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1.
J Mech Behav Biomed Mater ; 114: 104233, 2021 02.
Article in English | MEDLINE | ID: mdl-33302168

ABSTRACT

Total hip replacement is the most effective treatment for late stage osteoarthritis. However, adverse local tissue reactions (ALTRs) have been observed in patients with modular total hip implants. Although the detailed mechanisms of ALTRs are still unknown, fretting corrosion and the associated metal ion release from the CoCrMo femoral head at the modular junction has been reported to be a major factor. The purpose of this study is to increase the fretting corrosion resistance of the CoCrMo alloy and the associated metal ion release by applying hard coatings to the surface. Cathodic arc evaporation technique (arc-PVD) was used to deposit TiSiN and ZrN hard coatings on CoCrMo substrates. The morphology, chemical composition, crystal structures and residual stress of the coatings were characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy, and X-ray diffractometry. Hardness, elastic modulus, and adhesion of the coatings were measured by nano-indentation, nano-scratch test, and the Rockwell C test. Fretting corrosion resistance tests of coated and uncoated CoCrMo discs against Ti6Al4V spheres were conducted on a four-station fretting testing machine in simulated body fluid at 1Hz for 1 million cycles. Post-fretting samples were analyzed for morphological changes, volume loss and metal ion release. Our analyses showed better surface finish and lower residual stress for ZrN coating, but higher hardness and better scratch resistance for TiSiN coating. Fretting results demonstrated substantial improvement in fretting corrosion resistance of CoCrMo with both coatings. ZrN and TiSiN decreased fretting volume loss by more than 10 times and 1000 times, respectively. Both coatings showed close to 90% decrease of Co ion release during fretting corrosion tests. Our results suggest that hard coating deposition on CoCrMo alloy can significantly improve its fretting corrosion resistance and could thus potentially alleviate ALTRs in metal hip implants.


Subject(s)
Alloys , Hip Prosthesis , Corrosion , Humans , Materials Testing , Surface Properties
2.
Acta Biomater ; 62: 352-361, 2017 10 15.
Article in English | MEDLINE | ID: mdl-28818689

ABSTRACT

Integration of in vitro-formed cartilage on a suitable substrate to form tissue-engineered implants for osteochondral defect repair is a considerable challenge. In healthy cartilage, a zone of calcified cartilage (ZCC) acts as an intermediary for mechanical force transfer from soft to hard tissue, as well as an effective interlocking structure to better resist interfacial shear forces. We have developed biphasic constructs that consist of scaffold-free cartilage tissue grown in vitro on, and interdigitated with, porous calcium polyphosphate (CPP) substrates. However, as CPP degrades, it releases inorganic polyphosphates (polyP) that can inhibit local mineralization, thereby preventing the formation of a ZCC at the interface. Thus, we hypothesize that coating CPP substrate with a layer of hydroxyapatite (HA) might prevent or limit this polyP release. To investigate this we tested both inorganic or organic sol-gel processing methods, asa barrier coating on CPP substrate to inhibit polyP release. Both types of coating supported the formation of ZCC in direct contact with the substrate, however the ZCC appeared more continuous in the tissue formed on the organic HA sol gel coated CPP. Tissues formed on coated substrates accumulated comparable quantities of extracellular matrix and mineral, but tissues formed on organic sol-gel (OSG)-coated substrates accumulated less polyP than tissues formed on inorganic sol-gel (ISG)-coated substrates. Constructs formed with OSG-coated CPP substrates had greater interfacial shear strength than those formed with ISG-coated and non-coated substrates. These results suggest that the OSG coating method can modify the location and distribution of ZCC and can be used to improve the mechanical integrity of tissue-engineered constructs formed on porous CPP substrates. STATEMENT OF SIGNIFICANCE: Articular cartilage interfaces with bone through a zone of calcified cartilage. This study describes a method to generate an "osteochondral-like" implant that mimics this organization using isolated deep zone cartilage cells and a sol-gel hydroxyapatite coated bone substitute material composed of calcium polyphosphate (CPP). Developing a layer of calcified cartilage at the interface should contribute to enhancing the success of this "osteochondral-like" construct following implantation to repair cartilage defects.


