Prevalence, trend, and associated risk factors for cleft lip with/without cleft palate: a national study on live births from 2016 to 2021.

BACKGROUNDS: Cleft lip with or without cleft palate (CL/P) is the most common congenital craniofacial anomaly, including non-syndromic cleft lip with or without cleft palate and cleft palate only. Failure in the fusion of median and lateral nasal processes, the maxillary prominence, and soft tissues around the oral cavity can cause CL/P. Previously, the prevalence has been estimated to be 1 among every 1000 births in 2014 among American neonates and no other reports have been available since. Thus, this study aimed to calculate the prevalence and trend of isolated CL/P among American live births from 2016 to 2021 with its associated risk factors. METHODS AND MATERIALS: In this cross-sectional population-based retrospective study, we used live birth data provided by the National Center for Health Statistics (NCHS) from the Center for Disease Control and Prevention (CDC). We calculated the prevalence per 10,000 live births of isolated (non-syndromic) CL/P from 2016 to 2021. To examine risk factors for developing isolated CL/P, we used logistic regression modelling. RESULTS: The total prevalence per 10,000 births from 2016 to 2021 was 4.88 (4.79-4.97), for both sexes, and 5.96 (5.82-6.10) for males, and 3.75 (3.64-3.87) for females. The prevalence did not show any consistent linear decreasing or increasing pattern. We found significant association between increased odds of developing isolated CL/P among cases with 20 to 24 year-old mothers (OR = 1.07, 1.01-1.13, p = 0.013), mothers who smoked 11 to 20 cigarettes per day (OR = 1.46, 1.33-1.60, p <  0.001), mothers with extreme obesity (OR = 1.32, 1.21-1.43, p <  0.001), mothers with grade II obesity (OR = 1.32, 1.23-1.42, p <  0.001), mothers with pre-pregnancy hypertension (OR = 1.17, 1.04-1.31, p = 0.009), mothers with pre-pregnancy diabetes mellitus (OR = 1.96, 1.71-2.25, p <  0.001), and mothers who used assisted reproductive technology (OR = 1.40, 1.18-1.66, p <  0.001). CONCLUSIONS: Our findings suggest a minuscule increase, albeit insignificant, in the trend of CL/P prevalence from 2016 to 2021. Developing CL/P had greater odds among mothers with pre-pregnancy diabetes, smoking, obesity, and pre-pregnancy hypertension mothers along with mothers who used assisted reproductive technology. Isolated CL/P had the highest prevalence in non-Hispanic Whites, American Indian or Alaskan Native and Native Hawaiian and Other Pacific Islanders.

Application of biodegradable Patient-specific scaffolds for maxillofacial bone regeneration: a scoping review of clinical studies.

This study aimed to systematically review clinical studies in which biodegradable patient-specific scaffolds were used for bone regeneration. Studies in which biodegradable scaffolds were fabricated through computer-assisted design and computer-assisted manufacturing (CAD-CAM) procedures were included. Those that applied non-biodegradable materials or used biodegradable materials in a condensable powder or block form were excluded. Among a total of 26 included studies, 11 used customised allogeneic bone blocks, five used polycaprolactone (PCL)-containing scaffolds, four used hydroxyapatite (HA) scaffolds, and four biphasic calcium phosphate (BCP). The majority of the studies applied scaffolds for minor intraoral defects. All the large defects were reconstructed with polymer-containing scaffolds. Results of the included studies showed partial to complete filling of the defect following the application of biodegradable scaffolds. However, limited graft exposure was reported when using PCL, BCP, and HA scaffolds. Tissue engineering can be considered a potential method for the treatment of maxillofacial bone defects. However, more evidence is required, especially for the application of biodegradable scaffolds in large defects.

Addition of Bone-Marrow Mesenchymal Stem Cells to 3D-Printed Alginate/Gelatin Hydrogel Containing Freeze-Dried Bone Nanoparticles Accelerates Regeneration of Critical Size Bone Defects.

