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ABSTRACT: The conventional approach to biomaterial design and development typically focuses upon the mechanical and material properties with long-term objectives that include an inert host immune response and long-lasting mechanical and structural support. The emergence and interest in tissue engineering and regenerative medicine has driven the development of novel cell friendly biomaterials, materials with tailored degradation rates, materials with highly specific architectures and surfaces, and vehicles for delivery of bioactive molecules, among numerous other advancements. Each of these biomaterial developments support specific strategies for tissue repair and reconstruction. These advancements in biomaterial form and function,, combined with new knowledge of innate and acquired immune system biology, provide an impetus for re-examination of host-biomaterial interactions including host-biomaterial interface events, spatial and temporal patterns of in-vivo biomaterial remodeling, and related downstream functional outcomes. An examination of such issues is provided herein with a particular focus on macrophage polarization and its implications in tissue engineering and regenerative medicine.
Acta biomaterialia 10/2012; · 3.98 Impact Factor
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ABSTRACT: The host response to biomaterials has been studied for decades. Largely, the interaction of host immune cells, macrophages in particular, with implanted materials has been considered to be a precursor to granulation tissue formation, the classic foreign body reaction, and eventual encapsulation with associated negative impacts upon device functionality. However, more recently, it has been shown that macrophages, depending upon context dependent polarization profiles, are capable of affecting both detrimental and beneficial outcomes in a number of disease processes and in tissue remodeling following injury. Herein, the diverse roles played by macrophages in these processes are discussed in addition to the potential manipulation of macrophage effector mechanisms as a strategy for promoting site-appropriate and constructive tissue remodeling as opposed to deleterious persistent inflammation and scar tissue formation.
Biomaterials 03/2012; 33(15):3792-802. · 7.40 Impact Factor
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ABSTRACT: A device composed of extracellular matrix (ECM) was investigated as an inductive template in vivo for reconstruction of the temporomandibular joint (TMJ) disk after discectomy.
A scaffold material composed of porcine-derived ECM was configured to mimic the shape and size of the TMJ. This device was implanted in a canine model of bilateral TMJ discectomy. After discectomy, 1 side was repaired with an ECM scaffold material and the contralateral side was left empty as a control. At 6 months after implantation, the joint space was opened, the joints were evaluated for signs of gross pathologic degenerative changes, and newly formed tissue was excised for histologic, biochemical, and biomechanical analysis.
The results showed that implantation of an initially acellular material supported the formation of site-appropriate, functional host tissue that resembled that of the native TMJ disk. Furthermore, this prevented gross degenerative changes in the temporal fossa and mandibular condyle. No tissue formation and mild to severe gross pathologic changes were observed in the contralateral controls.
These results suggest that an ECM-based bioscaffold could represent an off-the-shelf solution for TMJ disk replacement.
Journal of oral and maxillofacial surgery: official journal of the American Association of Oral and Maxillofacial Surgeons 02/2012; 70(11):2656-68. · 1.58 Impact Factor
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ABSTRACT: Macrophages have been classified as having plastic phenotypes which exist along a spectrum between M1 (classically activated; pro-inflammatory) and M2 (alternatively activated; regulatory, homeostatic). To date, the effects of polarization towards an M1 or M2 phenotype have been studied largely in the context of response to pathogen or cancer. Recently, M1 and M2 macrophages have been shown to play distinct roles in tissue remodeling following injury. In the present study, the M1/M2 paradigm was utilized to examine the role of macrophages in the remodeling process following implantation of 14 biologically derived surgical mesh materials in the rat abdominal wall. In situ polarization of macrophages responding to the materials was examined and correlated to a quantitative measure of the observed tissue remodeling response to determine whether macrophage polarization is an accurate predictor of the ability of a biologic scaffold to promote constructive tissue remodeling. Additionally the ability of M1 and M2 macrophages to differentially recruit progenitor-like cells in vitro, which are commonly observed to participate in the remodeling of those ECM scaffolds which have a positive clinical outcome, was examined as a possible mechanism underlying the differences in the observed remodeling responses. The results of the present study show that there is a strong correlation between the early macrophage response to implanted materials and the outcome of tissue remodeling. Increased numbers of M2 macrophages and higher ratios of M2:M1 macrophages within the site of remodeling at 14 days were associated with more positive remodeling outcomes (r(2)=0.525-0.686, p<0.05). Further, the results of the present study suggest that the constructive remodeling outcome may be due to the recruitment and survival of different cell populations to the sites of remodeling associated with materials that elicit an M1 vs. M2 response. Both M2 and M0 macrophage conditioned media were shown to have higher chemotactic activities than media conditioned by M1 macrophages (p<0.05). A more thorough understanding of these issues will logically influence the design of next generation biomaterials and the development of regenerative medicine strategies for the formation of functional host tissues.
