Tissue-engineered meniscal constructs.
Humans; Biomechanical Phenomena; *Biocompatible Materials; *Tissue Engineering; Menisci; Tibial/*anatomy & histology/physiology
The medial and lateral menisci play important roles in knee biomechanics, kinematics, and stability. Unfortunately, these structures are prone to damage and, because of a tenuous blood supply, have great difficulty healing. Many interventions have been proposed for treatment of damaged meniscal tissue, but most surgical options are fraught with difficulties, from continued osteoarthritic degeneration to potential for disease transmission. The field of tissue engineering has made wide inroads into constructing meniscal tissue. Investigations involving collagenous tissue, meniscal fibrochondrocytes, chondrocytes, synthetic scaffolds, and gene therapy have all been reported in the literature. Despite these advances, however, more work needs to be done, including incorporating concepts and applications from other engineering disciplines, to potentiate the possibility of a tissue-engineered meniscus that approximates native tissue. In particular, the histologic, morphologic, and biomechanical properties of tissue-engineered meniscal constructs must be better understood to facilitate this goal.
Schoenfeld Andrew J; Landis William J; Kay David B
American journal of orthopedics (Belle Mead, N.J.)
2007
2007-11
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
The nature and role of periosteum in bone and cartilage regeneration.
Animals; Bone Regeneration/genetics/*physiology; Cartilage/*physiology; Cattle; Collagen Type I/genetics/metabolism; Collagen Type II/genetics/metabolism; Core Binding Factor Alpha 1 Subunit/genetics/metabolism; Gene Expression Regulation; Integrin-Binding Sialoprotein/genetics/metabolism; Mice; Nude; Periosteum/anatomy & histology/diagnostic imaging/*physiology; Prosthesis Implantation; Radiography; Tissue Engineering; Tissue Scaffolds
This study was undertaken to determine whether periosteum from different bone sources in a donor results in the same formation of bone and cartilage. In this case, periosteum obtained from the cranium and mandible (examples of tissue supporting intramembranous ossification) and the radius and ilium (examples of tissues supporting endochondral ossification) of individual calves was used to produce tissue-engineered constructs that were implanted in nude mice and then retrieved after 10 and 20 weeks. Specimens were compared in terms of their osteogenic and chondrogenic potential by radiography, histology, and gene expression levels. By 10 weeks of implantation and more so by 20 weeks, constructs with cranial periosteum had developed to the greatest extent, followed in order by ilium, radius, and mandible periosteum. All constructs, particularly with cranial tissue although minimally with mandibular periosteum, had mineralized by 10 weeks on radiography and stained for proteoglycans with safranin-O red (cranial tissue most intensely and mandibular tissue least intensely). Gene expression of type I collagen, type II collagen, runx2, and bone sialoprotein (BSP) was detectable on QRT-PCR for all specimens at 10 and 20 weeks. By 20 weeks, the relative gene levels were: type I collagen, ilium \textgreater\textgreater radial \textgreater/= cranial \textgreater/= mandibular; type II collagen, radial \textgreater ilium \textgreater cranial \textgreater/= mandibular; runx2, cranial \textgreater\textgreater\textgreater radial \textgreater mandibular \textgreater/= ilium; and BSP, ilium \textgreater/= radial \textgreater cranial \textgreater mandibular. These data demonstrate that the osteogenic and chondrogenic capacity of the various constructs is not identical and depends on the periosteal source regardless of intramembranous or endochondral ossification. Based on these results, cranial and mandibular periosteal tissues appear to enhance bone formation most and least prominently, respectively. The appropriate periosteal choice for bone and cartilage tissue engineering and regeneration should be a function of its immediate application as well as other factors besides growth rate.
Matsushima Seika; Isogai Noritaka; Jacquet Robin; Lowder Elizabeth; Tokui Taku; Landis William J
Cells, tissues, organs
2011
2011
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1159/000324642" target="_blank" rel="noreferrer noopener">10.1159/000324642</a>
Mineral deposition in the extracellular matrices of vertebrate tissues: identification of possible apatite nucleation sites on type I collagen.
