RESUMEN
PURPOSE: Osteoradionecrosis of the jaw is a therapy-resistant condition that may occur after treatment for head and neck cancer. The aim of this study was to investigate the incidence of osteoradionecrosis in patients with oropharyngeal cancer in relation to tooth extraction prior to radiation therapy. METHODS: Patients who had undergone radiation therapy for oropharyngeal cancer 5-10 years earlier were included and evaluated for the development of osteoradionecrosis (n = 75). RESULTS: Among the 75 patients, 62 had molar teeth present in the >50 Gy radiation field and 36 of those patients had teeth extracted prior to radiation therapy. Extraction of molars before radiotherapy significantly increased the risk of developing osteoradionecrosis (P < 0.05). There were no identifiable statistically significant correlations between the time from tooth extraction and the start of radiation therapy, the number of teeth in the radiation field, smoking habits, human papillomavirus-status, gender, age or tumor location and the development of osteoradionecrosis. CONCLUSION: Tooth extraction prior to radiation therapy increases the risk of developing osteoradionecrosis. For patients with good oral hygiene and absence of dental disease, avoidance of tooth extraction in the radiation field could therefore reduce the risk of complications.
Asunto(s)
Neoplasias de Cabeza y Cuello , Neoplasias Orofaríngeas , Osteorradionecrosis , Extracción Dental , Extracción Dental/efectos adversos , Neoplasias Orofaríngeas/complicaciones , Neoplasias Orofaríngeas/radioterapia , Osteorradionecrosis/complicaciones , Osteorradionecrosis/epidemiología , Enfermedades Maxilomandibulares/epidemiología , Enfermedades Maxilomandibulares/etiología , Neoplasias de Cabeza y Cuello/complicacionesRESUMEN
This article describes a woman in her forties who spontaneously developed facial pain 19 years after double-jaw orthognathic surgery. The focus of her pain was the left side of the face, including the temporomandibular joint (TMJ). Conservative treatment was initiated, including several occlusal splints, in addition to injections with local anesthesia, botulinum toxin, and corticosteroids, with limited effects. Surgical treatments with arthroscopy and discectomy, and ultimately a TMJ prosthesis, improved the patient's joint function but did not reduce pain. The question is whether the degenerated joint was due to progression of the original disease process or to multiple surgical procedures.
Asunto(s)
Prótesis Articulares , Trastornos de la Articulación Temporomandibular , Femenino , Humanos , Trastornos de la Articulación Temporomandibular/cirugía , Articulación Temporomandibular , Dolor Facial/etiología , Dolor Facial/cirugíaRESUMEN
A 55 year-old woman presented with an invasive hemangioma of the hard palate. One year after partial surgical excision there was no sign of recurrency or oroantral fistulation. No malignant cells could be identified.
RESUMEN
Resolvin D1 (RvD1) is a pro-resolving lipid mediator of inflammation, endogenously synthesized from omega-3 docosahexaenoic acid. The purpose of this study was to investigate the effect of RvD1 on bone regeneration using a rat calvarial defect model. Collagen 3D nanopore scaffold (COL) and Pluronic F127 hydrogel (F127) incorporated with RvD1 (RvD1-COL-F127 group) or COL and F127 (COL-F127 group) were implanted in symmetrical calvarial defects. After implantation, RvD1 was administrated subcutaneously every 7 days for 4 weeks. The rats were sacrificed at weeks 1 and 8 post-implantation. Tissue samples were analyzed by real-time reverse transcriptase-polymerase chain reaction and histology at week 1. Radiographical and histological analyses were done at week 8. At week 1, calvarial defects treated with RvD1 exhibited decreased numbers of inflammatory cells and tartrate-resistant acid phosphatase (TRAP) positive cells, greater numbers of newly formed blood vessels, upregulated gene expression of vascular endothelial growth factor and alkaline phosphatase, and downregulated gene expression of receptor activator of nuclear factor-κB ligand, interleukin-1ß and tumor necrosis factor-α. At week 8, the radiographical results showed that osteoid area fraction of the RvD1-COL-F127 group was higher than that of the COL-F127 group, and histological examination exhibited enhanced osteoid formation and newly formed blood vessels in the RvD1-COL-F127 group. In conclusion, this study showed that RvD1 enhanced bone formation and vascularization in rat calvarial defects.
