isolation-mesenchymal-stem-cells-from-human-alveolar-periosteum

Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells

Mesenchymal stem cells (MSCs) are present in a variety of tissues and can be differentiated into numerous cell types, including osteoblasts. Among the ...

Chang Gung University

Chang Gung Memorial Hospital

Taipei Medical University

Taipei Medical University Bilingual Education Center

This study was designed to analyze the post-rehabilitation occlusal function of subjects treated with complex mandibular resection and subsequently rehabilitated with fibula osteoseptocutaneous flaps, dental implants, and fixed prostheses utilizing the T-scan system.

Using the T-scan III system to analyze occlusal function in mandibular reconstruction patients: a pilot study.

Designed sockets prepared on the mandibles of nine Beagle dogs were divided into three groups: Calcitriol +Alloplast, Alloplast and Empty. Five of the nine dogs received Vit.D3 and calcium supplement (Vit.D/Ca group), while the other four dogs without supplements were assigned to Non-Vit.D/Ca group. After 4 weeks, the extent of vertical ridge resorption (VRR), bone density (density), new bone formation (NBF) and implant stability quotient (ISQ) were measured. Following systemic Vit.D/Ca administration, the Empty subgroup showed significant differences from the Calcitriol + Alloplast subgroup on variants NBF/Density/VRR and the Alloplast subgroup on items NBF/Density/ISQ/VRR. Alternatively, the Calcitriol + Alloplast subgroup revealed higher values of NBF/Density/ISQ (P < 0.001) and a lower VRR value (P = 0.001) than the Alloplast subgroup. Although there were no significant differences in NBF (P = 0.349), density (P = 0.796), ISQ (P = 0.577) and VRR (0.979) comparisons on alloplast treatment between the Vit.D/Ca and Non-Vit.D/Ca groups, local application with Calcitriol + Alloplast demonstrated better NBF/Density/ISQ (P = 0.02 to <0.001) effects than which of Alloplast subgroups. Consequently, the results showed that both systemic and local vitamin D3 treatment might accelerate bone regeneration in dogs. Within the using dose, systemic vitamin D3 treatment displayed a superior stimulating effect than local vitamin D3 application did. 

Association of vitamin D3 with alveolar bone regeneration in dogs.

Chemotherapy plays an important role in current cancer therapy; however, several problems remain unsolved on the issue of host-therapeutics interaction. The purpose of this study was to investigate the host responses after 5-flurouracil (5-FU) administration and to find the target genes and their relationship with other cytokines in the 5-FU-induced oral mucositis (OM) mouse model through transcriptomic analysis.

Comprehensive Assessment of Host Responses to 5-Fluorouracil-Induced Oral Mucositis through Transcriptomic Analysis.

During dental treatments, patients may experience negative emotions associated with the procedure. This study was conducted with the aim of using functional magnetic resonance imaging (fMRI) to visualize cerebral cortical stimulation among dental patients in response to auditory stimuli produced by ultrasonic scaling and power suction equipment. Subjects (n = 7) aged 23-35 years were recruited for this study. All were right-handed and underwent clinical pure-tone audiometry testing to reveal a normal hearing threshold below 20 dB hearing level (HL). As part of the study, subjects initially underwent a dental calculus removal treatment. During the treatment, subjects were exposed to ultrasonic auditory stimuli originating from the scaling handpiece and salivary suction instruments. After dental treatment, subjects were imaged with fMRI while being exposed to recordings of the noise from the same dental instrument so that cerebral cortical stimulation in response to aversive auditory stimulation could be observed. The independent sample confirmatory t-test was used. Subjects also showed stimulation in the amygdala and prefrontal cortex, indicating that the ultrasonic auditory stimuli elicited an unpleasant response in the subjects. Patients experienced unpleasant sensations caused by contact stimuli in the treatment procedure. In addition, this study has demonstrated that aversive auditory stimuli such as sounds from the ultrasonic scaling handpiece also cause aversive emotions. This study was indicated by observed stimulation of the auditory cortex as well as the amygdala, indicating that noise from the ultrasonic scaling handpiece was perceived as an aversive auditory stimulus by the subjects. Subjects can experience unpleasant sensations caused by the sounds from the ultrasonic scaling handpiece based on their auditory stimuli. 