Subject(s)
Cartilage , Durapatite , Materials Testing , Membranes, Artificial , Polyphosphates , Tissue Engineering/methods , Animals , Cartilage/injuries , Cartilage/metabolism , Cartilage/pathology , Cattle , Durapatite/chemistry , Durapatite/pharmacology , Extracellular Matrix/metabolism , Extracellular Matrix/pathology , Phase Transition , Polyphosphates/chemistry , Polyphosphates/pharmacology , Porosity
3.
J Biomed Mater Res B Appl Biomater ; 105(4): 874-884, 2017 05.
Article in English | MEDLINE | ID: mdl-26833448

ABSTRACT

This study investigates the characteristics of porous calcium polyphosphate particulates (CPPp) formed using two different processing treatments as bone void fillers in non- or minimally load-bearing sites. The two calcium polyphosphate particulate variants (grades) were formed using different annealing conditions during particulate preparation to yield either more slowly degrading calcium polyphosphate particulates (SD-CPPp) or faster degrading particulates (FD-CPPp) as suggested by a previous degradation study conducted in vitro (Hu et al., Submitted for publication 2016). The two CPPp grades were compared as bone void fillers in vivo by implanting particulates in defects created in rabbit femoral condyle sites (critical size defects). The SD-CPPp and FD-CPPp were implanted for 4- and 16-week periods. The in vivo study indicated a significant difference in amount of new bone formed in the prepared sites with SD-CPPp resulting in more new bone formation compared with FD-CPPp. The lower bone formation characteristic of the FD-CPPp was attributed to its faster degradation rate and resulting higher local concentration of released polyphosphate degradation products. The study results indicate the importance of processing conditions on preparing calcium polyphosphate particulates for potential use as bone void fillers in nonload-bearing sites. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 874-884, 2017.


Subject(s)
Bone Substitutes , Femur , Osteogenesis/drug effects , Polyphosphates , Animals , Bone Substitutes/chemistry , Bone Substitutes/pharmacology , Femur/injuries , Femur/metabolism , Femur/pathology , Polyphosphates/chemistry , Polyphosphates/pharmacology , Rabbits
4.
J Biomed Mater Res B Appl Biomater ; 101(6): 972-80, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23529933

ABSTRACT

Porous calcium polyphosphate (CPP) structures with 30 volume percent porosity and made by solid freeform fabrication (SFF) were implanted in rabbit femoral condyle sites for 6-wk periods. Two forms of SFF implants with different stacked layer orientation were made in view of prior studies reporting on anisotropic/orthotropic mechanical properties of structures so formed. In addition, porous CPP implants of equal volume percent porosity made by conventional sintering and machining methods were prepared. Bone ingrowth and in vivo degradation of the three different implant types were compared using back-scattered scanning electron microscopy (BS-SEM) of implant samples and quantitative analysis of the images. The results indicated bone ingrowth with all samples resulting in 30-40% fill of available porosity by bone within the 6-wk period. In the 6-wk in vivo period, approximately 7-9% loss of CPP by degradation had occurred.


Subject(s)
Bone Substitutes/chemistry , Polyphosphates/chemistry , Animals , Calcium Phosphates/chemistry , Femur/pathology , Femur/surgery , Male , Materials Testing , Microscopy, Electron, Scanning , Osseointegration , Porosity , Prostheses and Implants , Rabbits
5.
J Biomed Mater Res B Appl Biomater ; 101(1): 1-8, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23143776

ABSTRACT

Porous calcium polyphosphate (CPP) is being investigated for fabrication of novel biodegradable bone substitutes. In this study, porous CPP implants formed by conventional CPP powder packing and using a two-step sinter/anneal process was used to form 20 and 30 vol % porous samples displaying relatively high strength. These were implanted in rabbit femoral condyle sites to study their ability for secure fixation in prepared sites through bone ingrowth. Porous implants of 20 and 30 vol % porosity and displaying compressive strengths ~80 and 35 MPa, respectively, were used. Bone ingrowth sufficient to allow secure implant fixation was observed by 6 weeks (~19% bone ingrowth per available pore space for the 30 vol % and 13% for the 20 vol % porous implants). The results of the in vivo study suggest the potential usefulness of porous CPP as biodegradable bone substitutes/augments in high load-bearing skeletal regions.


