Chia-Ling Ko

Properties of osteoconductive biomaterials: Calcium phosphate cement with different ratios of platelet-rich plasma as identifiers

This study aims to evaluate further the performance of a platelet-rich plasma (PRP) additive incorporated with calcium phosphate bone cement (CPC) in vitro to prove its efficiency as bone graft substitutes and its compatibility to be incorporated into the CPC with other techniques in clinical restoration in vivo. The growth factor release ability and the osteogenic evaluation of PRP, CPC, and PRP/CPC testing groups with 5, 10, and 15 wt.% PRP were compared in vitro. Four groups were measured using non-decalcified staining methods in vivo, which include the testing group of 10 wt.% PRP/CPC selected from the evaluation in vitro, by using both the autograft with rabbit trabecular and CPC-only as comparison groups and the group without grafting material as the control sample. The results obtained through specimen immersion show that growth factor release and alkaline phosphatase activities after osteoprogenitor cell culture had a significantly better effect on 10 and 15 wt.% PRP/CPC than on the other groups in vitro. Analysis results suggest that PRP was still retained in the CPC matrix even after 32 days of immersion. The results in vivo show that the histology of the autograft bone and the control group without grafting material exhibited fibrous connective and adipose tissues, which obviously filled the created cavity even at nine weeks after the operation. Osteoregeneration was more successful in the PRP-additive group, which accumulated bone remodeling than in the other groups. In conclusion, CPC could be a potential carrier with adequate PRP additives that bear a therapeutic potential for enhanced bone tissue regeneration.

Calcium phosphate bone cement with 10 wt% platelet-rich plasma in vitro and in vivo.

OBJECTIVES The aim of this study was to evaluate the performance of a 10 wt% platelet-rich plasma (PRP) additive composite with calcium phosphate cement (CPC) in vitro and in vivo. METHODS The in vitro testing of modulus, the apatite conversion rate, morphology, cell and alkaline phosphatase (ALP) activities, and in vivo testing of histological examinations between two groups of 10 wt% PRP/CPC and CPC were characterised and compared. RESULTS Although the crystallite morphologies showed a retarded effect in the PRP/CPC group in vitro, the modulus results showed that the 10 wt% PRP/CPC group had a significant reduction in strength but had no significant changes in the relative conversion ratio of the apatite phase with CPC only. The osteogenic evaluation of ALP expression was significantly increased by the PRP additives group with stem cells (D1) cultured for different periods (2-32 days). Our histological examinations showed that greater remodelling and the phenomenon of isolated/detached CPC particles were significantly observed at 9 weeks after implantation when the 10 wt% PRP/CPC composite was used. CONCLUSION The results demonstrate that CPC may be a potential candidate as a carrier with PRP additives for bone regeneration.

Deriving fast setting properties of tetracalcium phosphate/ dicalcium phosphate anhydrous bone cement with nanocrystallites on the reactant surfaces

OBJECTIVE This study attempts to reveal how nanocrystallites on the ceramic surfaces of non-dispersive calcium phosphate cement (nd-CPC) participate in setting processes as compared with conventional CPC (c-CPC). METHODS The compositions and morphologies of CPC during the early setting reactions were studied with X-ray diffraction and a scanning transmission electron microscope equipped with an energy dispersive spectroscopy system. The pH values and dispersive properties of CPC during the early setting reactions were investigated as well as the compressive strength of nd-CPC after 24h of immersion with varying liquid to powder ratios. RESULTS The mechanical strength of nd-CPC was approximately 60MPa after a 24h immersion in simulate body solution with a P/L ratio between 3.3 and 4.2g/mL. The nanocrystallites on the particle surfaces of nd-CPC were shown to grow rapidly and provided interlocking sites that allowed rapid development of the apatite phase in the cement, and were also shown to be non-dispersive in solution as determined by an injection test of c-CPC. CONCLUSIONS The interlocking particles produced by whisker growth on the ceramic particles or new crystallites formed between the ceramic particles caused the cement to be non-dispersive in solution. The particles of reactants with nanocrystallites on surfaces also gave this cement the ability to be shaped easily as a paste during an operation or to be injected into a cavity.

Interaction of progenitor bone cells with different surface modifications of titanium implant.

