Sequential osseointegration from osseohealing to osseoremodeling - Histomorphological comparison of novel 3D porous and solid Ti-6Al-4V titanium implants.

In the present study, we analyzed the histological characteristics of osseointegration of an open-porous Ti-6Al-4V material that was produced in a space holder method creating a 3-D through-pores trabecular design that mimics the inhomogeneity and size relationships of trabecular bone in macro- as well as microstructure. Pairs of cylindrical implants with a porosity of 49 % and an average pore diameter of 400 µm (PI) or equal sized solid, corundum blasted devices (SI) as reference were bilaterally implanted press fit in the lateral condyles of 16 rabbits. Histological examination was performed after 4 weeks of short-term osseohealing and 12 weeks of mid-term osseoremodeling and we summarized the criteria for sequential osseointegration. After 4 weeks, osteoid had already been largely replaced by mineralized woven bone in both types of implants but was only represented to a greater extent in the deeper pores of PI. The cortical as well as trabecular region showed regular osseohealing with excessive and spatially undirected formation of immature woven bone. A dense bone mass was found in the cortical area, while in the trabecular region the bone mass was reduced distinctly, presenting large lacuna-like recesses and a demarcating trabecular structure. The pores near the implant surface contained more mineralized woven bone than the deeper pores. After 12 weeks, the osseoremodeling was largely completed with a physiological maturation to lamellar bone. The newly formed bone mass increased for PI and SI compared to the 4-week group and osteoid was only detectable in the deeper pores. The inhomogeneous trabecular design of the pores enables an excellent ingrowth of mineralized lamellar bone after remodeling to a pore depth of 1800 µm, which proves a functional load transfer from the surrounding bone into the implant. According to the concept of osseointegration by Branemark and Albrektsson, the histological evaluation confirms a successful, superior osseointegration of the presented porous properties improving long-term implant stability. The presented study protocol allows an excellent evaluation and comparison of the sequential osseointegration from short-term osseohealing to mid-term osseoremodeling.

Osseointegration of a novel 3D porous Ti-6Al-4V implant material - Histomorphometric analysis in rabbits.

Porous structure properties are known to conduct initial and long-term stability of titanium alloy implants. This study aims to assess the histomorphometric effect of a 3-D porosity in Ti-6Al-4V implants (PI) on osseointegration in comparison to solid Ti-6Al-4V implants (SI). The PI was produced in a spaceholder method and sintering and has a pore size of mean 400 µm (50 µm to 500 µm) and mimics human trabecular bone. Pairs of PI and equal sized SI as reference were bilaterally implanted at random in the lateral femoral condyle of 16 Chinchilla-Bastard rabbits. The animals were sacrificed after 4 and 12 weeks for histomorphometric analysis. The histomorphometric evaluation confirmed a successful short-term osseohealing (4 weeks) and mid-term osseoremodeling (12 weeks) for both types of implants. The total newly formed bone area was larger for PI than for SI after 4 and 12 weeks, with the intraporous bone area being accountable for the significant difference (p<0.05). A more detailed observation of bone area distribution revealed a bony accumulation in a radius of +/- 500 µm around the implant surface after remodeling. The bone-to-implant contact (BIC) increased significantly (p<0.05) from 4 to 12 weeks (PI 26.23 % to 42.68 %; SI 28.44 % to 47.47 %) for both types of implants. Due to different surface properties, however, PI had a significant (p<0.05) larger absolute osseous contact (mm) to the implant circumference compared to the SI (4 weeks: 7.46 mm vs 5.72 mm; 12 weeks: 11.57 mm vs 9.52 mm [PI vs. SI]). The regional influences (trabecular vs. cortical) on bone formation and the intraporous distribution were also presented. Conclusively, the porous structure and surface properties of PI enable a successful and regular osseointegration and enhance the bony fixation compared to solid implants under experimental conditions.

Generation of highly differentiated BHY oral squamous cell carcinoma multicellular spheroids.

Three-dimensional (3D) multicellular spheroids (MCS) are considered suitable models in cancer research and anticancer drug development. Although studying the complex tumour characteristics from all different degrees of malignancy is vital, MCS generation has only been described in a few moderately- and poorly differentiated oral squamous cell carcinoma (OSCC) cell lines. No previous study has demonstrated the MCS formation in a highly differentiated OSCC cell line. For the first time, the present study aimed to generate MCS from the highly differentiated OSCC cell line BHY. BHY spheroids were grown in three independent experiments in 96-well plates through the use of the liquid overlay technique. Although BHY cells are grow slowly and are difficult to culture, they formed compact MCS within 24 h. After 3 days of incubation, no further increase in spheroid size was observed. MCS were harvested, paraffin-embedded and 2 µm tissue sections were prepared for further analysis. The diameter and volume of each spheroid were determined. BHY MCS diameter ranged between 46.76 and 233.26 µm, with a volume range from 5.35×104-6.65×106 µm3. In conclusion, using the liquid overlay technique, the highly differentiated OSCC cell line BHY forms different sized spheroids, which may be used for further investigations.

Evaluation of EGFR protein expression by immunohistochemistry using H-score and the magnification rule: re-analysis of the SATURN study.

INTRODUCTION: The phase III SATURN study demonstrated that first-line maintenance erlotinib extended progression-free survival (PFS) and overall survival (OS) versus placebo in patients with advanced non-small cell lung cancer (NSCLC). Analysis of epidermal growth factor receptor (EGFR) expression by immunohistochemistry (IHC) found no significant interaction between EGFR IHC status and PFS (p = 0.63) or OS (p = 0.52). The FLEX study of first-line cetuximab plus chemotherapy demonstrated that EGFR IHC expression was predictive of improved OS with cetuximab when assessed by H-score with a magnification rule. This novel method was used to reassess samples from SATURN. METHODS: The H-score method assigned a score of 0-300 to each patient, based on the percentage of cells stained at different intensities viewed at various magnifications. The discriminatory threshold was set at 200, per the FLEX study, and existing samples were re-read and classed as low (H-score < 200) or high (≥200) EGFR expression. PFS and OS were re-analyzed based on these new classifications. RESULTS: In the overall and EGFR wild-type populations, erlotinib provided a consistent survival benefit versus placebo. Hazard ratios (HRs) in the overall population were similar between EGFR IHC-positive and -negative patients for median PFS (HR 0.68 [95% confidence interval (CI) 0.53-0.86] and 0.76 [95% CI 0.62-0.93], respectively) and OS (HR 0.80 [95% CI 0.62-1.05] and 0.80 [95% CI 0.64-1.01] for IHC-positive and IHC-negative, respectively). In the EGFR wild-type population, HRs were again similar between EGFR IHC-positive and -negative subpopulations for PFS (HR 0.69 [95% CI 0.51-0.95] and 0.84 [95% CI 0.63-1.12], respectively) and OS (HR 0.78 [95% CI 0.55-1.10] and 0.76 [95% CI 0.55-1.05], respectively). CONCLUSIONS: These data suggest that EGFR IHC does not have value as a marker to predict erlotinib benefit in the first-line maintenance setting for advanced NSCLC.