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1.
ERJ Open Res ; 7(3)2021 Jul.
Article in English | MEDLINE | ID: mdl-34476245

ABSTRACT

RATIONALE: COPD and smoking are characterised by pulmonary inflammation. 18F-fluorodeoxyglucose positron emission tomography/computed tomography (FDG PET/CT) imaging may improve knowledge of pulmonary inflammation in COPD patients and aid early development of novel therapies as an imaging biomarker. OBJECTIVES: To evaluate pulmonary inflammation, assessed by FDG uptake, in whole and regional lung in "usual" (smoking-related) COPD patients, alpha-1 antitrypsin deficiency (α1ATD) COPD patients, smokers without COPD and never-smokers using FDG PET/CT. Secondly, to explore cross-sectional associations between FDG PET/CT and systemic inflammatory markers in COPD patients and repeatability of the technique in COPD patients. METHODS: Data from two imaging studies were evaluated. Pulmonary FDG uptake (normalised Ki; nKi) was measured by Patlak graphical analysis in four subject groups: 84 COPD patients, 11 α1ATD-COPD patients, 12 smokers and 10 never-smokers. Within the COPD group, associations between nKi and systemic markers of inflammation were assessed. Repeatability was evaluated in 32 COPD patients comparing nKi values at baseline and at 4-month follow-up. RESULTS: COPD patients, α1ATD-COPD patients and smokers had increased whole lung FDG uptake (nKi) compared with never-smokers (0.0037±0.001, 0.0040±0.001, 0.0040±0.001 versus 0.0028±0.001 mL·cm-3·min-1, respectively, p<0.05 for all). Similar results were observed in upper and middle lung regions. In COPD participants, plasma fibrinogen was associated with whole lung nKi (ß=0.30, p=0.02) in multivariate analysis adjusted for current smoking, forced expiratory volume in 1 s % predicted, systemic neutrophils and C-reactive protein levels. Mean percentage difference in nKi between the baseline and follow-up was 3.2%, and the within subject coefficient of variability was 7.7%. CONCLUSIONS: FDG PET/CT has potential as a noninvasive tool to enable whole lung and regional quantification of FDG uptake to assess smoking- and COPD-related pulmonary inflammation.

2.
J Tissue Eng Regen Med ; 9(11): E27-38, 2015 Nov.
Article in English | MEDLINE | ID: mdl-23576328

ABSTRACT

Despite extensive research into cartilage tissue engineering (CTE), there is still no scaffold ideal for clinical applications. Various synthetic and natural polymers have been investigated in vitro and in vivo, but none have reached widespread clinical use. The authors investigate the potential of POSS-PCU, a synthetic nanocomposite polymer, for use in CTE. POSS-PCU is modified with silsesquioxane nanostructures that improve its biological and physical properties. The ability of POSS-PCU to support the growth of ovine nasoseptal chondrocytes was evaluated against a polymer widely used in CTE, polycaprolactone (PCL). Scaffolds with varied concentrations of the POSS molecule were also synthesized to investigate their effect on chondrocyte growth. Chondrocytes were seeded onto scaffold disks (PCU negative control; POSS-PCU 2%, 4%, 6%, 8%; PCL). Cytocompatibilty was evaluated using cell viability, total DNA, collagen and GAG assays. Chondrocytes cultured on POSS-PCU (2% POSS) scaffolds had significantly higher viability than PCL scaffolds (p < 0.001). Total DNA, collagen and sGAG protein were also greater on POSS-PCU scaffolds compared with PCL (p > 0.05). POSS-PCU (6% and 8% POSS) had improved viability and proliferation over an 18 day culture period compared with 2% and 4% POSS-PCU (p < 0.0001). Increasing the percentage of POSS in the scaffolds increased the size of the pores found in the scaffolds (p < 0.05). POSS-PCU has excellent potential for use in CTE. It supports the growth of chondrocytes in vitro and the POSS modification significantly enhances the growth and proliferation of nasoseptal chondrocytes compared with traditional scaffolds such as PCL.


Subject(s)
Cartilage/physiology , Nanocomposites/chemistry , Nanostructures/chemistry , Organosilicon Compounds/chemistry , Tissue Engineering/methods , Animals , Cartilage/metabolism , Cell Adhesion , Cell Proliferation , Cell Survival , Chondrocytes/cytology , Chondrocytes/metabolism , Collagen/chemistry , DNA/chemistry , Humans , L-Lactate Dehydrogenase/chemistry , Microscopy, Electron, Scanning , Nucleic Acids/chemistry , Polycarboxylate Cement/chemistry , Polymers/chemistry , Porosity , Regeneration , Sheep , Tissue Scaffolds/chemistry , Urethane/chemistry
3.
Methods Mol Biol ; 1001: 145-52, 2013.
Article in English | MEDLINE | ID: mdl-23494426

ABSTRACT

Autologous hydrogel manufacture is an exciting technique within the field of regenerative medicine. Fibrin is a protein with many biocompatible and regenerative features. The ability to generate fibrin scaffolds with the necessary matrix topography for cell integration, from a patient's autologous tissue, could improve the translation of many tissue engineering efforts from bench to clinical application. Here we describe the rapid extraction and production of fibrin hydrogels for development of organs, using a simple low-cost series of centrifugations and ethanol precipitation, which produces hydrogels of a more predictable amount and morphology.


