Biomedical subjects

Ioannis V Yannas

Publications and source records attributed to Ioannis V Yannas.

10 recordsLinked to original sources

PubMed2005

Similarities and differences between induced organ regeneration in adults and early foetal regeneration.

At least three organs (skin, peripheral nerves and the conjunctiva) have been induced to regenerate partially in adults following application of porous, degradable scaffolds with highly specific structure (templates). Templates blocked contraction and scar formation by inducing a reduction in the density of contractile fibroblasts (probably myofibroblasts) and by preventing these cells to organize themselves appropriately in the wound. In contrast, during early foetal healing, myofibroblasts were absent and wounds did not close by contraction but rather by spontaneous regeneration. The adult regenerative process has so far led to imperfect recovery of the physiological anatomy of skin (skin appendages were missing), while early foetal healing has led to apparently complete restoration. Furthermore, the mechanism of the adult regenerative process involves thwarting of myofibroblast function while, during early foetal healing, differentiation of myofibroblasts has not yet occurred. The data suggest that induced organ regeneration in the adult is the result of partial reversion to early foetal healing. If so, the adult may conceal a foetal response that may be subject to activation following application of highly active scaffolds or of other substances or cells.

Adult↗

PubMed2005

Fabricating tubular scaffolds with a radial pore size gradient by a spinning technique.

A novel fabrication process has been developed to produce collagen-based, porous tubular scaffolds to facilitate the study of myofibroblast migration during peripheral nerve regeneration; however, this fabrication technique offers broader appeal for the production of a variety of tubular structures without the use of a complicated mold system. A collagen-glycosaminoglycan (CG) suspension in acetic acid was spun in a cylindrical copper mold about its longitudinal axis at variable angular velocities and for different times, resulting in variable relative sedimentation of the CG content towards the mold outer edge; after the specified spinning time, the spinning mold was placed into a bath of liquid nitrogen where the CG suspension was rapidly frozen. Due to the rapid solidification, the CG content remained sedimented while an interconnected network of ice crystals formed throughout. Sublimation of the frozen mass removed the solvent (acetic acid) content, producing a porous, tubular structure defined by sedimentation and ice crystal nucleation processes. A porous, tubular scaffold with a sharply defined inner tube wall can be produced; further, increasing the spinning time and/or spinning velocity increases the sedimentation effect leading to the production of a hollow tube with a larger inner diameter. The tube walls display a radially aligned pore structure, even in cases where sedimentation was not sufficient to produce a hollow tube. A gradient of porosity along the tube radius was also observed in cases of extreme sedimentation: the pore structure of the external portion of the tube wall had a larger solid volume fraction and a smaller mean pore size compared to the internal portion of the tube. This tubular structure may allow preferential cell migration from the inner tube lumen towards the outer tube edge while blocking cell entrance into the tube through its outer surface due to increased scaffold relative density and decreased pore size.

Collagen↗

PubMed2005

Peripheral nerve regeneration.

The nerve chamber model has dominated the experimental study of peripheral nerve (PN) regeneration with animal models as well as in several clinical applications, such as the treatment of paralysis of limbs following severe trauma. The two stumps resulting from nerve transection are inserted inside a tubular chamber made from one of several materials, occasionally filled with various substances, and the quality of the reconnected nerve is assayed. Recent use of methods for data reduction has led to generation of a large normalized database from independent investigations. Methods for data normalization (reduction) are based on systematic use of the critical axon elongation, Lc, the gap length between the transected stumps at which the frequency of reconnection is just 50% for a given configuration. Four theories are compared for their ability to explain the normalized data. Although the neurotrophic and contact guidance theories explain some of the data, combined use of the more recent microtube theory and pressure cuff theory appears capable of explaining a much larger data set. PN regeneration appears to be upregulated by chamber configurations that facilitate formation of basement membrane microtubes about 10-20 microm in diameter, comprising linear columns of Schwann cells surrounded by basement membrane, into which axons elongate and eventually become myelinated. Regeneration is downregulated by experimental configurations that permit formation of a contractile cell (myofibroblast) capsule around the regenerating nerve that appears to restrict growth of a nerve trunk by application of circumferential mechanical forces. These two processes work competitively to regulate nerve regeneration in the chamber model.

Axons↗

PubMed2005

Facts and theories of induced organ regeneration.

