Benefits and Biosafety of Use of 3D-Printing Technology for Titanium Biomedical Implants: A Pilot Study in the Rabbit Model.

BACKGROUND: Titanium has been used in osteosynthesis for decades and its compatibility and safety is unquestioned. Studies have shown that there is release and collection of titanium in the organ systems with little note of toxicity. The gold standard is considered to be titanium osteosynthesis plate produced by milling methods. The use of customized titanium plates produced with 3D printing, specifically direct metal laser sintering, have found increasing use in recent years. It is unknown how much titanium is released in these printed titanium implants, which is known to be potentially porous, depending on the heat settings of the printer. We hypothesize that the amount of titanium released in printed titanium implants may be potentially more or equal compared to the gold standard, which is the implant produced by milling. METHODS: We studied the biosafety of this technology and its products by measuring serum and organ titanium levels after implantation of 3D-printed versus traditionally fabrication titanium plates and screws in a pilot study using the rabbit model. A total of nine rabbits were used, with three each in the control, milled and printed titanium group. The animals were euthanized after six months. Serum and organs of the reticuloendothelial system were harvested, digested and assayed for titanium levels. RESULTS: Organ and serum titanium levels were significantly higher in rabbit subjects implanted with titanium implants (milled and printed) compared to the control group. However, there was no significant difference in organ and serum titanium levels of subjects implanted with milled and traditionally fabricated titanium implants. CONCLUSIONS: The biosafety of use of 3D-printed titanium implants and traditionally fabricated titanium implants are comparable. With this in mind, 3D-printed custom implants can not only replace, but will very possibly surpass traditionally fabricated titanium implants in the mode and extent of use.

Investigation of trefoil factor expression in saliva and oral mucosal tissues of patients with oral squamous cell carcinoma.

OBJECTIVES: The aims of our study were to determine levels of trefoil factor (TFF) peptides in saliva and oral mucosal tissues from patients with oral squamous cell carcinoma (OSCC), and to evaluate whether individual members of TFFs (TFF1, TFF2, and TFF3) might act as biomarkers of disease. MATERIALS AND METHODS: Saliva samples were from 23 healthy subjects and 23 OSCC patients. Tissue samples were collected from 32 normal oral mucosa (NOM) and 32 OSCC biopsy specimens. ELISA and immunohistochemical methods were used to evaluate the expression of TFF1, TFF2, and TFF3 in saliva and oral mucosal tissues, respectively. RESULTS: Expression of TFF2 and TFF3 in oral mucosal tissues of OSCC patients was strongly downregulated when compared to healthy subjects (p < 0.001 and p = 0.002, respectively). However, there were no differences in levels of salivary TFF concentrations between OSCC patients and healthy subjects. CONCLUSIONS: The present study extends previous observations, demonstrating the reduction of TFF2 and TFF3 expression in oral mucosal tissues of OSCC patients. CLINICAL RELEVANCE: These findings suggest the clinical significance of TFF2 and TFF3 molecules as negative markers of tumor progression in OSCC. Quantification of TFF levels in saliva may not be optimal in terms of diagnostic or predictive value for OSCC derived from oral mucosa.

Bacterial cellulose adhesive composites for oral cavity applications.

Topical approaches to oral diseases require frequent dosing due to limited retention time. A mucoadhesive drug delivery platform with extended soft tissue adhesion capability of up to 7 days is proposed for on-site management of oral wound. Bacterial cellulose (BC) and photoactivated carbene-based bioadhesives (PDz) are combined to yield flexible film platform for interfacing soft tissues in dynamic, wet environments. Structure-activity relationships evaluate UV dose and hydration state with respect to adhesive strength on soft tissue mimics. The bioadhesive composite has an adhesion strength ranging from 7 to 17 kPa and duration exceeding 48 h in wet conditions under sustained shear forces, while other mucoadhesives based on hydrophilic macromolecules exhibit adhesion strength of 0.5-5 kPa and last only a few hours. The work highlights the first evaluation of BC composites for mucoadhesive treatments in the buccal cavity.

Evaluation of decellularized tilapia skin as a tissue engineering scaffold.