A 3D-printed biodegradable hydrogel, consisting of alginate, gelatin, and freeze-dried bone allograft nanoparticles (npFDBA), is developed as a scaffold for enhancing cell adhesion, proliferation, and osteogenic differentiation when combined with rat bone marrow mesenchymal stem cells (rBMSCs). This composite hydrogel is intended for the regeneration of critical-sized bone defects using a rat calvaria defect model. The behavior of rBMSCs seeded onto the scaffold is evaluated through scanning electron microscope, MTT assays, and quantitative real-time PCR. In a randomized study, thirty rats are assigned to five treatment groups: 1) rBMSCs-loaded hydrogel, 2) rBMSCs-loaded FDBA microparticles, 3) hydrogel alone, 4) FDBA alone, and 5) an empty defect serving as a negative control. After 8 weeks, bone regeneration is assessed using H&E, Masson's trichrome staining, and immunohistochemistry. The 3D-printed hydrogel displays excellent adhesion, proliferation, and differentiation of rBMSCs. The rBMSCs-loaded hydrogel exhibits comparable new bone regeneration to the rBMSCs-loaded FDBA group, outperforming other groups with statistical significance (P-value < 0.05). These findings are corroborated by Masson's trichrome staining and osteocalcin expression. The rBMSCs-loaded 3D-printed hydrogel demonstrates promising potential for significantly enhancing bone regeneration, surpassing the conventional clinical approach (FDBA).

Bilayer scaffolds/membranes for bone tissue engineering applications: A systematic review.

OBJECTIVE: This systematic review evaluates the purpose, materials, physio-mechanical, and biological effects of bilayer scaffolds/membranes used for bone tissue engineering applications. METHODS: A comprehensive electronic search of English-language literature from 2012 to October 2022 was conducted in PubMed, Scopus, ScienceDirect, and Google Scholar online databases according to the PRISMA 2020 guidelines. The quality of animal studies was evaluated through the SYRCLE's risk of bias tool. RESULTS: A total of 77 studies were sought for retrieval, and 39 studies met the inclusion criteria. According to the synthesis results, most bilayers had a dense barrier layer that prevented connective tissue penetration and a loose osteogenic layer that supported cell migration and osteogenesis. PLGA, PCL, and chitosan were the most common polymers in the barrier layers, while the most utilized polymers in osteogenic layers were PLGA and gelatin. Electrospinning and solvent casting were the most common fabrication methods to design the bilayer structures. Many studies reported higher biological results for bilayers compared to their single layers. Also, fabricated bilayers' in vitro osteogenesis and in vivo new bone formation were significantly superior or at least comparable to the frequently used commercial membranes. CONCLUSION: 1) Bilayers with two distinct layers and different materials, porosities, mechanical properties, and biological behavior can significantly improve heterogeneous bone regeneration; 2) the addition of ceramics and/or drugs to the osteogenic layer enhances the osteogenic properties of the bilayers; 3) fabrication method and pore size of the layers play an important role in determining the mechanical and biological behavior of them.

Tissue buccal fat pad-stromal vascular fraction as a safe source in maxillofacial bone regeneration: A clinical pilot study.

AIM: The purpose of this study was to examine the biological properties of the buccal fat pad (BFP)-derived tissue stromal vascular fraction (tSVF) in vitro and compare them with BFP-derived cellular SVF (cSVF). Furthermore, a clinical pilot study assessed the safety of using BFP-derived tSVF for maxillofacial bone regeneration. MATERIALS AND METHODS: This study was performed in two sections: 1) experimental section: BFP tissue was harvested from three healthy donors, and then cSVF and tSVF were isolated by enzymatic and mechanical methods to assess their biological properties and 2) clinical section: Ten patients with maxillofacial bone defects were enrolled according to eligibility criteria and offered two options for surgery, including autologous BFP-tSVF (n = 5) and autologous bone grafting (n = 5), to evaluate safety after a year of follow-up. RESULTS: The BFP-tSVF exhibited high cell viability and various cell surface markers, including CD45, CD31, and CD34. There was no population-doubling time and multilineage differentiation capacity compared with BFP-cSVF. BFP-tSVF is safe because of the lack of intervention-related adverse events reported in donor and surgery sites during a one-year period. In addition, cell therapy was feasible because it can be performed during surgery and requires little preparation time. Patients in the ABG group experienced pain and tenderness in the iliac crest, leading to dissatisfaction and complications. CONCLUSION: The experimental results confirmed that the cells isolated from BFP-tSVF have stemness properties similar to BFP-cSVF. Clinical evaluation also indicated that this cellular product could be used safely to regenerate maxillofacial bone defects.

