Showing papers in "Journal of Biomedical Materials Research in 1998"
TL;DR: The process of bacterial adhesion to biomaterial surfaces is a complicated process affected by many factors, including the characteristics of the bacteria themselves, the target material surface, and the environmental factors, such as the presence of serum proteins or bactericidal substances.
Abstract: This article reviews the mechanisms of bacterial adhesion to biomaterial surfaces and the factors affecting the adhesion The process of bacterial adhesion includes an initial physicochemical interaction phase (phase one) and a late molecular and cellular interaction phase (phase two), which is a complicated process affected by many factors, including the characteristics of the bacteria themselves, the target material surface, and the environmental factors, such as the presence of serum proteins or bactericidal substances
1,090 citations
TL;DR: Photopolymerized crosslinked networks of poly(ethylene glycol; PEG) diacrylate (MW 8000) were derivitized throughout their bulk with Arg-Gly-Asp (RGD)-containing peptide sequences to promote spreading of human foreskin fibroblasts over 24 h.
Abstract: Photopolymerized crosslinked networks of poly(ethylene glycol; PEG) diacrylate (MW 8000) were derivitized throughout their bulk with Arg-Gly-Asp (RGD)-containing peptide sequences. Incorporation was achieved by functionalizing the amine terminus of the peptide with an acrylate moiety, thereby enabling the adhesion peptide to copolymerize rapidly with the PEG diacrylate upon photoinitiation. PEG diacrylate hydrogels derivitized with RGD peptide at surface concentrations ranging from 0.001 to 1 pmol/cm2 were studied in vitro for their ability to promote spreading of human foreskin fibroblasts over 24 h. Hydrogels not derivitized with peptides were poor substrates for adhesion, permitting spreading of only 5% of the seeded cells. When immobilized with no spacer arm, both RGD and RDG (inactive control) supported spreading of approximately 50% and approximately 15% of cells at 1 and 0.1 pmol/cm2 surface concentrations respectively; lower concentrations did not promote spreading. When a MW 3400 PEG spacer arm was incorporated between the hydrogel and the peptide linkage, incorporation of 1 pmol/cm2 RGD promoted 70% spreading whereas RDG at the same concentration did not promote spreading. In addition, when cells were seeded in serum-free medium, only RGD peptides incorporated with a spacer arm were able to promote spreading. Thus peptide incorporated into PEG 8000 diacrylate hydrogels without a spacer arm nonspecifically mediated cell spreading whereas incorporation via a MW 3400 PEG spacer arm was required to permit cell spreading to be specifically mediated.
978 citations
TL;DR: It is concluded that fewer proteins are adsorbed and their original conformation is not changed on polymer surfaces that possess a high free water fraction.
Abstract: The amount of plasma protein adsorbed on a phospholipid polymer having a 2-methacryloyloxyethyl phosphorylcholine (MPC) moiety was reduced compared to the amount of protein adsorbed onto poly[2-hydroxyethyl methacrylate (HEMA)], poly[n-butyl methacrylate (BMA)], and BMA copolymers with acrylamide (AAm) or N-vinyl pyrrolidone (VPy) moieties having a hydrophilic fraction. To clarify the reason for the reduced protein adsorption on the MPC polymer, the water structure in the hydrated polymer was examined with attention to the free water fraction. Hydration of the polymers occurred when they were immersed in water. The differential scanning calorimetric analysis of these hydrated polymers revealed that the free water fractions in the poly(MPC-co-BMA) and poly(MPC-co-n-dodecyl methacrylate) with a 0.30 MPC mole fraction were above 0.70. On the other hand, the free water fractions in the poly(HEMA), poly(AAm-co-BMA), and poly(VPy-co-BMA) were below 0.42. The conformational change in proteins adsorbed on the MPC polymers and poly(HEMA) were determined using ultraviolet and circular dichroism spectroscopic measurements. Proteins adsorbed on poly(HEMA) changed considerably, but those adsorbed on poly(MPC-co-BMA) with a 0.30 MPC mole fraction differed little from the native state. We concluded from these results that fewer proteins are adsorbed and their original conformation is not changed on polymer surfaces that possess a high free water fraction.
950 citations
TL;DR: This novel process, a combination of high pressure gas foaming and particulate leaching techniques, allows one to fabricate matrices with a well controlled porosity and pore structure, and avoids the potential negatives associated with the use of high temperatures and/or organic solvents in biomaterials processing.
Abstract: Engineering tissues utilizing biodegradable polymer matrices is a promising approach to the treatment of a number of diseases. However, processing techniques utilized to fabricate these matrices typically involve organic solvents and/or high temperatures. Here we describe a process for fabricating matrices without the use of organic solvents and/or elevated temperatures. Disks comprised of polymer [e.g., poly (D,L-lactic-co-glycolic acid)] and NaCl particles were compression molded at room temperature and subsequently allowed to equilibrate with high pressure CO2 gas (800 psi). Creation of a thermodynamic instability led to the nucleation and growth of gas pores in the polymer particles, resulting in the expansion of the polymer particles. The polymer particles fused to form a continuous matrix with entrapped salt particles. The NaCl particles subsequently were leached to yield macropores within the polymer matrix. The overall porosity and level of pore connectivity were regulated by the ratio of polymer/salt particles and the size of salt particles. Both the compressive modulus (159+/-130 kPa versus 289+/-25 kPa) and the tensile modulus (334+/-52 kPa versus 1100+/-236 kPa) of the matrices formed with this approach were significantly greater than those formed with a standard solvent casting/particulate leaching process. The utility of these matrices was demonstrated by engineering smooth muscle tissue in vitro with them. This novel process, a combination of high pressure gas foaming and particulate leaching techniques, allows one to fabricate matrices with a well controlled porosity and pore structure. This process avoids the potential negatives associated with the use of high temperatures and/or organic solvents in biomaterials processing.
