RESUMO
BACKGROUND: The conditioned medium from human dermal fibroblasts (dermal fibroblast-conditioned medium; DFCM) contains a diverse array of secretory proteins, including growth factors and wound repair-promoting proteins. Angiogenesis, a crucial process that facilitates the infiltration of inflammatory cells during wound repair, is induced by a hypoxic environment and inflammatory cytokines. METHODS: In this study, we conducted a comprehensive bioinformatic analysis of 337 proteins identified through proteomics analysis of DFCM. We specifically focused on 64 DFCM proteins with potential involvement in angiogenesis. These proteins were further classified based on their characteristics, and we conducted a detailed analysis of their protein-protein interactions. RESULTS: Gene Ontology protein classification categorized these 64 DFCM proteins into various classes, including metabolite interconversion enzymes (N = 11), protein modifying enzymes (N = 10), protein-binding activity modulators (N = 9), cell adhesion molecules (N = 6), extracellular matrix proteins (N = 6), transfer/carrier proteins (N = 3), calcium-binding proteins (N = 2), chaperones (N = 2), cytoskeletal proteins (N = 2), RNA metabolism proteins (N = 1), intercellular signal molecules (N = 1), transporters (N = 1), scaffold/adaptor proteins (N = 1), and unclassified proteins (N = 9). Furthermore, our protein-protein interaction network analysis of DFCM proteins revealed two distinct networks: one with medium confidence level interaction scores, consisting of 60 proteins with significant connections, and another at a high confidence level, comprising 52 proteins with significant interactions. CONCLUSIONS: Our bioinformatic analysis highlights the presence of a multitude of secretory proteins in DFCM that form significant protein-protein interaction networks crucial for regulating angiogenesis. These findings underscore the critical roles played by DFCM proteins in various stages of angiogenesis during the wound repair process.
Assuntos
Angiogênese , Pele , Humanos , Meios de Cultivo Condicionados/farmacologia , Cicatrização , Biologia ComputacionalRESUMO
BACKGROUND: Human dermal fibroblasts secrete diverse proteins that regulate wound repair and tissue regeneration. METHODS: In this study, dermal fibroblast-conditioned medium (DFCM) proteins potentially regulating nerve restoration were bioinformatically selected among the 337 protein lists identified by quantitative liquid chromatography-tandem mass spectrometry. Using these proteins, protein-protein interaction network analysis was conducted. In addition, the roles of DFCM proteins were reviewed according to their protein classifications. RESULTS: Gene Ontology protein classification categorized these 57 DFCM proteins into various classes, including protein-binding activity modulator (N = 11), cytoskeletal protein (N = 8), extracellular matrix protein (N = 6), metabolite interconversion enzyme (N = 5), chaperone (N = 4), scaffold/adapter protein (N = 4), calcium-binding protein (N = 3), cell adhesion molecule (N = 2), intercellular signal molecule (N = 2), protein modifying enzyme (N = 2), transfer/carrier protein (N = 2), membrane traffic protein (N = 1), translational protein (N = 1), and unclassified proteins (N = 6). Further protein-protein interaction network analysis of 57 proteins revealed significant interactions among the proteins that varied according to the settings of confidence score. CONCLUSIONS: Our bioinformatic analysis demonstrated that DFCM contains many secretory proteins that form significant protein-protein interaction networks crucial for regulating nerve restoration. These findings underscore DFCM proteins' critical roles in various nerve restoration stages during the wound repair process.
Assuntos
Biologia Computacional , Fibroblastos , Regeneração Nervosa , Mapas de Interação de Proteínas , Humanos , Fibroblastos/metabolismo , Regeneração Nervosa/fisiologia , Mapas de Interação de Proteínas/fisiologia , Meios de Cultivo Condicionados , Cicatrização/fisiologia , Células Cultivadas , Espectrometria de Massas em Tandem , Derme/citologia , Derme/metabolismoRESUMO
Background: Skin cooling during laser or radiofrequency (RF) treatments is a method to minimize thermal damage to the epidermis, reduce pain, and decrease post-treatment downtime. We evaluated the effect of parallel contact cooling (PCC) on RF-induced thermal reactions in minipig skin in vivo after bipolar microneedling RF treatment. Methods: RF treatments were administered at frequencies of 0.5, 1, and 2 MHz with single (500 ms), six (1000 ms), and ten (5000 ms) sub-pulse packs to minipig skin with or without PCC. Subsequently, thermometric imaging and histology were used to analyze skin reactions to RF. Results: Thermometric images showed that PCC promptly lowered skin temperature in the RF-treated area, with this effect persisting for over 60s. Regardless of the PCC, RF treatments lasting for 500 ms with a single pulse pack resulted in peri-electrode coagulative necrosis (PECN) zones and inter-electrode non-necrotic thermal reaction (IENT) zones in the dermis. In contrast, treatment lasting 5000 ms with 10 sub-pulse packs produced distinct IENT without notable PECN over a wide dermal area. Skin specimens obtained at 1 h and 3, 7, and 14 days after PCC-assisted RF treatments showed a higher degree of thermal tissue reactions in the deeper dermal regions compared to those after RF treatments without PCC. Conclusion: PCC-assisted RF treatment, utilizing an invasive bipolar microneedling device, enhanced RF-induced skin reactions in the mid to deep dermis while preserving the epidermis and upper papillary dermis from excessive thermal tissue injury.