation highlight its potential as a therapeutic target for addressing age-related skin wound healing.
Asunto(s)
Proliferación Celular , Ciclina D1 , Fibroblastos , ARN Largo no Codificante , Ciclina D1/metabolismo , Ciclina D1/genética , Fibroblastos/metabolismo , Fibroblastos/citología , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Animales , Ratones , Piel/metabolismo , Piel/citología , MicroARNs/genética , MicroARNs/metabolismo , Células Cultivadas , Envejecimiento de la Piel/genética , Dermis/citología , Dermis/metabolismo , Senescencia Celular/genética , Regulación de la Expresión Génica , Cicatrización de Heridas/genética , Envejecimiento/genéticaRESUMEN
Hair follicle (HF) regeneration during wound healing continues to present a significant clinical challenge. Dermal papilla cell-derived exosomes (DPC-Exos) hold immense potential for inducing HF neogenesis. However, the accurate role and underlying mechanisms of DPC-Exos in HF regeneration in wound healing remain to be fully explained. This study, represents the first analysis into the effects of DPC-Exos on fibroblasts during wound healing. Our findings demonstrated that DPC-Exos could stimulate the proliferation and migration of fibroblasts, more importantly, enhance the hair-inducing capacity of fibroblasts. Fibroblasts treated with DPC-Exos were capable of inducing HF neogenesis in nude mice when combined with neonatal mice epidermal cells. In addition, DPC-Exos accelerated wound re-epithelialization and promoted HF regeneration during the healing process. Treatment with DPC-Exos led to increased expression levels of the Wnt pathway transcription factors ß-catenin and Lef1 in both fibroblasts and the dermis of skin wounds. Specifically, the application of a Wnt pathway inhibitor reduced the effects of DPC-Exos on fibroblasts and wound healing. Accordingly, these results offer evidence that DPC-Exos promote HF regeneration during wound healing by enhancing the hair-inducing capacity of fibroblasts and activating the Wnt/ß-catenin signaling pathway. This suggests that DPC-Exos may represent a promising therapeutic strategy for achieving regenerative wound healing.
Asunto(s)
Proliferación Celular , Exosomas , Fibroblastos , Folículo Piloso , Ratones Desnudos , Regeneración , Vibrisas , Vía de Señalización Wnt , Cicatrización de Heridas , beta Catenina , Animales , Ratones , Fibroblastos/metabolismo , Exosomas/metabolismo , Vibrisas/fisiología , beta Catenina/metabolismo , Dermis/metabolismo , Movimiento Celular , Factor de Unión 1 al Potenciador Linfoide/metabolismoRESUMEN
Wound healing is a complex process that is still not fully understood despite extensive research. To address this, we aimed to design and characterize a standardized porcine model for the evaluation of wound healing, dressings, cell therapies, and pharmaceutical agents. Using a standardized approach, we examined the wound healing process in 1.2 mm-deep dermatome wounds at defined positions in 11 female pigs. Unlike previous studies that have only described/analyzed selected punch biopsies, we performed and described histological analyses along the complete wound length using quantitative morphometric methods. All animals remained fully healthy following surgery and showed no signs of infection. Our histopathological evaluation using a predetermined grading score and quantitative manual morphometry demonstrated the impact of different tissue sampling methods, sampling sites, and residual dermis thickness on wound healing. Our study presents a reproducible model for wound healing evaluation and demonstrates the usefulness of porcine models for assessing dermal and epidermal wound healing. The use of histological analyses over the complete wound length provides advantages over previous studies, leading to the possibility of a deeper understanding of the wound healing process. This model could potentially facilitate future research on novel wound dressings and local wound healing therapies.
