109 research outputs found

    Molecule Microscopy

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    Contains reports on two research projects.Joint Services Electronics Program (Contract DAABO7-74-C-0630)National Institutes of Health (Grant 5 PO1 HL14322-04

    Electron Optics

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    Contains reports on one research project.Joint Services Electronics Program (Contract DAAB07-74-C-0630

    Collagen Fibrillogenesis in Tissues, in Solution and from Modeling: A Synthesis

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    Collagen fibril formation has been studied in tissues by light and electron microscopy; in solution by light scattering and microscopy; and from modeling based on the amino acid sequence of type I collagen. Taken together these studies indicate that collagen fibril assembly involves a stepwise formation of intermediate aggregates in which each intermediate is formed from earlier aggregates. In this sequence, monomeric collagen contributes only to the formation of early aggregates; and fibrils grow in length by the addition of intermediate aggregates to the end of a subfibril and in width by lateral wrapping of subfibrils. Modeling based on amino acid sequence data of possible intermolecular charge-charge interactions indicate 2 different kinds, one which promotes linear aggregation and the other which promotes lateral aggregation. The effects of different colla-gens and coprecipitants such as glycoproteins and proteoglycans can begin to be explained by their influence on the character of intermediate subassemblies. Ultrastructural data from 2 tissues, embryonic cornea and tendon, indicate that the site of fibril growth and assembly is at the cell surface

    Collagen Deposition During Wound Repair

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    Collagen fiber diameters, amount of birefringent collagen (brightness) and birefringence retardation were measured in implanted collagen-based sponges containing hyaluronic acid (HA) and fibronectin (FN). In the presence of HA and FN, increased number of fibroblasts and brightness were observed 6 days after wounding. Increased brightness in the presence of HA and FN reflected increased deposition of oriented collagen fibers. From days 9 to 12, increased fiber diameters were similar in implanted collagen-based sponges with or without HA and FN. Increased birefringence retardation in sponges containing HA and FN was consistent with increased packing density of collagen fibers observed by scanning electron microscopy. Our results suggest that HA and FN are effective in promoting fibroblast movement into a collagen sponge and deposition of collagen fibers during the early phases of wound healing. Use of a collagen-based sponge containing HA and FN may enhance collagen deposition in situations where healing is compromised as in the case of dermal ulcers

    Achilles Tendon Replacement by a Collagen Fiber Prosthesis: Morphological Evaluation of Neotendon Formation.

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    Reconstituted type I collagen was processed into fibers which were subsequently severely dehydrated and cyanamide cross-linked. Fibers prepared by this method were stronger and more resistant to degradation than uncrosslinked fibers. When used as a tendon replacement prosthesis, morphological events occurred which were observed by light, scanning, transmission electron microscopy and electron histochemistry. Resorption was the initial host response to the prosthesis and involved gradual biodegradation. Formation of a host-replacement tendon was the second response. Increased collagen fibril diameters and a transition in the proteoglycan/collagen fibril interactions occurred in the newly developing connective tissue between 3 and 10 weeks post-implantation. These extracellular matrix transitions were major events occurring during wound healing and led to the assembly of a mature connective tissue. When used as a tendon prosthesis, these collagen fibers rapidly resorb while allowing simultaneous formation of aligned connective tissue. The fibers may have other applications in the fields of Orthopaedic Surgery, Neurosurgery and Biomaterials Research

    Fibroblast and Epidermal Cell-Type I Collagen Interactions: Cell Culture and Human Studies