Asunto(s)
Ácidos Docosahexaenoicos , Cráneo , Factor A de Crecimiento Endotelial Vascular , Animales , Ratas , Regeneración Ósea , Ácidos Docosahexaenoicos/farmacología , Cráneo/metabolismoRESUMEN
OBJECTIVE: The objective of this study was to compare the outcome of surgical and conservative treatment approaches for medication-related osteonecrosis of the jaw. STUDY DESIGN: Publications in Medline, The Cochrane Library, EMBASE, and PubMed (non-indexed articles) and by Health Technology Assessment organizations were searched. Quality of evidence in primary studies were assessed using Grading of Recommendations Assessment, Development and Evaluation (GRADE) and the level of bias in systematic reviews by a measurement tool to assess systematic reviews (AMSTAR). RESULTS: Quality assessment identified 3 primary studies with moderate GRADE score. Moderate risk of bias was found in 7 systematic reviews and low risk of bias in 3. Nine studies were included in the meta-analysis, where 62.1% healing was reported after surgical treatment (144 of 232 included patients) and 28.8% healing was reported after conservative treatment (38 of 132 included patients). Moderate heterogeneity was found among the included studies (P = .02). The overall odds ratio for resolution of osteonecrosis after surgical versus conservative treatment was 1.25 (95% confidence interval, 0.24-2.26) and was not statistically significant. CONCLUSION: Slightly better outcomes are reported after surgical treatment, in particular for advanced disease stages, but there is a lack of standardized treatment protocols and outcome measures. Overall, the quality of evidence is poor, and the majority of studies have a low evidence certainty rating and high risk of bias.
Asunto(s)
Tratamiento Conservador , Osteonecrosis , Humanos , Cicatrización de HeridasRESUMEN
Injection phobia and dental anxiety can, in severe cases, lead to avoidance of necessary treatment. The aim of this pilot study was to investigate self-reported injection phobia and dental anxiety among individuals with tattoos and/or piercings. The Injection Phobia Scale-Anxiety (IPSA) short form and the Modified Dental Anxiety Scale (MDAS) questionnaires were applied. Both the total IPSA and MDAS scores were significantly higher for individuals with tattoos in comparison with a control group (P < 0.001), suggesting a need for anxiety-reducing measures and facilitated treatment for this group of patients.
Asunto(s)
Trastornos Fóbicos , Tatuaje , Ansiedad al Tratamiento Odontológico/etiología , Humanos , Proyectos Piloto , Autoinforme , Encuestas y Cuestionarios , Tatuaje/efectos adversosRESUMEN
Gelatin has emerged as a biocompatible polymer with high printability in scaffold-based tissue engineering. The aim of the current study was to investigate the potential of genipin-crosslinked 3D printed gelatin scaffolds for temporomandibular joint (TMJ) cartilage regeneration. Crosslinking with genipin increased the stability and mechanical properties, without any cytotoxic effects. Chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSC) on the scaffolds were compared to cell pellets and spheres. Although hBMSC seeded scaffolds showed a lower expression of chondrogenesis-related genes compared to cell pellets and spheres, they demonstrated a significantly reduced expression of collagen (COL) 10, suggesting a decreased hypertrophic tendency. After 21 days, staining with Alcian blue and immunofluorescence for SOX9 and COL1 confirmed the chondrogenic differentiation of hBMSC on genipin-crosslinked gelatin scaffolds. In summary, 3D printed gelatin-genipin scaffolds supported the viability, attachment and chondrogenic differentiation of hBMSC, thus, demonstrating potential for TMJ cartilage regeneration applications.
