In Vivo Evaluation of Different Collagen Scaffolds in an Achilles Tendon Defect Model.

In the present study, a newly introduced bovine cross-linked collagen scaffold (test material) was investigated in vivo in an Achilles tendon defect model and compared to a commercially available porcine collagen scaffold (control material). In total, 28 male Sprague Dawley rats (about 400 g) were examined. The defined Achilles tendon defect of 5 mm of the right hind limb was replaced by one of the scaffold materials. After euthanasia, the hind limbs were transected for testing. Biomechanical evaluation was carried out via tensile testing (n = 8 each group, observation time: 28 days). Nonoperated tendons from the bilateral side were used as a control (native tendon, n = 4). For the histological evaluation, 12 animals were sacrificed at 14 and 28 days postoperatively (n = 3 each group and time point). Stained slices (Hematoxylin & Eosin) were evaluated qualitatively in terms of presence of cells and cell migration into scaffolds as well as structure and degradation of the scaffold. All transected hind limbs were additionally analyzed using MRI before testing to verify if the tendon repair using a collagen scaffold was still intact after the observation period. The maximum failure loads of both scaffold materials (test material: 54.5 ± 16.4 N, control: 63.1 ± 19.5 N) were in the range of native tendon (76.6 ± 11.6 N, p ≥ 0.07). The stiffness of native tendons was twofold higher (p ≤ 0.01) and the tear strength was approximately fivefold higher (p ≤ 0.01) compared to the repaired tendons with both scaffolds. Histological findings indicated that neither the test nor the control material induced inflammation, but the test material underwent a slower remodeling process. An overall repair failure rate of 48% was observed via MRI. The experimental data of the newly developed test material showed similar outcomes compared to the commercially available control material. The high repair failure rate indicated that MRI is recommended as an auxiliary measurement tool to validate experimental data.

Comparison of different suture techniques for Achilles tendon repair in rat model using collagen scaffolds.

PURPOSE: Tendon injury is an increasing problem in orthopedic and trauma surgery due to aging of the population and increased activity demands. Many rodent animal models are used in order to evaluate tendon reconstruction. Although tendon reruptures are a well- -known clinical problem, the outcomes of tendon repair in animal models are rarely discussed in the literature. The goal of the present experimental study was to compare the primary fixation stability of three suture techniques for repair of Achilles tendon defects in a rat model using a collagen scaffold. METHODS: Cadaveric left hind limbs of Sprague-Dawley rats were prepared with an Achilles tendon defect of 3 mm and rejoined using a collagen scaffold. Three suture configurations (simple, simple stitch with additional framing suture, and modified Mason-Allen stitch; n = 5 each) underwent tensile testing until complete failure was observed. RESULTS: Under a load of a mean value of 6.6 N, the failure load of simple stitches was the significantly lowest ( p < 0.01). Both, modified Mason-Allen stitches and simple stitches with additional framing suture showed a mean failure load of more than 14 N. Regardless of the suture technique, most of the samples showed failure of tendon due to suture tear-out. The suture material as well as the scaffold remained mostly intact. CONCLUSIONS: Although simple end-to-end suture techniques are common in the literature, stitches with more suture strands should be preferred. Using techniques like an additional framing suture or modified Mason-Allen stitch, maximum failure load can be doubled and the risk of tendon rerupture may be decreased within in vivo testing.

Biomechanical, Biochemical, and Cell Biological Evaluation of Different Collagen Scaffolds for Tendon Augmentation.

Tendon augmentation is increasingly clinically relevant due to rising amount of tendon ruptures because of the aging and more demanding population. Therefore, newly developed scaffolds based on bovine epoxide stabilized collagen maintaining the native fibril-like collagen structure were characterized and compared to two commercially available porcine collagen scaffolds. For biomechanical testing (ultimate load, ultimate stress, stiffness, and elastic modulus), bovine collagen scaffolds were hydrated and compared to reference products. Cell viability and proliferation were assessed by seeding human primary fibroblasts on each collagen-based scaffold and cultured over various time periods (3 d, 7 d, and 14 d). Live/dead staining was performed and metabolic cell activity (WST-1 assay) was measured. Biochemical degradability was investigated by enzymatic digestion. The bovine collagen scaffold showed significantly enhanced biomechanical properties. These persisted over different rehydration times. Cell biological tests revealed that the bovine collagen scaffolds support reproducible cell colonization and a significant increase in the number of viable cells during cultivation. The results are comparable with the viability and proliferation rate of cells grown on porcine reference materials. With regard to biochemical degradability, all tested materials showed comparable resistance to enzymatic degradation in vitro. Due to imitating the natural tendon structure the new scaffold material is supposed to provide beneficial effects in future clinical application.