Acta biomaterialia 12/2011; 8(3):978-87. · 3.98 Impact Factor
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ABSTRACT: Bioprosthetic devices, constructed from a variety of materials, are routinely implanted in a variety of anatomical locations. Essential to their success is the formation of a non-destructive interface with the host tissue and appropriate tissue remodelling. Traditionally, the main method of assessing the host-material interface has been qualitative histological evaluation, using pattern recognition and comparative assessment to identify changes in the normal tissue architecture that are characteristic of scar tissue. In the present study, the recently developed technique of multispectral imaging was used to revisit a little-described histological stain, Herovici's polychrome, which is capable of distinguishing between types I and III collagen. Combined, these techniques allowed quantification of collagen content and distribution of collagen types within a tissue sample. Samples of rat tail and human scar tissue were used to optimize the staining, while comparison with immunolabelled samples was used to develop a reproducible quantification system, based on the specific colour profiles for types I and III collagen. Finally the remodelling of rat abdominal wall defects repaired with crosslinked or non-crosslinked extracellular matrix scaffolds derived from porcine urinary bladder was assessed with this technique. Compared to standard histological assessment, the combination of multispectral imaging and Herovici's polychrome staining presents a quick, simple, reliable technique that can provide accurate quantification of tissue remodelling and specifically identify the expression and distribution of types I and III collagen. Copyright © 2011 John Wiley & Sons, Ltd.
Journal of Tissue Engineering and Regenerative Medicine 11/2011; · 3.28 Impact Factor
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ABSTRACT: A device consisting of powdered porcine urinary bladder extracellular matrix (UBM-ECM) encapsulated within sheets of the same material was investigated as a scaffold for temporomandibular joint (TMJ) meniscus reconstruction.
Five dogs underwent unilateral resection of the native meniscus and replacement with a UBM-ECM device. Necropsies were performed at 3, 4, 8, 12, and 24 weeks. Two additional dogs underwent bilateral resection of the meniscus with replacement with a UBM-ECM device on 1 side, leaving the contralateral side empty as a control. Necropsies were performed at 24 weeks for bilaterally treated animals.
Macroscopically, the UBM-ECM implants were remodeled rapidly and were indistinguishable from newly deposited host tissue at all time points. Microscopically, remodeling was characterized by a dense infiltration of predominantly CD68(+) mononuclear cells and smooth muscle actin-positive fibroblast-like cells at early time points changing with time to a sparse population of smooth muscle actin-negative spindle-shaped cells resembling those of the native fibrocartilaginous TMJ meniscus. Furthermore, the remodeling process showed deposition of predominantly type I collagen, the density and organization of which resembled those of the native meniscus by the 24-week time point. Ingrowth of calsequestrin-positive skeletal muscle tissue was also observed at the periphery of the remodeled UBM-ECM device and was similar to that found at the attachment site of the native meniscus to the surrounding soft tissues. Histologic results were identical for samples excised from both unilaterally and bilaterally treated animals. No adverse changes in the articulating surfaces of the condyle or fossa were observed in UBM-ECM-implanted joints. In the bilaterally treated animals, the unimplanted control side was characterized by degeneration and pitting of the articulating surfaces of both the condyle and the fossa, with disorganized bands of fibrous connective tissue observed within the joint space.