Asunto(s)
Articulación Temporomandibular , Andamios del Tejido , Cartílago , Gelatina , Humanos , Iridoides , Impresión TridimensionalRESUMEN
Total joint prostheses are a viable treatment option after removal of malignancies invading the temporomandibular joint, even when adjuvant radiation therapy is required.
RESUMEN
OBJECTIVE: In the present study, we assessed the rate of complications and morbidity after mandibular setback with bilateral intraoral vertical ramus osteotomy (IVRO). STUDY DESIGN: In total, 133 patients were included. The prevalence of neurosensory disturbance (NSD), surgical site infection (SSI), and other complications were registered 2 months and 1 year after surgery. The correlations between complications and age, sex, American Society of Anesthesiologists classification, body mass index, blood loss, and operative time were evaluated. RESULTS: NSD was reported for 6.8% of the patients (9 of 133) 2 months after surgery (3.8% of the operated sites). The prevalence was significantly higher in female patients (P < .05). Two patients described persistent unilateral reduced sensibility after 1 year (1.5%). In total, 0.8% of the operated sites (2 of 266) had persistent NSD after 1 year. None of the patients required prolonged hospitalization, and 95.5% (127 of 133) were discharged the day after surgery. None of the patients experienced severe bleeding, and only 1 patient developed SSI. There were no significant correlations between patient-specific or intraoperative parameters evaluated and registered complications. CONCLUSIONS: This study shows that IVRO is a safe surgical technique associated with a low complication rate. IVRO can be an alternative technique for mandibular setback in patients who can tolerate postoperative maxillomandibular fixation.
Asunto(s)
Prognatismo , Cefalometría , Estudios de Cohortes , Femenino , Humanos , Mandíbula , Osteotomía Sagital de Rama Mandibular/efectos adversos , Estudios RetrospectivosRESUMEN
Hormonal changes in the menstrual cycle may cause autoimmune progesterone-induced stomatitis. This case illustrates that insertion of a hormone spiral can be a treatment option to reduce mucosal lesions and symptoms.
Asunto(s)
Granuloma de Cuerpo Extraño/diagnóstico , Enfermedades de los Labios/diagnóstico , Mucosa Bucal/patología , Sarcoidosis/complicaciones , Femenino , Granuloma de Cuerpo Extraño/inmunología , Granuloma de Cuerpo Extraño/patología , Granuloma de Cuerpo Extraño/cirugía , Humanos , Enfermedades de los Labios/inmunología , Enfermedades de los Labios/patología , Enfermedades de los Labios/cirugía , Persona de Mediana Edad , Mucosa Bucal/inmunología , Mucosa Bucal/cirugía , Sarcoidosis/inmunología , Resultado del TratamientoAsunto(s)
Detergentes/toxicidad , Hidróxidos/toxicidad , Enfermedades de los Labios/inducido químicamente , Enfermedades de la Boca/inducido químicamente , Úlceras Bucales/inducido químicamente , Compuestos de Potasio/toxicidad , Enfermedades de la Lengua/inducido químicamente , Adulto , Humanos , Masculino , Ilustración MédicaRESUMEN
Tissue engineering for fibrocartilage regeneration using mesenchymal stromal cells (MSC) and biomaterial scaffolds is emerging as a promising strategy, but inhibiting vascularization to prevent endochondral ossification is important to develop stable implants. The objective of this study was to investigate the effect of angiostatin on inhibition of angiogenesis and promotion of chondrogenesis by collagen scaffolds with or without MSC implanted subcutaneously in rats. One scaffold from the following groups was implanted in each animal: Collagen scaffolds only, scaffolds functionalized with angiostatin, scaffolds loaded with MSC and scaffolds functionalized with angiostatin and loaded with MSC. The various scaffolds were harvested after 2 and 8 weeks for histological analysis, Real-time quantitative polymerase chain reaction (RT-qPCR) and immunofluorescence quantification. Results demonstrated significantly decreased expression of inflammatory (interleukin 1 alpha and beta) and angiogenic genes (platelet and endothelial cell adhesion molecule 1) in scaffolds functionalized with angiostatin after 2 weeks in vivo. Histologically, after 8 weeks, the scaffolds with angiostatin had less inflammatory cells and more collagen matrix formation, but no fibrocartilage formation was detected. Thus, although angiostatin suppressed angiogenesis, it did not stimulate ectopic chondrogenesis in tissue engineered constructs in vivo.
