RESUMO
OBJECTIVE: The success of engineered tissues continues to be limited by time to vascularization and perfusion. Recently, we described a simple microsurgical approach, termed micropuncture (MP), which could be used to rapidly vascularize an adjacently placed scaffold from the recipient macrovasculature. Here we studied the long-term persistence of the MP-induced microvasculature. METHODS: Segmental 60 µm diameter MPs were created in the recipient rat femoral artery and vein followed by coverage with a simple Type 1 collagen scaffold. The recipient vasculature and scaffold were then wrapped en bloc with a silicone sheet to isolate intrinsic vascularization. Scaffolds were harvested at 28 days post-implantation for detailed analysis, including using a novel artificial intelligence (AI) approach. RESULTS: MP scaffolds demonstrated a sustained increase of vascular density compared to internal non-MP control scaffolds (p < 0.05) secondary to increases in both vessel diameters (p < 0.05) and branch counts (p < 0.05). MP scaffolds also demonstrated statistically significant increases in red blood cell (RBC) perfused lumens. CONCLUSIONS: This study further highlights that the intrinsic MP-induced vasculature continues to persist long-term. Its combination of rapid and stable angiogenesis represents a novel surgical platform for engineered scaffold and graft perfusion.
Assuntos
Inteligência Artificial , Alicerces Teciduais , Animais , Ratos , Punções , Silicones , Engenharia Tecidual , AngiogêneseRESUMO
Tissue loss, irrespective of etiology, often requires extensive reconstruction. In many instances, the need exceeds what current treatments and technologies modern medicine can offer. Tissue engineering has made immense strides within the past few decades due to advances in biologics, biomaterials, and manufacturing. The convergence of these three domains has created limitless potential for future surgical care. Unfortunately, there still exists a disconnect on how to best implant these 'replacement parts' and care for the patient. It is therefore vital to develop paradigms for the integration of advanced surgical and tissue engineering technologies. This paper explores the convergence between tissue engineering and reconstructive surgery. We will describe the clinical problem of tissue loss, discuss currently available solutions, address limitations, and propose processes for integrating surgery and tissue engineering, thereby ushering in the era of regenerative surgery.
Assuntos
Materiais Biocompatíveis , Engenharia Tecidual , Humanos , Materiais Biocompatíveis/uso terapêutico , Alicerces TeciduaisRESUMO
Salmonellosis is one of the most common and widely distributed foodborne diseases. A sensitive and robust detection method of Salmonella Typhimurium (S. Typhimurium) in food can critically prevent a disease outbreak. In this work, the use of phage displayed peptides was explored for the detection of S. Typhimurium. A phage-displayed random dodecapeptide library was subjected to biopanning against lipopolysaccharide (LPS) of S. Typhimurium. The peptide NFMESLPRLGMH (pep49) derived from biopanning displayed a high affinity (25.8nM) for the LPS of S. Typhimurium and low cross-reactivity with other strains of Salmonella and related Gram-negative bacteria. Molecular insights into the interaction of pep49 with the LPS of S. Typhimurium was gleaned using atomistic molecular dynamics simulations and docking. It was deduced that the specificity of pep49 with S. Typhimurium LPS originated from the interactions of pep49 with abequose that is found only in the O-antigen of S. Typhimurium. Further, pep49 was able to detect S. Typhimurium at a LOD of 10(3) CFU/mL using ELISA, and may be a potential cost efficient alternative to antibodies.
Assuntos
Lipopolissacarídeos , Biblioteca de Peptídeos , Salmonella typhimurium , Limite de Detecção , Lipopolissacarídeos/química , Lipopolissacarídeos/metabolismo , Simulação de Acoplamento Molecular , Tipagem Molecular , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Salmonella typhimurium/química , Salmonella typhimurium/isolamento & purificação , Salmonella typhimurium/metabolismoRESUMO
Microfluidic lab-on-a-chip presents an ideal solution for bacterial sensing and identification due to its advantages like large surface-to-volume ratio, requirement of low sample volume and multiplexing possibility. The present work deals with the development of an immunosensor chip using circular microchannels fabricated directly with microdimensional copper wire and permanent magnet for capture of Fe(3)O(4) magnetic nanoparticle (MNP) conjugate. The MNP facilitate capture of the antigen in a confined space and hence, enhanced fluorescence signal for detection. The multiplexed microfluidic chip permits visual detection and quantification of waterborne pathogens viz. Escherichia coli and Salmonella typhimurium simultaneously. CdTe quantum dots (QDs) with different emission wavelengths were conjugated with anti-E. coli and anti-S. typhimurium antibodies for concurrent fluorescence detection. The present technique provides an inexpensive yet powerful tool to image and quantify pathogens at low numbers with passage of large sample volumes.
Assuntos
Escherichia coli/isolamento & purificação , Microfluídica/métodos , Salmonella typhimurium/isolamento & purificação , Microbiologia da Água , Anticorpos Antibacterianos/imunologia , Escherichia coli/imunologia , Nanopartículas Metálicas , Microscopia de Fluorescência , Pontos Quânticos , Salmonella typhimurium/imunologiaRESUMO
Microchannel is basic functional component of microfluidic chip and every step-forward of its construction technique has been receiving concern all over the world. The present work describes a novel, rapid and simple fabrication technique for building 3D microchannels in poly(dimethyl siloxane) (PDMS) elastomer. These microchannels were used for rapid detection of antigens (E. coli) by quantum dot (QD) based approach. Luminescent QD (CdTe) were synthesized by aqueous method and characterized using high resolution transmission electron microscopy (HRTEM), fluorescence spectroscopy and X-ray diffraction (XRD). The QDs were functionalized with anti-E. coli antibodies for immuno-detection. The reported process allowed easier and faster method of fabrication of circular 3D micochannels and demonstrated their potential use in an immuno-biosensor device.