Subject(s)
Bone Substitutes , Calcium Phosphates , Animals , Microscopy, Electron, Scanning , Rabbits
6.
Tissue Eng Part A ; 18(11-12): 1282-92, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22429075

ABSTRACT

Clinical utilization of tissue-engineered cartilage constructs has been limited by their inferior mechanical properties compared to native articular cartilage. A number of strategies have been investigated to increase the accumulation of major extracellular matrix components within in vitro-formed cartilage, including the administration of growth factors and mechanical stimulation. In this study, the anabolic effect of inorganic polyphosphates, a linear polymer of orthophosphate residues linked by phosphoanhydride bonds, was demonstrated in both chondrocyte cultures and native articular cartilage cultured ex vivo. Compared to untreated controls, polyphosphate treatment of three-dimensional primary chondrocyte cultures induced increased glycosaminoglycan and collagen accumulation in a concentration- and chain length-dependent manner. This effect was transient, because chondrocytes express exopolyphosphatases that hydrolyze polyphosphate. The anabolic effect of polyphosphates was accompanied by a lower rate of DNA increase within the chondrocyte cultures treated with inorganic polyphosphate. Inorganic polyphosphate enhances cartilage matrix accumulation and is a promising approach to improve the quality of tissue-engineered cartilage constructs.


Subject(s)
Cartilage, Articular/drug effects , Cartilage, Articular/physiology , Chondrogenesis/drug effects , Polyphosphates/pharmacology , Tissue Engineering , Acid Anhydride Hydrolases/metabolism , Animals , Cattle , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrogenesis/genetics , DNA/metabolism , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Gene Expression Regulation/drug effects , Polyphosphates/administration & dosage , Time Factors
7.
Acta Biomater ; 8(4): 1603-15, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22222151

ABSTRACT

A major challenge for cartilage tissue engineering remains the proper integration of constructs with surrounding tissues in the joint. Biphasic osteochondral constructs that can be anchored in a joint through bone ingrowth partially address this requirement. In this study, a methodology was devised to generate a cell-mediated zone of calcified cartilage (ZCC) between the in vitro-formed cartilage and a porous calcium polyphosphate (CPP) bone substitute in an attempt to improve the mechanical integrity of that interface. To do so, a calcium phosphate (CaP) film was deposited on CPP by a sol-gel process to prevent the accumulation of polyphosphates and associated inhibition of mineralization as the substrate degrades. Cartilage formed in vitro on the top surface of CaP-coated CPP by deep-zone chondrocytes was histologically and biochemically comparable to that formed on uncoated CPP. Furthermore, the mineral in the ZCC was similar in crystal structure, morphology and length to that formed on uncoated CPP and native articular cartilage. The generation of a ZCC at the cartilage-CPP interface led to a 3.3-fold increase in the interfacial shear strength of biphasic constructs. Improved interfacial strength of these constructs may be critical to their clinical success for the repair of large cartilage defects.


Subject(s)
Calcification, Physiologic/drug effects , Calcium Phosphates/pharmacology , Cartilage/drug effects , Cartilage/physiology , Chondrogenesis/drug effects , Polyphosphates/pharmacology , Shear Strength/drug effects , Animals , Bone Substitutes/pharmacology , Cartilage/cytology , Cartilage/ultrastructure , Cattle , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/drug effects , Chondrocytes/metabolism , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Gene Expression Regulation/drug effects , Microscopy, Electron, Scanning , Solutions
8.
Acta Biomater ; 7(4): 1788-96, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21185409

ABSTRACT

This study addresses the mechanical properties of calcium polyphosphate (CPP) structures formed by stacked layers using a powder-based solid freeform fabrication (SFF) technique. The mechanical properties of the 35% porous structures were characterized by uniaxial compression testing for compressive strength determination and diametral compression testing to determine tensile strength. Fracture cleavage surfaces were analyzed using scanning electron microscopy. The effects of the fabrication process on the microarchitecture of the CPP samples were also investigated. Results suggest that the orientation of the stacked layers has a substantial influence on the mechanical behavior of the SFF-made CPP samples. The samples with layers stacked parallel to the mechanical compressive load are 48% stronger than those with the layers stacked perpendicular to the load. However, the samples with different stacking orientations are not significantly different in tensile strength. The observed anisotropic mechanical properties were analyzed based on the physical microstructural properties of the CPP structures.


Subject(s)
Calcium Phosphates/chemistry , Mechanical Phenomena , Polyphosphates/chemistry , Anisotropy , Compressive Strength , Materials Testing/methods , Microscopy, Electron, Scanning , Particle Size , Porosity , Powders , Surface Properties , Tensile Strength , X-Ray Microtomography
9.
J Biomed Mater Res B Appl Biomater ; 94(1): 178-86, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20524193

