Clinical evaluation of implants placed within or beyond the boundaries of the alveolar ridge preservation: a retrospective case series with 10 weeks of observations in 28 patients.

PURPOSE: This study investigated the effect of implant vertical positioning within alveolar ridge preservation (ARP) sites on implant stability quotient (ISQ) values, which were measured 10 weeks post-implantation. METHODS: Patients who underwent ARP using collagenized deproteinized bovine bone mineral, followed by implant placement in the posterior area, were divided into 2 groups: the within-ARP group and the beyond-ARP group. In the within-ARP group, osteotomy and implant placement occurred within the ARP boundary. In contrast, in the beyond-ARP group, these procedures were performed beyond the ARP boundary, incorporating 3 mm of pristine bone at the implant's apex. Bone quality was assessed by tactile sense, and both insertion torque during implant surgery and ISQ values at 10 weeks post-implant surgery were measured. Multiple linear regression analysis and Pearson correlation analysis were used to explore the relationship between insertion torque and ISQ values. RESULTS: In total, 30 ARP sites in 28 patients were analyzed. There was no significant difference in bone quality, as determined by tactile sense, between the within-ARP and beyond-ARP groups. At the time of implant placement, the beyond-ARP group exhibited a higher insertion torque (33.33±13.39 Ncm) compared to the within-ARP group (17.08±11.17 Ncm). However, the ISQ values were similar between the 2 groups 10 weeks after implant placement. A positive correlation between insertion torque and ISQ values was confirmed at 10 weeks post-implant. CONCLUSIONS: The engagement of pristine bone may facilitate high insertion torque during the placement of implants in ARP sites. Nevertheless, by 10 weeks post-implantation, the ISQ values were found to be comparable, irrespective of the implant's position.

A randomized controlled trial of immediate implant placement comparing hydroxyapatite nano-coated and uncoated sandblasted/acid-etched implants using a digital surgical guide.

PURPOSE: This study evaluated the implant stability, volumetric changes, and patient-reported outcome measures (PROMs) of hydroxyapatite (HA) nano-coated sandblasted/acid-etched (SLA) implants compared to uncoated SLA implants. METHODS: Forty patients were recruited and randomly allocated to HA nano-coated SLA group (test, n = 20) and uncoated SLA group (control, n = 20) using single-blinded/block randomization. Implants were immediately placed in maxillary posterior region using a digital surgical guide. Insertion torque and implant stability quotient (ISQ) were measured at implant surgery and 1, 2, 3, and 4 months postoperatively. Intraoral scans, PROMs and soft tissue inflammation data were collected, and multivariable linear regression analysis of ISQ was performed. RESULTS: In total, 48 implants (test; n = 24, control; n = 24) in 37 patients (test; n = 19, control; n = 18) were analyzed. Despite no significant between-group difference at surgery, the test group showed higher ISQ values than the control group at 2 (76.53 ± 4.17 vs. 71.32 ± 4.79, p < 0.01), 3 (77.45 ± 4.41 vs. 73.85 ± 4.69, p < 0.05), and 4 months (79.08 ± 2.96 vs. 73.43 ± 3.52, p < 0.0001) postoperatively. There were no significant differences in linear and volumetric changes, PROMs, and soft tissue inflammation analysis between two groups. The ISQ at implant surgery was influenced by age and diabetes mellitus (DM) at the implant level and DM and predicted total bone-to-implant contact area at the patient level. CONCLUSION: HA nano-coated SLA implants promoted favorable immediate implants stability during early osseointegration phase compared to uncoated SLA implants, but displayed similar dimensional changes, PROMs, and soft tissue inflammation outcomes. TRIAL REGISTRATION: Clinical Research Information Service (CRIS), KCT0006364. Registered 21 July 2021, https://cris.nih.go.kr/cris/search/detailSearch.do?seq=24221&search_page=L .

The cumulative survival rate of dental implants with micro-threads: a long-term retrospective study.

PURPOSE: This study aimed to evaluate the long-term cumulative survival rate (CSR) of dental implants with micro-threads in the neck over a 10-year follow-up period and to examine the factors influencing the survival rate of dental implants. METHODS: This retrospective study was based on radiographic and dental records. In total, 151 patients received 490 Oneplant® dental implants with an implant neck micro-thread design during 2006-2010 in the Department of Periodontology of Seoul National University Dental Hospital. Implant survival was evaluated using Kaplan-Meier analysis. Cox proportional hazard regression analysis was used to identify the factors influencing implant failure. RESULTS: Ten out of 490 implants (2.04%) failed due to fixture fracture. The CSR of the implants was 97.9%, and no significant difference was observed in the CSR between external- and internal-implant types (98.2% and 97.6%, respectively, P=0.670). In Cox regression analysis, 2-stage surgery significantly increased the risk of implant failure (hazard ratio: 4.769, P=0.039). There were no significant differences in influencing factors, including sex, age, implant diameter, length, fixture type, location, surgical procedure, bone grafting, and restoration type. CONCLUSIONS: Within the limitations of this retrospective study, the micro-thread design of the implant neck was found to be favorable for implant survival, with stable clinical outcomes.

