Analysis and monitoring of single HaCaT cells using volumetric Raman mapping and machine learning

No explorer reached a pole without a map, no chef served a meal without tasting, and no surgeon implants untested devices. Higher accuracy maps, more sensitive taste buds, and more rigorous tests increase confidence in positive outcomes. Biomedical manufacturing necessitates rigour, whether developing drugs or creating bioengineered tissues [1]–[4]. By designing a dynamic environment that supports mammalian cells during experiments within a Raman spectroscope, this project provides a platform that more closely replicates in vivo conditions. The platform also adds the opportunity to automate the adaptation of the cell culture environment, alongside spectral monitoring of cells with machine learning and three-dimensional Raman mapping, called volumetric Raman mapping (VRM). Previous research highlighted key areas for refinement, like a structured approach for shading Raman maps [5], [6], and the collection of VRM [7]. Refining VRM shading and collection was the initial focus, k-means directed shading for vibrational spectroscopy map shading was developed in Chapter 3 and exploration of depth distortion and VRM calibration (Chapter 4). “Cage” scaffolds, designed using the findings from Chapter 4 were then utilised to influence cell behaviour by varying the number of cage beams to change the scaffold porosity. Altering the porosity facilitated spectroscopy investigation into previously observed changes in cell biology alteration in response to porous scaffolds [8]. VRM visualised changed single human keratinocyte (HaCaT) cell morphology, providing a complementary technique for machine learning classification. Increased technical rigour justified progression onto in-situ flow chamber for Raman spectroscopy development in Chapter 6, using a Psoriasis (dithranol-HaCaT) model on unfixed cells. K-means-directed shading and principal component analysis (PCA) revealed HaCaT cell adaptations aligning with previous publications [5] and earlier thesis sections. The k-means-directed Raman maps and PCA score plots verified the drug-supplying capacity of the flow chamber, justifying future investigation into VRM and machine learning for monitoring single cells within the flow chamber

The use of vibrational spectroscopy to study the pathogenesis multiple sclerosis and other neurological conditions

Spectroscopy techniques are valuable tools in biomedical research and have been used extensively in the study of disease. However, neurological conditions such as multiple sclerosis (MS) have received little attention and the available spectroscopy studies are limited, both in overall numbers of patients studied and the disease samples considered. MS is a complex immune-mediated disease, with variable clinical courses and limited therapeutic options. This review aims to summarize current literature in the area, demonstrating how spectroscopy techniques can provide valuable information to inform and advance research into the most common neurological condition affecting young adults

Advances of Proteomic Sciences in Dentistry

Applications of proteomics tools revolutionized various biomedical disciplines such as genetics, molecular biology, medicine, and dentistry. The aim of this review is to highlight the major milestones in proteomics in dentistry during the last fifteen years. Human oral cavity contains hard and soft tissues and various biofluids including saliva and crevicular fluid. Proteomics has brought revolution in dentistry by helping in the early diagnosis of various diseases identified by the detection of numerous biomarkers present in the oral fluids. This paper covers the role of proteomics tools for the analysis of oral tissues. In addition, dental materials proteomics and their future directions are discussed

Generation of Pearl/Calcium Phosphate Composite Particles and Their Integration into Porous Chitosan Scaffolds for Bone Regeneration

Bone tissue engineering using osteoconductive scaffolds holds promise for regeneration, with pearl powder gaining interest for its bioactive qualities. This study used freeze drying to create chitosan (CS) scaffolds with pearl/calcium phosphate (p/CaP) powders, mimicking bone tissue structurally and compositionally. Characterization included scanning electron microscopy (SEM) and mechanical testing. X-ray diffraction (XRD) Fourier-transform infrared–photoacoustic photo-acoustic sampling (FTIR−PAS), and FTIR- attenuated total reflectance (FTIR-ATR) were used to characterize p/CaP. In vitro tests covered degradation, cell activity, and SEM analysis. The scaffolds showed notable compressive strength and modulus enhancements with increasing p/CaP content. Porosity, ranging from 60% to 90%, decreased significantly at higher pearl/CaP ratios. Optimal cell proliferation and differentiation were observed with scaffolds containing up to 30 wt.% p/CaP, with 30 wt.% pearl powder and 30 wt.% p/CaP yielding the best results. In conclusion, pearl/calcium phosphate chitosan (p/CaP_CS) composite scaffolds emerged as promising biomaterials for bone tissue engineering, combining structural mimicry and favourable biological responses

Evaluation of the Effect of Nano and Micro Hydroxyapatite Particles on the Impact Strength of Acrylic Resin: In Vitro Study