ABSTRACT

The formation of biodegradable implants for use in osteosynthesis has been a major goal of biomaterials research for the past 2-3 decades. Self-reinforced polylactide systems represent the most significant success of this research to date, however, with elastic constants up to 12-15 GPa at best, they fail to provide the initial stiffness required of devices for stabilizing fractures of major load-bearing bones. Our research has investigated the use of calcium polyphosphate (CPP), an inorganic polymer in combination with polyvinyl-urethane carbonate (PVUC) organic polymers for such applications. Initial studies indicated that composite samples formed as interpenetrating phase composites (IPC) exhibited suitable as-made strength and stiffness, however, they displayed a rapid loss of properties when exposed to in vitro aging. An investigation to determine the mechanism of this accelerated in vitro degradation for the IPCs as well as to identify possible design changes to overcome this drawback was undertaken using a model IPC system. It was found that strong interfacial strength and minimal swelling of the PVUC are very important for obtaining and maintaining appropriate mechanical properties in vitro.


Subject(s)
Calcium Phosphates/chemistry , Carbonates , Polyphosphates , Polyvinyls , Urethane , Absorbable Implants , Biocompatible Materials/chemistry , Biocompatible Materials/metabolism , Calcium Phosphates/metabolism , Carbonates/chemistry , Carbonates/metabolism , Elasticity , Fracture Fixation, Internal/methods , Humans , Materials Testing , Polyphosphates/chemistry , Polyphosphates/metabolism , Polyvinyls/chemistry , Polyvinyls/metabolism , Resin Cements/chemistry , Resin Cements/metabolism , Shear Strength , Stress, Mechanical , Tensile Strength , Urethane/chemistry , Urethane/metabolism , Weight-Bearing
10.
Acta Biomater ; 6(8): 3302-9, 2010 Aug.
Article in English | MEDLINE | ID: mdl-20188870

ABSTRACT

A major challenge to the successful clinical application of bioengineered cartilage remains its integration to surrounding tissues upon implantation. One way to address this consists of generating biphasic constructs composed of articular cartilage formed in vitro on the top surface and integrated with the porous sub-surface of a bone substitute material - in the case of this study, calcium polyphosphate (CPP). To improve the mechanical integrity of the cartilage-bone substitute interface, attempts have been made to generate a zone of calcified cartilage (ZCC) within the CPP-cartilage interface, thereby mimicking the native joint architecture. The purpose of this work was to establish the effects of the degradation products of CPP on cartilage calcification in order to explain the observed positioning of a ZCC away from the interface junction. It was determined that polyphosphate released from the CPP accumulates within in vitro-grown cartilage and inhibits cartilage calcification in a concentration and chain length (i.e. molecular weight) dependent manner. It was found that this effect is transient as chondrocytes express exopolyphosphatases which hydrolyze polyphosphate to release orthophosphate. Hence, the generation of biphasic constructs with a properly located ZCC will require tailoring of CPP substrates with lower degradation rates or the upregulation of exopolyphosphatases by chondrocytes.


Subject(s)
Bone Substitutes/pharmacology , Calcification, Physiologic/drug effects , Cartilage/drug effects , Cartilage/physiology , Phosphates/pharmacology , Phosphoric Acids/pharmacology , Polymers/pharmacology , Animals , Calcium/metabolism , Cattle , Chondrocytes/cytology , Chondrocytes/drug effects , Phosphoric Acids/chemistry , Polymers/chemistry
11.
J Biomed Mater Res A ; 94(1): 122-9, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20128012

ABSTRACT

We have shown previously that cyclic compression of newly forming bioengineered cartilage in vitro results in improved tissue formation via changes in expression of matrix metalloproteases, such as, MT1-MMP (membrane type metalloprotease), and increased synthesis of matrix molecules. Several studies have suggested an association between MT1-MMP and integrins, which are known to influence cell shape. Thus, the objectives of this study were to determine the effect of compressive mechanical stimulation on cell shape and the role of integrins and MT1-MMP in mediating these changes and influencing matrix accumulation. Bovine articular chondrocytes were grown on the surface of a porous ceramic substrate for 72 h and then cyclically compressed for 30 min. Scanning electron microscopy and morphometric analysis demonstrated that compression induced a rapid, transient increase in chondrocyte spreading by 10 min, followed by a retraction to prestimulated size within 6 h. This was associated with increased accumulation of newly synthesized proteoglycans, as determined by quantification of radioisotope incorporation. Blocking the alpha5beta1 integrin, or its beta1 subunit, inhibited cell spreading and resulted in a partial inhibition of compression-induced increase in matrix accumulation. Knockdown of MT1-MMP expression partially inhibited cell retraction and resulted in a reduced matrix accumulation as well. These results suggest that chondrocyte spreading and retraction following cyclic compression in vitro regulates matrix accumulation. Understanding the mechanisms that regulate chondrocyte mechanotransduction may ultimately lead to the design of improved repair tissue for cartilage damage. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.