Asunto(s)
Condrogénesis , Células Madre Mesenquimatosas , Angiostatinas , Animales , Colágeno , Ratas , Andamios del TejidoRESUMEN
Two wood-derived cellulose nanofibril (CNF) porous scaffolds were prepared by TEMPO-oxidation and carboxymethylation. The effects of these scaffolds on the production of inflammatory cytokines by human macrophage-like cells (U937) was profiled in vitro after 1 and 3â¯days and in subcutaneous tissues of rats after 4 and 30â¯days, using PCR and Multiplex arrays. Tissue culture plates (TCP) and gelatin scaffolds served as controls in vitro and in vivo respectively. After 3â¯days in vitro, there was no significant difference between the effects of CNF scaffolds and TCP on the production of chemokines/growth factors and pro-inflammatory cytokines. At day 4 in vivo there was significantly higher gene expression of the anti-inflammatory IL-1Ra in the CNF scaffolds than the gelatin scaffold. Production of IL-1ß, IL-6, MCP-1, MIP-1α CXCL-1 and M-CSF was significantly less than in the gelatin, demonstrating an early mild inflammatory response. At day 30, both CNF scaffolds significantly stimulated the production of the anti-inflammatory cytokine IL-10. Unlike gelatin, neither CNF scaffold had degraded 180â¯days post-implantation. The slow degradation of CNF scaffolds resulted in a foreign body reaction, with high production of IL-1ß, IL-2, TNF-α, IFN-Ï, MCP-1, MIP-1α, M-CSF, VEGF cytokines and expression of MMP-9 gene. The surface chemistry of the CNF scaffolds elicited a modest effect on cytokine production and did not shift the inflammatory profile in vitro or in vivo. The decisive role in development of the foreign body reaction was the slow degradation of the CNF scaffolds.
Asunto(s)
Celulosa/química , Inflamación/etiología , Nanoestructuras/efectos adversos , Andamios del Tejido/efectos adversos , Madera/química , Animales , Proliferación Celular , Celulosa/efectos adversos , Citocinas/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Implantes Experimentales , Inflamación/inducido químicamente , Inflamación/metabolismo , Ensayo de Materiales/métodos , Nanoestructuras/química , Ratas Wistar , Andamios del Tejido/química , Células U937RESUMEN
The aim is to evaluate the effect of modifying poly[(l-lactide)-co-(ε-caprolactone)] scaffolds (PLCL) with nanodiamonds (nDP) or with nDP+physisorbed BMP-2 (nDP+BMP-2) on in vivo host tissue response and degradation. The scaffolds are implanted subcutaneously in Balb/c mice and retrieved after 1, 8, and 27 weeks. Molecular weight analysis shows that modified scaffolds degrade faster than the unmodified. Gene analysis at week 1 shows highest expression of proinflammatory markers around nDP scaffolds; although the presence of inflammatory cells and foreign body giant cells is more prominent around the PLCL. Tissue regeneration markers are highly expressed in the nDP+BMP-2 scaffolds at week 8. A fibrous capsule is detectable by week 8, thinnest around nDP scaffolds and at week 27 thickest around PLCL scaffolds. mRNA levels of ALP, COL1α2, and ANGPT1 are significantly upregulating in the nDP+BMP-2 scaffolds at week 1 with ectopic bone seen at week 8. Even when almost 90% of the scaffold is degraded at week 27, nDP are observable at implantation areas without adverse effects. In conclusion, modifying PLCL scaffolds with nDP does not aggravate the host response and physisorbed BMP-2 delivery attenuates inflammation while lowering the dose of BMP-2 to a relatively safe and economical level.