835 citations
TL;DR: Whether this no-load healing period is validated by the experimental literature is examined, which suggests that there is a critical threshold of micromotion above which fibrous encapsulation prevails over osseointegration, and suggestions are made for the earliest loading time that achieves osseointedegration.
Abstract: A significant no-load healing period is the generally accepted prerequisite for osseointegration in dental implantology. The aim of this article was to examine whether this no-load healing period is validated by the experimental literature. In vivo histological data was scrutinized to identify the effect of early loading protocols on the bone-implant interface. Several loading modes were identified. They were categorized into groups according to implant design and the type of prosthetic reconstruction, and by their ability to introduce a distinct magnitude of motion at the interface. Specific histologic responses of early loaded implants (i.e., fibrous repair or osseointegration) were suggested to be directly related to the specific combinations of the above parameters. Early loading per se was not found to be detrimental to osseointegration. Specifically, only excessive micromotion was directly implicated in the formation of fibrous encapsulation. The literature suggests that there is a critical threshold of micromotion above which fibrous encapsulation prevails over osseointegration. This critical level, however, was not zero micromotion as generally interpreted. Instead, the tolerated micromotion threshold was found to lie somewhere between 50 and 150 microns. Suggestions are made for the earliest loading time that achieves osseointegration.
754 citations
TL;DR: Results are consistent with earlier studies on SLA implants and suggest that this surface promotes greater osseous contact at earlier time points compared to TPS-coated implants.
Abstract: Many dental clinical implant studies have fo- cused on the success of endosseous implants with a variety of surface characteristics. Most of the surface alterations have been aimed at achieving greater bone-to-implant con- tact as determined histometrically at the light microscopic level. A previous investigation in non-oral bone under short- term healing periods (3 and 6 weeks) indicated that a sand- blasted and acid-etched titanium (SLA) implant had a greater bone-to-implant contact than did a comparably- shaped implant with a titanium plasma-sprayed (TPS) sur- face. In this canine mandible study, nonsubmerged implants with a SLA surface were compared to TPS-coated implants under loaded and nonloaded conditions for up to 15 months. Six foxhound dogs had 69 implants placed in an alternating pattern with six implants placed bilaterally in each dog. Gold crowns that mimicked the natural occlusion were fabricated for four dogs. Histometric analysis of bone contact with the implants was made for two dogs after 3 months of healing (unloaded group), 6 months of healing (3 months loaded), and after 15 months of healing (12 months loaded). The SLA implants had a significantly higher (p < 0.001) percentage of bone-to-implant contact than did the TPS implants after 3 months of healing (72.33 ± 7.16 versus 52.15 ± 9.19; mean ± SD). After 3 months of loading (6 months of healing) no significant difference was found be- tween the SLA and TPS surfaced implants (68.21 ± 10.44 and 78.18 ± 6.81, respectively). After 12 months of loading (15 months of healing) the SLA implants had a significantly greater percentage (p < 0.001) of bone-to-implant contact than did the TPS implants (71.68 ± 6.64 and 58.88 ± 4.62, respectively). No qualitative differences in bone tissue were observed between the two groups of implants nor was there any difference between the implants at the clinical level. These results are consistent with earlier studies on SLA im- plants and suggest that this surface promotes greater osse- ous contact at earlier time points compared to TPS-coated implants. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 1-11, 1998.
655 citations
TL;DR: Preadsorption of the model surfaces with bovine serum albumin resulted in a pattern of cell attachment very similar to that observed following preadsorption with dilute serum, suggesting an important role for BSA in regulating cell attachment to biomaterials exposed to complex biological media.
Abstract: Understanding the relationships between material surface properties, adsorbed proteins, and cellular responses is essential to designing optimal material surfaces for implantation and tissue engineering. In this study, we have prepared model surfaces with different functional groups to provide a range of surface wettability and charge. The cellular responses of attachment, spreading, and cytoskeletal organization have been studied following preadsorption of these surfaces with dilute serum, specific serum proteins, and individual components of the extracellular matrix. When preadsorbed with dilute serum, cell attachment, spreading, and cytoskeletal organization were significantly greater on hydrophilic surfaces relative to hydrophobic surfaces. Among the hydrophilic surfaces, differences in charge and wettability influenced cell attachment but not cell area, shape, or cytoskeletal organization. Moderately hydrophilic surfaces (20-40 degree water contact angle) promoted the highest levels of cell attachment. Preadsorption of the model surfaces with bovine serum albumin (BSA) resulted in a pattern of cell attachment very similar to that observed following preadsorption with dilute serum, suggesting an important role for BSA in regulating cell attachment to biomaterials exposed to complex biological media.
617 citations
TL;DR: Long-term aging in water caused a reduction in the KIc, independent of composition, but had little effect on other properties, suggesting limited degradation of composites in water.