Subject(s)
Chondrocytes , Extracellular Matrix/metabolism , Integrin alpha5beta1/metabolism , Mechanotransduction, Cellular/physiology , Animals , Cattle , Cell Shape , Cells, Cultured , Chondrocytes/cytology , Chondrocytes/metabolism , Compressive Strength , Matrix Metalloproteinase 14/metabolism , Proteoglycans/metabolism , Stress, Mechanical
12.
J Biomed Mater Res B Appl Biomater ; 93(2): 510-9, 2010 May.
Article in English | MEDLINE | ID: mdl-20162726

ABSTRACT

Solid freeform fabrication (SFF) enables the fabrication of anatomically shaped porous components required for formation of tissue engineered implants. This article reports on the characterization of a three-dimensional-printing method, as a powder-based SFF technique, to create reproducible porous structures composed of calcium polyphosphate (CPP). CPP powder of 75-150 microm was mixed with 10 wt % polyvinyl alcohol (PVA) polymeric binder, and used in the SFF machine with appropriate settings for powder mesh size. The PVA binder was eliminated during the annealing procedure used to sinter the CPP particles. The porous SFF fabricated components were characterized using scanning electron microscopy, micro-CT scanning, X-ray diffraction, and mercury intrusion porosimetry. In addition, mechanical testing was conducted to determine the compressive strength of the CPP cylinders. The 35 vol % porous structures displayed compressive strength on average of 33.86 MPa, a value 57% higher than CPP of equivalent volume percent porosity made through conventional gravity sintering. Dimensional deviation and shrinkage analysis was conducted to identify anisotropic factors required for dimensional compensation during SFF sample formation and subsequent sintering. Cell culture studies showed that the substrate supported cartilage formation in vitro, which was integrated with the top surface of the porous CPP similar to that observed when chondrocytes were grown on CPP formed by conventional gravity sintering methods as determined histologically and biochemically.


Subject(s)
Biocompatible Materials , Calcium Phosphates , Cartilage/cytology , Chondrocytes/cytology , Tissue Engineering/methods , Tissue Scaffolds , Animals , Cattle , Cells, Cultured , Materials Testing/methods , Polyvinyl Alcohol , Porosity
13.
Spine J ; 10(2): 174-83, 2010 Feb.
Article in English | MEDLINE | ID: mdl-20006559

ABSTRACT

BACKGROUND CONTEXT: Degeneration of the intervertebral disc (IVD) involves structural changes in the annulus fibrosus (AF), which could alter the mechanical forces imposed on the nucleus pulposus (NP) tissue. This could contribute to degenerative changes that occur in the NP. PURPOSE: The purpose of the study was to determine whether circumferential constraint affects anabolic and catabolic gene expression, biochemical composition, and mechanical properties of NP tissue. STUDY DESIGN: Nucleus pulposus cells were isolated from bovine caudal IVD and allowed to form tissue for a period of two weeks. The effect of no, intermediate, or high circumferential constraint on biochemical composition (cellularity and proteoglycan and collagen synthesis), gene expression, and compressive mechanical properties was evaluated. RESULTS: Increasing the rigidity of circumferential constraint surrounding in vitro formed NP tissue resulted in decreased gene expression of aggrecan and type II collagen and increased expression of MMP-1 and ADAMTS-5. This was associated with decreased accumulation of extracellular matrix and a deterioration of the compressive mechanical properties of the tissue. CONCLUSIONS: As increased circumferential constraint can have a significant negative effect on the composition and quality of NP tissue and this raises the possibility that the AF may contribute to the degenerative or age-related alterations that occur in the NP. Further study in a functional spinal unit is required to validate this.


Subject(s)
Intervertebral Disc/metabolism , Stress, Mechanical , ADAM Proteins/genetics , ADAM Proteins/metabolism , Aggrecans/genetics , Aggrecans/metabolism , Analysis of Variance , Animals , Cattle , Cells, Cultured , Collagen Type II/genetics , Collagen Type II/metabolism , Gene Expression , Intervertebral Disc/physiopathology , Matrix Metalloproteinase 1/genetics , Matrix Metalloproteinase 1/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction
14.
Tissue Eng Part A ; 15(12): 3789-98, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19537959

