Effect of cell-seeded hydroxyapatite scaffolds on rabbit radius bone regeneration.

Highly porous hydroxyapatite (HA) scaffolds were developed as bone graft substitutes using a template coating process, characterized, and seeded with bone marrow-derived mesenchymal stem cells (BMSCs). To test the hypothesis that cell-seeded HA scaffolds improve bone regeneration, HA scaffolds without cell seeding (HA-empty), HA scaffolds with 1.5 × 10(4) BMSCs (HA-low), and HA scaffolds with 1.5 × 10(6) BMSCs (HA-high) were implanted in a 10-mm rabbit radius segmental defect model for 4 and 8 weeks. Three different fluorochromes were administered at 2, 4, and 6 weeks after implantation to identify differences in temporal bone growth patterns. It was observed from fluorescence histomorphometry analyses that an increased rate of bone infiltration occurred from 0 to 2 weeks (p < 0.05) of implantation for the HA-high group (2.9 ± 0.5 mm) as compared with HA-empty (1.8 ± 0.8 mm) and HA-low (1.3 ± 0.2 mm) groups. No significant differences in bone formation within the scaffold or callus formation was observed between all groups after 4 weeks, with a significant increase in bone regenerated for all groups from 4 to 8 weeks (28.4% across groups). Although there was no difference in bone formation within scaffolds, callus formation was significantly higher in HA-empty scaffolds (100.9 ± 14.1 mm(3) ) when compared with HA-low (57.8 ± 7.3 mm(3) ; p ≤ 0.003) and HA-high (69.2 ± 10.4 mm(3) ; p ≤ 0.02) after 8 weeks. These data highlight the need for a better understanding of the parameters critical to the success of cell-seeded HA scaffolds for bone regeneration.

A cellular perspective to bioceramic scaffolds for bone tissue engineering: the state of the art.

A vast number of manufacturing techniques have been employed in the last five years to manufacture three dimensional (3D) calcium phosphate (CaP) scaffolds, with the intention to replicate the architecture of native bone as well as to repair and restore bone function. Design features such as architectural control and sintering temperature and their impact on scaffold performance is presented in this review. In vitro cell responses to bioceramic scaffolds and their in vivo performances have been enhanced. Current frontiers of active research on HA scaffolds have included the relationship between fluid flow and mechanotransduction as well as cell signaling pathways that induce endothelial cell recruitment and angiogenesis. Additionally, current research has focused on a better understanding of cell signaling and its environmental cues. The availability of non-invasive and non-destructive quantitative imaging modalities has also become critical in aiding the characterization of scaffolds and predicting scaffold performance. It is thus anticipated that further knowledge gained from this research will allow the overall advancement of scaffolds that can be clinically used to restore large bone defects.

Plasma surface modification of poly(D,L-lactic acid) as a tool to enhance protein adsorption and the attachment of different cell types.

We have studied the influence of oxygen radio frequency glow discharge (RfGD) on the surface and bulk properties of poly(D,L-lactic acid) (PDLLA) and the effect of this surface modification on both protein adsorption and bone cell behavior. PDLLA films were characterized before and after plasma surface modification by water contact angle, surface energy, and adhesion tension of water as well as by scanning electron microscopy (SEM), X-ray electron spectroscopy (XPS), and Fourier transform infra-red (FTIR) spectroscopy. RfGD-films showed an increase in hydrophilicity and surface energy when compared with untreated films. Surface morphological changes were observed by SEM. Chemical analysis indicated significant differences in both atomic percentages and oxygen functional group. Protein adsorption was evaluated by combining solute depletion and spectroscopic techniques. Bovine serum albumin (BSA), fibronectin (FN), vitronectin (VN), and fetal bovine serum (FBS) were used in this study. RfGD-treated surfaces adsorbed more BSA and FN from single specie solutions than FBS that is a more complex, multi-specie solution. MG63 osteoblast-like cells and primary cultures of fetal rat calvarial (FRC) cells were used to assess both the effect of RfGD treatment and protein adsorption on cell attachment and proliferation. In the absence of preadsorbed proteins, cells could not distinguish between treated and untreated surfaces, with the exception of MG63 cells cultured for longer periods of time. In contrast, the adsorption of proteins increased the cells' preference for treated surfaces, thus indicating a crucial role for adsorbed proteins in mediating the response of osteogenic cells to the RfGD-treated PDLLA surface.

Antibacterial and osteogenic properties of silver-containing hydroxyapatite coatings produced using a sol gel process.

