Mandibular condylar process remodeling in rats with different bite-altering devices.

The objective was to compare different dental splint models and materials for inducing abnormal loading on the gross morphology and histological appearance of the mandibular condylar processes of Sprague Dawley rats. Three different types of dental splints (resin molar, aluminum incisor, stainless-steel incisor) were placed unilaterally to induce occlusal perturbation for 4 weeks. At that time, mandibular condylar processes were assessed by gross appearance and histology. Quantitative measurements were also conducted on the hematoxylin and eosin images for condyle shape. The results showed that although the condylar cartilage was affected by all splint types, the resin molar splint was associated with the most extensive mandibular condylar process remodeling, which was primarily a slant (skewness) of the lateral aspect of the condylar process. Additionally, quantitative measurements on the histological specimens demonstrated that the split and tilt angle of the left (ipsilateral) condylar processes in the resin molar group (124.8 ± 12.7° and 104.1 ± 12.7°, respectively) increased significantly (p < 0.05) when compared to right (contralateral) condylar processes (104.7 ± 5.8°and 91.6 ± 4.4°, respectively). However, no changes were noted on the thickness of the fibrocartilage layer at medial, central, and lateral regions of the condylar process. Another major finding is the high variability of morphology of the naïve animals. Future studies will assess the impact of longer durations of splinting, age, and sex on the remodeling of the mandibular condylar process, allowing for the development of diagnostics and therapies.

Regenerative Potential of Mandibular Condyle Cartilage and Bone Cells Compared to Costal Cartilage Cells When Seeded in Novel Gelatin Based Hydrogels.

The field of temporomandibular joint (TMJ) condyle regeneration is hampered by a limited understanding of the phenotype and regeneration potential of cells in mandibular condyle cartilage. It has been shown that chondrocytes derived from hyaline and costal cartilage exhibit a greater chondro-regenerative potential in vitro than those from mandibular condylar cartilage. However, our recent in vivo studies suggest that mandibular condyle cartilage cells do have the potential for cartilage regeneration in osteochondral defects, but that bone regeneration is inadequate. The objective of this study was to determine the regeneration potential of cartilage and bone cells from goat mandibular condyles in two different photocrosslinkable hydrogel systems, PGH and methacrylated gelatin, compared to the well-studied costal chondrocytes. PGH is composed of methacrylated poly(ethylene glycol), gelatin, and heparin. Histology, biochemistry and unconfined compression testing was performed after 4 weeks of culture. For bone derived cells, histology showed that PGH inhibited mineralization, while gelatin supported it. For chondrocytes, costal chondrocytes had robust glycosaminoglycan (GAG) deposition in both PGH and gelatin, and compression properties on par with native condylar cartilage in gelatin. However, they showed signs of hypertrophy in gelatin but not PGH. Conversely, mandibular condyle cartilage chondrocytes only had high GAG deposition in gelatin but not in PGH. These appeared to remain dormant in PGH. These results show that mandibular condyle cartilage cells do have innate regeneration potential but that they are more sensitive to hydrogel material than costal cartilage cells.

Poly (glycerol sebacate) elastomer supports bone regeneration by its mechanical properties being closer to osteoid tissue rather than to mature bone.

Mechanical load influences bone structure and mass. Arguing the importance of load-transduction, we investigated the mechanisms inducing bone formation using an elastomeric substrate. We characterized Poly (glycerol sebacate) (PGS) in vitro for its mechanical properties, compatibility with osteoprogenitor cells regarding adhesion, proliferation, differentiation under compression versus static cultures and in vivo for the regeneration of a rabbit ulna critical size defect. The load-transducing properties of PGS were compared in vitro to a stiffer poly lactic-co-glycolic-acid (PLA/PGA) scaffold of similar porosity and interconnectivity. Under cyclic compression for 7days, we report focal adhesion kinase overexpression on the less stiff PGS and upregulation of the transcription factor Runx2 and late osteogenic markers osteocalcin and bone sialoprotein (1.7, 4.0 and 10.0 folds increase respectively). Upon implanting PGS in the rabbit ulna defect, histology and micro-computed tomography analysis showed complete gap bridging with new bone by the PGS elastomer by 8weeks while minimal bone formation was seen in empty controls. Immunohistochemical analysis demonstrated the new bone to be primarily regenerated by recruited osteoprogenitors cells expressing periostin protein during early phase of maturation similar to physiological endochondral bone development. This study confirms PGS to be osteoconductive contributing to bone regeneration by recruiting host progenitor/stem cell populations and as a load-transducing substrate, transmits mechanical signals to the populated cells promoting differentiation and matrix maturation toward proper bone remodeling. We hence conclude that the material properties of PGS being closer to osteoid tissue rather than to mineralized bone, allows bone maturation on a substrate mechanically closer to where osteoprogenitor/stem cells differentiate to develop mature load-bearing bone. SIGNIFICANCE OF SIGNIFICANCE: The development of effective therapies for bone and craniofacial regeneration is a foremost clinical priority in the mineralized tissue engineering field. Currently at risk are patients seeking treatment for craniofacial diseases, traumas and disorders including birth defects such as cleft lip and palate, (1 in 525 to 714 live births), craniosynostosis (300-500 per 1,000,000 live births), injuries to the head and face (20 million ER visits per year), and devastating head and neck cancers (8000 deaths and over 30,000 new cases per year). In addition, approximately 6.2 million fractures occur annually in the United States, of which 5-10% fail to heal properly, due to delayed or non-union [1], and nearly half of adults aged 45-65 have moderate to advanced periodontitis with associated alveolar bone loss, which, if not reversed, will lead to the loss of approximately 6.5 teeth/individual [2]. The strategies currently available for bone loss treatment largely suffer from limitations in efficacy or feasibility, necessitating further development and material innovation. Contemporary materials systems themselves are indeed limited in their ability to facilitate mechanical stimuli and provide an appropriate microenvironment for the cells they are designed to support. We propose a strategy which aims to leverage biocompatibility, biodegradability and material elasticity in the creation of a cellular niche. Within this niche, cells are mechanically stimulated to produce their own extracellular matrix. The hypothesis that mechanical stimuli will enhance bone regeneration is supported by a wealth of literature showing the effect of mechanical stimuli on bone cell differentiation and matrix formation. Using mechanical stimuli, to our knowledge, has not been explored in vivo in bone tissue engineering applications. We thus propose to use an elastomeric platform, based on poly(glycerol sebacate (PGS), to mimic the natural biochemical environment of bone while enabling the transmission of mechanical forces. In this study we report the material's load-transducing ability as well as falling mechanically closer to bone marrow and osteoid tissue rather than to mature bone, allowed osteogenesis and bone maturation. Defying the notion of selecting bone regeneration scaffolds based on their relative mechanical comparability to mature bone, we consider our results in part novel for the new application of this elastomer and in another fostering for reassessment of the current selection criteria for bone scaffolds.