Human amygdala activation by the sound produced during dental treatment: A fMRI study.

Clinical conclusions in studying the stability and osseointegration of mandibular implants positioned using the bone expansion techniques are conflicting and limited.

Implant Stability Quotients of Osteotome Bone Expansion and Conventional Drilling Technique for 4.1 mm Diameter Implant at Posterior Mandible.

Clinical decisions regarding the stability and osseointegration of mandibular implants positioned using the bone expansion techniques are conflicting and limited. The objective was to evaluate the stability of implants placed using 2 surgical techniques, selected according to the initial width of the mandibular posterior edentulous ridge, with D3 bone density, during a 12-week period. Fifty-eight implants in 33 patients were evaluated. Thirty-two implants in 24 patients were positioned using the osteotome expansion technique, and 26 fixtures in 17 patients were installed using the conventional drilling technique. The implant stability quotient values were recorded at weeks 0, 1, 2, 3, 4, 6, 8, 10, and 12 postsurgery and evaluated using analysis of variance, independent, and paired t tests. Calibrated according to the stability reading of a 3.3-mm diameter implant, the osteotome expansion group was associated with a lower bone density than the conventional group (64.96 ± 6.25 vs 68.98 ± 5.06, P = .011). The osteotome expansion group achieved a comparable primary stability (ISQb-0, P = .124) and greater increases in secondary stability (ISQb-12, P = .07) than did the conventional technique. A D3 quality ridge with mild horizontal deficiency is expandable by using the osteotome expansion technique. Although the 2 groups presented similar implant stability quotient readings during the study period, the osteotome expansion technique showed significant improvement in secondary stability. The healing patterns for these techniques are therefore inconsistent.

Developing Stability of Posterior Mandibular Implants Placed With Osteotome Expansion Technique Compared With Conventional Drilling Techniques.

This study characterized alveolar periosteum-derived mesenchymal stem cells (P-MSCs) and examined the hypothesis that 1,25-(OH)D (calcitriol) exerts osteoinductive effects on P-MSCs. The mRNA expressions of alkaline phosphatase (ALP), bone sialoprotein (BSP), core-binding factor alpha-1 (CBFA1), collagen-1 (Col-1), osteocalcin (OCN), and vitamin D receptor (VDR) were assessed after incubation with calcitriol for 2 weeks. Vitamin C as positive control (Vit. C-p) increased ALP and CBFA1 mRNA expression at both 1 and 2 weeks and increased BSP and Col-1 mRNA expression only at the first week. A concentration of 10 M calcitriol enhanced ALP, CBFA1, Col-1, and OCN mRNA expression at both weeks and BSP mRNA expression at the first week. Furthermore, 10 M calcitriol increased the mRNA expressions of all compounds at both weeks, except that of CBFA1 at the first week. 10 M calcitriol and Vit. C-p enhanced ALP activity at the second and third weeks. The results revealed that 10, 10, and 10 M calcitriol induced osteoinduction in alveolar P-MSCs by increasing ALP, CBFA1, Col-1, and OCN mRNA expression. A 10 M calcitriol yielded a higher mRNA expression than Vit. Cp on VDR and OCN mRNA expression at both weeks and on Col-1 mRNA at the second week.

Potential Osteoinductive Effects of Calcitriol on the m-RNA of Mesenchymal Stem Cells Derived from Human Alveolar Periosteum.