Poly (methylmethacrylate) is considered the basis material for a denture base. However, such substance has some drawbacks such as poor impact resistance, which is thought to be the primary cause of fracture of denture base resins. The purpose of the study was to determine how Nano and Micro hydroxyapatite particles affected the impact strength of acrylic resin. Thirty specimens were constructed of heat-cured acrylic resin and were divided into three groups: Ten specimens for the control, 10 for 1%nano hydroxyapatite, and 10 for micro hydroxyapatite. Acrylic samples were subjected to an impact strength test via a Charpy-type. Data were then analysed using SPSS v20. The ANOVA test was used for comparison among the groups. Highly statistically significant differences among all studied groups (P-value <0.0001). Both 5% Micro hydroxyapatite and 1% Nano hydroxyapatite had a higher mean value than the control. Incorporating Nano and Micro hydroxyapatite into PMMA improved the impact strength of acrylic resins

Developing A Synthetic Composite Membrane For Cleft Palate Repair

An oronasal fistula is a passage between the oral and nasal cavity. Currently, surgical procedures use mucosal flaps or collagen grafts to make a barrier between oral and nasal cavities. Our aim was to develop a cell-free synthetic repair material for closure of nasal fistulas. We surface functionalized electrospun polyurethane (PU) and poly-L-lactic acid (PLLA) and composite polymer (PU-PLLA) membranes with acrylic acid through plasma polymerization. Membranes were treated in a layer-by-layer approach to develop highly charged electrostatic layer that could bind heparin as a pro-angiogenic glycosaminoglycan. The properties were evaluated through physical, chemical, and mechanical characterization techniques. Cytotoxicity was tested with MC3T3 pre-osteoblast cell lines for 3, 7, and 14 days, and vasculogenesis was assessed by implantation into the chorio-allantoic membrane in chick embryos for 7 days. In vivo biocompatibility was assessed by subcutaneous implantation in rats for 1, 3, and 6 weeks. The membranes consisted of random fibers of PLLA-PU with fiber diameters of 0.47 and 0.12 μm, respectively. Significantly higher cell proliferation and migration of MC3T3 cells at 3, 7, and 14 days were shown on plasma-coated membranes compared with uncoated membranes. Further, it was found that plasma-coated membranes were more angiogenic than controls. In vivo implantation of membranes in rats did not reveal any gross toxicity to the materials, and wound healing was comparable with the native tissue repair (sham group). We therefore present a plasma-functionalized electrospun composite polymer membrane for use in the treatment of fistulas. These membranes are flexible, non-cytotoxic, and angiogenic, and we hope it should lead to permanent closure of oronasal fistula

An Experimental Channel Capacity Analysis of Cooperative Networks Using Universal Software Radio Peripheral (USRP)

Cooperative communication (CC) is one of the best solutions to overcome channel fading and to improve channel capacity. However, most of the researchers evaluate its performance based on mathematical modeling or by simulations. These approaches are often unable to successfully capture many real-world radio signal propagation problems. Hardware based wireless communication test-bed provides reliable and accurate measurements, which are not attainable through other means. This research work investigates experimental performance analysis of CC over direct communication (DC) in the lab environment. The experimental setup is built using Universal Software Radio Peripheral (USRP) and Laboratory Virtual Instrument Engineering Workbench (LabVIEW). A text message is transmitted by using Phase Shift Keying (PSK) modulation schemes. The setup uses amplify and forward (AF) relaying mode and two time slot transmission protocols. The maximum ratio combining (MRC) technique is used for combining SNR at the receiver. Channel capacity analysis is performed in order to evaluate the performance of CC over DC with and without obstacle. Moreover, optimal position of the relay is also analyzed by varying the position of the relay. Extensive experiments are carried out in the lab environment to evaluate the performance of the system for different hardware setups. The results reveal that cooperative communication attains significant improvement in terms of channel capacity of the system

Ceramic Stereolithography of Bioactive Glasses: Influence of Resin Composition on Curing Behavior and Green Body Properties

Herein we report on the preparation of a bioactive glass (BAG)-based photocurable resin for the additive manufacturing of BAG scaffolds with high filler loadings. The preparation of glass/ceramics resins for stereolithography with high filler loading is always a challenge, especially for fillers with a high refractive index variance. Various photocurable resin compositions with and without bioactive glass fillers have been investigated to see the influence of bioactive glass on physical properties of the resin and resulting green body. The effect of concentration of monomers, reactive diluent, light absorber (Sudan orange G dye), photoinitiator (PI), non-reactive diluent, and fillers (BAG) on rheology and photocuring behavior of the resin and tomography of the resulting 3D structures have been investigated. The BAG contents affect the rheology of resin and influence the rate of the polymerization reaction. The resin compositions with 55–60% BAG, 10% PEG-200 (diluent), 1% of PI and 0.015% of the dye were found to be suitable compositions for the stereolithographic fabrication. A higher percentage of PI caused over-curing, while a higher amount of dye decreased the cure depth of the resin. The micro-computed tomography (µ-CT) and scanning electron microscopic (SEM) images of the resulting green bodies display a relatively dense glass scaffold without any visible cracks and good interlayer connection and surface finishing. These properties play an important role in the mechanical behavior of 3D scaffolds. This study will be helpful to prepare high density glass/ceramic slurries and optimize their printing properties