Preosteoblast Adhesion and Viability Study of Freeze-Dried Bovine Bone Block Scaffold Coated with Human Umbilical Cord Mesenchymal Stem Cell Secretome.

OBJECTIVES: Combining a three-dimensional scaffold with growth factors before implantation is one method used to increase scaffold bioactivity in bone tissue engineering. The mesenchymal stem cell (MSC)-conditioned medium (CM), called secretome, contains many proteins and growth factors required for tissue repair and growth. This study evaluated the bioactivity of a bovine bone scaffold combined with the secretome of human umbilical cord MSCs (hUC-MSCs) by analyzing MC3T3-E1 cell adhesion and viability on the scaffold. MATERIALS AND METHODS: This in vitro laboratory study evaluated the effect of hUC-MSC secretome applied to bovine bone scaffolds processed using various techniques on MC3T3-E1 cell adhesion and viability. The three experimental groups included deproteinized bovine bone mineral-secretome (DBBM-CM), freeze-dried bovine bone-secretome (FDBB-CM), and decellularized FDBB-CM, whereas the control group was treated with DBBM alone. The cell adhesion test was performed using the centrifugation method after 6 and 24 hours, whereas the cell viability test was conducted using the trypan blue exclusion method after 24, 48, and 72 hours. Cell attachment was visualized after 4',6-diamidino-2-phenylindole staining and viewed under inverted fluorescence microscopy. STASTICAL ANALYSIS: Statistical analyses were performed using one-way analysis of variance, followed by a post hoc test in cases of significant differences. RESULTS: Statistical analyses showed significantly greater adhesion of the preosteoblasts to the FDBB-CM scaffold at 6 hours (p = 0.002). The results of the adhesion test at 24 hours and the viability tests at all observation times showed no significant differences (p > 0.05). This study found that the average MC3T3-E1 cell adhesions and viabilities were highest for the FDBB-CM and DBBM-CM scaffolds. DBBM scaffolds with the secretome had better cell adhesion and viability than those without the secretome. CONCLUSION: The addition of MSC secretome increased bovine bone scaffold bioactivity especially in DBBM and FDBB scaffolds.

Effect of α-Mangostin on Interleukin-11 and Runt-related Transcription Factor-2 Gene Expression on Cell Line Osteoblast Cultures Induced with Lipopolysaccharide.

Background: Loss of permanent teeth after tooth extraction without replacement of missing teeth can result in impaired masticatory, esthetic, phonetic functions, and impaired balance of the masticatory organ in the mouth. Therefore, a method is needed to inhibit the alveolar bone resorption process so that the dimensions of the tooth socket can be maintained vertically or horizontally until the time of implant placement, which is called the socket preservation procedure. α-mangostin is known to have a potential anti-inflammatory effect and most likely can be used as a potential therapeutic agent to inhibit bone resorption caused by posttooth extraction inflammatory processes. Aims: The aim of the study was to determine the effect on the inflammatory process and osteogenesis on osteoblast cell line culture by induction with lipopolysaccharide (LPS) and α-mangostin. Materials and Methods: This was an in vitro laboratory experimental study on mouse osteoblast cell line culture. The treatment was given with LPS, α-mangostin, and combination on osteogenic medium, using the same concentration for all concentrates. The sample will then be processed and analyzed using the real-time polymerase chain reaction. Results: The highest interleukin-11 (IL-11) gene expression was found in α-mangostin treatment, but there was no significant difference in IL-11 expression between the study groups. The highest runt-related transcription factor-2 (RUNX-2) gene expression was found in a group that received induction with LPS and α-mangostin, and from these results, it was found that there was a significant difference in RUNX-2 expression between the study groups. Conclusions: LPS and α-mangostin can increase osteogenesis in osteoblast cell culture in the osteogenic medium.

Osteoinductive and Osteogenic Capacity of Freeze-Dried Bovine Bone Compared to Deproteinized Bovine Bone Mineral Scaffold in Human Umbilical Cord Mesenchymal Stem Cell Culture: An In Vitro Study.