TMJ disc removal: comparison between pre-clinical studies and clinical findings.

The debate continues surrounding the use of disc removal (discectomy) as the primary surgical treatment for patients suffering from severe temporomandibular joint disorders. Furthermore, the effectiveness of pre-clinical animal models for predicting the response of the joint to discectomy in humans remains uncertain. This review compares the results of animal models with the most recent clinical findings while also focusing on investigations that use imaging as an assessment tool. After a review of the literature from well-established animal studies to clinical findings, it was found that the results of animal models for discectomy corresponded to the clinical findings seen in patients. Overall, there is adaptive remodeling or degeneration of the TMJ following discectomy. Additionally, there is some reduction in pain but with various amounts of dysfunction remaining following disc removal. Noteworthy, in the most recent clinical studies, imaging was not used as an outcome to assess the success of discectomy at preventing further joint degeneration.

Effects of ascorbic acid concentration on the tissue engineering of the temporomandibular joint disc.

The temporomandibular joint (TMJ) disc is a specialized fibrocartilaginous tissue. When the disc becomes an obstacle and becomes damaged, surgeons have no choice but to perform a discectomy. Tissue engineering may provide a novel treatment modality for TMJ disorder patients who undergo discectomy. No studies have been conducted on the most favourable media for TMJ disc cells. The objective of the current study was to examine the effects on biochemical and biomechanical properties of varying ascorbic acid concentrations (0, 25, or 50 microg/ml) on TMJ disc cells seeded on non-woven PGA scaffolds. The ascorbic acid concentration of the 25 microg/ml group resulted in more effective cell seeding of the scaffolds, with 1.53 million cells per construct, by comparison with the 0 and 50 microg/ml groups which had 1.20 million and 1.32 million cells per scaffold respectively. At week 4, the 25 microg/ml group had a higher collagen content than the 0 microg/ml group, with 30.4 +/- 2.7 and 24.9 +/- 3.3 microg of collagen per construct respectively. The 25 microg/ml group had a higher aggregate modulus than the 50 microg/ml group, with values of 6.1 +/- 1.3 and 4.0 +/- 0.9 kPa respectively at week 4. The results of this study indicate that the use of 25 microg/ml of ascorbic acid in culture media is effective for the tissue engineering of the TMJ disc, significantly outperforming media without or with 50 microg/ml of ascorbic acid.

Evaluation of three growth factors in combinations of two for temporomandibular joint disc tissue engineering.

Tissue engineering of the temporomandibular joint disc could be a great value in treatments that require discectomy. Potential benefit has been found in the use of three growth factors: insulin-like growth factor-I, basic fibroblast growth factor and transforming growth factor-beta1 in maintaining disc-like tissue in culture. In the present study, these three growth factors were combined in pairs and tested at two different concentrations over a 6-week period. All combinations of the growth factors appear to be beneficial since only three-quarters of the control constructs (without growth factors) retained mechanical integrity, compared with the majority of constructs exposed to growth factors. Importantly, the concentrations of the presented growth factors had a significant impact on the cellularity of constructs at Week 6. When a high concentration of the two growth factors was used, at least twice as many cells remained in the constructs compared with controls.

Biochemical analysis of the porcine temporomandibular joint disc.

Tissue engineering can be a boon in treating lesions of the disc in the temporomandibular joint (TMJ). Unfortunately, little is known about its biochemical content, so we analysed the discs of six slaughtered pigs. We measured the content and distribution of total DNA, glycosaminoglycan, and collagen. The mean (S.D.) content of DNA was 0.14% (0.08%) of the dry weight, of glycosaminoglycan 0.96% (0.39%), and of collagen 68.2% (14.5%). There were no significant differences from top to bottom, but from front to back the smallest concentration of glycosaminoglycan was in the posterior band, and the highest concentration of collagen was in the intermediate zone. The concentrations of DNA and glycosaminoglycan were higher in the medial than in the lateral area of the disc.