Induced organ regeneration is de novo synthesis of a physiological, or nearly physiological, organ at the same anatomical site as the organ that is being replaced. Regeneration of skin, peripheral nerves and the conjunctiva, described in this chapter, have been accomplished using biologically active scaffolds (regeneration templates) seeded with epithelial cells; devices for regeneration of the first two organs are in clinical use. There is substantial empirical evidence that templates induce regeneration by blocking contraction, the major mechanism for closure of severe wounds in adults. Templates appear to function by interfering with normal myofibroblast function well as by acting as temporary configurational guides for synthesis of new stroma that resembles that of the organ under replacement. The combined evidence supports a theory which predicts that selective blocking of the adult healing response uncovers the latent fetal response to injury and leads to organ regeneration. An independent theory suggests that loss of regenerative potential during the mammalian fetal-adult transition is associated with simultaneous acquisition of individual immunocompetence.

Adaptation, Physiological↗

PubMed2004

Synthesis of tissues and organs.

Symbolism that describes the synthetic processes for chemical compounds has been used to describe, in qualitative terms, the synthesis of tissues and organs at the correct anatomical site. The synthetic process is summarized in the reaction diagram, a shorthand representation of the reactants, reactor, and products. Analysis of a large number of independent protocols has led to identification of the simplest synthetic pathways for two organs that have been studied extensively: skin and peripheral nerves. These apparently irreducible reaction diagrams for the two organs are not only simple but surprisingly similar, a fact suggesting the existence of general rules for the synthesis of other organs as well. The only two reactants that are required are an active scaffold (a macromolecular network synthesized as a highly porous analogue of the extracellular matrix) and a seeding of epithelial cells of the organ being synthesized. Scaffolds possessed biological (regenerative) activity provided that they were capable of blocking the contraction process that leads to closure of the injured site. Such activity requires that the density of ligands for binding of contractile cells on the scaffold maintains a sufficiently high level over the period of synthesis.

Animals↗

PubMed2004

Selection of biomaterials for peripheral nerve regeneration using data from the nerve chamber model.

Peripheral nerve regeneration has been studied in a variety of animal models. Of these, the nerve chamber model has clearly dominated. It has been used to generate a large base of data that, however, cannot be analyzed usefully due to lack of standardization of experimental conditions and assays. Lack of standardization of critical experimental parameters of the model has, however, greatly limited the opportunity to compare directly data from independent investigators; as a result, progress in understanding conditions for optimal nerve regeneration has been stunted. In this article, we provide an overview of the major experimental parameters that must be controlled in order to generate data from independent investigators that can be compared directly (normalized data). Such parameters include the gap length, animal species, and the identity of assays used to evaluate the product of the regenerative process. Use of the recently introduced concept of critical axon elongation, the gap length at which the probability of axonal outgrowth (reinnervation) across the gap is 50%, leads to generation of a normalized database that includes data from several independent investigators. Conclusions are drawn about the relative efficacy of the various biomaterials and devices employed. Nerve chamber configurations that had the highest regenerative activity were those in which the tube wall comprised collagen and certain synthetic biodegradable polymers rather than silicone, and was cell-permeable rather than protein-permeable. In addition, the following tube fillings showed very high regenerative activity: suspensions of Schwann cells; a solution either of acidic or basic fibroblast growth factor; insoluble ECM substrates rather than solutions or gels; polyamide filaments oriented along the tube axis; highly porous, insoluble analogs of the ECM with specific structure and controlled degradation rate.

Absorbable Implants↗

PubMed2004

Influence of freezing rate on pore structure in freeze-dried collagen-GAG scaffolds.

The cellular structure of collagen-glycosaminoglycan (CG) scaffolds used in tissue engineering must be designed to meet a number of constraints with respect to biocompatibility, degradability, pore size, pore structure, and specific surface area. The conventional freeze-drying process for fabricating CG scaffolds creates variable cooling rates throughout the scaffold during freezing, producing a heterogeneous matrix pore structure with a large variation in average pore diameter at different locations throughout the scaffold. In this study, the scaffold synthesis process was modified to produce more homogeneous freezing by controlling of the rate of freezing during fabrication and obtaining more uniform contact between the pan containing the CG suspension and the freezing shelf through the use of smaller, less warped pans. The modified fabrication technique has allowed production of CG scaffolds with a more homogeneous structure characterized by less variation in mean pore size throughout the scaffold (mean: 95.9 microm, CV: 0.128) compared to the original scaffold (mean: 132.4 microm, CV: 0.185). The pores produced using the new technique appear to be more equiaxed, compared with those in scaffolds produced using the original technique.

Biocompatible Materials↗

PubMedDate not supplied

Contractile forces generated by articular chondrocytes in collagen-glycosaminoglycan matrices.