Asunto(s)
Proteína Morfogenética Ósea 2/química , Nanodiamantes/química , Poliésteres/química , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/metabolismo , Angiopoyetina 1/genética , Angiopoyetina 1/metabolismo , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/metabolismo , Materiales Biocompatibles/farmacología , Proteína Morfogenética Ósea 2/metabolismo , Regeneración Ósea/fisiología , Huesos/diagnóstico por imagen , Huesos/fisiología , Quimiocinas/deficiencia , Quimiocinas/metabolismo , Citocinas/genética , Citocinas/metabolismo , Femenino , Inyecciones Subcutáneas , Ratones , Ratones Endogámicos BALB C , Neovascularización Fisiológica , Prótesis e Implantes , Piel/metabolismo , Piel/patología , Regulación hacia Arriba/efectos de los fármacos , Microtomografía por Rayos XRESUMEN
It has been suggested that the effect of implanted cells on the local environment is important when selecting the appropriate cell type for tissue regeneration. Our aim was to compare the local tissue response to implanted human mesenchymal stem cells (MSC) and human umbilical vein endothelial cells (EC). MSC and EC were cultured in poly(L-lactide-co-1,5-dioxepan-2-one) scaffolds for 1 week in a bioreactor system, after which they were implanted subcutaneously in NOD/SCID mice. After 3 weeks, scaffolds were retrieved, and the mRNA expression of selected genes involved in hypoxia and inflammation was examined by real-time reverse transcription polymerase chain reaction and correlated with immunofluorescent staining for corresponding proteins. The Toll-like receptor signaling pathway was examined by superarray hybridization. The expression of 53 angiogenesis-related proteins was investigated by a proteome profiler angiogenesis antibody array kit. Vascularization was quantified using immunohistochemistry for CD31. The expression of hypoxia-inducible factors and biomarkers for angiogenesis was more strongly upregulated in response to implanted EC than to MSC, suggesting a higher sensitivity to low oxygen tension among EC. Hypoxic signaling was increased after implantation of EC compared with MSC, leading to a prolonged acute inflammatory phase that promoted ingrowth of vascular cells and establishment of the circulation. Inflammatory cytokines were also differently expressed at the gene and protein levels in the two experimental groups, resulting in altered recruitment of acute and chronic inflammatory cells. The end result of these differences was increased vessel formation within the constructs in the EC group.
Asunto(s)
Células Endoteliales de la Vena Umbilical Humana , Células Madre Mesenquimatosas , Neovascularización Fisiológica , Andamios del Tejido/química , Animales , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Ratones , Ratones Endogámicos NOD , Ratones SCIDRESUMEN
Constructs intended for bone tissue engineering (TE) are influenced by the initial cell seeding density. Therefore, the objective of this study was to determine the effect of bone marrow stromal stem cells (BMSCs) density loaded onto copolymer scaffolds on bone regeneration. BMSCs were harvested from rat's bone marrow and cultured in media with or without osteogenic supplements. Cells were seeded onto poly(l-lactide-co-ε-caprolactone) [poly(LLA-co-CL)] scaffolds at two different densities: low density (1 × 10(6) cells/scaffold) or high density (2 × 10(6) cells/scaffold) using spinner modified flasks and examined after 1 and 3 weeks. Initial attachment and spread of BMSC onto the scaffolds was recorded by scanning electron microscopy. Cell proliferation was assessed by DNA quantification and cell differentiation by quantitative real-time reverse transcriptase-polymerized chain reaction analysis (qRT-PCR). Five-millimeter rat calvarial defects (24 defects in 12 rats) were implanted with scaffolds seeded with either low or high density expanded with or without osteogenic supplements. Osteogenic supplements significantly increased cell proliferation (p < 0.001). Scaffolds seeded at high cell density exhibited higher mRNA expressions of Runx2 p = 0.001, Col1 p = 0.001, BMP2 p < 0.001, BSP p < 0.001, and OC p = 0.013. More bone was formed in response to high cell seeding density (p = 0.023) and high seeding density with osteogenic medium (p = 0.038). Poly (LLA-co-CL) scaffolds could be appropriate candidates for bone TE. The optimal number of cells to be loaded onto scaffolds is critical for promoting Extracellular matrix synthesis and bone formation. Cell seeding density and osteogenic supplements may have a synergistic effect on the induction of new bone.