ABSTRACT

The inferior biomechanical properties of in vitro-formed tissue remain a significant obstacle in bioengineering articular cartilage tissue. We have previously shown that cyclic compression (30 minutes, 1 kPa, 1 Hz) of chondrocytes isolated from full-thickness cartilage can induce greater matrix synthesis, although articular cartilage is composed of different subpopulations of chondrocytes, and their individual contribution to enhanced tissue formation has not been fully characterized. This study examines the contribution of chondrocyte subpopulations to this response. Bovine articular chondrocytes were isolated from superficial to mid zones (SMZs) or deep zones (DZs), placed in three-dimensional culture, and subjected to cyclic compression. DZ chondrocytes on calcium polyphosphate substrates formed thicker tissue than those from SMZs. Compression increased matrix accumulation in SMZ chondrocytes while decreasing accumulation in DZ chondrocytes. The SMZ and DZ chondrocytes also differed in their type 1 membrane-bound matrix metalloproteinase (MMP) and MMP-13 expression, enzymes that play a crucial role in mediating the response to mechanical stimulation. In addition, the duration of the culture period was important in determining the DZ response, raising the possibility that matrix accumulation plays a role in the response to stimulation. Understanding the cellular response to mechanical stimulation during tissue formation will facilitate our understanding of tissue growth and allow for further optimization of cartilage tissue formation in vitro.


Subject(s)
Cartilage, Articular/cytology , Stress, Mechanical , Animals , Biomarkers/metabolism , Cattle , Chondrocytes/cytology , Chondrocytes/enzymology , Collagen/genetics , Collagen/metabolism , Compressive Strength , DNA/metabolism , Extracellular Matrix/metabolism , Gene Expression Regulation , Matrix Metalloproteinase 13/genetics , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 14/genetics , Matrix Metalloproteinase 14/metabolism , Proteoglycans/metabolism
15.
Int J Oral Maxillofac Implants ; 23(3): 544-50, 2008.
Article in English | MEDLINE | ID: mdl-18700381

ABSTRACT

PURPOSE: The aim of this study was to obtain histometric measurements of bone and peri-implant mucosal tissue contact with implants of 2 sintered porous-surfaced designs. The "short-collar" design had a collar height (smooth coronal region) of 0.75 mm, while the "long-collar" model had a smooth coronal region of 1.8 mm. MATERIALS AND METHODS: Implants (2 per side) were placed in healed mandibular extraction sites of 4 beagle dogs using a submerged technique. After 4 weeks of healing, they were uncovered and used to support fixed partial dentures for a 9-month period. After sacrifice, specimens were retrieved and nondemineralized sections were examined histometrically to determine the most coronal bone-to-implant contact (first BIC) using the microgap as a reference and standard mucosal parameters of "biologic width." RESULTS: Significant (P = .001) differences in first BIC were found between designs (1.97 mm for long-collar versus 1.16 mm for short-collar implants) for posteriorly located implants but not for anteriorly located ones (1.21 mm versus 1.38 mm; P = .40). If crestal bone loss involved sintered surface, fibrous connective tissue ingrowth was observed to replace lost bone. No significant differences in peri-implant mucosal measurements (total peri-implant mucosal thickness; length of the epithelial component of this mucosa, and thickness of the connective tissue component) were detected between implant designs. CONCLUSIONS: Results suggest that "biologic width" accommodation drives initial crestal bone loss with sintered porous-surfaced implants. Histometric data obtained for bone contact showed no significant differences between the long- and short-collar implant designs.


Subject(s)
Dental Implants/adverse effects , Gingiva/anatomy & histology , Alveolar Bone Loss/etiology , Animals , Bone Remodeling , Dental Implantation, Endosseous , Dental Prosthesis Design , Dogs , Porosity
16.
Biomaterials ; 29(16): 2477-89, 2008 Jun.
Article in English | MEDLINE | ID: mdl-18314188

ABSTRACT

Chondrocyte behaviour has been shown previously to be influenced by the architecture of the substrate on which the cells are grown. Chondrocytes cultured on fully porous titanium alloy substrates showed greater spreading and more matrix accumulation when compared to cells grown on porous-coated substrates with solid bases. We hypothesized that these features developed because of differences in fluid-induced shear stresses due to substrate architecture and that integrins mediate these responses. Computational fluid dynamics analyses predicted that cells on fully porous substrates experience time-dependent shear stresses that differ from those experienced by cells on porous-coated substrates with solid bases where media flow-through is restricted. To validate this model, the seeding protocol was modulated to affect fluid flow and this affected cell spreading and matrix accumulation as predicted. Integrin blocking experiments revealed that alpha5beta1 integrins regulated cell shape under these two conditions and when cell spreading was prevented the increased accumulation of collagen and proteoglycans by chondrocytes seeded on fully porous substrates did not occur. Identifying the substrate-induced mechanical and molecular mechanisms that influence chondrocyte behaviour and tissue formation may ultimately lead to the formation of a tissue that more closely resembles natural articular cartilage.