Results of this study suggest that the UBM-ECM device provides an effective interpositional material while serving as an inductive template for reconstruction of the TMJ meniscus.
Journal of oral and maxillofacial surgery: official journal of the American Association of Oral and Maxillofacial Surgeons 06/2011; 69(12):e488-505. · 1.58 Impact Factor
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ABSTRACT: Inflammation and airway remodeling occur in a variety of airway diseases. Modeling aspects of the inflammatory and fibrotic processes following repeated exposure to particulate matter may provide insights into a spectrum of airway diseases, as well as prevention/treatment strategies. An agent-based model (ABM) was created to examine the response of an abstracted population of inflammatory cells (nominally macrophages, but possibly including other inflammatory cells such as lymphocytes) and cells involved in remodeling (nominally fibroblasts) to particulate exposure. The model focused on a limited number of relevant interactions, specifically those among macrophages, fibroblasts, a pro-inflammatory cytokine (TNF-α), an anti-inflammatory cytokine (TGF-β1), collagen deposition, and tissue damage. The model yielded three distinct states that were equated with (1) self-resolving inflammation and a return to baseline, (2) a pro-inflammatory process of localized tissue damage and fibrosis, and (3) elevated pro- and anti-inflammatory cytokines, persistent tissue damage, and fibrosis outcomes. Experimental results consistent with these predicted states were observed in histology sections of lung tissue from mice exposed to particulate matter. Systematic in silico studies suggested that the development of each state depended primarily upon the degree and duration of exposure. Thus, a relatively simple ABM resulted in several, biologically feasible, emergent states, suggesting that the model captures certain salient features of inflammation following exposure of the lung to particulate matter. This ABM may hold future utility in the setting of airway disease resulting from inflammation and fibrosis following particulate exposure.
Mathematical biosciences 03/2011; 231(2):186-96. · 1.30 Impact Factor
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Biomaterials 01/2011; 32(1):316-9. · 7.40 Impact Factor
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ABSTRACT: The host response, and in particular the innate immune response, is critical to the successful application of tissue engineering
to the reconstruction of injured or missing tissues. Cell-based, scaffold-based, and signal molecule-based strategies are
utilized in regenerative medicine and each of these approaches elicits a distinct host immune response that has a significant
impact upon the downstream outcome. Modulation, but not suppression of the immune component of wound healing appears to be
essential for constructive remodeling of tissues and organs. Promotion of a pro-wound healing and anti-inflammatory response,
and avoidance of the foreign body reaction is associated with a constructive functional remodeling outcome. While macrophages
play a pivotal role in this response, other immune cells and the interactions between all cell types involved in tissue remodeling
are also clearly important. The objective of this chapter is to provide an overview of the host response to biomaterials including
both the pro-inflammatory and resultant foreign body reaction, and the pro-wound healing, anti-inflammatory response that
is associated with constructive remodeling.
12/2010: pages 353-375;
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ABSTRACT: Decellularization of tissues and organs is a successful platform technology for creating scaffolding materials for tissue engineering and regenerative medicine. It has been suggested that the success of these materials upon implantation is due to the molecular signals provided by the remaining scaffold extracellular matrix (ECM) components presented to probing cells in vivo as they repopulate the surface. For this study, decellularized matrices were created from esophagus, bladder, and small intestine harvested from adult male Fischer 344 rats. The three decellularized matrices (each originating from source tissues which included an epithelial lining on their luminal surfaces) were immunostained for collagen IV and laminin to determine basement membrane retention. Scanning electron micrographs of the surfaces were used to provide insight into the surface topography of each of the decellularized tissues. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to generate high-resolution mass spectra for the surfaces of each scaffold. This surface-sensitive technique allows for detailed molecular analysis of the outermost 1-2 nm of a material and has been applied previously to thin protein films and secreted ECM proteins on poly(N-isopropyl acrylamide) (polyNIPAAM) surfaces. To extract trends from within the complex ToF-SIMS dataset, a multivariate analysis technique, principal component analysis (PCA), was employed. Using this method, a molecular fingerprint of each surface was created and separation was seen in the PCA scores between the decellularized esophagus and the decellularized small intestine samples. The PCA scores for the decellularized bladder sample fell between the previous two decellularized samples. Protein films of common extracellular matrix constituents (collagen IV, collagen I, laminin, and Matrigel) were also investigated. The PCA results from these protein films were used to develop qualitative hypotheses for the relationship of the key fragments identified from the PCA of the decellularized ECMs.