Surface Coating of Polyurethane Films with Gelatin, Aspirin and Heparin to Increase the Hemocompatibility of Artificial Vascular Grafts.

Purpose: A hemocompatible substrate can offer a wonderful facility for nitric oxide (NO) production by vascular endothelial cells in reaction to the inflammation following injuries. NO inhibits platelet aggregation this is especially critical in small-diameter vessels. Methods: The substrate films were made of polyurethane (PU) in a casting process and after plasma treatments, their surface was chemically decorated with polyethylene glycol (PEG) 2000, gelatin, gelatin-aspirin, gelatin-heparin and gelatin-aspirin-heparin. The concentrations of these ingredients were optimized in order to achieve the biocompatible values and the resulting modifications were characterized by water contact angle and Fourier transform infra-red (FTIR) assays. The values of NO production and platelet adhesion were then examined. Results: The water contact angle of the modified surface was reduced to 26±4∘ and the newly developed hydrophilic chemical groups were confirmed by FTIR. The respective concentrations of 0.05 mg/ml and 100 mg/mL were found to be the IC50 values for aspirin and heparin. However, after the surface modification with aspirin, the bioactivity of the substrate increased in compared to the other experimental groups. In addition, there was a synergistic effect between these reagents for NO synthesis. While, heparin inhibited platelet adhesion more than aspirin. Conclusion: Because of the highly hydrophilic nature of heparin, this reagent was hydrolyzed faster than aspirin and therefore its influence on platelet aggregation and cell growth was greater. Taken together, the results give the biocompatible concentrations of both biomolecules that are required for endothelial cell proliferation, NO synthesis and platelet adhesion.

3D-printed MgO nanoparticle loaded polycaprolactone ß-tricalcium phosphate composite scaffold for bone tissue engineering applications: In-vitro and in-vivo evaluation.

Magnesium (Mg) plays an important role in controlling bone apatite structure and density and is a potential bioactive material in repairing critical-sized bone defects. In this study, we aimed to evaluate the effect of adding NanoMgO to polycaprolactone/beta-tricalcium phosphate (PCL/ß-TCP) scaffolds on bone regeneration. Novel 3D-printed porous PCL/ß-TCP composite scaffolds containing 10% nanoMgO were fabricated by fused deposition modeling (FDM) and compared with PCL/ß-TCP (1:1) scaffolds (control). The morphology and physicochemical properties of the scaffolds were characterized by ATR-FTIR, XRD, scanning electron microscope-energy dispersive X-ray analysis (SEM-EDX), transmission-electron-microscopy (TEM), water contact angle, and compressive strength tests and correlated to its cytocompatibility and osteogenic capacity in-vitro. To evaluate in-vivo osteogenic capacity, bone-marrow-derived stem cell (BMSC)-loaded scaffolds were implanted into 8 mm rat critical-sized calvarial defects for 12 weeks. The hydrophilic scaffolds showed 50% porosity (pore size = 504 µm). MgO nanoparticles (91.5 ± 27.6 nm) were homogenously dispersed and did not adversely affect BMSCs' viability and differentiation. Magnesium significantly increased elastic modulus, pH, and degradation. New bone formation (NBF) in Micro-CT was 30.16 ± 0.31% and 23.56 ± 1.76% in PCL/ß-TCP/nanoMgO scaffolds with and without BMSCs respectively, and 19.38 ± 2.15% and 15.75 ± 2.24% in PCL/ß-TCP scaffolds with and without BMSCs respectively. Angiogenesis was least remarkable in PCL/ß-TCP compared with other groups (p < .05). Our results suggest that the PCL/ß-TCP/nanoMgO scaffold is a more suitable bone substitute compared to PCL/ß-TCP in critical-sized calvarial defects.

Periosteum as a covering vascular flap in posterior mandibular augmentation: A retrospective cohort study.