Effect of polydeoxyribonucleotide on early bone formation in lateral bone augmentation with immediate implant placement: an experimental in vivo study.

This study investigated early bone formation using collagenated biphasic calcium phosphate (CBCP) with or without polynucleotide (PDRN). Third (P3) or fourth (P4) premolars of six male beagle dogs were extracted and 5-mm-high dehiscence defects were created, followed by 3D-printed implant placement. The buccal bone defects were grafted with (i) CBCP and collagen membrane or (ii) CBCP soaked in polydeoxyribonucleotide (CBCP/PDRN) and collagen membrane. Samples of the experimental sites were harvested at 2- and 6-weeks post-surgery. The specimens were evaluated with radiologic and histomorphometric analysis. No significant differences were found between the CBCP and CBCP/PDRN groups in the micro-CT analysis at 2 or 6 weeks. No significant differences were observed in bone-to-implant contact (BIC) or bone area fraction occupancy (BAFO) in buccal augmented and lingual non-augmented areas. In the qualitative analysis, the new bone (NB) area and NB proportion in buccal augmented areas showed significantly higher values in the CBCP/PDRN group than in the CBCP group at 2 and 6 weeks. Peri-implant buccal dehiscence defects with immediate 3D-printed implant placement were corrected using a collagen membrane and CBCP or CBCP/PDRN. PDRN might have the potential to facilitate early bone formation with sufficient stability over time in dehiscence defects.

Minimally Invasive Regenerative Surgery by Peri-implant Excision and Regenerative Surgery (PERS) in Peri-implantitis: One-year Result on Three Cases.

This case series presents three patients undergoing minimally invasive regenerative surgery for peri-implantitis using peri-implant excision and regenerative surgery (PERS). No report of a resolved inflammatory state with peri-implant bone loss following nonsurgical treatment was included in this case report. After the suprastructure of the implant was disconnected, a peri-implant circular incision was made to remove inflammatory tissue. The combination decontamination method was conducted using a chemical agent and a mechanical device. After copious irrigation with normal saline, collagenated demineralized bovine bone mineral was applied to fill the peri-implant defect. The suprastructure of the implant was connected following the PERS procedure. The three patients with peri-implantitis that underwent successfully PERS procedures suggest that surgical intervention is a feasible approach to obtaining proper peri-implant bone filling of 3.42 ± 1.08 mm. However, this novel technique should be investigated in a larger sample size to determine its reliability and validity.

The effect of hard-type crosslinked hyaluronic acid with particulate bone substitute on bone regeneration: positive or negative?

PURPOSE: The role of hard-type crosslinked hyaluronic acid (HA) with particulate bone substitutes in bone regeneration for combined inlay-onlay grafts has not been fully investigated. We aimed to evaluate the effect of hard-type crosslinked HA used with bone substitute in terms of new bone formation and space maintenance. METHODS: A 15-mm-diameter round defect was formed in the calvaria of 30 New Zealand White rabbits. All animals were randomly assigned to 1 of 3 groups: the control group (spontaneous healing without material, n=10), the biphasic calcium phosphate (BCP) graft group (BCP, n=10), and the BCP graft with HA group (BCP/HA, n=10). The animals were evaluated 4 and 12 weeks after surgery. Half of the animals from each group were sacrificed at 4 and 12 weeks after surgery. Samples were evaluated using micro-computed tomography, histology, and histomorphometry. RESULTS: The BCP group showed higher bone volume/tissue volume (BV/TV) values than the control and BCP/HA groups at both 4 and 12 weeks. The BCP and BCP/HA groups showed higher bone surface/tissue volume (BS/TV) values than the control group at both 4 and 12 weeks. The BCP group showed higher BS/TV values than the control and BCP/HA groups at both 4 and 12 weeks. No statistically significant difference in newly formed bone was found among the 3 groups at 4 weeks. The BCP group showed significantly higher new bone formation than the BCP/HA group at 12 weeks. CONCLUSIONS: Hard-type crosslinked HA did not show a positive effect on new bone formation and space maintenance. The negative effect of hard-type crosslinked HA may be due to the physical properties of HA that impede osteogenic potential.

Impact of lattice versus solid structure of 3D-printed multiroot dental implants using Ti-6Al-4V: a preclinical pilot study.