Since bacterial infection is a rising complication following the wide use of implant, there is considerable attention on the effect of implant surface properties on bacterial adhesion. In this study, the effect of silver (Ag) doped hydroxyapatite (HA) coatings on initial antibacterial adhesion and osteoblast cell proliferation and differentiation was investigated. Using a sol-gel process, HA coatings doped with 1 wt % AgNO(3) (AgHA1.0) and 1.5 wt % Ag (AgHA1.5) were prepared. Coated surfaces were characterized using X-ray diffraction (XRD) and contact angles measurements. The initial bacteria adhesion was evaluated using a RP12 strain of Staphylococcus epidermidis (ATCC 35984) and the Cowan I strain of Staphylococcus aureus, whereas osteoblast proliferation and differentiation were evaluated using human embryonic palatal mesenchyme cells (HEPM), an osteoblast precursor cell line. In this study, XRD analysis of all surfaces indicated peaks corresponding to HA. Contact angles for AgHA surfaces were observed to be significantly lower when compared to HA surfaces. In vitro initial bacterial adhesion study indicated a significantly reduced number of S. epidermidis and S. aureus on AgHA surfaces when compared to HA surface. The use of HEPM cells indicated no significant difference in double-stranded DNA (dsDNA) production between all surfaces. Additionally, no differences in alkaline phosphatase specific activity were observed between HA and AgHA1.0 surfaces. Overall, it was concluded that AgHA1.0 has the similar biological activity as HA, with respect to bone cell proliferation and differentiation. In addition, the AgHA1.0 was also concluded to have the ability to minimize the initial bacteria adhesion. (c) 2007 Wiley Periodicals, Inc. J Biomed Mater Res, 2007.

Characterization of biomimetic calcium phosphate on phosphorylated chitosan films.

This study examined the effect of chitosan degree of deacetylation (DDA), concentration of simulated body fluid (SBF), and mineralization time on the composition, structure, and crystallinity of calcium phosphate (CaP) biomimetically deposited on chitosan and on osteoblast cell growth. Phosphorylated chitosan films of 92.3%, 87.4%, and 80.6% DDA were soaked in SBF (1.0x or 1.5x) for 7, 14, or 21 days. Scanning electron microscopy revealed that CaP precipitated from 1.5x SBF had a porous, granular morphology; while the coatings precipitated in 1.0x SBF were smoother and more uniform. X-ray diffraction showed that films mineralized in 1.0x SBF were amorphous, while films mineralized in 1.5x SBF for 21 days exhibited crystalline peaks similar to hydroxyapatite, with the most crystalline peaks seen on 92.3% DDA chitosan. When mineralized films were placed in cell media for 14 days, more calcium phosphate precipitated onto all films, and the most calcium phosphate was found on 92.3% DDA films mineralized in 1.5x SBF. After seven days of osteoblast culture, there were approximately three times as many cells (based on DNA measurements, p < 0.05) on 92.3% DDA films soaked in 1.0x SBF for seven or 21 days than on 80.6% DDA films soaked in 1.0x SBF for any length of time or any films soaked in 1.5x SBF. The DDA of chitosan, concentration of SBF and mineralization time affect the structure of and biological response to chitosan/biomimetic CaP films, and these factors must be considered when designing new materials to be used in orthopaedic and dental/craniofacial implant applications.

Modulating bone cells response onto starch-based biomaterials by surface plasma treatment and protein adsorption.

The effect of oxygen-based radio frequency glow discharge (rfGD) on the surface of different starch-based biomaterials (SBB) and the influence of proteins adsorption on modulating bone-cells behavior was studied. Bovine serum albumin, fibronectin and vitronectin were used in single and complex protein systems. RfGD-treated surfaces showed to increase in hydrophilicity and surface energy when compared to non-modified SBB. Biodegradable polymeric blends of cornstarch with cellulose acetate (SCA; 50/50wt%), ethylene vinyl alcohol (SEVA-C; 50/50wt%) and polycaprolactone (SPCL; 30/70wt%) were studied. SCA and SCA reinforced with 10% hydroxyapatite (HA) showed the highest degree of modification as result of the rfGD treatment. Protein and control solutions were used to incubate with the characterized SBB and, following this, MG63 osteoblast-like osteosarcoma cells were seeded over the surfaces. Cell adhesion and proliferation onto SCA was found to be enhanced for non-treated surfaces and on SCA+10%HA no alteration was brought up by the plasma modification. Onto SCA surfaces, BSA, FN and VN single solutions improved cell adhesion, and this same effect was found upscaled for ternary systems. In addition, plasma treated SEVA-C directed an increase in both adhesion and proliferation comparing to non-treated surfaces. Even though adhesion onto treated and untreated SPCL was quite similar, plasma modification clearly promoted MG63 cells proliferation. Regarding MG63 cells morphology it was shown that onto SEVA-C surfaces the variation of cell shape was primarily defined by the protein system, while onto SPCL it was mainly affected by the plasma treatment.