Changes in the physical and chemical properties of Ti surfaces can be attributed to cell performance, which improves surface biocompatibility. The cell proliferation, mineralization ability, and gene expression of progenitor bone cells (D1 cell) were compared on five different Ti surfaces, namely, mechanical grinding (M), electrochemical modification through potentiostatic anodization (ECH), sandblasting and acid etching (SLA), sandblasting, hydrogen peroxide treatment, and heating (SAOH), and sandblasting, alkali heating, and etching (SMART). SAOH treatment produced the most hydrophilic surface, whereas SLA produced the most hydrophobic surface. Cell activity indicated that SLA and SMART produced significantly rougher surfaces and promoted D1 cell attachment within 1 day of culturing, whereas SAOH treatment produced moderate roughness (Ra=1.26μm) and accelerated the D1 cell proliferation up to 7 days after culturing. The ECH surface significantly promoted alkaline phosphatase (ALP) expression and osteocalcin (OCN) secretion in the D1 cells compared with the other surface groups. The ECH and SMART-treated Ti surfaces resulted in maximum ALP and OCN expressions during the D1 cell culture. SLA, SAOH, and SMART substrate surfaces were rougher and exhibited better cell metabolic responses during the early stage of cell attachment, proliferation, and morphologic expressions within 1 day of D1 cell culture. The D1 cells cultured on the ECH and SMART substrates exhibited higher differentiation, and higher ALP and OCN expressions after 10 days of culture. Thus, the ECH and SMART treatments promote better ability of cell mineralization in vitro, which demonstrate their great potential for clinical use.

Roughened titanium surfaces with silane and further RGD peptide modification in vitro

The strategy to achieve osteoregeneration of dental implants during early-stage regeneration is strongly related to surface conditions for achieving highly successful effects after implantation. Surface modifications, namely, mechanical ground, silanization, bonded and sandblasted with pentasequence Gly-Arg-Gly-Asp-Ser (GRGDS) peptide, and acid-etched with Arg-Gly-Asp (RGD) peptide, were compared for their ability to support cell attachment, proliferation, and differentiation on titanium surfaces. The characteristics and comparative in vitro bio-interactions toward osteoprogenitor cells were tested in the four groups with various surface modifications. Compared with the other groups, the sandblasted and acid-etched, and silane with subsequent RGD peptide modified surfaces had the smallest wetting angle, absence of a significant cell viability difference, and largest quantity of alkaline phosphatase production during the expressions of early-stage cell differentiation. The method of synthesizing GRGDS peptides on roughened titanium surfaces has the potential to provide a combination of early bone regeneration and implant of long-term anchored capabilities.

Osteoregenerative capacities of dicalcium phosphate-rich calcium phosphate bone cement.

Calcium phosphate cement (CPC) is a widely used bone substitute. However, CPC application is limited by poor bioresorption, which is attributed to apatite, the stable product. This study aims to systematically survey the biological performance of dicalcium phosphate (DCP)-rich CPC. DCP-rich CPC exhibited a twofold, surface-modified DCP anhydrous (DCPA)-to-tetracalcium phosphate (TTCP) molar ratio, whereas conventional CPC (c-CPC) showed a onefold, surface unmodified DCPA-to-TTCP molar ratio. Cell adhesion, morphology, viability, and alkaline phosphatase (ALP) activity in the two CPCs were examined with bone cell progenitor D1 cultured in vitro. Microcomputed tomography and histological observation were conducted after CPC implantation in vivo to analyze the residual implant ratio and new bone formation rate. D1 cells cultured on DCP-rich CPC surfaces exhibited higher cell viability, ALP activity, and ALP quantity than c-CPC. Histological evaluation indicated that DCP-rich CPC showed lesser residual implant and higher new bone formation rate than c-CPC. Therefore, DCP-rich CPC can improve bioresorption. The newly developed DCP-rich CPC exhibited potential therapeutic applications for bone reconstruction.

Biphasic products of dicalcium phosphate-rich cement with injectability and nondispersibility

In this study, a calcium phosphate cement was developed using tetracalcium phosphate and surface-modified dicalcium phosphate anhydrous (DCPA). This developed injectable bone graft substitute can be molded to the shape of the bone cavity and set in situ through the piping system that has an adequate mechanical strength, non-dispersibility, and biocompatibility. The materials were based on the modified DCPA compositions of calcium phosphate cement (CPC), where the phase ratio of the surface-modified DCPA is higher than that of the conventional CPC for forming dicalcium phosphate (DCP)-rich cement. The composition and morphology of several calcium phosphate cement specimens during setting were analyzed via X-ray diffractometry and transmission electron microscopy coupled with an energy dispersive spectroscopy system. The compressive strength of DCP-rich CPCs was greater than 30MPa after 24h of immersion in vitro. The reaction of the CPCs produced steady final biphasic products of DCPs with apatite. The composites of calcium phosphate cements derived from tetracalcium phosphate mixed with surface-modified DCPA exhibited excellent mechanical properties, injectability, and interlocking forces between particles, and they also featured nondispersive behavior when immersed in a physiological solution.