*Calcification; Animals; Apatites/*metabolism; Calcium/metabolism; Collagen Type I/chemistry/*metabolism; Extracellular Matrix/*metabolism; Humans; Microfibrils/chemistry; Models; Molecular; Phosphates/metabolism; Physiologic; Vertebrates/*metabolism
The possible means by which type I collagen may mediate mineralization in normal vertebrate bone, tendon, dentin and cementum as well as in pathological mineral formation are not fully understood. One consideration in this regard is that the structure of the protein is somehow important in binding calcium and phosphate ions in a stereochemical configuration conducive to nucleation of apatite crystals. In the present study, type I collagen, packed in a quarter-staggered arrangement in two dimensions and a quasi-hexagonal model of microfibrillar assembly in three dimensions, has been examined in terms of several of its charged amino acid residues. These included glutamic and aspartic acid, lysine, arginine, hydroxylysine and histidine, whose positions along the three alpha-chain axes of the collagen molecule were determined with respect to each other. It was found that the locations of these residues specified sites uniquely suited as potential apatite nucleation centers following binding of calcium and phosphate ions. From this analysis, it would appear that type I collagen provides a template of charged amino acid residues that dictates ion binding critical to subsequent nucleation events for mineral formation in vertebrate tissues.
Landis William J; Silver Frederick H
Cells, tissues, organs
2009
2009
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1159/000151454" target="_blank" rel="noreferrer noopener">10.1159/000151454</a>
Tissue engineering models of human digits: effect of periosteum on growth plate cartilage development.
*Models; *Tissue Engineering; Animals; Articular/*growth & development; Biological; Cartilage; Cattle; Experimental; Fingers/diagnostic imaging; Growth Plate/cytology/*growth & development; Humans; Implants; Male; Mice; Nude; Periosteum/*physiology; Radiography
Tissue-engineered middle phalanx constructs of human digits were investigated to determine whether periosteum wrapped partly about model midshafts mediated cartilage growth plate formation. Models were fabricated by suturing ends of polymer midshafts in a human middle phalanx shape with polymer sheets seeded with heterogeneous chondrocyte populations from bovine articular cartilage. Half of each midshaft length was wrapped with bovine periosteum. Constructs were cultured, implanted in nude mice for up to 20 weeks, harvested and treated histologically to assess morphology and cartilage proteoglycans. After 20 weeks of implantation, chondrocyte-seeded sheets adjacent to periosteum-wrapped midshaft halves established cartilage growth plates resembling normal tissue in vivo. Sheets adjacent to midshafts without periosteum had disorganized cells and no plate formation. Proteoglycans were present at both midshaft ends. Periosteum appears to guide chondrocytes toward growth plate cartilage organization and tissue engineering provides means for carefully examining construct development of this tissue.
Landis William J; Jacquet Robin; Lowder Elizabeth; Enjo Mitsuhiro; Wada Yoshitaka; Isogai Noritaka
Cells, tissues, organs
2009
2009
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1159/000151432" target="_blank" rel="noreferrer noopener">10.1159/000151432</a>
Development of bone and cartilage in tissue-engineered human middle phalanx models.
*Models; Aggrecans/genetics/metabolism; Animals; Biological; Bone Development/drug effects/*physiology; Calcium Phosphates/pharmacology; Cartilage/cytology/drug effects/*growth & development; Cattle; Chondrocytes/cytology/drug effects/metabolism; Collagen Type II/genetics/metabolism; Durapatite/pharmacology; Electron; Experimental; Finger Phalanges/cytology/diagnostic imaging/drug effects/*physiology; Gene Expression Profiling; Gene Expression Regulation/drug effects; Humans; Implants; Integrin-Binding Sialoprotein; Mice; Microscopy; Paraffin Embedding; Periosteum/cytology/drug effects; Polyesters/pharmacology; Radiography; Scanning; Sialoglycoproteins/genetics/metabolism; Tissue Engineering/*methods; Tissue Scaffolds/chemistry
Human middle phalanges were tissue-engineered with midshaft scaffolds of poly(L-lactide-epsilon-caprolactone) [P(LA-CL)], hydroxyapatite-P(LA-CL), or beta-tricalcium phosphate-P(LA-CL) and end plate scaffolds of bovine chondrocyte-seeded polyglycolic acid. Midshafts were either wrapped with bovine periosteum or left uncovered. Constructs implanted in nude mice for up to 20 weeks were examined for cartilage and bone development as well as gene expression and protein secretion, which are important in extracellular matrix (ECM) formation and mineralization. Harvested 10- and 20-week constructs without periosteum maintained end plate cartilage but no growth plate formation. They also consisted of chondrocytes secreting type II collagen and proteoglycan, and they were composed of midshaft regions devoid of bone. In all periosteum-wrapped constructs at like times, end plate scaffolds held chondrocytes elaborating type II collagen and proteoglycan and cartilage growth plates resembling normal tissue. Chondrocyte gene expression of type II collagen, aggrecan, and bone sialoprotein varied depending on midshaft composition, presence of periosteum, and length of implantation time. Periosteum produced additional cells, ECM, and mineral formation within the different midshaft scaffolds. Periosteum thus induces midshaft development and mediates chondrocyte gene expression and growth plate formation in cartilage regions of phalanges. This work is important for understanding developmental principles of tissue-engineered phalanges and by extension those of normal growth plate cartilage and bone.