Characterization of chitosan films and effects on fibroblast cell attachment and proliferation.

Chitosan has been researched for implant and wound healing applications. However, there are inconsistencies in reports on the tissue and fibroblast responses to chitosan materials. These inconsistencies may be due to variations in chitosan material characteristics. The aim of this study was to correlate fibroblast responses with known chitosan material characteristics. To achieve this aim, chitosan was characterized for degree of deacetylation (DDA), molecular weight (MW), residual protein and ash contents, and then solution cast into films and characterized for hydrophilicity by water contact angle. The films were seeded with normal human dermal fibroblasts and the number of attached cells was evaluated for after 30 min. Cell proliferation was evaluated over 5 days. This study found no relationship between DDA, contact angle, cell attachment, and or proliferation. General trends were observed for increasing proliferation with increasing residual ash content and decreasing residual protein. These data indicate that chitosan characteristics other than DDA may be important to their biological performance.

In vitro anti-bacterial and biological properties of magnetron co-sputtered silver-containing hydroxyapatite coating.

Bacterial infection after implant placement is a significant rising complication. In order to reduce the incidence of implant-associated infections, several biomaterial surface treatments have been proposed. In this study, the effect of in vitro antibacterial activity and in vitro cytotoxicity of co-sputtered silver (Ag)-containing hydroxyapatite (HA) coating was evaluated. Deposition was achieved by a concurrent supply of 10 W to the Ag target and 300 W to the HA target. Heat treatment at 400 degrees C for 4 h was performed after 3 h deposition. X-ray diffraction, contact angles measurements, and surface roughness were used to characterize the coating surfaces. The RP12 strain of Staphylococcus epidermidis (ATCC 35984) and the Cowan I strain of Staphylococcus aureus were used to evaluate the antibacterial activity of the Ag-HA coatings, whereas human embryonic palatal mesenchyme cells, an osteoblast precursor cell line, were used to evaluate the in vitro cytotoxicity of the coatings. X-ray diffraction analysis performed in this study indicated peaks corresponding to Ag and HA on the co-sputtered Ag-HA surfaces. The contact angles for HA and Ag-HA surfaces were observed to be significantly lower when compared to Ti surfaces, whereas no significant difference in surface roughness was observed for all groups. In vitro bacterial adhesion study indicated a significantly reduced number of S. epidermidis and S. aureus on Ag-HA surface when compared to titanium (Ti) and HA surfaces. In addition, no significant difference in the in vitro cytotoxicty was observed between HA and Ag-HA surfaces. Overall, it was concluded that the creation of a multifunctional surface can be achieved by co-sputtering the osteoconductive HA with antibacterial Ag.

Effects of dissolved calcium and phosphorous on osteoblast responses.

The dissolution behavior of hydroxyapatite (HA) and its effect on the initial cellular response is of both fundamental and clinical importance. In this study, plasma-sprayed HA coatings were characterized by X-ray diffraction and Fourier transform infrared spectroscopy (FTIR). Calcium (Ca) and inorganic phosphorous (Pi) ions released from plasma-sprayed HA coatings within 3 weeks were measured by flame atomic absorption and colorimetrically molybdenum blue complex, respectively. To investigate the effect of dissolution of HA coatings on osteoblast response, additional Ca and Pi were added into the cell culture media to simulate the dissolution concentrations. Human embryonic palatal mesenchyme cells, an osteoblast precursor cell line, were used to evaluate the biological responses to enhanced Ca and Pi media over 2 weeks. Osteoblast differentiation and mineralization were measured by alkaline phosphatase-specific assay and 1,25 (OH)2 vitamin D3 stimulated osteocalcin production. The coatings exhibited an HA-type structure. FTIR indicated the possible presence of carbonates on the coatings. A dissolution study indicated a continual increase in Ca and Pi over time. In the cell culture study, enhanced osteoblast differentiation occurred in the presence of additional Ca concentration in the cell culture media. However, additional Pi concentration in the cell culture media was suggested to slow down osteoblast differentiation and mineralization.