OBJECTIVE: To analyze the impact of creating periosteal vascular flaps on the amount of bone augmentation following inlay bone grafting (IBG) and cortical autogenous tenting (CAT). MATERIALS AND METHODS: This was a retrospective cohort study enrolling a sample cohort of patients presented to a private clinic in 2015 and 2019 for posterior mandibular ridge augmentation before dental implant placement. The predictor variables were surgical methods: CAT vs. CAT in conjunction with periosteal flap (CATP) vs. IBG vs. IBG in conjunction with periosteal flap (IBGP). The primary outcome variables were supra bundle bone (SBB) superior to the inferior alveolar canal (ΔH) and crestal width difference (ΔW) at a 4-month follow-up. Appropriate statistics were computed at 0.05 significance level. RESULTS: A total of 29 cases (10 males and 19 females) with a mean age of 57.96±7.14 years were included. A total of 33 sites were augmented through CATP, 16 sites through IBGP, 33 sites through CAT, and 11 sites through IBG techniques. All patients healed uneventfully without permanent neurosensory changes, and adequate horizontal (ΔW:3.33±0.71 mm) and vertical (ΔH:5.10±2.04 mm) bone dimensions were restored that allowed implant placement. Using periosteal vascular flaps significantly increased bone augmentation in both vertical and horizontal dimensions (P < 0.01). CONCLUSION: Periosteal vascular flaps can increase the efficacy of mandibular augmentation techniques and decrease post-surgical complications.

Comparative evaluation of the effects of three hydraulic calcium silicate cements on odontoblastic differentiation of human dental pulp stem cells: an in vitro study.

OBJECTIVE: The study aimed to compare the response of human dental pulp stem cells (hDPSCs) towards three hydraulic calcium silicate cements (HCSCs) by measuring cytotoxicity and expression of dentinogenic genes. METHODOLOGY: Dental pulps of five impacted mandibular third molars were extirpated as a source for hDPSCs. Next to culturing, hDPSCs were subjected to fluorescence-activated cell sorting after the third passage to validate stemness of the cells. Human DPSCs were exposed to diluted supernatants of OrthoMTA (OMTA), Biodentine (BD) and Calcium-Enriched Mixture (CEM) at concentrations 10, 25, 50 and 100% at the first, third and fifth day of culture. Then, cells were exposed to 10% concentrations supernatant of HCSCs to determine DSPP and DMP1 gene expression, using a quantitative polymerase-chain reaction. Data were analyzed using one-way and three-way ANOVA, followed by Tukey post hoc statistical tests. RESULTS: Optimal cell proliferation was observed in all groups, regardless of concentration and time-point. HCSC supernatants were non-cytotoxic to hDPSCs at all three time-points, except for 100% Biodentine on day five. On day seven, OMTA group significantly upregulated the expression of DSPP and DMP1 genes. On day 14, expression of DMP1 and DSPP genes were significantly higher in BD and OMTA groups, respectively. CONCLUSION: Biodentine significantly upregulated DMP1 gene expression over 14 days, whereas CEM was associated with only minimal expression of DSPP and DMP1 .

Application of CAD-CAM Technologies for Maxillofacial Bone Regeneration: A Narrative Review of the Clinical Studies.

The application of regenerative methods in treating maxillofacial defects can be categorized as functional bone regeneration in which scaffolds without protection are used and in-situ bone regeneration in which a protected healing space is created to induce bone formation. It has been shown that functional bone regeneration can reduce surgical time and obviate the necessity of autogenous bone grafting. However, studies mainly focused on applying this method to reconstruct minor bone effects, and more investigation concerning the large defects is required. In terms of in situ maxillofacial bone regeneration with the help of CAD-CAM technologies, the present data have suggested feasible mesh rigidity, perseverance of the underlying space, and apt augmentative results with CAD-CAM-based individualized Ti meshes. However, complications, including dehiscence and mesh exposure, coupled with consequent graft loss, infection and impeded regenerative rates have also been reported.

A smart magnetic hydrogel containing exosome promotes osteogenic commitment of human adipose-derived mesenchymal stem cells.