Wada Yoshitaka; Enjo Mitsuhiro; Isogai Noritaka; Jacquet Robin; Lowder Elizabeth; Landis William J
Tissue engineering. Part A
2009
2009-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1089/ten.TEA.2009.0078" target="_blank" rel="noreferrer noopener">10.1089/ten.TEA.2009.0078</a>
Histochemical analyses of tissue-engineered human menisci.
*Tissue Scaffolds; Animals; Cell Differentiation/physiology; Cell Proliferation; Cell Shape/physiology; Cells; Chondrocytes/*cytology/*metabolism/transplantation; Chondrogenesis/physiology; Collagen/metabolism; Cultured; Fibrocartilage/cytology/metabolism/transplantation; Graft Survival/physiology; Heterologous/methods; Histocytochemistry; Humans; Male; Menisci; Mice; Middle Aged; Nude; Phenotype; Polyglycolic Acid/pharmacology/therapeutic use; Tibial/*cytology/*metabolism/transplantation; Tissue Engineering/*methods; Transplantation
The field of tissue engineering remains one of the least explored areas of current meniscal research but holds great promise. In this investigation, meniscal fibrochondrocytes were isolated from fresh human meniscal tissue and seeded onto synthetic polyglycolic acid (PGA) scaffolds. Constructs were implanted into the dorsal subcutaneous space of athymic nude mice. Control scaffolds, devoid of meniscal cells, were simultaneously implanted in additional mice. Constructs were harvested over 12 weeks and treated with a variety of histochemical stains to analyze general specimen morphology, cellular viability and proliferation, and collagen secretion. Results indicate that meniscal fibrochondrocyte proliferation increased over the time of implantation with cellular consolidation occurring as the PGA scaffolding was progressively hydrolyzed. Collagen production also increased over time. There were favorable similarities between constructs and human meniscal controls in terms of cellular morphology, phenotypic expression, and collagen production. These initial findings demonstrate procedures supporting proliferation of meniscal fibrochondrocytes, expression of fibrochondral phenotype, and the formation of putative meniscal tissue.
Schoenfeld Andrew J; Jacquet Robin; Lowder Elizabeth; Doherty Alison; Leeson Mark C; Landis William J
Connective tissue research
2009
1905-7
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1080/03008200902721887" target="_blank" rel="noreferrer noopener">10.1080/03008200902721887</a>
Analysis of osteopontin in mouse growth plate cartilage by application of laser capture microdissection and RT-PCR.
*Reverse Transcriptase Polymerase Chain Reaction; Animals; Articular/chemistry/cytology; Cartilage; Chondrocytes/chemistry; Gene Expression; Growth Plate/*chemistry/cytology; Inbred C57BL; Laser Therapy; Messenger/metabolism; Mice; Microdissection/*methods; Newborn; Non-programmatic; Osteopontin; RNA; Sialoglycoproteins/*analysis/genetics; Tibia
Gene expression of osteopontin (OPN) has been investigated in mice by application of laser capture microdissection (LCM) and reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. LCM permits individual cells to be isolated ("captured") from tissue sections for molecular analyses. In this study, chondrocytes were captured from growth plate zones in frozen sections of tibiae from 1-11-day-old postnatal mice. RNA was extracted from cells, DNAse-treated, and reverse-transcribed. cDNA was amplified by PCR and OPN mRNA was revealed on agarose gels. Whole cartilage and brain (a positive control) from the same animals also were examined. Reactions containing no RT were negative controls, and 18S rRNA standardized expressed message from captured cells. RT-PCR analysis of laser-captured whole cartilage showed a general qualitative loss of OPN mRNA as animal age increased. Youngest mice gave equivalent OPN expression over all laser-microdissected cartilage zones. For 7-11 day-old mice, OPN expression was qualitatively greatest in resting and lowest in hypertrophic regions of cartilage. Expression of OPN correlated with mineral in the tissue suggests that OPN functionally may inhibit normal vertebrate growth plate mineralization, and its loss with increasing tissue maturation appears permissive to mineral development.