Decellularized bovine and porcine tissues have been used as scaffolds to support tissue regeneration but inherit religious restrictions and risks of disease transmission to humans. Decellularized marine tissues are seen as attractive alternatives due to their similarity to mammalian tissues, reduced biological risks, and less religious restrictions. The aim of this study was to derive an acellular scaffold from the skin of tilapia and evaluate its suitability as a tissue engineering scaffold. Tilapia skin was treated with a series of chemical and enzymatic treatments to remove cellular materials. The decellularized tilapia skin (DTS) was then characterized and evaluated in vitro and in vivo to assess its biological compatibility. The results indicated that the decellularization process removed 99.6% of the DNA content from tilapia skin. The resultant DTS was shown to possess a high denaturation temperature of 68.1 ± 1.0°C and a high Young's modulus of 56.2 ± 14.4 MPa. The properties of DTS were also compared against those of crosslinked electrospun tilapia collagen membrane, another form of tilapia-derived collagen scaffold. In vitro studies revealed that both DTS and crosslinked electrospun tilapia collagen promoted cellular metabolic activity, differentiation, and mineralization of murine preosteogenic MC3T3-E1 cells. The rat calvarial defect model was used to evaluate the in vivo performance of the scaffolds, and both scaffolds did not induce hyperacute rejections. Furthermore, they enhanced bone regeneration in the critical defect compared with the sham control. This study suggests that tilapia-derived scaffolds have great potential in tissue engineering applications.

Application of BMP-Bone Cement and FGF-Gel on Periodontal Tissue Regeneration in Nonhuman Primates.

The ultimate challenge of tissue engineering research is the translation of experimental knowledge into clinical application. In the preclinical testing phase of any new therapy, animal models remain the gold standard. Therefore, the methodological choice of a suitable model is critical to meet the requirements for a safe clinical application of the developed treatment. For instance, we have shown in rats that the application of calcium phosphate cement (CPC)/propylene glycol alginate (PGA) with bone morphogenetic protein (BMP)-2 or fibroblast growth factor (FGF)-2 resulted in the regeneration of periodontal defects. However, it is debated whether using small models form a predictive method for translation to larger species. At the same time, the 3R framework is encouraged as guiding principles of the ethical use of animal testing. Therefore, based on the successful rat study, the objective of this study was to further investigate the periodontal regenerative efficacy of the CPC/BMP and PGA/FGF system in a periodontal defect model with a low number of nonhuman primates (NHPs). Three Macaca fascicularis-overstocked from breeding for other purposes-were used (reuse of animals and appropriateness of the experimental animal species according to 3R framework). Three-wall periodontal defects were surgically created in the mandible. In total, 10 defects were created and distributed over two groups: (1) control group: PGA+CPC (n = 5) and (2) experimental group: PGA/FGF+CPC/BMP (n = 5). After 3 months, tissue regeneration was evaluated by histomorphometry and radiographic measurements. Data showed that epithelial downgrowth, cementum, and ligament regeneration were significantly enhanced in the experimental group compared with the control group (n = 5; p = 0.013, p = 0.028, and p = 0.018, respectively). However, the amount of newly formed bone did not differ (p = 0.146). Overall, as a translational proof-of-principle study, the hybrid periodontal regenerative method of CPC/BMP+PGA/FGF promoted periodontal regeneration in NHPs. This study warrants the application of CPC/BMP/PGA/FGF in clinical trials. Impact Statement This study validated an earlier successful periodontal regeneration strategy from a rat model into a few spare nonhuman primates (NHPs). The hybrid periodontal regenerative method of calcium phosphate cement (CPC)/bone morphogenetic protein (BMP)-2/propylene glycol alginate (PGA)/fibroblast growth factor (FGF)-2 promoted periodontal regeneration in NHPs, which corroborated the previous rat results. This translational approach was a very practical option and thus reduced the number and species of experimental animals in translational research. These results found in NHPs indicate a consistent conclusion with the earlier findings in the rat model. It further warrants the application of CPC/BMP-2+PGA/FGF-2 in human clinical trials.

Bone Response to Porous Poly(methyl methacrylate) Cement Loaded with Hydroxyapatite Particles in a Rabbit Mandibular Model.

The aim of the current study was to evaluate bone formation and tissue response to porous poly(methyl methacrylate) (PMMA) cement with or without hydroxyapatite (HA) in a rabbit mandibular model. Therefore, 14 New Zealand White rabbits were randomly divided into two groups of seven according to the designed study end points of 4 and 12 weeks. For each rabbit, two decorticated defects (6 mm in height and 10 mm in width for each) were prepared at both sides of the mandible. Subsequently, the defects were filled with, respectively, porous PMMA and porous PMMA-HA cement. After reaching the designated implantation period, the rabbits were euthanized and the mandibles were retrieved for histological analysis. Results showed that both porous PMMA and porous PMMA-HA supported bone repair. Neither of the bone cements caused significant inflammation to nerve or other surrounding tissues. After implantation of 12 weeks, majority of the porosity was filled with newly formed bone for both cements, which supports the concept that a porous structure within PMMA can enhance bone ingrowth. Histomorphometrical evaluation, using histological grading scales, demonstrated that, at both implantation times, the presence of HA in the PMMA enhanced bone formation. Bone was always in direct contact with the HA particles, while intervening fibrous tissue was present at the PMMA-bone interface. On the basis of results, it was concluded that injectable porous PMMA-HA cement might be a good candidate for craniofacial bone repair, which should be further evaluated in a more clinically relevant large animal model.