Subject(s)
Fibrin/chemistry , Hydrogels/chemical synthesis , Regeneration/physiology , Regenerative Medicine/methods , Tissue Scaffolds/chemistry , Centrifugation , Chemical Precipitation , Ethanol , Humans
4.
Ann Plast Surg ; 71(2): 238-44, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23429216

ABSTRACT

Currently, the gold standard for reconstruction after rhinectomy or severe trauma to the nose, includes transposition of autologous mucosal flaps plus autologous cartilage grating and coverage using a skin flap. Difficulties with this approach arise where; cartilage and mucosa harvested from autologous donor sites is insufficient to achieve a passable aesthetic and functional reconstruction. Skin flaps are often bulky, poor color matches with hair follicles that reduce the aesthetic quality of the reconstruction. We suggest that tissue engineering could be a source of functional replacement tissues for nasal reconstructive surgery. However, the advancement of such an approach is dependent on the dissemination of scientific information into the clinical community, regarding the engineering of tissues such as mucosa, skin, and cartilage. This paper therefore reviews how the tissue engineering strategies available for producing clinically viable tissues could help resolve issues around reconstructing the human nose.


Subject(s)
Guided Tissue Regeneration/methods , Rhinoplasty/methods , Tissue Engineering/methods , Guided Tissue Regeneration/instrumentation , Humans , Nasal Cartilages/surgery , Nasal Mucosa/surgery , Organ Culture Techniques , Tissue Scaffolds
5.
J Surg Res ; 181(1): 41-8, 2013 May 01.
Article in English | MEDLINE | ID: mdl-22819310

ABSTRACT

BACKGROUND: Advancements in cartilage tissue engineering have the potential to ameliorate facial and joint reconstructive surgery as we know it. The translation of in vitro models of cartilage regeneration into clinical scenarios is the next phase of cartilage tissue engineering research. To engineer larger, more robust, and clinical relevant constructs, a great number of viable chondrocytic cells are needed. However, there is a paucity of literature concerning the most favorable method of chondrocyte isolation. Isolation methods are inconsistent, resulting in small yields and poor cell quality, and thus unreliable neocartilage formation. This study aimed to optimize the chondrocyte isolation protocol to give a maximum yield with optimal cell viability for the engineering of large cartilaginous constructs such as the human nose and ear. METHODS: We employed several enzymes (pronase, dispase, hyaluronidase, and collagenase), enzyme concentrations, and digest lengths to digest freshly harvested ovine nasoseptal cartilage. We used automated trypan blue live/dead staining, immunofluorescent labeling of CD44, collagenase II, collagenase I, and Aggrecan, and alamarBlue to assess cell yield and viability. RESULTS: Incubation length in enzymatic solutions had the greatest effect on cell viability, whereas concentrations of enzymes had a lesser effect. Isolated cells maintained their expression of chondrocyte-specific cell surface markers. CONCLUSIONS: The optimum incubation period was 10 h using collagenase at a 0.2% (w/v) solution. An average of 1-1.5 × 10(6) cells could be harvested per gram of cartilage using this method.


Subject(s)
Cell Separation/methods , Chondrocytes/cytology , Tissue Engineering/methods , Animals , Cell Count , Chondrocytes/chemistry , Collagenases/metabolism , Fluorescent Antibody Technique , Humans , Male , Sheep
6.
J Tissue Eng Regen Med ; 6(10): 757-68, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22095677

ABSTRACT

Cartilage tissue engineering is a rapidly progressing area of regenerative medicine with advances in cell biology and scaffold engineering constantly being investigated. Many groups are now capable of making neocartilage constructs with some level of morphological, biochemical, and histological likeness to native human cartilage tissues. The application of this useful technology in articular cartilage repair is well described in the literature; however, few studies have evaluated its application in head and neck reconstruction. Although there are many studies on auricular cartilage tissue engineering, there are few studies regarding cartilage tissue engineering for complex nasal reconstruction. This study therefore highlighted the challenges involved with nasal reconstruction, with special focus on nasal cartilage tissue, and examined how advancements made in cartilage tissue engineering research could be applied to improve the clinical outcomes of total nasal reconstructive surgery.


Subject(s)
Cartilage, Articular/surgery , Nasal Cartilages/surgery , Rhinoplasty/methods , Tissue Engineering/methods , Humans
7.
Development ; 138(13): 2807-21, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21652653

ABSTRACT

The embryonic head mesoderm gives rise to cranial muscle and contributes to the skull and heart. Prior to differentiation, the tissue is regionalised by the means of molecular markers. We show that this pattern is established in three discrete phases, all depending on extrinsic cues. Assaying for direct and first-wave indirect responses, we found that the process is controlled by dynamic combinatorial as well as antagonistic action of retinoic acid (RA), Bmp and Fgf signalling. In phase 1, the initial anteroposterior (a-p) subdivision of the head mesoderm is laid down in response to falling RA levels and activation of Fgf signalling. In phase 2, Bmp and Fgf signalling reinforce the a-p boundary and refine anterior marker gene expression. In phase 3, spreading Fgf signalling drives the a-p expansion of MyoR and Tbx1 expression along the pharynx, with RA limiting the expansion of MyoR. This establishes the mature head mesoderm pattern with markers distinguishing between the prospective extra-ocular and jaw skeletal muscles, the branchiomeric muscles and the cells for the outflow tract of the heart.


Subject(s)
Body Patterning/physiology , Head/embryology , Mesoderm/embryology , Mesoderm/metabolism , Animals , Body Patterning/genetics , Chick Embryo , Chickens , Gene Expression Regulation, Developmental/genetics , Gene Expression Regulation, Developmental/physiology , In Situ Hybridization
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