Objectives: Exosomes, as nano-sized extracellular vehicles acting as cell-to-cell communicators, are novel promising therapeutics in the area of bone tissue engineering. Moreover, magnetic nanoparticles, whose integration with other appropriate components is viewed as an intriguing approach to strengthen bone tissue engineering efficacy. We investigated the effect of magnetic enriched with exosomes on osteogenic differentiation. Materials and Methods: Exosomes were isolated from human adipose-derived mesenchymal stem cells by Exo-spin™ kit (MSC-EX). Alginate (Alg) scaffold containing 1% (w/w) cobalt ferrite nanoparticles (CoFe2O4) was produced. MSC-EX were gently loaded onto Alg and Alg-cobalt ferrite (Alg-CF) scaffolds yielding Alg-EX and Alg-CF-EX scaffolds. The effects of MSC-Ex and magnetic hydrogel composite under an external static magnetic field (SMF) on proliferation and differentiation of MSCs were evaluated by alkaline phosphatase (ALP) activity measurement, alizarin red staining, and energy dispersive X-ray (EDX) analysis. Results: Our results showed that Alg and Alg-CF scaffolds were not only cytotoxic but also supported AdMSCs proliferation. MSC-EX loading of the scaffolds enhanced AdMSCs proliferation significantly. According to the results, Alg-CF-EX scaffolds under magnetic stimulation exhibited the most potent effect on osteogenic differentiation of cultured AdMSCs as evidenced by higher ALP activity and mineralization. Conclusion: We provided evidence that the combination of Alg hydrogel, CFNPs, and MSC-EX resulted in the construction of a bone tissue-engineering scaffold that highly supports the osteogenic commitment of MSCs.

Effects of Lipopolysaccharide from Porphyromonas gingivalis and Escherichia coli on Gene Expression Levels of Toll-like Receptors and Inflammatory Cytokines in Human Dental Pulp Stem Cells.

BACKGROUND: Periodontal diseases originate from a group of oral inflammatory infections initiated by oral pathogens. Among these pathogens, Gram-negative bacteria such as p. gingivalis play a major role in chronic periodontitis. P. gingivalis harbours lipopolysaccharide (LPS) which enables it to attach to TLR2. OBJECTIVES: Evaluating the effects of P. gingivalis and E. coli LPS on the gene expression of TLRs and inflammatory cytokines in human dental pulp stem cells (hDPSCs). METHODS: We evaluated the expression level of TLR2, TLR4, IL-6, IL-10, and 1L-18 in hDPSCs treated with 1µg/mL of P. gingivalis lipopolysaccharide and E. coli LPS at three different exposure times using Real-time RT-PCR. RESULT: The test group treated with P. gingivalis LPS showed a high level of TLR4 expression in 24 hours exposure period and the lowest expression in 48 hours of exposure time. In the case of IL-10, the lowest expression was in the 24 hours exposure period. Although in the E.coli LPS treated group, IL-10 showed the highest expression in 24 and lowest in 48 hours exposure period. Moreover, IL-18 in P. gingivalis LPS treated group showed a significant difference between 6, 24, and 48-time periods of exposure, but not in the E. coli LPS treated group. CONCLUSION: Both types of LPS stimulate inflammation through TLR4 expression. P. gingivalis LPS performs more potentially than E. coli in terms of stimulating inflammation at the first 24 hours of exposure. Nevertheless, our study confirmed that increasing P. gingivalis and/or the E.coli LPS exposure time, despite acting as an inflammatory stimulator, apparently showed anti-inflammatory properties.

Orthognathic Surgery for Management of Gummy Smile.

Excessive gingival show is mainly caused by hypermobility of the upper lip, altered passive eruption, gingival hyperplasia, and bony maxillary vertical excess. Orthognathic surgery is the optimal treatment option for patients with moderate and severe vertical maxillary excess. Surrounding anatomic structures and soft tissue changes such as alternation in the nasal morphology confine the amount of impaction. Therefore, Le Fort 1 may be performed in conjunction with horseshoe osteotomy or partial turbinectomy. The possible necessity of further mandibular orthognathic surgeries and chin repositioning has to be considered. No common major complication and long-term relapse have been reported for maxillary impaction.

Gnathodiaphyseal dysplasia with a novel genetic variant in a large family from Iran.