OBJECTIVE: Freeze-dried bovine bone scaffold (FDBB) or decellularized FDBB (dc-FDBB) was developed as an ideal scaffold with osteoinductive properties. This research aims to compare the osteoinductive properties marked by the expression of runt-related transcription factor-2 (RUNX2) and Osterix (OSX) and the osteogenic capacity of these scaffolds imbued with human umbilical cord mesenchymal stem cells (hUCMSCs). MATERIALS AND METHODS: This study was performed in five experimental groups: a negative control group (C-) of hUCMSCs with a normal growth medium, a positive control group (C + ) of hUCMSCs with an osteogenic medium, experimental group 1 (E1) with an FDBB conditioned medium (CM), and experimental group 2 (E2) with a dc-FDBB-CM, and a third experimental group (E3) consisting of a DBBM-CM. Alizarin red staining was performed to qualitatively assess osteoinductive capacity. RUNX2 and OSX expression was quantified using real-time quantification polymerase chain reaction with two replications on day six (D6) and day 12 (D12) as fold changes. RESULTS: This experiment revealed that hUCMSCs were positively expressed by CD73, CD90, and CD105 but were not expressed by CD34. Alizarin red staining showed that E1 had the most calcium deposition on D6 and D12, followed by E3 and then E2 The RUNX2 and OSX expression was higher in E1 but this difference was not significant. The OSX expression in E1,E2,E3 was lower on D12 and C+ of OSX had the highest expression. There was a significant difference of fold change measured between all groups (p < 0.05), and there was no significant difference between any of the groups treated with OSX and RUNX2 on D6 and D12. CONCLUSION: FDBB osteoinduction and osteogenic capacity were higher when compared with DBBM and dc-FDBB.

Decision-Making Criteria of Odontectomy or Surgical Exposure in Impacted Maxillary Canine Based on Treatment Difficulty Index Modification.

OBJECTIVE: Canine impaction is a difficult condition to treat, and it usually necessitates a combination of surgical exposure and orthodontic traction or surgical extraction. An accurate assessment of the maxillary canine's position can help determine the severity of the impaction, the difficulty of therapy, and the treatment's prognosis. MATERIALS AND METHODS: A total of 55 impacted canines were studied and selected retrospectively. Difficulty indexes were used to measure the severity of impaction with pretreatment panoramic radiographs. STATISTICAL ANALYSIS: Pearson correlation was used to test the validity of the difficulty index modification score. Regression statistical analysis was used to evaluate any correlation between total scoring from each index with surgical treatment. RESULTS: The validity test on the variable modification index score showed a valid value (p = 0.000). According to both treatment difficulty and modification index, odontectomy group showed higher mean of total scoring than surgical exposure group. Treatment difficulty and modification index showed a significant correlation with surgical treatment (p = 0.003 and p = 0.001). CONCLUSIONS: The higher the severity of canine impaction, the greater is the possibility of odontectomy than surgical exposure. Both indexes can consider to be used in determining surgical treatment planning.

Analyses of Bone Regeneration Capacity of Freeze-Dried Bovine Bone and Combined Deproteinized-Demineralized Bovine Bone Particles in Mandibular Defects: The Potential Application of Biological Forms of Bovine-Bone Filler.

OBJECTIVE: This study aimed to evaluate bone regeneration capacity of FDBX granules compared to composite DBBM/DFDBX granules for filling of bone defect in rabbit mandible. MATERIAL AND METHODS: Critical size defects were created in 45 rabbits' mandible. The defect in the control group is left untreated, while in other groups the defects were filled with FDBX granules and composite DBBM/DFDBX granules, respectively. Specimens were collected at 2, 4, and 8 weeks for histology and immunohistochemical analyses. Significant difference is set at p-value < 0.05. RESULTS: The osteoblast-osteoclast quantification, osteoblast expression of Runx2, alkaline phosphatase, collagen-I, and osteocalcin, and osteoclast expression of receptor activator of NF-kB ligand (RANKL) and osteoprotegerin (OPG) in FDBX groups were statistically comparable (p > 0.05) with the composite group, while OPG/RANKL ratio, bone healing scores, and trabecular area were significantly higher (p < 0.05) in the composite compared to FDBX group. CONCLUSION: Composite DBBM/DFDBX granules, within the limitation of this study, has better bone forming capacity than FDBX granules for filling of bone defects in the mandible.

Stability of Tissue Augmented with Deproteinized Bovine Bone Mineral Particles Associated with Implant Placement in Anterior Maxilla.