Landis William J; Jacquet Robin; Hillyer Jennifer; Zhang Jean
Connective tissue research
2003
1905-06
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1080/03008200390152052" target="_blank" rel="noreferrer noopener">10.1080/03008200390152052</a>
The structure and function of normally mineralizing avian tendons.
Animals; Biomechanical Phenomena; Birds/*anatomy & histology/*physiology; Minerals/*metabolism; Non-programmatic; Tendons/anatomy & histology/*cytology/*physiology
The leg tendons of certain avian species normally calcify. The gastrocnemius, or Achilles, tendon of the domestic turkey, Meleagris gallopavo, is one such example. Its structure and biomechanical properties have been studied to model the adaptive nature of this tendon to external forces, including the means by which mineral deposition occurs and the functional role mineralization may play in this tissue. Structurally, the distal rounded, thick gastrocnemius bifurcates into two smaller proximal segments that mineralize with time. Mineral deposition occurs at or near the bifurcation, proceeding in a distal-to-proximal direction along the segments toward caudal and medial muscle insertions of the bird hip. Mineral formation appears mediated first by extracellular matrix vesicles and later by type I collagen fibrils. Biomechanical analyses indicate lower tensile strength and moduli for the thick distal gastrocnemius compared to narrow, fan-shaped proximal segments. Tendon mineralization here appears to be strain-induced, the muscle forces causing matrix deformation leading conceptually to calcium binding through the exposure of charged groups on collagen, release of sequestered calcium by proteoglycans, and increased diffusion. Functionally, the mineralized tendons limit further tendon deformation, reduce tendon strain at a given stress, and provide greater load-bearing capacity to the tissue. They also serve as important and efficient elastic energy storage reservoirs, increasing the amount of stored elastic energy by preventing flexible type I collagen regions from stretching and preserving muscle energy during locomotion of the animals.
Landis William J; Silver Frederick H
Comparative biochemistry and physiology. Part A, Molecular & integrative physiology
2002
2002-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/s1095-6433(02)00248-9" target="_blank" rel="noreferrer noopener">10.1016/s1095-6433(02)00248-9</a>
The potential of tissue engineering in orthopedics.
*Artificial Limbs; *Bioprosthesis; *Orthopedics; Animals; Cartilage/growth & development; Finger Joint/growth & development; Fingers/growth & development; Humans; Mice; Periosteum/growth & development; Tendons/growth & development; Tissue Engineering/*methods
This article presents models of human phalanges and small joints developed by tissue engineering. Biodegradable polymer scaffolds support growth of osteoblasts, chondrocytes, and tenocytes after implantation of the models in athymic mice. The cell-polymer constructs are vascularized by the host mice, form new bone, cartilage, and tendon with characteristic gene expression and protein synthesis and secretion, and maintain the shape of human phalanges with joints. The study demonstrates critical progress in the design and fabrication of bone, cartilage, and tendon by tissue engineering and the potential of this field for human clinical orthopedic applications.
Landis William J; Jacquet Robin; Hillyer Jennifer; Zhang Jean; Siperko Lorraine; Chubinskaya Susan; Asamura Shinichi; Isogai Noritaka
The Orthopedic clinics of North America
2005
2005-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ocl.2004.06.006" target="_blank" rel="noreferrer noopener">10.1016/j.ocl.2004.06.006</a>
Effects of FGF-2 and OP-1 in vitro on donor source cartilage for auricular reconstruction tissue engineering.