Abstract: The purpose of this study was to evaluate the long-term effect of aging in water on the physical properties of experimental composites having systematically controlled differences in degree of conversion (DC), filler volume fraction (Vf), and percentage of silane-treated fillers. Composites were made with a 50% Bis-GMA:50% TEGDMA light-cured resin and a 1-2 microm (average size) strontium glass filler (+ 5 wt% SiO2 microfiller). For composites A-E, the DC was varied from 56-66% by changing the curing time; for D and F-I, the Vf was varied from 28-62 vol%; and for D and J-M, the percent of fillers with a silane coupling agent (gamma-MPS) was varied from 20-100%. Fracture toughness (KIc), flexure strength (FS), elastic modulus (E), and hardness (KHN) were tested after soaking in water at 37 degrees C for 1 day, 6 months, 1 year, and 2 years. The KIc was reduced 20-30% for all composites after 6 months, with minimal changes thereafter. The FS was reduced for several composites at 6 months, but only those with poor cure (A and B) were lower at 2 years than they were initially. The E was not reduced for most composites. Hardness was reduced for most composites after 6 months, but many returned to their original levels at 2 years. Long-term aging in water caused a reduction in the KIc, independent of composition, but had little effect on other properties, suggesting limited degradation of composites in water.
589 citations
TL;DR: Results indicate that the linkage of the aminoalkylsilane to the oxidized surface is stable and that bound proteins such alkaline phosphatase and albumin retain their enzymatic activity and antigenicity, respectively.
Abstract: The surface of implantable biomaterials is in di- rect contact with the host tissue and plays a critical role in determining biocompatibility. In order to improve the inte- gration of implants, it is desirable to control interfacial re- actions such that nonspecific adsorption of proteins is mini- mized and tissue-healing phenomena can be controlled. In this regard, our goal has been do develop a method to func- tionalize oxidized titanium surfaces by the covalent immo- bilization of bioactive organic molecules. Titanium first was chemically treated with a mixture of sulfuric acid and hy- drogen peroxide to eliminate surface contaminants and to produce a consistent and reproducible titanium oxide sur- face layer. An intermediary aminoalkylsilane spacer mol- ecule was then covalently linked to the oxide layer, followed by the covalent binding of either alkaline phosphatase or albumin to the free terminal NH2 groups using glutaralde- hyde as a coupling agent. Surface analyses following coating procedures consisted of X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and atomic force microscopy (AFM). Enzymatic activity of coupled al- kaline phosphatase was assayed colorimetrically, and sur- face coverage by bound albumin was evaluated by SEM visualization of colloidal gold immunolabeling. Our results indicate that the linkage of the aminoalkylsilane to the oxi- dized surface is stable and that bound proteins such alkaline phosphatase and albumin retain their enzymatic activity and antigenicity, respectively. The density of immunolabel- ing for albumin suggests that the binding and surface cov- erage obtained is in excess of what would be expected for inducing biological activity. In conclusion, this method of- fers the possibility of covalently linking selected molecules with known biological activity to oxidized titanium surfaces in order to guide and promote the tissue healing that occurs during implant integration in bone and soft tissues. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 324-335, 1998.
495 citations
TL;DR: The cytotoxicity of 43 metal salts were evaluated by the colony formation method using two kinds of cultured cells and the effects of the difference in valence numbers of metallic elements in the salts on cytot toxicity were examined.
Abstract: Metallic biomaterials are generally used for replacement of structural components of the human body such as bones, joints, and tooth roots. When they are implanted inside a body, metallic biomaterials may corrode and/or wear, releasing metal ions and debris which may have toxic effects on tissues and organs. Since it is important for biomaterials to have no toxicity against a living body, a systematic and quantitative evaluation of the cytotoxicity of metallic elements is required for the development of new metallic biomaterials with superior biocompatibility. In this study, the cytotoxicity of 43 metal salts were evaluated by the colony formation method using two kinds of cultured cells. The effects of the difference in valence numbers of metallic elements in the salts on cytotoxicity were examined. The cytotoxicity of the salts of metallic elements' oxo acids was also investigated. As a result, the intensity of metal salts' cytotoxicity tends to be quite similar between MC3T3-E1 and L929 (the correlation coefficient of metal salts' IC50s is 0.82). The intensity of metal salts' cytotoxicity depends on the kinds of metallic elements, their chemical states, and concentrations. The IC50 of the highest toxic salt is 1.36 x 10(-6) mol L-1, which differs four orders of magnitude from the IC50 of the lowest toxic salt. K2Cr2O7, CdCl2, VCl3, AgNO3, HgCl2, SbCl3, BeSO4, and InCl3 are high toxic salts in which IC50s are smaller then 10(-5) mol L-1 for both or either of the cell lines. HgCl, Tl(NO3)3, GaCl3, CuCl2, MnCl2, CoCl2, ZnCl2, NiCl2, SnCl2, IrCl4, TlNO3, CuCl, RhCl3, Pb(NO3)2, Cr(NO3)3 and Bi(NO3)3 are relatively high toxic salts in which IC50s are smaller than 10(-4) mol L-1 for both or either cell lines.
492 citations
TL;DR: There was no cell type identified which was consistently less or more sensitive to the toxic effects of the tested compounds than the others, and for several of the highly cytotoxic composite components, less cytot toxic alternatives are available.