Subject(s)
Cell Shape , Chondrocytes/cytology , Integrin alpha5beta1/physiology , Shear Strength , Alloys , Animals , Cattle , Cells, Cultured , Chondrocytes/ultrastructure , Collagen/physiology , Extracellular Matrix/physiology , Microscopy, Electron, Scanning , Models, Biological , Porosity , Proteoglycans/physiology , Titanium
17.
Spine (Phila Pa 1976) ; 33(4): 356-65, 2008 Feb 15.
Article in English | MEDLINE | ID: mdl-18277865

ABSTRACT

STUDY DESIGN: In vitro-formed bovine nucleus pulposus (NP) tissues were used as a model for tumor necrosis factor-alpha (TNF-alpha) induced NP degeneration. OBJECTIVE: To elucidate the signal transduction mechanisms regulating TNF-alpha induced matrix metalloproteinase (MMP) activity. SUMMARY OF BACKGROUND DATA: TNF-alpha is thought to contribute to the pathophysiology of intervertebral disc (IVD) degeneration by up-regulating MMPs, such as MMP-2. MMP-2 has been implicated in influencing disease progression and in the induction of neovascularization. METHODS: In vitro-formed bovine NP tissues were treated with TNF-alpha to examine its effect on MMP-2 gene and protein levels and activity. The effect of TNF-alpha on membrane type (MT)1-MMP, an activator of MMP-2, was also assessed. MT1-MMP functional activation by TNF-alpha was confirmed using promoter-reporter luciferase constructs. Immunoblots and electrophoretic mobility shift assays were used to examine the expression and DNA binding activity of transcription factors known to regulate transcriptional activation of MT1-MMP. RESULTS: TNF-alpha treatment induced MMP-2 gelatinase activity, which occurred in the absence of any change in MMP-2 gene or protein expression, but did correlate with increased MT1-MMP mRNA and protein levels. Up-regulation of MMP-2 activity was dependent on the ERK-MAPK pathway. ERK-1/2 activation up-regulated early growth factor (Egr-1) expression and its DNA binding activity to the MT1-MMP promoter. There was no effect on Sp-1 binding activity. Reporter constructs demonstrated that TNF-alpha induced MT1-MMP transcriptional activation and that this response was dependant on ERK MAPK and Egr-1. CONCLUSION: TNF-alpha induced MMP-2 gelatinase activity correlated with induction of MT1-MMP and not MMP-2 expression. MMP-2 activation was dependent on the ERK-MAPK pathway. As ERK also appeared to regulate MT1-MMP production, this suggests that TNF-alpha induction of MMP-2 gelatinase activity may be regulated by MT1-MMP. These findings elucidate the regulation of gelatinase activity and identify a mechanism whereby TNF-alpha may contribute to matrix degradation in NP tissue.


Subject(s)
Intervertebral Disc/enzymology , Matrix Metalloproteinase 14/metabolism , Matrix Metalloproteinase 2/metabolism , Tumor Necrosis Factor-alpha/physiology , Animals , Cattle , Electrophoretic Mobility Shift Assay , Extracellular Signal-Regulated MAP Kinases/physiology , Gene Expression , Immunoblotting , In Vitro Techniques , Luciferases/genetics , Matrix Metalloproteinase 14/genetics , Matrix Metalloproteinase 2/genetics , Reverse Transcriptase Polymerase Chain Reaction , Up-Regulation
18.
Eur Cell Mater ; 13: 66-73; discussion 73-4, 2007 Apr 12.
Article in English | MEDLINE | ID: mdl-17429796

ABSTRACT

The development of tissue engineered cartilage is a promising new approach for the repair of damaged or diseased tissue. Since it has proven difficult to generate cartilaginous tissue with properties similar to that of native articular cartilage, several studies have used mechanical stimuli as a means to improve the quantity and quality of the developed tissue. In this study, we have investigated the effect of multi-axial loading applied during in vitro tissue formation to better reflect the physiological forces that chondrocytes are subjected to in vivo. Dynamic combined compression-shear stimulation (5% compression and 5% shear strain amplitudes) increased both collagen and proteoglycan synthesis (76 +/- 8% and 73 +/- 5%, respectively) over the static (unstimulated) controls. When this multi-axial loading condition was applied to the chondrocyte cultures over a four week period, there were significant improvements in both extracellular matrix (ECM) accumulation and the mechanical properties of the in vitro-formed tissue (3-fold increase in compressive modulus and 1.75-fold increase in shear modulus). Stimulated tissues were also significantly thinner than the static controls (19% reduction) suggesting that there was a degree of ECM consolidation as a result of long-term multi-axial loading. This study demonstrated that stimulation by multi-axial forces can improve the quality of the in vitro-formed tissue, but additional studies are required to further optimize the conditions to favour improved biochemical and mechanical properties of the developed tissue.