Influence of post-deposition heating time and the presence of water vapor on sputter-coated calcium phosphate crystallinity.

Extensive research suggested that calcium phosphate (CaP) coatings on titanium implants are essential for early bone response. However, the characterization of CaP crystallinity and the means to control coating crystallinity are not well-established. In this study, the effect of a 400 degrees C heat treatment for 1, 2, or 4 hours, and in the presence or absence of water vapor, on CaP crystallinity was investigated. Scanning electron microscopy indicated dense as-sputtered coatings. Increase in coating crystallinity was observed to be consistent with the increasing number of PO(4) peaks observed as a result of different heat treatments. In addition, x-ray diffraction analyses indicated amorphous as-sputtered coatings, whereas crystalline CaP coatings in the range of 0-85% were observed after different post-deposition heat treatments. It was concluded that the presence of water vapor and post-deposition heat treatment time significantly affect the crystallinity of CaP coatings, which may ultimately affect bone healing.

Osteoblast precursor cell attachment on heat-treated calcium phosphate coatings.

The influence of properties of calcium phosphate (CaP) coatings on bone cell activity and bone-implant osseointegration is not well-established. This study investigated the effects of characterized CaP coatings of various heat treatments on osteoblast response. It was hypothesized that heat treatments of CaP coatings alter the initial osteoblast attachment. The 400 degrees C heat-treated coatings were observed to exhibit poor crystallinity and significantly greater phosphate or apatite species compared with as-sputtered and 600 degrees C heat-treated coatings. Similarly, human embryonic palatal mesenchyme (HEPM) cells, an osteoblast precursor cell line, seeded on 400 degrees C heat-treated coatings, exhibited significantly greater cell attachment compared with Ti surfaces, as-sputtered coatings, and 600 degrees C heat-treated coatings. The HEPM cells on Ti surfaces and heat-treated coatings were observed to attach through filopodia, and underwent cell division, whereas the cells on as-sputtered coatings displayed fewer filopodia extensions and cell damage. Analysis of the data suggested that heat treatment of CaP coatings affects cell attachment.

Effect of plasma-glow discharge as a sterilization of titanium surfaces.

In this study, in vitro osteoblast responses to glow-discharged, commercially pure titanium (Ti) surfaces were investigated. It was hypothesized that the glow-discharge treatment would be an effective sterilization procedure for Ti implantations before implantation. The Ti surfaces were prepared by grinding to 600 grits followed by cleaning. These were then divided into two groups, with one group being the control and the other group undergoing glow-discharge treatment using oxygen. Human embryonic palatal mesenchyme cells, an osteoblast precursor, were used to evaluate the cell responses to glow-discharged and control Ti surfaces. It was observed from this study that protein production and osteocalcin production on both surfaces exhibited no significant differences during the 10-day study. Similarly, no significant differences were observed for alkaline phosphatase (ALP) specific activity during the first 7 days of incubation. However, at day 10, the ALP specific activity for control Ti surfaces was significantly higher than the ALP activity for the glow-discharged surface. Overall, this study suggested that the use of glow discharge as an alternative sterilization procedure for medical and dental implants did not inhibit osteoblast phenotypic expression.

Characterization and dissolution behavior of sputtered calcium phosphate coatings after different postdeposition heat treatment temperatures.

There is a lack of correlation between specific properties of hydroxyapatite coating surfaces, osseointegration processes, and implant success. The aim of this study was to evaluate the relationship between well-characterized structural and chemical properties of radio-frequency sputtered calcium phosphate (CaP) coatings and their dissolution behavior. Sputtered CaP coatings were evaluated as-sputtered (non-heat treated) or after 1 hour of postsputter heat treatments at 400 degrees C or 600 degrees C. All coatings were characterized by X-ray diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and contact angle measurement. The dissolution behavior of CaP coatings in the presence and absence of proteins was also investigated. It was observed from this study that as-sputtered CaP coatings were amorphous. The 400 degrees C heat-treated CaP coatings exhibited low crystallinity (1.9% +/- 0.4%), whereas the 600 degrees C heat-treated CaP coatings were highly crystalline (67.0% +/- 2.4%). The increase of Ca/P ratio, PO4/HPO4 ratio, and the number of PO4 peaks were observed to be consistent with the increase in heating temperature and the degrees of coating crystallinity. Phosphorus ions released from CaP coatings decreased with the increase of crystallinity of CaP coatings. In addition, immersion of CaP coatings in media containing proteins resulted in an increase in P ions released as compared with coatings immersed in media without proteins. It was concluded that the degree of CaP coating crystallinity can be controlled by varying the postdeposition heat-treatment temperature. It was also concluded that, aside from coating crystallinity, dissolution and reprecipitation of the coatings can be controlled by knowing the presence of proteins in the media and PO4/HPO4 ratio within the coatings.

