This review is directed at understanding how neuronal death occurs in two distinct insults, global ischemia and focal ischemia. These are the two principal rodent models for human disease. Cell dea...

Ischemic Cell Death in Brain Neurons

The repair of wounds is one of the most complex biological processes that occur during human life. After an injury, multiple biological pathways immediately become activated and are synchronized to respond. In human adults, the wound repair process commonly leads to a non-functioning mass of fibrotic tissue known as a scar. By contrast, early in gestation, injured fetal tissues can be completely recreated, without fibrosis, in a process resembling regeneration. Some organisms, however, retain the ability to regenerate tissue throughout adult life. Knowledge gained from studying such organisms might help to unlock latent regenerative pathways in humans, which would change medical practice as much as the introduction of antibiotics did in the twentieth century.

/pdf/wound-repair-and-regeneration-1rzgi7hifn.pdf

Wound repair and regeneration

Fibroblasts can condense a hydrated collagen lattice to a tissue-like structure 1/28th the area of the starting gel in 24 hr. The rate of the process can be regulated by varying the protein content of the lattice, the cell number, or the con- centration of an inhibitor such as Colcemid. Fibroblasts of high population doubling level propagated in vitro, which have left the cell cycle, can carry out the contraction at least as efficiently as cycling cells. The potential uses of the system as an immu- nologically tolerated "tissue" for wound hea ing and as a model for studying fibroblast function are discussed.

Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative

Scaffolds represent important components for tissue engineering. However, researchers often encounter an enormous variety of choices when selecting scaffolds for tissue engineering. This paper aims to review the functions of scaffolds and the major scaffolding approaches as important guidelines for selecting scaffolds and discuss the tissue-specific considerations for scaffolding, using intervertebral disc as an example.

Scaffolding in tissue engineering: general approaches and tissue-specific considerations.

Collagen molecules are structural macro-molecules of the extracellular matrix that include in their structure one or several domains that have a characteristic triple helical conformation. They have been classified by types that define distinct sets of polypeptide chains that can form homo- and heterotrimeric assemblies. All the collagen molecules participate in supramolecular aggregates that are stabilized in part by interactions between triple helical domains. Fourteen collagen types have been defined so far. They form a wide range of structures. Most notable are 1) fibrils that are found in most connective tissues and are made by alloys of fibrillar collagens (types I, II, III, V, and XI) and 2) sheets constituting basement membranes (type IV collagen), Descemet's membrane (type VIII collagen), worm cuticle, and organic exoskeleton of sponges. Other collagens, present in smaller quantities in tissues, play the role of connecting elements between these major structures and other tissue components. The f...

https://faseb.onlinelibrary.wiley.com/doi/pdf/10.1096/fasebj.5.13.1916105

Collagen family of proteins.

Background Ocular trauma or disease may lead to severe corneal opacification and, consequently, severe loss of vision as a result of complete loss of corneal epithelial stem cells. Transplantation of autologous corneal stem-cell sources is an alternative to allograft transplantation and does not require immunosuppression, but it is not possible in many cases in which bilateral disease produces total corneal stem-cell deficiency in both eyes. We studied the use of autologous oral mucosal epithelial cells as a source of cells for the reconstruction of the corneal surface. Methods We harvested 3-by-3-mm specimens of oral mucosal tissue from four patients with bilateral total corneal stem-cell deficiencies. Tissue-engineered epithelial-cell sheets were fabricated ex vivo by culturing harvested cells for two weeks on temperature-responsive cell-culture surfaces with 3T3 feeder cells that had been treated with mitomycin C. After conjunctival fibrovascular tissue had been surgically removed from the ocular surfa...

https://www.nejm.org/doi/pdf/10.1056/NEJMoa040455

Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium.

Type IX collagen, containing molecules of the three distinct polypeptides alpha 1(IX), alpha 2(IX), and alpha 3(IX), is an interesting hybrid extracellular matrix component in cartilage and eye tissues, with the properties of both a proteoglycan and a collagen. The alpha 1 (IX) chain has two forms, as a result of the tissue-specific utilization of two alternative promoters; the alpha 2(IX) chain carries a covalently attached glycosaminoglycan side chain. We have introduced a gene construct controlled by a tissue-specific promoter/enhancer and expressing a truncated alpha 1(IX) chain into mice. Examination of the offspring of two different founders revealed pathological changes similar to osteoarthritis in the articular cartilage of knee joints. In addition, mice homozygous for the transgene developed mild chondrodysplasia (i.e., mild dwarfism, anterior tonguing in the vertebral bodies, and ophthalmopathy). The relative ratio of transgene product to the endogenous alpha 1(IX) chain was approximately one in homozygotes and less than one in heterozygotes. Therefore, the phenotypic severity correlated well with the level of transgene expression. These findings suggest that mutations in type IX collagen genes may cause certain forms of osteoarthritis and chondrodysplasia in humans.

https://www.pnas.org/content/pnas/90/7/2870.full.pdf

Osteoarthritis associated with mild chondrodysplasia in transgenic mice expressing alpha 1(IX) collagen chains with a central deletion

Type V collagen and type I collagen were obtained from human placentas by pepsin treatment, followed by salt fractionation. The precipitates formed at 37°c from a mixed solution of type V collagen and type I collagen, reacted with antibodies to either type V collagen or type I collagen. The precipitates seen by electron microscopy were fine flexible fibrils, with a D-periodic banding pattern. The average diameter of hybrid fibrils was smaller than 50nm, when the proportion of type V collagen exceeded that of type I collagen. Type V collagen directly interacts with type I collagen in forming hybrid fibrils, resulting in limitation of the growth of type I collagen fibrils into thicker fibrils. We propose that the fibrils with a predominant type V collagen content may occur in the pericellular environment of various tissues, as a basic structure in connecting basal laminae with interstitial collagen fibrils.

In vitro Formation of Hybrid Fibrils of Type v Collagen and Type I Collagen Limited Growth of Type I Collagen Into Thick Fibrils by Type V Collagen

Fibroblasts were cultured within FITC-prelabeled type I collagen. Cells initially round in shape protruded processes, and began to collect the fibrils into their vicinity. Repeated protrusion and withdrawal of cell processes was observed. Consequently, condensed fluorescence was observed on the elongated bipolar cells stained with rhodamine-phalloidin. Concomitantly with these events, the gel began to contract in overall size, with an increase of fluorescence intensity. Scanning electron micrographs of the contracted gel showed a disproportional distribution of collagen fibrils: a highly condensed region surrounding cell bodies and a moderately condensed region. A major portion of condensed fibrils may have been derived from reconstituted collagen fibrils, since fibroblasts within collagen gel synthesized little collagen. When the gel adhered to glass tightly, so that overall contraction was prevented, the fluorescence in a range of scores of micrometers from the cells disappeared owing to depletion of fibrils by the cells. The combined spaces with null fluorescence in total under repressed contraction corresponded well to the reduction in volume due to gel contraction. It seems likely that the fibril condensation onto the cells causes the overall gel contraction.

Eijiro Adachi

Papers

Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium.

Osteoarthritis associated with mild chondrodysplasia in transgenic mice expressing alpha 1(IX) collagen chains with a central deletion

In vitro Formation of Hybrid Fibrils of Type v Collagen and Type I Collagen Limited Growth of Type I Collagen Into Thick Fibrils by Type V Collagen

Condensation of collagen fibrils to the direct vicinity of fibroblasts as a cause of gel contraction

Growth rate of human fibroblasts is repressed by the culture within reconstituted collagen matrix but not by the culture on the matrix.