The purpose of this study was to relate the proportions of bone-supported root length of a 2D view into the amount of a 3D bone-attached root surface area (BA-RSA) by using a dental laser scanner examination. White-light 3D scanning technology was used to probe 36 maxillary and 35 mandibular single-rooted premolars. The bone-supported height (BSH) and BA-RSA at designated levels (95-25%) were compared using statistical t tests. The 100% BSH and BA-RSA of the maxillary/mandibular premolars were 12.6 ± 1.60 mm/13.45 ± 1.47 mm (p < 0.05) and 220.78 ± 35.31 mm/199.51 ± 26.33 mm (p < 0.01), respectively. Approximately 79-80%, 59-60%, and 35-36% premolars 2D BSH remained in comparison to 75%, 50%, and 25% 3D BA-RSA preservation, respectively. However, corresponding to a 75%, 50%, and 25% 2D BSH reserve, premolars retained 67-68%, 39-41%, and 15-17% 3D BA-RSA, respectively. When taking 1.0 mm connective tissue attachment into account, 60% 3D BA-RSA and 50% 2D BSH loss were noted at the 5.1-5.4 mm clinical attachment level. Assigning a periodontal prognosis and determining the severity of periodontitis for premolars with alveolar bone loss based on 3D's or 2D's measurement is inconsistent.

Decreased Amount of Supporting Alveolar Bone at Single-Rooted Premolars Is Under Estimated by 2D Examinations.

This study aimed to identify predictors of success rate of mesenchymal stem cell (MSC) isolation from human deciduous teeth pulp. A total of 161 deciduous teeth were extracted at the dental clinic of Chang Gung Memorial Hospital. The MSCs were isolated from dental pulps using a standard protocol. In total, 128 colonies of MSCs were obtained and the success rate was 79.5%. Compared to teeth not yielding MSCs successfully, those successfully yielding MSCs were found to have less severe dental caries (no/mild-to-moderate/severe: 63.3/24.2/12.5% versus 12.5/42.4/42.4%,  < 0.001) and less frequent pulpitis (no/yes: 95.3/4.7% versus 51.5/48.5%,  < 0.001). In a multivariate regression model, it was confirmed that the absence of dental caries (OR = 4.741, 95% CI = 1.564-14.371,  = 0.006) and pulpitis (OR = 9.111, 95% CI = 2.921-28.420,  < 0.001) was significant determinants of the successful procurement of MSCs. MSCs derived from pulps with pulpitis expressed longer colony doubling time than pulps without pulpitis. Furthermore, there were higher expressions of proinflammatory cytokines, interleukin- (IL-) 6 and monocyte chemoattractant protein- (MCP-) 1,  < 0.01, and innate immune response [toll-like receptor 1 (TLR1) and TLR8,  < 0.05; TLR2, TLR3, and TLR6,  < 0.01] in the inflamed than noninflamed pulps. Therefore, a carious deciduous tooth or tooth with pulpitis was relatively unsuitable for MSC processing and isolation.

Isolation of Mesenchymal Stem Cells from Human Deciduous Teeth Pulp.

Micro-computed tomography (micro-CT) was applied to elucidate the relationship between the three-dimensional (3D) root surface area (RSA) and two-dimensional (2D) crown-to-root ratio (CRR) of extracted teeth to classify the periodontitis and assign a periodontal/prosthetic prognosis. A total of 31 maxillary and 35 mandibular single-rooted human premolars were examined. The amount of periodontal support on the basis of 3D RSA and 2D root length (RL) at CRRs of 1:1, 5:4, 3:2, and 2:1 were analyzed. Both maxillary and mandibular premolars demonstrated a nonsignificant RSA percentage at the evaluated CRRs. The coronal 21%-22% 2D RL and the 26%-28% 3D RSA bone loss apical to the cemento-enamel junction corresponded to a CRR of 1:1, relating to mild-moderate periodontitis. The coronal 30%-31% 2D RL and the 41%-42% 3D RSA bone loss corresponded to a CRR of 5:4, correlating to severe periodontitis. More severe clinical attachment loss (CAL) was observed in the 3D RSA measurement than in the 2D RL measurement at the evaluated CRRs. The amount of CAL at the CRR of 1:1 was inadequate to assess the severity of periodontitis on the basis of the 2D RL and 3D RSA measurements.

Inconsistency in the Crown-to-Root Ratios of Single-Rooted Premolars Measured by 2D and 3D Examinations.