Biomaterials 11/2010; 32(1):137-43. · 7.40 Impact Factor
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Janet E Reing, Bryan N Brown,
Kerry A Daly,
John M Freund,
Thomas W Gilbert,
Susan X Hsiong,
Alexander Huber,
Karen E Kullas,
Stephen Tottey,
Matthew T Wolf,
Stephen F Badylak
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ABSTRACT: Biologic materials from various species and tissues are commonly used as surgical meshes or scaffolds for tissue reconstruction. Extracellular matrix (ECM) represents the secreted product of the cells comprising each tissue and organ, and therefore provides a unique biologic material for selected regenerative medicine applications. Minimal disruption of ECM ultrastructure and content during tissue processing is typically desirable. The objective of this study was to systematically evaluate effects of commonly used tissue processing steps upon porcine dermal ECM scaffold composition, mechanical properties, and cytocompatibility. Processing steps evaluated included liming and hot water sanitation, trypsin/SDS/TritonX-100 decellularization, and trypsin/TritonX-100 decellularization. Liming decreased the growth factor and glycosaminoglycan content, the mechanical strength, and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for all). Hot water sanitation treatment decreased only the growth factor content of the ECM (p ≤ 0.05). Trypsin/SDS/TritonX-100 decellularization decreased the growth factor content and the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). Trypsin/Triton X-100 decellularization also decreased the growth factor content of the ECM but increased the ability of the ECM to support in vitro cell growth (p ≤ 0.05 for both). We conclude that processing steps evaluated in the present study affect content, mechanical strength, and/or cytocompatibility of the resultant porcine dermal ECM, and therefore care must be taken in choosing appropriate processing steps to maintain the beneficial effects of ECM in biologic scaffolds.
Biomaterials 11/2010; 31(33):8626-33. · 7.40 Impact Factor
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Bryan N Brown,
John M Freund,
Li Han,
J Peter Rubin,
Janet E Reing,
Eric M Jeffries,
Mathew T Wolf,
Stephen Tottey,
Christopher A Barnes,
Buddy D Ratner,
Stephen F Badylak
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ABSTRACT: Extracellular matrix (ECM)-based scaffold materials have been used successfully in both preclinical and clinical tissue engineering and regenerative medicine approaches to tissue reconstruction. Results of numerous studies have shown that ECM scaffolds are capable of supporting the growth and differentiation of multiple cell types in vitro and of acting as inductive templates for constructive tissue remodeling after implantation in vivo. Adipose tissue represents a potentially abundant source of ECM and may represent an ideal substrate for the growth and adipogenic differentiation of stem cells harvested from this tissue. Numerous studies have shown that the methods by which ECM scaffold materials are prepared have a dramatic effect upon both the biochemical and structural properties of the resultant ECM scaffold material as well as the ability of the material to support a positive tissue remodeling outcome after implantation. The objective of the present study was to characterize the adipose ECM material resulting from three methods of decellularization to determine the most effective method for the derivation of an adipose tissue ECM scaffold that was largely free of potentially immunogenic cellular content while retaining tissue-specific structural and functional components as well as the ability to support the growth and adipogenic differentiation of adipose-derived stem cells. The results show that each of the decellularization methods produced an adipose ECM scaffold that was distinct from both a structural and biochemical perspective, emphasizing the importance of the decellularization protocol used to produce adipose ECM scaffolds. Further, the results suggest that the adipose ECM scaffolds produced using the methods described herein are capable of supporting the maintenance and adipogenic differentiation of adipose-derived stem cells and may represent effective substrates for use in tissue engineering and regenerative medicine approaches to soft tissue reconstruction.
