This study was undertaken to evaluate the effect of post-hydrothermal treatment on the biological responses of the plasma-sprayed hydroxyapatite (HA)-coated Ti-6Al-4V implant system both in vitro and in vivo. After hydrothermal treatment, the HA coating (HAC) shows the high mechanical strength and indices-of-crystallinity, denser microstructure, lower concentrations of amorphous and impurity phases, when compared with the as-sprayed HAC. The in vitro cell-culture studies, using UMR106 osteoblast-like cell, demonstrated no signifiacnt cell growth on both surface of as-sprayed and hydrothermal-treated HACs during 10-day culture. The in vivo studies, using the transcortical implant model in the femora of goats, evaluated the histological responses of two coatings. After 6 week of implantation, using backscattered electron images, no substantial histological variations in the extents of new bone apposition and new bone healing between the two HACs were observed. However, the as-sprayed HAC, owing to the dissolution induced the granular particles dissociated from the HAC, showed the statically lower extent of new bone apposition than hydrothermal-treated HAC at 12 weeks. The results suggest that hydrothermal treatment could be used to improve the mechanical strength, crystallinity, and phase composition of HAC, which are important factors of long-term fixation and stability of implant. Besides, the treated HAC could also achieve the initial fixation of implant in clinical use.

1. The results of spermatozoa assessment by the WST-8 (2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H-tetrazolium, monosodium salt) assay, flow cytometry (FC) or computer-assisted sperm analysis (CASA) were compared. 2. Different live/killed ratios of cockerel semen were serially diluted to 120, 60, and 30 × 10 cells/ml, and each sample was analysed by (1) WST-8 assay at 0, 10, 20, 30, 40, 50, 60 min, (2) viability with FC, and (3) motility with CASA. 3. The WST-8 reduction rate was closely correlated with spermatozoa viability and motility. The optimal semen concentration for the WST-8 assay was 120 × 10 cells/ml, and the standard curves for spermatozoa viability and motility predictions, respectively, were y = 162.8x + 104.96 (R = 0.9594) after 60 min of incubation and y = 225.09x + 96.299 (R = 0.8475) after 40 min of incubation. 4. It was concluded that the WST-8 assay is useful for the practical evaluation of cockerel spermatozoa viability and motility. Compared to FC and CASA, the WST-8 assay does not require expensive and complex instrumentation in the lab. Furthermore, one well of the WST-8 reaction can be used to predict spermatozoa viability and motility at the same time, which all lead it to be efficient and economical for semen quality assessment.