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Regeneration of the dermis does not occur spontaneously in the adult mammal. The epidermis is regenerated spontaneously provided there is a dermal substrate over which it can migrate. Certain highly porous, crosslinked collagen-glycosaminoglycan copolymers have induced partial morphogenesis of skin when seeded with dermal and epidermal cells and then grafted on standard, full-thickness skin wounds in the adult guinea pig. A mature epidermis and a nearly physiological dermis, which lacked hair follicles but was demonstrably different from scar, were regenerated over areas as large as 16 cm2. These chemical analogs of extracellular matrices were morphogenetically active provided that the average pore diameter ranged between 20 and 125 lsm, the resistance to degradation by collagenase exceeded a critical limit, and the density of autologous dermal and epidermal cells inoculated therein was >5 X 104 cells per cm2 of wound area. Unseeded copolymers with physical structures that were within these limits delayed the onset of wound contraction by about 10 days but did not eventually prevent it. Seeded copolymers not only delayed contraction but eventually arrested and reversed it while new skin was being regenerated. The data identify a model extracellular matrix that acts as if it were an insoluble growth factor with narrowly specified physicochemical structure, functioning as a transient basal lamina during'morphogenesis of skin.Throughout development, extracellular matrices (ECMs) are continuously being remodeled-i.e., synthesized, degraded, and resynthesized (1)(2)(3)(4)(5). Healing of a deep skin wound also requires remodeling of an ECM-the basal lamina (basement membrane) between the epidermis and the dermis (2). ECMs are largely insoluble and nondiffusible, and they confer stiffness and strength to multicellular systems (1, 2). During remodeling, the ECM necessarily suffers degradation of macromolecular chains, a process that dramatically reduces the insolubility of the ECM and impairs its role as mechanical reinforcement of a multicellular system undergoing development. It is not clear just how the resistance of the ECM to degradation affects its role during morphogenesis.In physical terms, ECMs can be described as macromolecular networks that are covalently crosslinked and are highly swollen in extracellular fluid. Accordingly, the physical structure of an ECM can be characterized initially by specifying the volume fraction 'of macromolecular components (swelling ratio), the average diameter of pores in the highly swollen network, the density ofcrosslinks tying chains to each other, and the degree of crystallinity present. This model leads to questions such as the following ones. Is it necessary for a developmentally active ECM to persist as an undegraded, crosslinked macromolecular network (and, therefore, remain insoluble and nondiffusible) over a critical time scale? Is it necessary for such an ECM to contain pores of a critical size? We have answered these questions in a preliminary way by use of we...

No abstract

Pertinent issues of collagen antigenicity and immunogenicity are concisely reviewed as they relate to the design and application of biomedical devices. A brief discussion of the fundamental concepts of collagen immunochemistry is presented, with a subsequent review of documented clinical responses to devices containing reconstituted soluble or solubilized collagen. The significance of atelocollagen, concerns regarding collagen-induced autoimmunity, and other relevant topics are also addressed in the context of current understanding of the human immune response to collagen.

Individuals who suffer extensive loss of skin, commonly in fires, are acutely ill, in danger of succumbing either to massive infection of to severe fluid loss. Patients who survive these early threats must often cope with problems of rehabilitation arising from deep, disfiguring scars and crippling contractures. In this report we describe the physiocochemical, biochemical, and mechanical considerations that form the basis for two-stage design of a membrane useful as an experimental wound closure. Stage I of the design, applicable to short-term acute use, calls for a membrane which displaces efficiently air pockets from a carefully prepared woundbed, free of weak boundary layers, and maintains the moisture flux through the wound at an optimal level. Optimization of the surface energy, modulus of elasticity, energy to fracture and moisture permeability of the membrane are among the essential attributes of Stage I design. Stage 2 of the design, applicable to long-term, chronic use, focuses on a nonantigenic membrane which performs as a biodegradable template for synthesis of neodermal tissue. A survey of candidate materials suggests reasons for selection of a porous, crosslinked collagen-glycosaminoglycan coprecipitate as the chemical basis of an evolving design which was initiated 10 years ago. Over the past several years a set of membranes has been iteratively designed on this basis and has been used to cover satisfactorily large experimental full-thickness skin wounds in guinea pigs. Such membranes have effectively protected these wounds from infection and fluid loss for over 25 days without rejection and without requiring change or other invasive manipulation. When appropriately designed for the purpose, the membranes have also strongly retarded wound contraction and have become replaced by newly synthesized, stable connective tissue. Several rules relating the molecular structure and morphology of these membranes to cellular response of adjacent tissue have also been derived. This report is the first in a series which details the methodology of preparation and the record of performance.

Prompt and long-term closure of full-thickness skin wounds is guinea pigs and humans is achieved by applying a bilayer polymeric membrane. The membrane comprises a top layer of a silicone elastomer and a bottom layer of a porous cross-linked network of collagen and glycosaminoglycan. The bottom layer can be seeded with a small number of autologous basal cells before grafting. No immunosuppression is used and infection, exudation, and rejection are absent. Host tissue utilizes the sterile membrane as a culture medium to synthesize neoepidermal and neodermal tissue. A functional extension of skin over the entire wound area is formed in about 4 weeks.

No abstract

Cell migration plays a critical role in a wide variety of physiological and pathological phenomena as well as in scaffold-based tissue engineering. Cell migration behavior is known to be governed by biochemical stimuli and cellular interactions. Biophysical processes associated with interactions between the cell and its surrounding extracellular matrix may also play a significant role in regulating migration. Although biophysical properties of two-dimensional substrates have been shown to significantly influence cell migration, elucidating factors governing migration in a three-dimensional environment is a relatively new avenue of research. Here, we investigate the effect of the three-dimensional microstructure, specifically the pore size and Young's modulus, of collagen-glycosaminoglycan scaffolds on the migratory behavior of individual mouse fibroblasts. We observe that the fibroblast migration, characterized by motile fraction as well as locomotion speed, decreases as scaffold pore size increases across a range from 90 to 150 mum. Directly testing the effects of varying strut Young's modulus on cell motility showed a biphasic relationship between cell speed and strut modulus and also indicated that mechanical factors were not responsible for the observed effect of scaffold pore size on cell motility. Instead, in-depth analysis of cell locomotion paths revealed that the distribution of junction points between scaffold struts strongly modulates motility. Strut junction interactions affect local directional persistence as well as cell speed at and away from the junctions, providing a new biophysical mechanism for the governance of cell motility by the extracellular microstructure.