Thermal cycling effects on adhesion of resin-bovine enamel junction among different composite resins

Thermal cycling is used to mimic the changes in oral cavity temperature experienced by composite resins when used clinically. The purpose of this study is to assess the thermal cycling effects of in-house produced composite resin on bonding strength. The dicalcium phosphate anhydrous filler surfaces are modified using nanocrystals and silanization (w/NP/Si). The resin is compared with commercially available composite resins Filtek Z250, Z350, and glass ionomer restorative material GIC Fuji-II LC (control). Different composite resins were filled into the dental enamel of bovine teeth. The bond force and resin-enamel junction graphical structures of the samples were determined after thermal cycling between 5 and 55°C in deionized water for 600 cycles. After thermal cycling, the w/NP/Si 30wt%, 50wt% and Filtek Z250, Z350 groups showed higher shear forces than glass ionomer GIC, and w/NP/Si 50wt% had the highest shear force. Through SEM observations, more of the fillings with w/NP/Si 30wt% and w/NP/Si 50wt% groups flowed into the enamel tubule, forming closed tubules with the composite resins. The push-out force is proportional to the resin flow depth and uniformity. The push-out tubule pore and resin shear pattern is the most uniform and consistent in the w/NP/Si 50wt% group. Accordingly, this developed composite resin maintains great mechanical properties after thermal cycling. Thus, it has the potential to be used in a clinical setting when restoring non-carious cervical lesions.

Characterization of the aspects of osteoprogenitor cell interactions with physical tetracalcium phosphate anchorage on titanium implant surfaces

Well-designed implants are used not only to modify the geometry of the implant but also to change the chemical properties of its surfaces. The present study aims to assess the biofunctional effects of tetracalcium phosphate (TTCP) particles as a physical anchor on the implant surface derived through sandblasting. The characteristics of the surface, cell viability, and alkaline phosphatase (ALP) activity toward osteoprogenitor cells (D1) were obtained. D1 cells were cultured on a plain surface that underwent sandblasting and acid etching (SLA) (control SLA group) and on different SLA surfaces with different anchoring TTCP rates (new test groups, M and H). The mean anchoring rates were 57% (M) and 74% (H), and the anchored thickness was estimated to range from 12.6μm to 18.3μm. Compared with the control SLA surface on Ti substrate, the new test groups with different TTCP anchoring rates (M and H) failed to improve cell proliferation significantly but had a well-differentiated D1 cell phenotype that enhanced ALP expression in the early stage of cell cultures, specifically, at day 7. Results suggest that the SLA surface with anchored TTCP can accelerate progenitor bone cell mineralization. This study shows the potential clinical application of the constructed geometry in TTCP anchorage on Ti for dental implant surface modification.

Thermal cycling effect of dicalcium phosphate-reinforced composites on auto-mineralized dental resin

The mineralizing capabilities of surface-modified dicalcium phosphate anhydrous (DCPA), reinforced and treated with nanocrystals and capped with silane, in composite resins were analyzed via thermal cycling. We compared two light-curable composites that were mixed at filler-to-resin mass ratios of 30/70 and 50/50. The strengths, elastic moduli, and topographical structures of the samples were determined after thermal cycling between 5 and 55°C in deionized water for 600 and 2400 cycles. Silane-capped particles decreased the strength but enhanced the mineralizing capability of the composites. Nanocrystal-treated filler surfaces significantly increased the strength and moduli of the composites after 600 thermal cycles. However, these values declined after 2400 thermal cycles. The nanocrystal-treated filler surfaces prevented the reduction in strength before and after 2400 thermal cycles. Prior to silane capping, the nanocrystal-treated DCPA filler surfaces exhibited good mineralization capability without compromising strength. The potential for barrier generation through mineralization yielded positive effects and prevented micro-leakages.