RESUMEN
Cell adhesion to the extracellular matrix and its natural outcome of cell spreading, along with the maintenance of barrier activity, are essential behaviors of epithelial cells, including retinal pigment epithelium (RPE). Disruptions in these characteristics can result in severe vision-threatening diseases such as diabetic macular edema and age-related macular degeneration. However, the precise mechanisms underlying how RPE cells regulate their barrier integrity and cell spreading are not fully understood. This study aims to elucidate the relative importance of upper glycolytic components in governing these cellular behaviors of RPE cells. Electric Cell-Substrate Impedance Sensing (ECIS) technology was utilized to assess in real-time the effects of targeting various upper glycolytic enzymes on RPE barrier function and cell spreading by measuring cell resistance and capacitance, respectively. Specific inhibitors used included WZB117 for Glut1 inhibition, Lonidamine for Hexokinase inhibition, PFK158 for PFKFB3/PFK axis inhibition, and TDZD-8 for Aldolase inhibition. Additionally, the viability of RPE cells was evaluated using a lactate dehydrogenase (LDH) cytotoxicity assay. The most significant decrease in electrical resistance and increase in capacitance of RPE cells were observed due to dose-dependent inhibition of Glut1 using WZB117, as well as Aldolase inhibition with TDZD-8. LDH level analysis at 24-72 h post-treatment with WZB117 (1 and 10 µM) or TDZD-8 (1 µM) showed no significant difference compared to the control, indicating that the disruption of RPE functionality was not attributed to cell death. Lastly, inhibition of other upper glycolytic components, including PFKFB3/PFK with PFK158 or Hexokinase with Lonidamine, did not significantly affect RPE cell behavior. This study provides insights into the varied roles of upper glycolytic components in regulating the functionality of RPE cells. Specifically, it highlights the critical roles of Glut1 and Aldolase in preserving barrier integrity and promoting RPE cell adhesion and spreading. Such understanding will guide the development of safe interventions to treat RPE cell dysfunction in various retinal disorders.
Asunto(s)
Glucólisis , Epitelio Pigmentado de la Retina , Epitelio Pigmentado de la Retina/metabolismo , Epitelio Pigmentado de la Retina/efectos de los fármacos , Epitelio Pigmentado de la Retina/citología , Glucólisis/efectos de los fármacos , Humanos , Transportador de Glucosa de Tipo 1/metabolismo , Hexoquinasa/metabolismo , Línea Celular , Supervivencia Celular/efectos de los fármacos , Células Epiteliales/metabolismo , Células Epiteliales/efectos de los fármacos , Adhesión Celular/efectos de los fármacos , Impedancia Eléctrica , Fosfofructoquinasa-2/metabolismo , Fosfofructoquinasa-2/antagonistas & inhibidoresRESUMEN
BACKGROUND: This review seeks to investigate the clinically relevant bone graft materials in single-level transforaminal lumbar interbody fusion (TLIF) procedures as defined by (1) primary outcomes (ie, fusion rates and complication rates) and (2) patient-reported outcomes (ie, visual analog scale [VAS] and Oswestry disability index [ODI]). Because of the advantages in stimulating bone growth, autologous bone grafts such as the iliac crest bone graft (ICBG) have been the gold standard. Numerous alternatives to ICBG have been introduced. Understanding the risks and benefits of bone graft options is vital to optimizing patient care. METHODS: A PubMed search was performed for all clinical studies published between January 2008 and March 2023 that referenced the single-level TLIF procedure as well as one of the following grafts: autograft, allograft, bone morphogenetic protein (BMP), demineralized bone matrix, or mesenchymal stem cells (MSCs). Case studies and reports were excluded. RESULTS: Twenty-eight studies met the inclusion criteria. Studies from the PubMed search demonstrated similarly high fusion rates across nearly all graft materials, the lone exception being MSCs, which showed lower fusion rates. ICBG grafts experienced higher rates of postoperative graft site pain. The BMP graft material had high rates of radiculitis, heterogeneous ossification, and vertebral osteolysis. Patients saw an overall improvement in VAS and ODI scores with all graft materials. CONCLUSION: Local autografts and ICBG have been the most studied. Fusion rates during single-level TLIF were similar across all graft materials except MSCs. Patient-reported pain levels improved after TLIF surgery regardless of the type of grafts used. While BMP implants have shown promising benefits, they have introduced a new array of complications not normally seen in ICBG implants. The study is limited by the lack of evidence of certain graft materials as well as nonuniformity in metrics evaluating the efficacy of graft materials.