Abstract: It was the purpose of this investigation to determine the cytotoxic effects (ED50 concentrations) of 35 monomers or additives identified in commercial dental resin composites. Monolayers of permanent 3T3 cells and three primary human fibroblast types derived from oral tissues (gingiva, pulp, and periodontal ligament) were used as test systems. All substances were tested in concentrations ranging from 0.01 to 5.0 mM. In general, ED50 values varied from 0.06 to > 5 mM. Within the groups of co(monomers), initiators, and cointiators, severe (e.g., Bis-GMA, UDMA, DMBZ, and DMDTA) or moderate (HEMA, BEMA, CQ, DMPT, and DMAPE) cytotoxic effects could be evaluated. Within the group of reaction/decomposition products, only moderate or slight effects were found (ED50: 0.7 to > 5 mM). The inhibitor BHT, the contaminant TPSb, and the photostabilizer HMBP, however, were highly cytotoxic in all cell cultures. In addition, the ED50 values of DBPO and HMBP significantly varied (0.43-3.8 mM, respectively, and 0.44-3.07 mM) with the applied cell culture. Our comprehensive screening shows that for several of the highly cytotoxic composite components, less cytotoxic alternatives are available. Furthermore, there was no cell type identified which was consistently less or more sensitive to the toxic effects of the tested compounds than the others. Primary human periodontal ligament and pulp fibroblasts, however, were found to be more sensitive than 3T3 and gingival fibroblasts to alterations from most tested substances.
TL;DR: Transmission electron microscopy was used to study metal wear particles that were either in situ in cells or had been extracted from the cells by a new technique based on enzymatic tissue digestion to explain the less intense tissue reaction around metal on metal total hip replacements.
Abstract: The less intense tissue reaction around metal on metal total hip replacements (THRs) compared to metal on polyethylene (PE) THRs may be explained by the differences in the characteristics of metal wear particles. In this study, transmission electron microscopy was used to study metal wear particles that were either in situ in cells or had been extracted from the cells by a new technique based on enzymatic tissue digestion. The tissues were obtained from 13 patients undergoing revision of metal on metal THRs with cobalt-chromium-molybdenum (CoCrMo) bearing couples. Most of the CoCrMo wear particles were smaller than 50 nm (range 6-834 nm) and round to oval in shape with irregular boundaries. This size range is considerably smaller than that reported for PE particles. While even a small volume of metal wear will produce high numbers of particles, the apparently less severe local tissue reaction to metal particles may be due to the possibility that corrosion, dissolution, and dissemination of metal particles may result in fewer local biological effects than the long-term retention of PE particles in the periprosthetic tissues.
TL;DR: Results indicate that the geometry of the interconnected porous structure in PPHAP and coral-HAP create spaces for vasculature that lead to osteogenesis while the smooth structure and close contact of particles in SPHAP inhibit vascular formation and proliferation of mesenchymal cells, preventing bone and cartilage formation.
Abstract: Bone morphogenetic protein (BMP) is known to require a suitable carrier to induce ectopic bone formation in vivo. Hydroxyapatite ceramics have been reported to be effective in some forms but ineffective in others as a carrier of BMP-induced bone formation. In this study we compare three geometrically different forms of hydroxyapatite to examine their functions as carriers of BMP-induced bone formation. A fraction containing all the active BMPs (BMP cocktail) was partially purified from a 4M guanidine extract from bovine bone by a three-step chromatographic procedure. The BMP cocktail was combined with each of three forms of hydroxyapatite--solid particles (SPHAP), porous particles (PPHAP), and coral-replicated porous tablets (coral-HAP)--and implanted subcutaneously into rats. Both the PPHAP and coral-HAP systems induced osteogenesis 2 weeks after implantation, as evidenced by morphological and biochemical observations. Details of the osteogenetic process were followed by double-fluorescence labeling in the coral-HAP system to confirm bone formation on the surface of hydroxyapatite. However, there was no evidence of osteogenesis or chondrogenesis in the SPHAP system. The results indicate that the geometry of the interconnected porous structure in PPHAP and coral-HAP create spaces for vasculature that lead to osteogenesis while the smooth structure and close contact of particles in SPHAP inhibit vascular formation and proliferation of mesenchymal cells, preventing bone and cartilage formation. It was concluded that the geometrical structure in hydroxyapatite ceramics that induces vasculature is crucial as a carrier for BMP-induced bone formation.
TL;DR: BoneSource-hydroxyapatite cement is a new self-setting calcium phosphate cement biomaterial whose unique and innovative physical chemistry coupled with enhanced biocompatibility make it useful for craniofacial skeletal reconstruction.
Abstract: BoneSource-hydroxyapatite cement is a new self-setting calcium phosphate cement biomaterial. Its unique and innovative physical chemistry coupled with enhanced biocompatibility make it useful for craniofacial skeletal reconstruction. The general properties and clinical use guidelines are reviewed. The biomaterial and surgical applications offer insight into improved outcomes and potential new uses for hydroxyapatite cement systems.
TL;DR: Polymer concepts regarding bone tissue engineering are discussed and recent advances of the laboratory on guided bone regeneration using biodegradable polymer scaffolds are reviewed.