Asunto(s)
Regeneración Ósea/efectos de los fármacos , Células Madre Mesenquimatosas/citología , Polímeros/farmacología , Andamios del Tejido/química , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/efectos de los fármacos , Recuento de Células , Proliferación Celular/efectos de los fármacos , Células Cultivadas , ADN/metabolismo , Masculino , Células Madre Mesenquimatosas/efectos de los fármacos , Microscopía Electrónica de Rastreo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas Endogámicas Lew , Reacción en Cadena en Tiempo Real de la Polimerasa , Cráneo/diagnóstico por imagen , Cráneo/patología , Microtomografía por Rayos XRESUMEN
A low dose of 1µg rhBMP-2 was immobilised by four different functionalising techniques on recently developed poly(l-lactide)-co-(ε-caprolactone) [(poly(LLA-co-CL)] scaffolds. It was either (i) physisorbed on unmodified scaffolds [PHY], (ii) physisorbed onto scaffolds modified with nanodiamond particles [nDP-PHY], (iii) covalently linked onto nDPs that were used to modify the scaffolds [nDP-COV] or (iv) encapsulated in microspheres distributed on the scaffolds [MICS]. Release kinetics of BMP-2 from the different scaffolds was quantified using targeted mass spectrometry for up to 70days. PHY scaffolds had an initial burst of release while MICS showed a gradual and sustained increase in release. In contrast, NDP-PHY and nDP-COV scaffolds showed no significant release, although nDP-PHY scaffolds maintained bioactivity of BMP-2. Human mesenchymal stem cells cultured in vitro showed upregulated BMP-2 and osteocalcin gene expression at both week 1 and week 3 in the MICS and nDP-PHY scaffold groups. These groups also demonstrated the highest BMP-2 extracellular protein levels as assessed by ELISA, and mineralization confirmed by Alizarin red. Cells grown on the PHY scaffolds in vitro expressed collagen type 1 alpha 2 early but the scaffold could not sustain rhBMP-2 release to express mineralization. After 4weeks post-implantation using a rat mandible critical-sized defect model, micro-CT and Masson trichrome results showed accelerated bone regeneration in the PHY, nDP-PHY and MICS groups. The results demonstrate that PHY scaffolds may not be desirable for clinical use, since similar osteogenic potential was not seen under both in vitro and in vivo conditions, in contrast to nDP-PHY and MICS groups, where continuous low doses of BMP-2 induced satisfactory bone regeneration in both conditions. The nDP-PHY scaffolds used here in critical-sized bone defects for the first time appear to have promise compared to growth factors adsorbed onto a polymer alone and the short distance effect prevents adverse systemic side effects.