RESUMEN
BACKGROUND: Retinal degenerative diseases such as diabetic retinopathy and diabetic macular edema are characterized by impaired retinal endothelial cells (RECs) functionality. While the role of glycolysis in glucose homeostasis is well-established, its contributions to REC barrier assembly and cell spreading remain poorly understood. This study aimed to investigate the importance of upper glycolytic components in regulating the behavior of human RECs (HRECs). METHODS: Electric cell-substrate impedance sensing (ECIS) technology was employed to analyze the real-time impact of various upper glycolytic components on maintaining barrier functionality and cell spreading of HRECs by measuring cell resistance and capacitance, respectively. Specific inhibitors were used: WZB117 to inhibit Glut1/3, lonidamine to inhibit hexokinases, PFK158 to inhibit the PFKFB3-PFK axis, and TDZD-8 to inhibit aldolases. Additionally, the viability of HRECs was evaluated using the lactate dehydrogenase (LDH) cytotoxicity assay. RESULTS: The most significant reduction in electrical resistance and increase in capacitance of HRECs resulted from the dose-dependent inhibition of PFKFB3/PFK using PFK158, followed by aldolase inhibition using TDZD-8. LDH level analysis at 24- and 48-hours post-treatment with PFK158 (1 µM) or TDZD-8 (1 and 10 µM) showed no significant difference compared to the control, indicating that the disruption of HRECs functionality was not attributed to cell death. Conversely, inhibiting Glut1/3 with WZB117 had minimal impact on HREC behavior, except at higher concentrations (10 µM) and prolonged exposure. Lastly, inhibiting hexokinase with lonidamine did not noticeably alter HREC cell behavior. CONCLUSION: This study illustrates the unique impacts of components within upper glycolysis on HREC functionality, emphasizing the crucial role of the PFKFB3/PFK axis in regulating HREC behavior. Understanding the specific contributions of each glycolytic component in preserving normal REC functionality will facilitate the development of targeted interventions for treating endothelial cell dysfunction in retinal disorders while minimizing effects on healthy cells.
Asunto(s)
Retinopatía Diabética , Edema Macular , Humanos , Retinopatía Diabética/metabolismo , Células Endoteliales/metabolismo , Transportador de Glucosa de Tipo 1/metabolismo , Edema Macular/metabolismo , Retina/metabolismo , Glucosa/farmacología , Glucosa/metabolismoRESUMEN
PURPOSE: With the recent increase in popularity of electronic cigarette use in the United States, its harmful effects are not only limited to smoke inhalation, but also to the possibility of e-cigarette device malfunction. The purpose of this review is to characterize oromaxillofacial trauma secondary to electronic cigarette device explosion. METHODS: For this systematic review, PubMed and Embase were searched in October 2019 using the following search terms: e-cigarette burns, e-cigarette injury, and e-cigarette explosions, which yielded 400 studies. Basic science research, animal studies, non-English studies, and reports of non-oromaxillofacial injuries were excluded. Study subject demographics, mechanism of trauma, injury type, treatment, and sequelae were recorded and analyzed. RESULTS: Of all studies, 20 studies met inclusion criteria, including 14 case reports and 6 case series, with a total of 21 study subjects. For cases that reported sex, 100% were male (20) with a mean age of 29.5 years. Most common lacerations and/or burns involved the lips (10/21), tongue (8/21), soft palate and/or hard palate (4/21), and nose (5/21). Thirteen subjects underwent surgeries including oral-maxillofacial surgery or dental implants (7/13), bone graft repair (3/13), open reduction and internal fixation for preservation of sinus outflow tracts (2/13), foreign body removal from the cervical spine (1/13), and iridectomy (1/13). Reported complications included bone loss secondary to traumatic fracture, tinnitus and hearing loss, lip paralysis secondary to persistent edema, major depressive disorder/ post-traumatic stress disorder, persistent sinusitis, photophobia, and bilateral axillary and hand contractures. CONCLUSIONS: Electronic cigarette device malfunction and explosion carries great risk for acute oromaxillofacial trauma that may be disfiguring. With the increasing popularity of electronic cigarette use, clinicians and patients should be advised regarding dangers of electronic cigarette use.