Abstract: Traumatic injuries, cancer treatment, and congenital abnormalities are often associated with abnormal bone shape or segmental bone loss Restoration of normal structure and function in these cases requires replacement of the missing bone that may be accomplished by surgical transfer of natural tissue from an uninjured location elsewhere in the body However, this procedure is limited by availability, adequate blood supply, and secondary deformities at the donor site One strategy to overcome these problems is to develop living tissue substitutes based on synthetic biodegradable polymers Three methods of bone regeneration using biodegradable polymers are being studied in our laboratory: tissue induction, cell transplantation, and fabrication of vascularized bone flaps Injectable polymers are used for filling skeletal defects and guiding bone tissue growth Their main advantage is minimizing the surgical intervention or the severity of the surgery Polymer-cell constructs also hold great promise in the field of tissue engineering They provide a scaffold on which cells grow and organize themselves As the cells begin to secrete their own extracellular matrix, the polymer degrades and is eventually eliminated from the body, resulting in completely natural tissue replacement Bone flaps can be fabricated ectopically into precise shapes and sizes With an attached vascular supply, these flaps can be transferred into areas deficient in vascularity This article discusses polymer concepts regarding bone tissue engineering and reviews recent advances of our laboratory on guided bone regeneration using biodegradable polymer scaffolds
TL;DR: In this paper, surface roughness has been shown to affect differentiation and local factor production of MG63 osteoblast-like cells, and the effect of roughness on cell response to systemic hormones has been examined.
Abstract: Surface roughness has been shown to affect differentiation and local factor production of MG63 osteoblast-like cells. This study examined whether surface roughness alters cellular response to circulating hormones such as 1 alpha,25-(OH)2D3. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then were machined and acid-etched (MA). Ti disks also were sandblasted (SB), sandblasted and acid etched (CA), or plasma sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were: PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on standard tissue culture polystyrene (plastic) or the Ti surfaces and then treated for 24 h with either 10(-8) M or 10(-7) M 1 alpha,25-(OH)2D3 or vehicle (control). Cellular response was measured by assaying cell number, cell layer alkaline phosphatase specific-activity, and the production of osteocalcin, latent (L) TGF beta, and PGE2. Alkaline phosphatase activity was affected by surface roughness; as the surface became rougher, the cells showed a significant increase in alkaline phosphatase activity. Addition of 1 alpha,25-(OH)2D3 to the cultures caused a dose-dependent stimulation of alkaline phosphatase activity that was synergistic with the effect caused by surface roughness alone. 1 alpha,25-(OH)2D3 also caused a synergistic increase in osteocalcin production as well as local factor (LTGF beta and PGE2) production on the rougher CA, SB, and PS surfaces, but it had no effect on the production on smooth surfaces. The inhibitory effect of surface roughness on cell number was not affected by 1 alpha,25-(OH)2D3 except on the SB surface. 1 alpha,25-(OH)2D3 decreased cell number, increased alkaline phosphatase activity and osteocalcin production, and had no effect on LTGF beta or PGE2 production by MG63 cells grown on tissue culture polystyrene. These data suggest that bone cell response to systemic hormones is modified by surface roughness and that surface roughness increases the responsiveness of MG63 cells to 1 alpha,25-(OH)2D3. They also suggest that the endocrine system is actively involved in normal bone healing around implants.
TL;DR: Results demonstrate that optimization of biomaterial-cell interactions provides a strategy for increasing the initial cell seeding density for the engineering of tissues of high cell density.
Abstract: A procedure for surface hydrolysis of poly(glycolic acid) (PGA) meshes was developed to increase cell seeding density and improve attachment of vascular smooth muscle cells. Hydrolysis of PGA in 1N NaOH transformed ester groups on the surface of PGA fibers to carboxylic acid and hydroxyl groups. After hydrolysis, the polymer scaffold retained its original gross appearance and dimensions while the fiber diameter decreased. A plot of fiber diameter versus the hydrolysis time showed a linear relationship, with a rate of decrease in fiber diameter of 0.65 microm/min. The molecular weight and thermal properties of the polymer did not change significantly following surface hydrolysis. In cell seeding experiments, surface-hydrolyzed mesh was seeded with more than twice as many cells as unmodified PGA mesh. Vascular smooth muscle cells attached to the surface-hydrolyzed PGA mesh both as individual cells and as cell aggregates while only cell aggregates were observed on the unmodified mesh. Control experiments indicated that adsorption of serum proteins onto the surface-hydrolyzed PGA fibers was correlated with the increase in cell seeding density. These results demonstrate that optimization of biomaterial-cell interactions provides a strategy for increasing the initial cell seeding density for the engineering of tissues of high cell density.
TL;DR: The benzyl ester of hyaluronic acid (HYAFF 11), a recently developed semisynthetic resorbable material, is promising for the use of this scaffold for tissue engineering of cartilage replacements.
Abstract: The aim of this study was to investigate the possibility of using the benzyl ester of hyaluronic acid (HYAFF® 11), a recently developed semisynthetic resorbable material, as a scaffold for the culture of human nasoseptal chondrocytes in tissue-engineering procedures of cartilage reconstruction. Different techniques such as immunohistochemistry, scanning electron microscopy, and confocal laser scanning microscopy were used to study the behavior, morphology, and phenotype expression of the chondrocytes, which were initially expanded and then seeded on the material. The nonwoven cell carrier allowed good viability and adhesivity of the cells without any surface treatment with additional substances. Furthermore, the cultured cells expressed cartilage-specific collagen type II, indicating that they were able to redifferentiate within the scaffold of HYAFF® 11 and were able to retain a chondrocyte phenotype even after a long period of in vitro conditions. Nevertheless, the expression of collagen type I, which was produced by dedifferentiated or incompletely redifferentiated chondrocytes, was noticeable. Additional data were obtained by subcutaneous implantation of samples seeded with human cells in the in vivo model of the athymic nude mouse. The results after 1 month revealed the development of tissue similar to hyaline cartilage. This study is promising for the use of this scaffold for tissue engineering of cartilage replacements. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 42, 172–181, 1998.