Tissue-engineered mandibular bone reconstruction for continuity defects: a systematic approach to the literature.

BACKGROUND: Despite significant surgical advances over the last decades, segmental mandibular bone repair remains a challenge. In light of this, tissue engineering might offer a next step in the evolution of mandibular reconstruction. PURPOSE: The purpose of the present report was to (1) systematically review preclinical in vivo as well as clinical literature regarding bone tissue engineering for mandibular continuity defects, and (2) to analyze their effectiveness. MATERIALS AND METHODS: An electronic search in the databases of the National Library of Medicine and ISI Web of Knowledge was carried out. Only publications in English were considered, and the search was broadened to animals and humans. Furthermore, the reference lists of related review articles and publications selected for inclusion in this review were systematically screened. Results of histology data and amount of bone bridging were chosen as primary outcome variables. However, for human reports, clinical radiographic evidence was accepted for defined primary outcome variable. The biomechanical properties, scaffold degradation, and clinical wound healing were selected as co-outcome variables. RESULTS: The electronic search in the databases of the National Library of Medicine and ISI Web of Knowledge resulted in the identification of 6727 and 5017 titles, respectively. Thereafter, title assessment and hand search resulted in 128 abstracts, 101 full-text articles, and 29 scientific papers reporting on animal experiments as well as 11 papers presenting human data on the subject of tissue-engineered reconstruction of mandibular continuity defects that could be included in the present review. CONCLUSIONS: It was concluded that (1) published preclinical in vivo as well as clinical data are limited, and (2) tissue-engineered approaches demonstrate some clinical potential as an alternative to autogenous bone grafting.

The use of a polycaprolactone-tricalcium phosphate scaffold for bone regeneration of tooth socket facial wall defects and simultaneous immediate dental implant placement in Macaca fascicularis.

Bone regeneration and aesthetic outcomes may be compromised when immediate implants are placed at extraction sites with dehiscence defects. The aim of this study was to compare, in a monkey model, peri-implant bone regeneration and implant stability after immediate implant placement into tooth sockets with facial wall defects in two treatment groups. In eight control monkeys, the bony defect was reconstructed with autogenous particulate bone, whereas in 10 test monkeys a polycaprolactone-tricalcium phosphate (PCL-TCP) scaffold was used. The monkeys were sacrificed after 6 months and the specimens were analyzed by histology and histomorphometry. Better maintenance of facial bone contour was noted in the test group; however, bone regeneration was seen only at areas adjacent to a bony wall of the defect. The mean bone-to-implant contact was 27.6 ± 19.1% (control group) versus 6.8 ± 7.9% (test group). The mean bone area percentage was 11.8 ± 10.1% (control group) versus 6.8 ± 6.9% (test group). Implant survival was 100% at 6 months for both the groups. It was concluded that although the use of a PCL-TCP scaffold showed better maintenance of the alveolar contour as compared to autogenous particulate bone at 6 months, there was minimal bone regeneration within the defect.

Segmental mandibular bone reconstruction with a carbonate-substituted hydroxyapatite-coated modular endoprosthetic poly(ɛ-caprolactone) scaffold in Macaca fascicularis.

A bio-degradable scaffold incorporating osteoinductive factors is one of the alternative methods for achieving the regeneration of a mandibular bone defect. The current pilot study addressed such a bone reconstruction in a non-human primate model, Macaca fascicularis monkeys, with an engineered poly(ɛ-caprolactone) (PCL) scaffold, provided with a carbonate-substituted hydroxyapatite coating. The scaffolds were implanted into unilaterally created mandibular segmental defects in 24 monkeys. Three experimental groups were formed: (1) scaffolds with rhBMP-2 (n = 8), (2) scaffolds with autologous mixed bone marrow cells (n = 8), and (3) empty scaffolds as a control group (n = 8). Evaluation was based on clinical observation as well as micro-CT, mechanical, and histological analyses. Despite a high infection rate, the overall results showed that the currently designed PCL scaffolds had insufficient load-bearing capability, and complete bone union was not achieved after 6 months of implantation. Nevertheless, the group of PCL scaffolds loaded with rhBMP-2 showed evidence of bone-regenerative potential, in contrast to PCL with autologous mixed bone marrow cells and the control group.