Tissue Engineering Part C Methods 11/2010; 17(4):411-21. · 4.64 Impact Factor
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ABSTRACT: Extracellular matrix (ECM) scaffolds prepared from different tissue sources or using different methods have been demonstrated to have distinctive effects upon cell adhesion patterns and the ability to support and maintain differentiated phenotypes. It is unknown whether the molecular composition or the ultrastructure of the ECM plays a greater role in determining the phenotype of the cells with which it comes into contact. However, when implanted, the topology and ligand landscape of the material will determine the host molecules that bind and the type and behavior of cells that mediate the host response. Therefore, a comprehensive understanding of surface characteristics is essential in the design of scaffolds for specific clinical applications. The surface characteristics of ECM scaffolds derived from porcine urinary bladder, small intestine, and liver as well as the effects of two commonly used methods of chemical cross-linking upon UBM were investigated. Electron microscopy and time of flight secondary ion mass spectroscopy were used to examine the surface characteristics of the scaffolds. The results show that ECM scaffolds have unique morphologic and structural properties which are dependant on the organ or tissue from which the scaffold is harvested. Furthermore, the results show that the surface characteristics of an ECM scaffold are changed through chemical cross-linking.
Biomaterials 10/2009; 31(3):428-37. · 7.40 Impact Factor
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ABSTRACT: Naturally occurring porcine-derived extracellular matrix (ECM) has successfully been used as a biological scaffold material for site-specific reconstruction of a wide variety of tissues. The site-specific remodelling process includes rapid degradation of the scaffold, with concomitant recruitment of mononuclear, endothelial and bone marrow-derived cells, and can lead to the formation of functional skeletal and smooth muscle tissue. However, the temporal and spatial patterns of innervation of the remodelling scaffold material in muscular tissues are not well understood. A retrospective study was conducted to investigate the presence of nervous tissue in a rat model of abdominal wall reconstruction and a canine model of oesophageal reconstruction in which ECM scaffolds were used as inductive scaffolds. Evidence of mature nerve, immature nerve and Schwann cells was found within the remodelled ECM at 28 days in the rat body wall model, and at 91 days post surgery in a canine model of oesophageal repair. Additionally, a microscopic and morphological study that investigated the response of primary cultured neurons seeded upon an ECM scaffold showed that neuronal survival and outgrowth were supported by the ECM substrate. Finally, matricryptic peptides resulting from rapid degradation of the ECM scaffold induced migration of terminal Schwann cells in a concentration-dependent fashion in vitro. The findings of this study suggest that the reconstruction of tissues in which innervation is an important functional component is possible with the use of biological scaffolds composed of extracellular matrix.
Journal of Tissue Engineering and Regenerative Medicine 08/2009; 3(8):590-600. · 3.28 Impact Factor
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ABSTRACT: Recently, macrophages have been characterized as having an M1 or M2 phenotype based on receptor expression, cytokine and effector molecule production, and function. The effects of macrophage phenotype upon tissue remodeling following the implantation of a biomaterial are largely unknown. The objectives of this study were to determine the effects of a cellular component within an implanted extracellular matrix (ECM) scaffold upon macrophage phenotype, and to determine the relationship between macrophage phenotype and tissue remodeling. Partial-thickness defects in the abdominal wall musculature of Sprague-Dawley rats were repaired with autologous body wall tissue, acellular allogeneic rat body wall ECM, xenogeneic pig urinary bladder tissue, or acellular xenogeneic pig urinary bladder ECM. At 3, 7, 14, and 28 days the host tissue response was characterized using histologic, immunohistochemical, and RT-PCR methods. The acellular test articles were shown to elicit a predominantly M2 type response and resulted in constructive remodeling, while those containing a cellular component, even an autologous cellular component, elicited a predominantly M1 type response and resulted in deposition of dense connective tissue and/or scarring. We conclude that the presence of cellular material within an ECM scaffold modulates the phenotype of the macrophages participating in the host response following implantation, and that the phenotype of the macrophages participating in the host response appears to be related to tissue remodeling outcome.
Biomaterials 01/2009; 30(8):1482-91. · 7.40 Impact Factor