Micro-computed tomography (micro-CT) was employed to relate the root surface area (RSA) to the periodontal attachment levels (PALs) of extracted premolars to diagnose periodontitis. Single-rooted human maxillary and mandibular premolars 31 and 36, respectively, were surveyed by micro-CT and its associated software. RSA levels from the 1st to 10th mm, corono-apically, were analyzed using statistical t tests. The average root length (RL) and RSA of the maxillary and mandibular premolars were significantly different (p < 0.05). Both premolars demonstrated a non-significant RSA percentage comparison at the evaluated PALs. For the 30% coronal 2-D radiographic RL, the 3-D RSAs 3.77 mm and 3.99 mm apical to the cementoenamel junction (CEJ) were 39.48% and 40.65% for maxillary and mandibular premolars, respectively. At the 15% coronal 2-D RL, the 3-D RSA 2 mm apical to the CEJ of the premolars was approximately 21%. At the 50% coronal 2-D RL level, approximately 62% coronal 3-D RSA and 6.5 mm RL decreased. The amount of decrease of the RSA attachment is significant in every 2-mm measurement for both premolars. Sampling periodontal microbial pathogens based on the condition of 2-D radiographic bone and clinical attachment losses without considering 3-D RSA is potentially inadequate and may underestimate the severity of the periodontitis.

Incompatible amount of 3-D and 2-D periodontal attachments on micro-CT scanned premolars.

The human external auditory canal can become deformed when the mandible moves, and this changes the sound field in the external auditory canal. This study measured the sound field gain in the external auditory canal while varying mouth-opening in three levels. The mandible was fixed at the 1/3, the 2/3, and the maximal mouth-opening levels. Seven 65-dB tones of 200, 500, 1000, 2000, 4000, 6000, and 8000 Hz, which are the sound pressure level and frequency range when people are talking at a normal level, were adopted as the sound stimulus to measure sound field gains at 5, 10, 15, and 20 mm to the interior of the external auditory canal. The results show that, with the exception of the 1.25 dB decrease from 12.96 to 11.71 dB at a depth of 5 mm with a stimulus at 8000 Hz, the differences in the sound field gain at the other depths and stimulus frequencies were within 1 dB and were not statistically significant. These results suggest that mouth-opening level has no effect on the measurement of the sound field in the external auditory canal.

Article	Year
Using the T-scan III system to analyze occlusal function in mandibular reconstruction patients: a pilot study. Biomedical journal\| PubMed ID: 25163500	2015
Association of vitamin D3 with alveolar bone regeneration in dogs. Journal of cellular and molecular medicine\| PubMed ID: 25753943	2015
Comprehensive Assessment of Host Responses to 5-Fluorouracil-Induced Oral Mucositis through Transcriptomic Analysis. PloS one\| PubMed ID: 26266941	2015
Human amygdala activation by the sound produced during dental treatment: A fMRI study. Noise & health\| PubMed ID: 26356376	2015
Implant Stability Quotients of Osteotome Bone Expansion and Conventional Drilling Technique for 4.1 mm Diameter Implant at Posterior Mandible. Clinical implant dentistry and related research\| PubMed ID: 27665748	2017
Developing Stability of Posterior Mandibular Implants Placed With Osteotome Expansion Technique Compared With Conventional Drilling Techniques. The Journal of oral implantology\| PubMed ID: 28048965	2017
Potential Osteoinductive Effects of Calcitriol on the m-RNA of Mesenchymal Stem Cells Derived from Human Alveolar Periosteum. BioMed research international\| PubMed ID: 28105418	2016
Decreased Amount of Supporting Alveolar Bone at Single-Rooted Premolars Is Under Estimated by 2D Examinations. Scientific reports\| PubMed ID: 28367999	2017
Isolation of Mesenchymal Stem Cells from Human Deciduous Teeth Pulp. BioMed research international\| PubMed ID: 28377925	2017
Inconsistency in the Crown-to-Root Ratios of Single-Rooted Premolars Measured by 2D and 3D Examinations. Scientific reports\| PubMed ID: 29184103	2017
Incompatible amount of 3-D and 2-D periodontal attachments on micro-CT scanned premolars. PloS one\| PubMed ID: 29518113	2018
Effect of mouth-opening levels on sound field gain in the ear canal. The Journal of the Acoustical Society of America\| PubMed ID: 29604713	2018

Hsiang-Hsi Hong

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