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    Fibroblast and epidermal cell-type I collagen sponge interactions were studied in cell culture as well as in humans. In cell culture, fibroblasts were observed to migrate and proliferate throughout a type I collagen sponge containing either hyaluronic acid (HA) or fibronectin (FN). Fibroblasts accumulated in the center of the pores in sponges containing HA and appeared to surround themselves with newly synthesized extracellular matrix. In sponges containing FN, fibroblasts attached to and elongated along the collagen fibers of the sponge. In the absence of FN or HA protein synthesis of fibroblasts appeared to be inhibited by the presence of the type I collagen sponge. Epidermal cells grown on plastic or on type I collagen, formed sheets. Epidermal cells grown on a collagen sponge morphologically appeared different than cells grown on plastic. The type I collagen matrix studied in cell culture was applied to dermal wounds of patients with pressure ulcers in order to evaluate its effect on dermal wound healing. The areas of ulcers treated for 6 weeks with a type I collagen sponge decreased by about 40% compared with no change in the areas of untreated controls. Preliminary results suggest that a type I collagen sponge is a biocornpatible substrate with fibroblasts and epidermal cells and may be effective in enhancing healing of chronic skin ulcers

    Clinical Applications of Electron Microscopy in the Analysis of Collagenous Biomaterials

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    Scanning and transmission electron microscopy are of clinical value in assessing the interaction between biomaterials and ingrowing tissues. Ultrastructural information allows the clinician and biomaterials specialist to determine events occurring during wound healing and the biocompatibility of prosthetic devices. This paper reviews some of the experimental and clinical studies done in our laboratory on the use of natural and reconstituted collagen as replacements for connective tissues. Consideration is given to collagen flakes used for the treatment of dermal ulcers, a collagen fiber prosthesis used for tendon and ligament replacement, the effects of chemical preservatives on cartilage used for replacement of tissues during plastic surgery and the growth and orientation of nerve cells on reconstituted collagen fibers. Our results show that reconstituted collagen can be prepared into prosthetic devices which encourage cell attachment and orientation thereby facilitating healing of injured tissues. Furthermore chemical preservation of cartilagenous tissues kills chondrocytes resulting in eventual resorption by inflammatory cells

    Identification of the Vibrational Optical Coherence Tomography Corneal Cellular Peak

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    Purpose: Our team previously identified the presence of five corneal resonant frequency (RF) peaks in healthy volunteers using vibrational optical coherence tomography (VOCT). Prior studies have suggested that the ≤100 Hz RF peak represents the cellular element of tissue. The aim of this study was to confirm that this peak reflects the human corneal cellular component using VOCT and histological analysis. Methods: Two human research globes were obtained from the same donor, and VOCT measurements were collected from the full-thickness corneas. A microkeratome was then used to create serial-free corneal caps from each cornea, with VOCT performed on the residual stromal bed after each excision. All lamellar sections from both globes were sent for histological analysis to determine cellularity. Cell counts on the specimens were performed by two independent observers. Results: The average of the normalized ≤100 Hz peak values before lamellar sectioning was significantly higher than the average of this peak values after the first, second, and third cuts (P = 0.023), which was 33.9% less than before any cuts. The cell count values in the first slice were significantly higher than the average cell count values of the three deeper slices (P \u3c 0.001), and the cell count dropped 84.4% after the first slice was removed. Conclusions: The findings of this study suggest that the ≤100 Hz corneal peak identified by VOCT corresponds to the cellular component of the cornea. Translational relevance: This work furthers our understanding of the origin of the corneal ≤100 Hz peak identified using VOCT

    Setting our sights on infectious diseases

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    In May 2019, the Wellcome Centre for Anti-Infectives Research (WCAIR) at the University of Dundee, UK, held an international conference with the aim of discussing some key questions around discovering new medicines for infectious diseases and a particular focus on diseases affecting Low and Middle Income Countries. There is an urgent need for new drugs to treat most infectious diseases. We were keen to see if there were lessons that we could learn across different disease areas and between the preclinical and clinical phases with the aim of exploring how we can improve and speed up the drug discovery, translational, and clinical development processes. We started with an introductory session on the current situation and then worked backward from clinical development to combination therapy, pharmacokinetic/pharmacodynamic (PK/PD) studies, drug discovery pathways, and new starting points and targets. This Viewpoint aims to capture some of the learnings
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