BACKGROUND: Implant placement in defective anterior maxilla poses a great challenge regarding functional and aesthetic outcomes. Therefore, it requires predictable alveolar ridge augmentation. Deproteinized bovine bone mineral (DBBM) particle has commonly been used for bone grafting. However, it is associated with low resorption rates which potentially compromise the outcome of horizontal augmentation in conjunction with implant placement. AIMS: This study is aimed at evaluating the stability of tissue augmented with DBBM particle associated with implant placement in the anterior maxilla. MATERIALS AND METHODS: The inclusive criteria consist of patients being treated with guided bone regeneration (GBR) incorporating the use of DBBM particles with either a simultaneous or staged approach. The parameters analyzed include the implant survival rate, post-GBR clinical stability based on tissue resorption level, and the tissue stability between simultaneous and staged approaches. Statistical analysis using Mann-Whitney test is performed with significance determined at p value < 0.05. RESULTS: Seventeen patients with 23 implant placements satisfy the criteria for this study. Simultaneous approach is adopted in 18 (78.3%) implants and a staged approach in 5 (21.7%) implants. The implant survival rate is 100%. The evaluation of horizontal tissue stability reveals a low resorption level in 19 (82.6%) implants, while moderate and high resorption levels are found in 3 (13.0%) and 1 (4.3%) implants, respectively. The statistical analysis shows that the simultaneous approach produces significantly (p = 0.005) lower resorption level compared to the staged approach. CONCLUSION: Horizontal ridge augmentation using DBBM particles associated with implant placement in the anterior maxilla produces good clinical stability. The stability appears to be higher in the simultaneous approach compared to the staged approach.

Osteogenic Differentiation of Human Amniotic Mesenchymal Stem Cells in Chitosan-Carbonate Apatite Scaffold (In Vivo Study).

BACKGROUND: Studies of bone tissue engineering as a viable alternative to autogenous bone graft show promising results, although its mechanism and effectiveness remain only partially understood. PURPOSE: To explain the osteogenic differentiation of scaffold chitosan (Ch)-carbonate apatite (CA) in seeding with human amniotic mesenchymal stem cells (hAMSCs) on the regeneration of calvarial bone defects in rats. MATERIALS AND METHODS: Shitosan-Carbonate Apatite (Ch-CA) scaffold was created by means of a freeze-drying method. Twenty Wistar rats were randomly divided into two groups: control and treatment. Defects were created in the calvarial bone of each treatment group with a scaffold subsequently implanted. After 8 weeks, the rats were terminated for histology and immunohistochemistry examination. RESULTS: Expressions of vascular endothelial growth factor, bone morphogenetic protein2, Runt-related transcription factor 2 (RUNX2), and angiogenesis occurred earlier in the tissue-engineered group than that in the control group. An 8-week analysis also showed that the expression of RUNX2, alkaline phosphatase, osteocalcin, and collagen type 1 was at more elevated levels in the treatment group than that in the control group. CONCLUSION: These results showed that the combination of hAMSCs and Ch-CA scaffold may become one of the candidates for bone tissue engineering.

Human Umbilical Cord Mesenchymal Stem Cells Over Platelet Rich Fibrin Scaffold for Mandibular Cartilage Defects Regenerative Medicine

ABSTRACT Objective: To evaluate the regeneration of mandibular cartilage defect after implantation of human umbilical cord mesenchymal stem cells (hUCMSC) over platelet rich fibrin (PRF) as scaffold. Material and Methods: 20 male Wistar rats were randomly divided into four experimental groups consisting of: a control group featuring untreated mandibular defects (C), experimental groups whose mandibular defects were implanted with hUCMSC (E1), mandibular defects implanted with PRF (E2), mandibular defects implanted with hUCMSC and PRF scaffold (E3). The subjects were sacrificed after six weeks of observation for immunohistochemical examination in order to evaluate the expression of Ki67, Sox9, FGF 18, type 2 collagen, and aggrecan, in addition to histology examination to evaluate chondrocyte number and cartilage thickness. Data was analyzed with univariate analysis (ANOVA). Results: The implantation of hUCMSC and PRF scaffold proved capable of regenerating mandibular cartilage defect through the expression of FGF 18, Sox9, Ki67, chondrosis counts, type 2 collagen, aggrecan, and cartilage thickness. The regeneration were significantly higher in group E3. Conclusion: Human umbilical cord mesenchymal stem cells in platelet rich fibrin scaffold proved capable of regenerating mandibular cartilage defect.