TL;DR: The uniformity of the oxide layer, rather than its thickness and composition, seems to be the predominant factor to explain the corrosion resistance improvement of NiTi alloys.
Abstract: Because of its good radiopacity, superelasticity, and shape memory properties, nickel-titanium (NiTi) is a potential material for fabrication of stents because these properties can facilitate their implantation and precise positioning. However, in vitro studies of NiTi alloys report the dependence of alloy biocompatibility and corrosion behavior on surface conditions. Surface oxidation seems to be very promising for improving the corrosion resistance and biocompatibility of NiTi. In this work, we studied the effect on corrosion resistance and surface characteristics of electropolishing, heat treatment, and nitric acid passivation of NiTi stents. Characterization techniques such as potentiodynamic polarization tests, scanning electron microscopy, Auger electron spectroscopy, and X-ray photoelectron spectroscopy were used to relate corrosion behavior to surface characteristics and surface treatments. Results show that all of these surface treatments improve the corrosion resistance of the alloy. This improvement is attributed to the plastically deformed native oxide layer removal and replacement by a newly grown, more uniform one. The uniformity of the oxide layer, rather than its thickness and composition, seems to be the predominant factor to explain the corrosion resistance improvement.
TL;DR: It is demonstrated that genipin is an effective crosslinking reagent for biological tissue fixation and has a mechanical strength and resistance against enzymatic degradation comparable to the glutaraldehyde-fixed tissue.
Abstract: Bioprostheses derived from biological tissues must be chemically modified and subsequently sterilized before they can be implanted in humans. Various crosslinking reagents, including formaldehyde, glutaraldehyde, dialdehyde starch, and epoxy compound, have been used to chemically modify biological tissues. However, these synthetic crosslinking reagents are all highly (or relatively highly) cytotoxic. It is therefore desirable to provide a crosslinking reagent suitable for use in biomedical applications that is of low cytotoxicity and that forms stable and biocompatible crosslinked products. This study evaluates the feasibility of using a naturally occurring crosslinking reagent--genipin--to chemically modify biological tissues. Genipin and its related iridoid compounds, extracted from gardenia fruits, have been used in traditional Chinese medicine for the treatments of jaundice and various inflammatory and hepatic diseases. In this feasibility study, the cytotoxicity of genipin and the crosslinking characteristics of genipin-fixed biological tissues were investigated. Fresh porcine pericardia procured from a slaughterhouse were used as raw materials. Glutaraldehyde and an epoxy compound (ethylene glycol diglycidyl ether), which has been used extensively in developing bioprostheses, were used as controls. It was found that the cytotoxicity of genipin was significantly lower than that of glutaraldehyde and the epoxy compound. The amino acid residues in the porcine pericardium that may react with genipin were lysine, hydroxylysine, and arginine. Additionally, the genipin-fixed tissue had a mechanical strength and resistance against enzymatic degradation comparable to the glutaraldehyde-fixed tissue. This suggests that genipin can form stable crosslinked products. The results of this in vitro study demonstrate that genipin is an effective crosslinking reagent for biological tissue fixation.
TL;DR: Four calcium phosphate cement formulations were implanted in the rabbit distal femoral metaphysis and middiaphysis and mineralogy, chemical composition, and osseous implantation site of these calcium phosphates significantly affected their in vivo host response.
Abstract: Four calcium phosphate cement formulations were implanted in the rabbit distal femoral metaphysis and middiaphysis. Chemical, crystallographic, and histological analyses were made at 2, 4, and 8 weeks after implantation. When implanted into the metaphysis, part of the brushite cement was converted into carbonated apatite by 2 2 weeks. Some of the brushite cement was removed by mononuclear macrophages prior to its conversion into apatite. Osteoclastlike cell mediated remodeling was predominant at 8 weeks after brushite had converted to apatite. The same histological results were seen for brushite plus calcite aggregate cement, except with calcite aggregates still present at 8 weeks. However, when implanted in the diaphysis, brushite and brushite plus calcite aggregate did not convert to another calcium phosphate phase by 4 weeks. Carbonated apatite cement implanted in the metaphysis did not transform to another calcium phosphate phase. There was no evidence of adverse foreign body reaction. Osteoclastlike cell mediated remodeling was predominant at 8 weeks. The apatite plus calcite aggregate cement implanted in the metaphysis that was not remodeled remained as poorly crystalline apatite. Calcite aggregates were still present at 8 weeks. There was no evidence of foreign body reaction. Osteoclastlike cell remodeling was predominant at 8 weeks. Response to brushite cements prior to conversion to apatite was macrophage dominated, and response to apatite cements was osteoclast dominated. Mineralogy, chemical composition, and osseous implantation site of these calcium phosphates significantly affected their in vivo host response.
TL;DR: It is confirmed that osteoblast adhesion to substrates can be controlled selectively and significantly by immobilized peptides, and elucidates criteria and strategies for the design of proactive dental/orthopedic implant biomaterials.
