Osteocytes Influence on Bone Matrix Integrity Affects Biomechanical Competence at Bone-Implant Interface of Bioactive-Coated Titanium Implants in Rat Tibiae.

Osseointegration is a prerequisite for the long-term success of implants. Titanium implants are preferred for their biocompatibility and mechanical properties. Nonetheless, the need for early and immediate loading requires enhancing these properties by adding bioactive coatings. In this preclinical study, extracellular matrix properties and cellular balance at the implant/bone interface was examined. Polyelectrolyte multilayers of chitosan and gelatin or with chitosan and Hyaluronic acid fabricated on titanium alloy using a layer-by-layer self-assembly process were compared with native titanium alloy. The study aimed to histologically evaluate bone parameters that correlate to the biomechanical anchorage enhancement resulted from bioactive coatings of titanium implants in a rat animal model. Superior collagen fiber arrangements and an increased number of active osteocytes reflected a significant improvement of bone matrix quality at the bone interface of the chitosan/gelatin-coated titan implants over chitosan/hyaluronic acid-coated and native implants. Furthermore, the numbers and localization of osteoblasts and osteoclasts in the reparative and remodeling phases suggested a better cellular balance in the chitosan/Gel-coated group over the other two groups. Investigating the micro-mechanical properties of bone tissue at the interface can elucidate detailed discrepancies between different promising bioactive coatings of titanium alloys to maximize their benefit in future medical applications.

Immunohistochemical comparison of lateral bone augmentation using a synthetic TiO2 block or a xenogeneic graft in chronic alveolar defects.

OBJECTIVES: To evaluate osteogenic markers and alveolar ridge profile changes in guided bone regeneration (GBR) of chronic noncontained bone defects using a nonresorbable TiO2 block. MATERIALS AND METHODS: Three buccal bone defects were created in each hemimandible of eight beagle dogs and allowed to heal for 8 weeks before GBR. Treatment was assigned by block randomization: TiO2 block: TiO2 -scaffold and a collagen membrane, DBBM particulates: Deproteinized bovine bone mineral (DBBM) and a collagen membrane, Empty control: Only collagen membrane. Bone regeneration was assessed on two different healing timepoints: early (4 weeks) and late healing (12 weeks) using several immunohistochemistry markers including alpha-smooth muscle actin (α-SMA), osteopontin, osteocalcin, tartrate-resistant acid phosphatase, and collagen type I. Histomorphometry was performed on Movat Pentachrome-stained and Von Kossa/Van Gieson-stained sections. Stereolithographic (STL) models were used to compare alveolar profile changes. RESULTS: The percentage of α-SMA and osteopontin increased in TiO2 group after 12 weeks of healing at the bone-scaffold interface, while collagen type I increased in the empty control group. In the defect area, α-SMA decreased in the empty control group, while collagen type I increased in the DBBM group. All groups maintained alveolar profile from 4 to 12 weeks, but TiO2 group demonstrated the widest soft tissue contour profile. CONCLUSIONS: The present findings suggested contact osteogenesis when GBR is performed with a TiO2 block or DBBM particulates. The increase in osteopontin indicated a potential for bone formation beyond 12 weeks. The alveolar profile data indicated a sustained lateral increase in lateral bone augmentation using a TiO2 block and a collagen membrane, as compared with DBBM and a collagen membrane or a collagen membrane alone.

Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals.

Bone histology of decalcified or undecalcified samples depends on the investigation. However, in research each method provides different information to answer the scientific question. Decalcification is the first step after sample fixation and governs what analysis is later feasible on the sections. Besides, decalcification is favored for immunostaining and in situ hybridization. Otherwise, sample decalcification can be damaging to bone biomaterials implants that contains calcium or strontium. On the other hand, after decalcification mineralization cannot be assessed using histology or imaging mass spectrometry. The current study provides a solution to the hardship caused by material presence within the bone tissue. The protocol presents a possibility of gaining sequential and alternating decalcified and undecalcified sections from the same bone sample. In this manner, investigations using histology, protein signaling, in situ hybridization, and mass spectrometry on the same sample can better answer the intended research question. Indeed, decalcification of sections and grindings resulted in well-preserved sample and biomaterials integrity. Immunostaining was comparable to that of classically decalcified samples. The study offers a novel approach that incites correlative analysis on the same sample and reduces the number of processed samples whether clinical biopsies or experimental animals.