Subject(s)
Biomechanical Phenomena/methods , Cartilage/physiology , Chondrocytes/cytology , Tissue Engineering/methods , Animals , Biophysical Phenomena , Biophysics , Cattle , Cells, Cultured , Chondrocytes/metabolism , Collagen/chemistry , Compressive Strength , Culture Techniques , Extracellular Matrix/metabolism , Proteoglycans/chemistry , Stress, Mechanical
19.
Lasers Surg Med ; 39(3): 286-93, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17252579

ABSTRACT

BACKGROUND AND OBJECTIVE: Forming cartilage tissue in vitro that resembles native tissue is one of the challenges of cartilage tissue engineering. The aim of this study was to determine whether low-power laser stimulation would improve the formation of cartilage tissue in vitro. STUDY DESIGN/MATERIALS AND METHODS: Bovine articular chondrocytes were seeded on the top surface of porous calcium polyphosphate substrates. After 2 days, laser stimulation was applied daily at a wavelength of 650 nm using a laser diode with energy densities of either 1.75 or 3 J/cm(2) for 4 weeks. Proteoglycan and collagen synthesis and matrix content were determined. Cartilage tissue morphology was evaluated histologically. RESULTS: Histologically, there was no difference in the appearance or cellularity of the tissues that formed in the presence or absence of laser stimulation at either dosage. There were no differences in DNA content between treated and untreated constructs and live-dead assay confirmed that this treatment was not toxic to the cells. Laser stimulation at 3 J/cm(2) enhanced matrix synthesis resulting in significantly more tissue formation than laser stimulation at 1.75 J/cm(2) or untreated cultures. CONCLUSION: Short exposures to low-power laser stimulation using a laser diode with 3 J/cm(2) dose improves cartilage tissue formation.


Subject(s)
Cartilage, Articular/radiation effects , Lasers , Tissue Engineering , Animals , Biocompatible Materials , Calcium Phosphates , Cartilage, Articular/cytology , Cattle , Cell Survival , Chondrocytes/pathology , Extracellular Matrix/physiology
20.
Int J Oral Maxillofac Implants ; 22(6): 948-54, 2007.
Article in English | MEDLINE | ID: mdl-18271376

ABSTRACT

PURPOSE: The purpose of this study was to compare patterns of crestal bone remodeling with 2 sintered porous-surfaced dental implant designs during a 14-month functional period. MATERIALS AND METHODS: Two root-form press-fit dental implants were evaluated in healed extraction sites in dog mandibles. The standard (control) design was a press-fit implant with a 2-mm machined collar; the remainder of the implant had a sintered porous surface. The test or "hybrid" design had 3 coronal machined threads instead of a machined collar; the remainder of the implant had a sintered porous surface. RESULTS: Standardized radiographs indicated significantly less crestal bone loss (0.82 to 0.93 mm versus 1.45 to 1.5 mm) with the hybrid design and a slower approach toward an apparent steady state (12 to 14 months for the hybrid versus 7 months for the standard design). Morphometric assessment of back-scattered scanning electron micrographs confirmed that crestal bone loss was significantly less for the hybrid design on all but the lingual implant aspect. CONCLUSION: The addition of coronal threads to an implant relying on a sintered porous surface geometry for its long-term osseointegration reduced the extent of crestal bone loss compared to a machined collar region.


Subject(s)
Alveolar Bone Loss/prevention & control , Dental Implantation, Endosseous/adverse effects , Dental Implants, Single-Tooth/adverse effects , Dental Prosthesis Design , Osseointegration , Alveolar Bone Loss/etiology , Animals , Dogs , Follow-Up Studies , Jaw, Edentulous, Partially/rehabilitation , Jaw, Edentulous, Partially/surgery , Male , Mandible/surgery , Mandibular Diseases/etiology , Mandibular Diseases/prevention & control , Pilot Projects , Porosity , Surface Properties
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