Abstract: Proactive, “next generation” dental/orthopedic biomaterials must be designed rationally to elicit specific, timely, and desirable responses from surrounding cells/tissues; for example, such biomaterials should support and enhance osteoblast adhesion (a crucial function for anchorage-dependent cells). In the past, integrin-binding peptides have been immobilized on substrates to partially control osteoblast adhesion; the present study focused on the design, synthesis, and bioactivity of the novel peptide sequence Lys-Arg-Ser-Arg that selectively enhances heparan sulfate-mediated osteoblast adhesion mechanisms. Osteoblast, but not endothelial cell or fibroblast, adhesion was enhanced significantly (p < 0.05) on substrates modified with Lys-Arg-Ser-Arg peptides, indicating that these peptides may be osteoblast- or bone cell specific. Blocking osteoblast cell-membrane receptors with various concentrations of soluble Arg-Gly-Asp-Ser peptides did not inhibit subsequent cell adhesion on substrates modified with Lys-Arg-Ser-Arg peptides, providing evidence that osteoblasts interact with Arg-Gly-Asp-Ser and with Lys-Arg-Ser-Arg peptides via distinct (i.e., integrin- and proteoglycan-mediated) mechanisms, each uniquely necessary for osteoblast adhesion. The present study constitutes an example of rational design/selection of bioactive peptides, confirms that osteoblast adhesion to substrates can be controlled selectively and significantly by immobilized peptides, and elucidates criteria and strategies for the design of proactive dental/orthopedic implant biomaterials. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 371–377, 1998.
TL;DR: It can be said that microtextured surfaces influence the orientation of intracellular and extracellular proteins.
Abstract: The microfilaments and vinculin-containing attachment complexes of rat dermal fibroblasts (RDF) incubated on microtextured surfaces were investigated with confocal laser scanning microscopy (CLSM) and digital image analysis (DIA). In addition, depositions of bovine and endogenous fibronectin and vitronectin were studied. Smooth and microtextured silicone substrata were produced that possessed parallel surface grooves with a groove and ridge width of 2.0, 5.0, and 10.0 μm. The groove depth was approximately 0.5 μm. CLSM and DIA make it possible to visualize and analyze intracellular and extracellular proteins and the underlying surface simultaneously. It was observed that the microfilaments and vinculin aggregates of the RDFs on the 2.0 μm grooved substrata were oriented along the surface grooves after 1, 3, 5, and 7 days of incubation while these proteins were significantly less oriented on the 5.0 and 10.0 μm grooved surfaces. Vinculin was located mainly on the surface ridges on all textured surfaces. In contrast, bovine and endogenous fibronectin and vitronectin were oriented along the surface grooves on all textured surfaces. These proteins did not seem to be hindered by the surface grooves since many groove-spanning filaments were found on all the microgrooved surfaces. In conclusion, it can be said that microtextured surfaces influence the orientation of intracellular and extracellular proteins. Although results corroborate three earlier published hypotheses, they do not justify a specific choice of any one of these hypotheses. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 291–300, 1998.
TL;DR: The effects of two precipitation variables, namely temperature and bicarbonate ion concentration, on the morphology, phase composition, and calcium, phosphorus, and carbon contents of precipitated carbonate hydroxyapatite are documents.
Abstract: Carbonate substitution in the apatite crystal lattice can occur in either the hydroxyl or the phosphate sites, designated as A or B type, respectively, and previous investigations generally have described precipitated carbonate hydroxyapatite as being B type on the basis of infra red and X-ray data. This paper documents the effects of two precipitation variables, namely temperature and bicarbonate ion concentration, on the morphology, phase composition, and calcium, phosphorus, and carbon contents of precipitated carbonate hydroxyapatite. Variations in both temperature and bicarbonate concentration could yield either acicular or spheroidal crystals. X-ray diffraction and infra red spectroscopy indicated the presence of carbonate in the A site for low carbonate contents ( 4 wt%), the carbonate was located predominantly in the B site. On the basis of these observations and chemical analyses, a new AB carbonate substitution mechanism is proposed that better describes the experimental data than the B-type models used previously.
TL;DR: Collagen sponges containing hydroxyapatite remain one of the most appropriately evaluated biomaterials used for the restoration of periodontal pockets and bone defects.
Abstract: The induction of a calcifying matrix is of great interest in the restoration of bone defects. In a previous in vitro study we demonstrated that a collagen sponge constituted of type I collagen fibrils, chondroitin sulfates, and hydroxyapatite crystals induces an earlier and a more abundant synthesis of a new extracellular calcifying matrix than do other biomaterials such as collagen or hydroxyapatite alone. Bone mineral contains various amounts of magnesium ions, either adsorbed at the surface of apatite crystals or incorporated inside the crystal structure. Magnesium is known to reduce the degradation rate of tricalcium phosphate ceramics and to influence the crystallization of mineral substance. Thus we evaluated two sponges modified with different substituted apatites. The substituted low magnesium-containing apatite sample decreased the osteoinductive properties of the sponge whereas the substituted high magnesium-containing apatite sample had a toxic effect on bone cells and prevented the formation of any extracellular matrix. Such a toxic effect can be explained by the presence of large numbers of magnesium ions released into the culture medium even though at physiological level magnesium is able to promote bone mineralization and to control the growth of hydroxyapatite crystals. Thus collagen sponges containing hydroxyapatite remain one of the most appropriately evaluated biomaterials used for the restoration of periodontal pockets and bone defects.