BACKGROUND: Gnathodiaphyseal dysplasia (GDD) is an ultrarare autosomal dominant bone dysplasia characterized by cementoosseous lesions of the jawbones, bone fragility, frequent bone fractures at the young age, bowing of tubular bones, and diaphyseal sclerosis of long bones associated with generalized osteopenia. GDD is caused by point mutations in anoctamin-5 (ANO5) on chromosome 11p14.3. For the past few years, next generation sequencing (NGS) technology has facilitated the discovery of causative variants in genetically heterogeneous diseases. METHODS: In this study, exome sequencing (ES) was performed using the DNA sample of the proband. Family histories and clinical information were collected through comprehensive medical examination and genetic counseling. RESULTS: ES results identified a heterozygous variant, NM_213599.3:c.1078T>C(p.Cys360Arg) in the ANO5 gene. Sanger sequencing was performed to confirm the detected pathogenic variant in DNA samples of the entire family (except deceased individuals), which segregated with the disease within the family. Finally, in silico analysis was applied to test the pathogenicity of the variant using various online software. CONCLUSION: In summary, our investigation identified a novel pathogenic variant in the ANO5, responsible for gnathodiaphyseal dysplasia in a large Iranian family. Therefore, based on the present study, this variant can be helpful for diagnosis and effective management of GDD patients.

Safety of Intraparenchymal Injection of Allogenic Placenta Mesenchymal Stem Cells Derived Exosome in Patients Undergoing Decompressive Craniectomy Following Malignant Middle Cerebral Artery Infarct, A Pilot Randomized Clinical Trial.

Background: Malignant middle cerebral artery infarct (mMCAI) largely contributes to high mortality and physical disability among adults. Surviving individuals may not have proper outcomes and suffer from severe lasting disabilities. Utilization of stem cells and paracrine factor for regenerative purposes is considered as a potential strategy for patients with neurological deficits. While preclinical stroke studies have shown that mesenchymal stem cells (MSCs) reduce post-treatment neurological deficits and prevent disability and also promote recovery, few randomized clinical trials (RCT) have assessed exosome therapy in humans. Methods: In this RCT, we assessed the safety of intraparenchymal injection placenta MSC-derived Exosome in mMCAI patients with average age of 62 years between January, 2019, till September, 2020. The study was done in a single-center as an open-label RCT, with a 3-months follow-up. Primary outcomes assessed the safety and also disability indexes were followed. Results: Five mMCAI patients were included with mean NIHSS: 17.6 ± 5.02. The mean MRS was 3.25 ± 0.95 in three patients. No serious adverse events were observed. Hematoma or local reaction as excessive edema were not seen at the site of injection. Conclusions: Intraparenchymal implantation of MSC-EXO showed no post-interventional adverse effects in five ischemic stroke patients. It is proposed Local injection Exosome treatment following mMCAI can be safe and in future, it would be applied as a supportive, restorative and preventive treatment in patients who suffer from acute ischemic stroke and post ischemic disability.

3D Printed Composite Scaffolds in Bone Tissue Engineering: A Systematic Review.

OBJECTIVE: This study aimed to analyze the effect of fabrication factors on both biological and physico-chemical features of 3-dimensional (3D) printed composite scaffolds. METHOD: Electronic search was done according to the PRISMA guideline in PubMed and Scopus databases limited to English articles published until May 2021. Studies in which composite scaffolds were fabricated through computer-aided design and computer-aided manufacturing (CADCAM)- based methods were included. Articles regarding the features of the scaffolds fabricated through indirect techniques were excluded. RESULTS: Full text of 121 studies were reviewed, and 69 met the inclusion criteria. According to analyzed studies, PCL and HA were the most commonly used polymer and ceramic, respectively. Besides, the solvent-based technique was the most commonly used composition technique, which enabled preparing blends with high concentrations of ceramic materials. The most common fabrication method used in the included studies was Fused Deposition Modeling (FDM). The addition of bio-ceramics enhanced the mechanical features and the biological behaviors of the printed scaffolds in a ratio-dependent manner. However, studies that analyzed the effect of ceramic weight ratio showed that scaffolds with the highest ceramic content did not necessarily possess the optimal biological and non-biological features. CONCLUSION: The biological and physico-chemical behaviors of the scaffold can be affected by pre-printing factors, including utilized materials, composition techniques, and fabrication methods. Fabricating scaffolds with high mineral content as of the natural bone may not provide the optimal condition for bone formation. Therefore, it is recommended that future studies compare the efficiency of different kinds of biomaterials rather than different weight ratios of one type.

Prefabrication technique by preserving a muscular pedicle from masseter muscle as an in vivo bioreactor for reconstruction of mandibular critical-sized bone defects in canine models.