Quantification of osseointegration of plasma-polymer coated titanium alloyed implants by means of microcomputed tomography versus histomorphometry.

A common method to derive both qualitative and quantitative data to evaluate osseointegration of implants is histomorphometry. The present study describes a new image reconstruction algorithm comparing the results of bone-to-implant contact (BIC) evaluated by means of µCT with histomorphometry data. Custom-made conical titanium alloyed (Ti6Al4V) implants were inserted in the distal tibial bone of female Sprague-Dawley rats. Different surface configurations were examined: Ti6Al4V implants with plasma-polymerized allylamine (PPAAm) coating and plasma-polymerized ethylenediamine (PPEDA) coating as well as implants without surface coating. After six weeks postoperatively, tibiae were explanted and BIC was determined by µCT (3D) and afterwards by histomorphometry (2D). In comparison to uncoated Ti6Al4V implants demonstrating low BIC of 32.4% (histomorphometry) and 51.3% (µCT), PPAAm and PPEDA coated implants showed a nonsignificant increase in BIC (histomorphometry: 45.7% and 53.5% and µCT: 51.8% and 62.0%, resp.). Mean BIC calculated by µCT was higher for all surface configurations compared to BIC detected by histomorphometry. Overall, a high correlation coefficient of 0.70 (p < 0.002) was found between 3D and 2D quantification of BIC. The µCT analysis seems to be suitable as a nondestructive and accurate 3D imaging method for the evaluation of the bone-implant interface.

Evaluation of osseointegration of titanium alloyed implants modified by plasma polymerization.

By means of plasma polymerization, positively charged, nanometre-thin coatings can be applied to implant surfaces. The aim of the present study was to quantify the adhesion of human bone cells in vitro and to evaluate the bone ongrowth in vivo, on titanium surfaces modified by plasma polymer coatings. Different implant surface configurations were examined: titanium alloy (Ti6Al4V) coated with plasma-polymerized allylamine (PPAAm) and plasma-polymerized ethylenediamine (PPEDA) versus uncoated. Shear stress on human osteoblast-like MG-63 cells was investigated in vitro using a spinning disc device. Furthermore, bone-to-implant contact (BIC) was evaluated in vivo. Custom-made conical titanium implants were inserted at the medial tibia of female Sprague-Dawley rats. After a follow-up of six weeks, the BIC was determined by means of histomorphometry. The quantification of cell adhesion showed a significantly higher shear stress for MG-63 cells on PPAAm and PPEDA compared to uncoated Ti6Al4V. Uncoated titanium alloyed implants showed the lowest BIC (40.4%). Implants with PPAAm coating revealed a clear but not significant increase of the BIC (58.5%) and implants with PPEDA a significantly increased BIC (63.7%). In conclusion, plasma polymer coatings demonstrate enhanced cell adhesion and bone ongrowth compared to uncoated titanium surfaces.

In vivo monitoring of implant osseointegration in a rabbit model using acoustic sound analysis.

Implant osseointegration can currently only be assessed reliably post mortem. A novel method that relies on the principle of acoustic sound analysis was developed to enable examination of the longitudinal progress of osseointegration. The method is based on a magnetic sphere inside a hollow cylinder of the implant. By excitation using an external magnetic field, collision of the sphere inside the implant produces a sound signal. Custom-made titanium implants equipped thusly were inserted in each lateral femoral epicondyle of 20 New Zealand White Rabbits. Two groups were investigated: Uncoated, machined surface versus antiadhesive surface; and calcium phosphate-coated surface versus antiadhesive surface. The sound analysis was performed postoperatively and weekly. After 4 weeks, the animals were euthanized, and the axial pull-out strengths of the implants were determined. A significant increase in the central frequency was observed for the loose implants (mean pull-out strength 21.1 ± 16.9 N), up to 6.4 kHz over 4 weeks. In comparison, the central frequency of the osseointegrated implants (105.2 ± 25.3 N) dropped to its initial value. The presented method shows potential for monitoring the osseointegration of different implant surfaces and could considerably reduce the number of animals needed for experiments.