Asunto(s)
Proteína Morfogenética Ósea 2 , Andamios del Tejido , Animales , Proteína Morfogenética Ósea 2/administración & dosificación , Proteína Morfogenética Ósea 2/química , Proteína Morfogenética Ósea 2/genética , Proteína Morfogenética Ósea 2/metabolismo , Regeneración Ósea/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Humanos , Masculino , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Microesferas , Poliésteres/química , Ratas Sprague-Dawley , Transducción de SeñalRESUMEN
INTRODUCTION: Inflammation plays a crucial role in tissue regeneration, wound healing, and the success of tissue-engineered constructs. The aim of this study was to investigate the influence of human umbilical vein endothelial cells (ECs) on leukocyte transmigration when co-cultured with primary human bone marrow-derived multipotent stromal cells (MSCs). METHODS: MSCs with and without ECs were cultured in poly (L-lactide-co-1, 5-dioxepan-2-one) (poly (LLA-co-DXO)) scaffolds for 1 week in vitro in a bioreactor system, after which they were implanted subcutaneously in non-obese diabetic/severe combined immunodeficient mice. After 1 and 3 weeks, scaffolds were retrieved, and the mRNA expression of interleukin 1-beta (IL-1ß), IL-6, IL-10, hypoxia-inducible factor 1-alpha (HIF-1α), HIF-1ß, and mammalian target of rapamycin was examined by real-time reverse transcription-polymerase chain reaction. Furthermore, immunofluorescent staining was performed for IL-1ß, IL-6, neutrophils, and CD11b. In addition, Western blotting was done for IL-1ß and IL-6. Leukocyte transmigration genes and genes in Toll-like receptor pathways, expressed by MSCs cultured in vitro with or without ECs, were further investigated with a microarray dataset. RESULTS: In vitro, genes involved in leukocyte transmigration and Toll-like receptor pathways were clearly influenced by the addition of ECs. Platelet/endothelial cell adhesion molecule-1 (PECAM-1) and cadherin-5 (CDH5), both genes involved in leukocyte transmigration, were expressed significantly higher in the MSC/EC group. CONCLUSIONS: The recruitment of leukocytes into tissue-engineered constructs with MSCs is strongly influenced by the addition of ECs via activation of leukocyte transmigration and Toll-like receptor pathways.
Asunto(s)
Células Endoteliales de la Vena Umbilical Humana/metabolismo , Leucocitos/metabolismo , Células Madre Mesenquimatosas/metabolismo , Receptores Toll-Like/metabolismo , Migración Transendotelial y Transepitelial , Animales , Translocador Nuclear del Receptor de Aril Hidrocarburo/genética , Translocador Nuclear del Receptor de Aril Hidrocarburo/metabolismo , Cadherinas/genética , Cadherinas/metabolismo , Técnicas de Cocultivo , Medios de Cultivo Condicionados/farmacología , Células Endoteliales de la Vena Umbilical Humana/citología , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Interleucinas/genética , Interleucinas/metabolismo , Leucocitos/efectos de los fármacos , Leucocitos/fisiología , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos NOD , Ratones SCID , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/genética , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ingeniería de Tejidos , Andamios del Tejido , Receptores Toll-Like/genéticaRESUMEN
INTRODUCTION: Rapid establishment of functional blood vessels is a prerequisite for successful tissue engineering. During vascular development, endothelial cells (ECs) and perivascular cells assemble into a complex regulating proliferation of ECs, vessel diameter and production of extracellular matrix proteins. The aim of this study was to evaluate the ability of mesenchymal stem cells (MSCs) to establish an endothelial-perivascular complex in tissue-engineered constructs comprising ECs and MSCs. METHODS: Primary human ECs and MSCs were seeded onto poly(L-lactide-co-1,5-dioxepan-2-one) (poly(LLA-co-DXO)) scaffolds and grown in dynamic culture before subcutaneous implantation in immunocompromised mice for 1 and 3 weeks. Cellular activity, angiogenic stimulation and vascular assembly in cell/scaffold constructs seeded with ECs or ECs/MSCs in a 5:1 ratio was monitored with real-time RT-PCR, ELISA and immunohistochemical microscopy analysis. RESULTS: A quiescent phenotype of ECs was generated, by adding MSCs to the culture system. Decreased proliferation of ECs, in addition to up-regulation of selected markers for vascular maturation was demonstrated. Baseline expression of VEGFa was higher for MSCs compared with EC (P<0.001), with subsequent up-regulated VEGFa-expression for EC/MSC constructs before (P<0.05) and after implantation (P<0.01). Furthermore, an inflammatory response with CD11b+cells was generated from implantation of human cells. At the end of the 3 week experimental period, a higher vascular density was shown for both cellular constructs compared with empty control scaffolds (P<0.01), with the highest density of capillaries being generated in constructs comprising both ECs and MSCs. CONCLUSIONS: Induction of a quiescent phenotype of ECs associated with vascular maturation can be achieved by co-seeding with MSCs. Hence, MSCs can be appropriate perivascular cells for tissue-engineered constructs.