Subcutaneous tissue response to titanium, poly(ε-caprolactone), and carbonate-substituted hydroxyapatite-coated poly(ε-caprolactone) plates: a rabbit study.

The aim of this study was to evaluate the soft tissue response to poly(ε-caprolactone) (PCL) implants with and without carbonate-substituted hydroxyapatite (CHA) coating compared to the commonly used titanium alloy (Ti-6Al-4V)-machined surface. Experimental materials were implanted subcutaneously in New Zealand white rabbits for 5 weeks. The tissue attachment strength, as evaluated by a tissue peel test, histological and histomorphology analysis, as well as scanning electron microscopy were compared between groups. The peel test result revealed no statistically significant difference between groups. Histological analysis found fibrous capsule formation around all implant materials. The fibrous capsule around PCL implants with and without CHA coating was significantly thinner compared with the capsule thickness around the titanium implants. However, the inflammatory cells, as present at the fibrous capsule-implant interface, were found to be significantly lower in the Ti-group. In conclusion, the current data do not prove that PCL or PCL with a CHA coating results in a superior soft tissue response compared with a machined titanium implant.

Speech outcome and velopharyngeal function in cleft palate: comparison of Le Fort I maxillary osteotomy and distraction osteogenesis--early results.

OBJECTIVE: To compare speech outcome and velopharyngeal (VP) status of subjects with repaired cleft palate who underwent either conventional Le Fort I osteotomy or maxillary distraction osteogenesis to correct maxillary hypoplasia. DESIGN: Prospective randomized study with blind assessment of speech outcome and VP status. SUBJECTS: Twenty-two subjects were randomized into conventional Le Fort I osteotomy and Le Fort I distraction groups. All were native Chinese (Cantonese) speakers. METHOD: Perceptual judgment of resonance and nasal emission, study of VP structures by nasoendoscopy, and instrumental measurement by nasometry. Assessments were performed preoperatively and at 3 months postoperatively. MAIN OUTCOME MEASURES: Assessment of VP closure, perceptual rating of hypernasality and nasal emission, nasalance, and amount of maxillary advancement. RESULTS: There was no statistical difference in any of the outcome measures between the 10 subjects with conventional Le Fort I osteotomy and the 12 subjects with maxillary distraction: hypernasality (chi-square = 3.850, p = 0.221), nasal emission (chi-square = 0.687, p = 0.774), VP gap size (chi-square = 1.527, p = 0.635, and nasalance (t = -0.145, p = 0.886). There was no correlation between amount of maxillary advancement and any of the outcome measures (p = .05 for all). Changes in VP gap size and resonance are described. CONCLUSION: Results need to be interpreted with caution because of the small sample size and early follow-up. However, this study utilized an assessment protocol involving a variety of outcome measures and careful consideration of reliability factors, which can be a model for further and follow-up studies.

The effect of cranio-maxillofacial osteotomies and distraction osteogenesis on speech and velopharyngeal status: a critical review.

OBJECTIVES: To review the impact of maxillary advancement by orthognathic surgery and distraction osteogenesis on speech and velopharyngeal status based on the literature of the past 30+ years, to review the methods employed in previous studies to explain discrepancies in results, and to make recommendations for future studies. METHOD: Thirty-nine published articles on the effect of cranio-maxillofacial osteotomies and distraction osteogenesis on speech and velopharyngeal status were identified and were systematically analyzed. A total of 747 cases of cleft and noncleft patients were selected, including craniofacial deformities and syndromes mainly involving maxillary hypoplasia. RESULTS: Findings varied. Many studies found that surgery had no impact on speech and velopharyngeal status. Some reported worsening only in patients with preexisting velopharyngeal impairment or those with borderline velopharyngeal function before surgery. There was no clear difference in outcome between distraction and conventional osteotomy, although there have been few systematic comparisons. There was great variation among reviewed studies in the number of subjects, speech sample, number and type of listeners, speech outcome measures, and timing of postoperative assessment. Few studies employed reliability measures. CONCLUSION: None of the 39 reviewed studies compared conventional osteotomy and distraction by including both groups in a single study. Randomized controlled trials with adequate number of subjects and follow-up duration are needed.