Bone Morphogenetic Protein 7/*pharmacology; Cell Differentiation/drug effects; Cells; Child; Chondrocytes; Chondrocytes/*drug effects/physiology; Congenital Abnormalities/diagnosis/*surgery; Congenital Microtia; Cultured; Ear Cartilage/surgery; Ear/*abnormalities/surgery; FGF-2; Fibroblast Growth Factor 2/*pharmacology; Humans; Male; Microtia; OP-1; Reconstructive Surgical Procedures/*methods; Reference Values; Tissue Donors; Tissue engineering; Tissue Engineering/*methods
OBJECTIVE: Microtia is a congenital partial or total loss of the external ear with current treatment approaches involving autologous construction from costal cartilage. Alternatively, tissue engineering provides possible use of normal or microtia auricular chondrocytes harvested from patients. This study investigated effects in vitro of basic fibroblast growth factor (FGF-2) and osteogenic protein 1 (OP-1) on human pediatric normal and microtia auricular chondrocytes and their potential proliferation and differentiation for cellular expansion. A working hypothesis was that FGF-2 promotes proliferation and OP-1 maintains an auricular phenotype of these cells. METHODS: Two patients, one undergoing otoplasty and one an ear construction, yielded normal and microtia auricular chondrocytes, respectively. The two donor sets of isolated chondrocytes were equally divided into four experimental cell groups. These were controls without added growth factors and cells supplemented with FGF-2, OP-1 or FGF-2/OP-1 combined. Cells were cultured 3, 5, 7, and 10 days (3 replicates/time point), counted and assayed by RT-qPCR to determine elastin and types II and III collagen gene expression. RESULTS: Compared to control counterparts, normal and microtia chondrocytes with
Shasti Mark; Jacquet Robin; McClellan Phillip; Yang Julianne; Matsushima Seika; Isogai Noritaka; Murthy Ananth; Landis William J
International journal of pediatric otorhinolaryngology
2014
2014-03
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ijporl.2013.11.028" target="_blank" rel="noreferrer noopener">10.1016/j.ijporl.2013.11.028</a>
Analysis of connective tissues by laser capture microdissection and reverse transcriptase-polymerase chain reaction.
*Microdissection; Animals; Cell Separation/*methods; Chondrocytes/chemistry/cytology; Connective Tissue Cells/chemistry/*cytology; Gene Expression Profiling/*methods; Growth Plate/cytology; Inbred Strains; Lasers; Messenger/analysis/*isolation & purification; Methods; Mice; Newborn; Reverse Transcriptase Polymerase Chain Reaction; RNA; Tibia/cytology
Studies of gene expression from bone, cartilage, and other tissues are complicated by the fact that their RNA, collected and pooled for analysis, often represents a wide variety of composite cells distinct in individual phenotype, age, and state of maturation. Laser capture microdissection (LCM) is a technique that allows specific cells to be isolated according to their phenotype, condition, or other marker from within such heterogeneity. As a result, this approach can yield RNA that is particular to a subset of cells comprising the total cell population of the tissue. This study reports the application of LCM to the gene expression analysis of the cartilaginous epiphyseal growth plate of normal newborn mice. The methodology utilized for this purpose has been coupled with real-time quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) to quantitate the expression of certain genes involved in growth plate development and calcification. In this paper, the approaches used for isolating and purifying RNA from phenotypically specific chondrocyte populations of the murine growth plate are detailed and illustrate and compare both qualitative and quantitative RT-PCR results. The technique will hopefully serve as a guide for the further analysis of this and other connective tissues by LCM and RT-PCR.
Jacquet Robin; Hillyer Jennifer; Landis William J
Analytical biochemistry
2005
2005-02
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.ab.2004.09.033" target="_blank" rel="noreferrer noopener">10.1016/j.ab.2004.09.033</a>
Murine metapodophalangeal sesamoid bones: morphology and potential means of mineralization underlying function.
Aging/physiology; Animal Structures; Animals; Biomechanical Phenomena; Bone Development/*physiology; Calcification; Collagen Type II/metabolism; Electron; Extracellular Matrix/metabolism/ultrastructure; Fibrocartilage/physiology/ultrastructure; Forelimb/*anatomy & histology/diagnostic imaging/growth & development; Hindlimb/*anatomy & histology/diagnostic imaging/growth & development; Mice; Microscopy; Movement/physiology; Muscle; Physiologic/*physiology; Proteoglycans/metabolism; Radiography; Sesamoid Bones/*cytology/diagnostic imaging/growth & development; Skeletal/anatomy & histology/physiology; Species Specificity; Tendons/physiology/ultrastructure; Transmission
Normal murine metapodophalangeal sesamoid bones, closely associated with tendons, were examined in terms of their structure and mineralization with reference to their potential function following crystal deposition. This study utilized radiography, whole mount staining, histology, and conventional electron microscopy to establish a maturation timeline of mineral formation in 1- to
Doherty Alison H; Lowder Elizabeth M; Jacquet Robin D; Landis William J
Anatomical record (Hoboken, N.J. : 2007)
2010
2010-05
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1002/ar.21095" target="_blank" rel="noreferrer noopener">10.1002/ar.21095</a>