Asunto(s)
Modelos Animales de Enfermedad , Cicatrización de Heridas , Animales , Porcinos , Femenino , Piel/patología , Piel/lesiones , Dermis/patología , VendajesRESUMEN
Fibroblasts are among the most abundant cell types in the human body, playing crucial roles in numerous physiological processes, including the structural maintenance of the dermis, production of extracellular matrix components, and mediation of inflammatory responses. Despite their importance, fibroblasts remain one of the least characterized cell populations. The advent of single-cell analysis techniques, particularly single-cell RNA sequencing (scRNA-seq) and fluorescence-activated cell sorting (FACS), has enabled detailed investigations into fibroblast biology. In this study, we present an extensive analysis of fibroblast surface markers suitable for cell sorting and subsequent functional studies. We reviewed over three thousand research articles describing fibroblast populations and their markers, characterizing and comparing subtypes based on their surface markers, as well as their intra- and extracellular proteins. Our detailed analysis identified a variety of distinct fibroblast subpopulations, each with unique markers, characteristics dependent on their location, and the physiological or pathophysiological environment. These findings underscore the diversity of fibroblasts as a cellular population and could lead to the development of novel diagnostic and therapeutic tools.
Asunto(s)
Biomarcadores , Separación Celular , Fibroblastos , Citometría de Flujo , Fibroblastos/metabolismo , Fibroblastos/citología , Humanos , Separación Celular/métodos , Biomarcadores/metabolismo , Citometría de Flujo/métodos , Dermis/citología , Dermis/metabolismo , Análisis de la Célula Individual/métodos , Supervivencia Celular , AnimalesRESUMEN
Despite the remarkable advances of dermal fillers that reduce wrinkles caused by dermis thickness reduction, they still lack effective hydrogel systems that stimulate collagen generation along with injection convenience. Here, we develop a stem cell-derived extracellular vesicle (EV)-bearing thermosensitive hydrogel (EVTS-Gel) for effective in vivo collagen generation. The TS-Gel undergoes sol-gel transition at 32.6 °C, as demonstrated by the storage and loss moduli crossover. Moreover, the TS-Gel and the EVTS-Gel have comparable rheological properties. Both hydrogels are injected in a sol state; hence, they require lower injection forces than conventional hydrogel-based dermal fillers. When locally administered to mouse skin, the TS-Gel extends the retention time of EVs by 2.23 times. Based on the nature of the controlled EV release, the EVTS-Gel significantly inhibits the dermis thickness reduction caused by aging compared to the bare EV treatment for 24 weeks. After a single treatment, the collagen layer thickness of the EVTS-Gel-treated dermis becomes 2.64-fold thicker than that of the bare EV-treated dermis. Notably, the collagen generation efficacy of the bare EV is poorer than that of the EVTS-Gel of a 10× lesser dose. Overall, the EVTS-Gel shows potential as an antiaging dermal filler for in vivo collagen generation.
Asunto(s)
Colágeno , Dermis , Vesículas Extracelulares , Hidrogeles , Animales , Ratones , Dermis/metabolismo , Dermis/efectos de los fármacos , Colágeno/química , Hidrogeles/química , Hidrogeles/farmacología , Vesículas Extracelulares/química , Vesículas Extracelulares/metabolismo , Humanos , Células Madre/citología , Células Madre/metabolismo , Células Madre/efectos de los fármacos , Rellenos Dérmicos/química , Rellenos Dérmicos/farmacologíaRESUMEN
Due to the limitations of the current skin wound treatments, it is highly valuable to have a wound healing formulation that mimics the extracellular matrix (ECM) and mechanical properties of natural skin tissue. Here, a novel biomimetic hydrogel formulation has been developed based on a mixture of Agarose-Collagen Type I (AC) combined with skin ECM-related components: Dermatan sulfate (DS), Hyaluronic acid (HA), and Elastin (EL) for its application in skin tissue engineering (TE). Different formulations were designed by combining AC hydrogels with DS, HA, and EL. Cell viability, hemocompatibility, physicochemical, mechanical, and wound healing properties were investigated. Finally, a bilayered hydrogel loaded with fibroblasts and mesenchymal stromal cells was developed using the Ag-Col I-DS-HA-EL (ACDHE) formulation. The ACDHE hydrogel displayed the best in vitro results and acceptable physicochemical properties. Also, it behaved mechanically close to human native skin and exhibited good cytocompatibility. Environmental scanning electron microscopy (ESEM) analysis revealed a porous microstructure that allows the maintenance of cell growth and ECM-like structure production. These findings demonstrate the potential of the ACDHE hydrogel formulation for applications such as an injectable hydrogel or a bioink to create cell-laden structures for skin TE.