Contact angle, protein adsorption and osteoblast precursor cell attachment to chitosan coatings bonded to titanium.

Chitosan, a derivative of the bio-polysaccharide chitin, has shown promise as a bioactive material for implant, tissue engineering and drug-delivery applications. The aim of this study was to evaluate the contact angle, protein adsorption and osteoblast precursor cell attachment to chitosan coatings bonded to titanium. Rough ground titanium (Ti) coupons were solution cast and bonded to 91.2% de-acetylated chitosan (1 wt% chitosan in 0.2% acetic acid) coatings via silane reactions. Non-coated Ti was used as controls. Samples were sterilized by ethylene oxide gas prior to experiments. Contact angles on all surfaces were measured using water. 5 x 10(4) cells/ml of ATCC CRL 1486 human embryonic palatal mesenchyme (HEPM) cells, an osteoblast precursor cell line, were used for the cell attachment study. SEM evaluations were performed on cells attached to all surfaces. Contact angles and cell attachment on all surfaces were statistically analyzed using ANOVA. The chitosan-coated surfaces (76.4 +/- 5.1 degrees) exhibited a significantly greater contact angle compared to control Ti surfaces (32.2 +/- 6.1 degrees). Similarly, chitosan-coated surfaces exhibited significantly greater (P < 0.001) albumin adsorption, fibronectin adsorption and cell attachment, as compared to the control Ti surfaces. Coating chitosan on Ti surfaces decreased the wettability of the Ti, but increased protein adsorption and cell attachment. Increased protein absorption and cell attachment on the chitosan-coated Ti may be of benefit in enhancing osseointegration of implant devices.

Experimental studies on bone induction using low-molecular-weight poly (DL-lactide-co-glycolide) as a carrier for recombinant human bone morphogenetic protein-2.

An appropriate carrier acting as a slow delivery vehicle for the BMPs is required for maximal clinical effectiveness of these bone-inductive proteins. The purpose of this study was to evaluate a low-molecular-weight PLGA copolymer as a synthetic, biodegradable carrier for rhBMP-2 implantation in vivo. Two, 10, or 50 microg of recombinant human BMP-2 were mixed with 10 mg of a poly (DL-lactide-co-glycolide) (PLGA) 50:50 copolymer and implanted into the calf muscles of Wistar rats. Soft X-ray analysis and histologic examination indicated that new bone formation occurred at all rhBMP-2-implanted sites within 3 weeks after implantation. Correlation of rhBMP-2 concentration with the amount of bone induction was confirmed by specific alkaline phosphatase activity and calcium content assay. In vitro analysis indicated that 78.5% of the PLGA copolymer was degraded to smaller molecular weight material after 14 days in PBS solution. It is suggested that rhBMP-2 was released in an active form at the implant site during the degradation of the copolymer, resulting in the induction of new bone formation. Thus this low-molecular-weight PLGA copolymer material represents a promising delivery vehicle for BMPs, and possibly other growth factors, around dental and orthopedic implants.

Effect of poling conditions on growth of calcium phosphate crystal in ferroelectric BaTiO3 ceramics.

Recently, ceramic materials have been given a lot of attention as candidates for implant materials, since they possess biologically favorable characteristics for osseointegration. Among them, BaTiO3 (BTO) ceramics are ferroelectric and piezoelectric after poling treatments. However, little or no information is available on the poling condition of BTO and their effect on calcium phosphate (CaP) formation. In this study, the effect of poling conditions on the formation of CaP layer was investigated. It was observed from this study that CaP was formed on negatively charged BTO surfaces. An increase in Ca/P ratio to 1.67 was observed when the poling temperature was increased above the Curie temperature. On positively charged BTO, no CaP layer was observed.

Bone response to radio frequency sputtered calcium phosphate implants and titanium implants in vivo.