The objective of the study was to directly measure the force of contraction of adult articular chondrocytes and to examine their contractile behavior in collagen-glycosaminoglycan analogs of extracellular matrix by live cell imaging in vitro. The contractile forces generated by passages 2 and 3 adult canine articular chondrocytes were measured using a cell force monitor. The contractile behavior of the cells was also directly imaged as they were cultured in collagen-glycosaminoglycan scaffolds. Passage 2 cells seeded in a collagen-glycosaminoglycan scaffold were capable of generating a force of 0.3 nN/cell. Chondrocytes subcultured through a third passage generated a force twice that level, paralleling the increase in the alpha-smooth muscle actin (SMA) content of the cells as demonstrated by Western blot analysis. Treatment of passage 3 cells with staurosporine reduced the force of contraction by approximately one-half, reflecting the effects of this agent in reducing the SMA content of the cells and disrupting the microfilaments. These values compare with 1 nN previously reported for lapine dermal fibroblasts of passage 5-7, using the same apparatus. Direct live cell imaging documented the contractile behavior of the articular chondrocytes in the collagen-glycosaminoglycan matrix in the time frame in which the force was directly measured in the cell force monitor. This imaging demonstrated how the cells acted individually and in unison to buckle the collagen struts making up the matrix. Adult articular chondrocytes are capable of generating a SMA-enabled force of contraction sufficient to model extracellular matrix molecules.

Animals↗

PubMedDate not supplied

Evidence for sequential utilization of fibronectin, vitronectin, and collagen during fibroblast-mediated collagen contraction.

Contraction plays a major role in wound healing and is inevitably mediated through the mechanical interaction of fibroblast cytoskeleton and integrins with their extracellular matrix ligands. Cell-matrix attachment is critical for such events. In human dermal fibroblasts most such interactions are mediated by the beta1-type integrins. This study investigated the role played by key components in this system, notably fibronectin, vitronectin, and integrin subcomponents alpha2 and alpha5, which recognize collagen and fibronectin. Inhibition of adhesion through these ligands was studied either by antibody blocking or with fibronectin and/or vitronectin depletion. Functional effects of inhibition were monitored as force generation in collagen-glycosaminoglycan (IntegraTM) sponges, over 20 hours using a culture force monitor. Dose and time-course inhibition studies indicated that initial attachment and force generation (approx. 0-5 hours postseeding) was through fibronectin receptors and this was followed by vitronectin ligand and receptor utilization (4 hours onward). Utilization of the collagen integrin subcomponent alpha2 appeared to be increasingly important between 6 and 16 hours and dominant thereafter. Additionally, there was evidence for functional interdependence between the three ligand systems fibronectin, vitronectin, and collagen. We propose that there is a short cascade of sequential integrin-ligand interactions as cells attach to, extend through, and eventually contract their matrix. (WOUND REP REG 2002;10:-408)

Biocompatible Materials↗

PubMedDate not supplied

Formation of lung alveolar-like structures in collagen-glycosaminoglycan scaffolds in vitro.

The objective of this study was to investigate the histology of tissue formed when fetal rat lung cells were grown in a collagen-glycosaminoglycan (GAG) tissue-engineering scaffold. The goal was the formation of lung histotypic structures in the tissue-engineering scaffolds in vitro. Achieving this goal would facilitate future investigations of the effects of selected scaffold design parameters on processes that may underlie aspects of lung regeneration in vivo. Lung cells were obtained from Sprague-Dawley rats after 16 and 19 days of gestation. These dissociated cells were seeded into type I collagen-chondroitin 6-sulfate matrices, 8 mm in diameter by 2 mm in thickness, cross-linked and sterilized by dehydrothermal treatment. Approximately 28 million cells were seeded into each spongelike sample. Histological and immunohistochemical studies were performed at termination periods of 2 days and 1, 2, and 3 weeks. The enzymatically dissociated 19-day gestation fetal rat lung cells formed and maintained alveolar-like structures, 50-60 microm in diameter, in the collagen- GAG scaffold. A novel finding was that all of the cell-seeded scaffolds underwent cell-mediated contraction that appeared to be associated with the finding by immunohistochemistry of expression of alpha-smooth muscle actin in some cells. These results demonstrate the capability of dissociated lung cells to form lung histotypic structures in collagen-GAG tissue-engineering scaffolds in vitro. This culture system may be of value in facilitating exploration of strategies for preparing such scaffolds for the regeneration of lung tissue in vivo.

Actins↗