In vivo bioreactors serve as regenerative niches that improve vascularization and regeneration of bone grafts. This study has evaluated the masseter muscle as a natural bioreactor for ßTCP or PCL/ßTCP scaffolds, in terms of bone regeneration. The effect of pedicle preservation, along with sole, or MSC- or rhBMP2-combined application of scaffolds, has also been studied. Twenty-four mongrel dogs were randomly placed in six groups, including ßTCP, ßTCP/rhBMP2, ßTCP/MSCs, PCL/ßTCP, PCL/ßTCP/rhBMP2, and PCL/ßTCP/MSCs. During the first surgery, the scaffolds were implanted into the masseter muscle for being prefabricated. After 2 months, each group was divided into two subgroups prior to mandibular bone defect reconstruction; one with a preserved vascularized pedicle and one without. After 12 weeks, animals were euthanized, and new bone formation was evaluated using histological analysis. Histological analysis showed that all ß-TCP scaffold groups had resulted in significantly greater rates of new bone formation, either with a pedicle surgical approach or non-pedicle surgical approach, comparing to their parallel groups of ßTCP/PCL scaffolds (p ≤ .05). Pedicled ß-TCP scaffold groups that were treated with either rhBMP2 (48.443% ± 0.250%) or MSCs (46.577% ± 0.601%) demonstrated the highest rates of new bone formation (p ≤ .05). Therefore, masseter muscle can be used as a local in vivo bioreactor with potential clinical advantages in reconstruction of human mandibular defects. In addition, scaffold composition, pedicle preservation, and treatment with MSCs or rhBMP2, influence new bone formation and scaffold degradation rates in the prefabrication technique.

Effect of mesenchymal stem cells with platelet-rich plasma carriers on bone formation after rapid maxillary expansion: An Animal Study.

OBJECTIVE: To evaluate the effect of bone mesenchymal stem cells (BMSCs) with or without platelet-rich plasma (PRP) carriers on sutural new bone formation after rapid palatal expansion (RPE). SETTINGS AND SAMPLE POPULATION: Sixty male Wistar rats were used in this study. MATERIAL AND METHODS: All samples were subjected to 50cN of palatal expansion force for 7 days followed by 3 weeks of the retention period. The experimental groups received a single-dose injection of the specified solution at the time of retainer placement (BMSCs, PRP, BMSCs+PRP, normal saline). BMSCs used in this study were marked with the green fluorescent protein (GFP). New bone formation (NBF) in the sutural area was evaluated by µCT and occlusal radiography. In addition, semi-quantitative analyses were performed on histology images to analyse the quality of sutural bone, connective tissue and vascularization. Immunohistochemistry analyses were conducted for osteocalcin and collagen type I proteins. RESULTS: After the 21-day retention period, limited GFP marked cells were detected around the sutural area. Samples treated with BMSCs + PRP had the highest NBF and showed higher expression of collagen type I and osteocalcin. CONCLUSION: Injecting BMSCs + PRP may increase sutural bone density significantly. However, injecting BMSCs or PRP carriers alone did not affect sutural bone density.

Identification Osteogenic Signaling Pathways Following Mechanical Stimulation: A Systematic Review.

INTRODUCTION: It has been shown that mechanical forces can induce or promote osteogenic differentiation as well as remodeling of the new created bone tissues. To apply this characteristic in bone tissue engineering, it is important to know which mechanical stimuli through which signaling pathway has a more significant impact on osteogenesis. METHODS: In this systematic study, an electronic search was conducted using PubMed and Google Scholar databases. This study has been prepared and organized according to the preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines. Included studies were first categorized according to the in vivo and in vitro studies. RESULTS: Six types of mechanical stresses were used in these articles and the most commonly used mechanical force and cell source were tension and bone marrow-derived mesenchymal stem cells (BMMSCs), respectively. These forces were able to trigger twelve signaling pathways in which Wnt pathway was so prominent. CONCLUSION: 1) Although specific signaling pathways are induced through specific mechanical forces, Wnt signaling pathways are predominantly activated by almost all types of force/stimulation, 2) All signaling pathways regulate expression of RUNX2, which is known as a master regulator of osteogenesis, 3) In Tension force, the mode of force administration, i.e, continuous or noncontinuous tension is more important than the percentage of elongation.