The objective of this study was to evaluate the effect of radio frequency sputtered calcium phosphate (CaP) coatings of titanium (Ti) implants on the bond strength at the bone-implant interface and percent bone contact length. Cylindrical coated or noncoated implants (4.0-mm diameter by 8-mm long) were implanted for 3 and 12 weeks. At 3 weeks after implant placement, the ultimate interfacial strengths for as-deposited CaP-coated and heat-treated CaP-coated implants were 2.29 +/- 0.14 MPa and 1.28 +/- 0.04 MPa, respectively. These ultimate interfacial strength values at 3 weeks were statistically greater than the mean ultimate interfacial strength for control Ti implants (0.67 +/- 0.13 MPa). At 12 weeks after implant placement, no statistical differences in the mean ultimate interfacial strengths were observed between the as-deposited CaP-coated, heat-treated CaP-coated, and control Ti implants. Histomorphometric evaluation indicated greater percent bone contact lengths for the as-deposited CaP-coated implants compared with the heat-treated CaP-coated and control Ti implants 3 and 12 weeks after implant placement.

Effect of biofluid environment on the dissolution and flexural strength of calcium phosphate bone cements.

This study investigates the dissolution of calcium (Ca2+), phosphorus (P5+), and the transverse strength of commercially available calcium phosphate (CaP) bone cements after immersion in fetal bovine serum and a tissue fluid substitute. It was observed that although a continual increase in Ca2+ and P5+ dissolution was detected in all three test media throughout the 21-day period, no statistical difference existed in the overall Ca2+ release after incubating the cements in the three different media. However, P5+ release after immersion in Tris solution (0.37 +/- 0.02 microgram/mm2) and fetal bovine serum solution (0.347 +/- 0.06 microgram/mm2) was significantly higher when compared with P5+ released (0.03 +/- 0.002 microgram/mm2) in tissue fluid substitute. In addition, no significant difference in transverse strength was observed for samples immersed in the three solutions during the 21-day period. However, the transverse strength for immersed CaP cement bars at 37 degrees C was statistically greater than non-immersed bars set aside at room temperature for the 21-day period (7.78 +/- 1.82 N and 3.19 +/- 0.93 N, respectively). It was concluded from this study that the transverse strength of the CaP bone cements was not significantly affected by the dissolution process but by the temperature at which the bone cement was exposed.

Directed evolution of polymerase function by compartmentalized self-replication.

We describe compartmentalized self-replication (CSR), a strategy for the directed evolution of enzymes, especially polymerases. CSR is based on a simple feedback loop consisting of a polymerase that replicates only its own encoding gene. Compartmentalization serves to isolate individual self-replication reactions from each other. In such a system, adaptive gains directly (and proportionally) translate into genetic amplification of the encoding gene. CSR has applications in the evolution of polymerases with novel and useful properties. By using three cycles of CSR, we obtained variants of Taq DNA polymerase with 11-fold higher thermostability than the wild-type enzyme or with a >130-fold increased resistance to the potent inhibitor heparin. Insertion of an extra stage into the CSR cycle before the polymerase reaction allows its application to enzymes other than polymerases. We show that nucleoside diphosphate kinase and Taq polymerase can form such a cooperative CSR cycle based on reciprocal catalysis, whereby nucleoside diphosphate kinase produces the substrates required for the replication of its own gene. We also find that in CSR the polymerase genes themselves evolve toward more efficient replication. Thus, polymerase genes and their encoded polypeptides cooperate to maximize postselection copy number. CSR should prove useful for the directed evolution of enzymes, particularly DNA or RNA polymerases, as well as for the design and study of in vitro self-replicating systems mimicking prebiotic evolution and viral replication.

Osteoblast responses to as-deposited and heat treated sputtered CaP surfaces.

The clinical success of dental implants is governed by implant surfaces and bone cell responses that promote rapid osseointegration. The objective of this study was to evaluate the in vitro osteoblast cell response to heat treated and non-heat treated CaP coatings. In this study, the heat treated surfaces exhibited a poorly crystallized HA-type structure whereas the non-heat treated surface exhibited an amorphous structure. The heat treated CaP surfaces were observed to have a mean contact angle measurement of 57.95+/-0.95 degrees, whereas the non-heat treated CaP surfaces were observed to have a mean contact angle measurement of 44.6+/-0.3 degrees. From the in vitro cell culture study, the ATTC CRL 1486 human embryonic palatal mesenchyme (HEPM) cells displayed a similar protein production and hexosaminidase activity on the heat treated and non-heat treated CaP surfaces throughout the nine day experiment. However, the HEPM cells cultured on non-heat treated CaP surfaces were observed to have higher specific ALP activity after nine days' incubation compared to cells cultured on heat treated CaP surfaces. The higher specific ALP activity by cells on non-heat treated surfaces were suggested to be attributed to the lower degree of crystallinity and the lower contact angles observed in this study.