TL;DR: Bone formation and resorption characteristics of the ABS material were characterized in a canine femoral slot defect model and the autograft and ABS were associated with similar new bone growth and defect filling characteristics.
Abstract: The in vitro and in vivo properties of a novel, fully resorbable, apatitic calcium phosphate bone substitute (ABS) are described. The ABS was prepared from calcium phosphate precursors that were hydrated to form an injectable paste that hardens endothermically at 37 degrees C to form a poorly crystalline apatitic calcium phosphate (PCA). The PCA reaction product is stable in vivo as determined by FTIR and XRD analysis of rabbit intramuscular implants of ABS retrieved 4, 7, and 14 days postimplantation. Bone formation and resorption characteristics of the ABS material were characterized in a canine femoral slot defect model. Femoral slot defects in dogs were filled with either autologous bone implants or the ABS material. Sections of femoral bone defect site from animals sacrificed at 3, 4, 12, 26, and 52 weeks demonstrated that new bone formation proceeded similarly in both autograft and ABS filled slots. Defects receiving either material were filled with trabecular bone in the first 3 to 4 weeks after implantation; lamellar or cortical bone formation was well established by week 12. New bone formation in ABS filled defects followed a time course comparable to autologous bone graft filled defects. Histomorphometric evaluation of ABS resorption and new bone formation indicated that the ABS material was greater than 99% resorbed within 26 weeks; residual ABS occupied 0.36+/-0.36% (SEM, n = 4) of the original defect area at 26 weeks. Quantitatively and qualitatively, the autograft and ABS were associated with similar new bone growth and defect filling characteristics.
TL;DR: It is suggested that sintered carbonate apatites, which have characteristics that can be favorably compared with those of bone, especially with respect to its reactivity to acid media, would be useful as bioresorbable bone substitutes.
Abstract: The dissolution behavior of sintered carbonate apatite was investigated in a 10 mM/L acetic acid solution adjusted to pH 5.0 at 37°C, and compared to that of sintered hydroxyapatite and bone apatite for the purpose of establishing some similarities between the physicochemical dissolution of apatite biomaterials in vitro and their ability to be resorbed by osteoclasts in vivo. Both the sintered carbonate apatite and the bone apatite dissolved to an appreciable extent. Their solution compositions changed in an almost identical manner until toward the end of the reaction. The solution compositions for sintered carbonate apatite at 30 s was comparable with that for sintered hydroxyapatite at 3.8 days with respect to the degree of supersaturation, indicating that the former specimen is much more soluble than the latter specimen. Osteoclasts which were obtained from the long bones of 1-day-old neonatal rabbits resorbed bone and sintered carbonate apatite, but not sintered hydroxyapatite. These findings suggest that sintered carbonate apatites, which have characteristics that can be favorably compared with those of bone, especially with respect to its reactivity to acid media, would be useful as bioresorbable bone substitutes. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 39, 603–610, 1998.
TL;DR: The tissue response to a nano-hydroxyapatite/collagen composite implanted in a marrow cavity was investigated by histology and scanning electron microscopy and a Knoop microhardness test was performed to compare the mechanical behavior of the composite and bone.
Abstract: The tissue response to a nano-hydroxyapatite/collagen composite implanted in a marrow cavity was investigated by histology and scanning electron microscopy. A Knoop microhardness test was performed to compare the mechanical behavior of the composite and bone. The ultrastructural features of the composite, especially the carbonate-substituted hydroxyapatite with low crystallinity and nanometer size, made it a bone-resembling material. It was bioactive, as well as biodegradable. At the interface of the implant and marrow tissue, solution-mediated dissolution and giant cell mediated resorption led to the degradation of the composite. Interfacial bone formation by osteoblasts was also evident. The process of implant degradation and bone substitution was reminiscent of bone remodeling. The composite can be incorporated into bone metabolism instead of being a permanent implant. For lack of the hierarchical organization similar to that of bone, the composite exhibited an isotropic mechanical behavior. However, the resistance of the composite to localized pressure could reach the lower limit of that of the femur compacta.
TL;DR: Thermally processed hydroxyapatite coatings used on dental implants and hip prostheses for enhanced fixation may typically consist of a number of chemical and structural phases that affect coating performance and tissue attachment.
Abstract: Thermally processed hydroxyapatite coatings used on dental implants and hip prostheses for enhanced fixation may typically consist of a number of chemical and structural phases. These phases affect coating performance and tissue attachment. Hydroxyapatite was plasma sprayed to examine the phase evolution during processing. Coatings were examined with X-ray diffraction and elemental analysis. Results indicate that phase transformations are produced by (a) preferential removal of hydroxyl and phosphate leading to a change in melt composition, and (b) the high cooling rate due to the thermal spray process. Hydroxyl group removal promotes the amorphous phase and oxyapatite. Further heating produces a less viscous melt facilitating decomposition of hydroxyapatite to tricalcium and tetracalcium phosphate. Phosphate removal during flight produces a more calcium-rich melt preferring tetracalcium phosphate and calcium oxide formation. A proposed model shows the phase location within the lamellae of these coatings. Coating processes must thus prevent removal of hydroxide and phosphate during processing to maximize the hydroxyapatite content.