PURPOSE: Various studies have investigated 3-dimensional (3D)-printed implants using Ti-6Al-4V powder; however, multi-root 3D-printed implants have not been fully investigated. The purpose of this study was to explore the stability of multirooted 3D-printed implants with lattice and solid structures. The secondary outcomes were comparisons between the 2 types of 3D-printed implants in micro-computed tomographic and histological analyses. METHODS: Lattice- and solid-type 3D-printed implants for the left and right mandibular third premolars in beagle dogs were fabricated. Four implants in each group were placed immediately following tooth extraction. Implant stability measurement and periapical X-rays were performed every 2 weeks for 12 weeks. Peri-implant bone volume/tissue volume (BV/TV) and bone mineral density (BMD) were measured by micro-computed tomography. Bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO) were measured in histomorphometric analyses. RESULTS: All 4 lattice-type 3D-printed implants survived. Three solid-type 3D-printed implants were removed before the planned sacrifice date due to implant mobility. A slight, gradual increase in implant stability values from implant surgery to 4 weeks after surgery was observed in the lattice-type 3D-printed implants. The marginal bone change of the surviving solid-type 3D-printed implant was approximately 5 mm, whereas the value was approximately 2 mm in the lattice-type 3D-printed implants. BV/TV and BMD in the lattice type 3D-printed implants were similar to those in the surviving solid-type implant. However, BIC and BAFO were lower in the surviving solid-type 3D-printed implant than in the lattice-type 3D-printed implants. CONCLUSIONS: Within the limits of this preclinical study, 3D-printed implants of double-rooted teeth showed high primary stability. However, 3D-printed implants with interlocking structures such as lattices might provide high secondary stability and successful osseointegration.

The effect of 3-D printed polylactic acid scaffold with and without hyaluronic acid on bone regeneration.

BACKGROUND: Three- dimensional (3D) technology has been suggested to overcome several limitations in guided bone regeneration (GBR) procedures because 3D-printed scaffolds can be easily molded to patient-specific bone defect site. This study aimed to investigate the effect of 3-D printed polylactic acid (PLA) scaffolds with or without hyaluronic acid (HA) in a rabbit calvaria model. METHODS: A calvaria defect with a diameter of 15 mm was created in 30 New Zealand white rabbits. The rabbits were randomly allocated into three groups including no graft group (control, n = 10), 3D printed PLA graft group (3D-PLA, n = 10), and 3D printed PLA with hyaluronic acid graft group (3D-PLA/HA, n = 10). Five animals in each group were sacrificed at 4 and 12 weeks after surgery. Microcomputed tomography and histologic and histomorphometric analyses were performed. RESULTS: Over the whole examination period, no significant adverse reactions were observed. There were no statistically significant differences in bone volume (BV) /tissue volume (TV) among the three groups at 4 weeks. However, the highest BV/TV was observed in the 3D-PLA/HA group at 12 weeks. The new bone area for control, 3D-PLA, and 3D-PLA/HA showed no statistical differences at 4 weeks. However, the value was significantly higher in the 3D-PLA and 3D-PLA/HA groups compared to the control group at 12 weeks. CONCLUSION: The 3D printed PLA scaffolds was biocompatible and integrated well with bone defect margin. They were also provided the proper space for new bone formation. Therefore, 3D printed PLA/HA might be a potential tool to enhance bone augmentation.

A Case Report of a 2-Year Follow-Up of Minimally Invasive Surgery in Peri-Implantitis: Peri-Implant Excisional Procedure and Access Surgery.

This case report describes the treatment of peri-implantitis lesions through a minimally invasive surgical procedure using a peri-implant excisional procedure and access surgery (PEAS). The prosthesis was disconnected, and the peri-implant granulation tissue removed after a peri-implant circular incision. Chemical debridement with hydrogen peroxide on a cotton ball and then mechanical debridement with a rotary round titanium brush and tufted brush with titanium bristles were conducted. The surgical intervention was effective in arresting the peri-implantitis. No further radiographic bone loss was observed over the 2-year follow-up period. This technique effectively cleans the contaminated implant surface, minimizes surgical morbidity, and allows for prosthesis delivery on the day of surgery. However, further studies with a larger sample size are needed to identify the reliability and validity of this novel technique.

Identification of stemness and differentially expressed genes in human cementum-derived cells.

PURPOSE: Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. METHODS: Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. RESULTS: The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. CONCLUSIONS: Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs.

The impact of surface treatment in 3-dimensional printed implants for early osseointegration: a comparison study of three different surfaces.

3D printing technology has been gradually applied to various areas. In the present study, 3D-printed implants were fabricated with direct metal laser sintering technique for a dental single root with titanium. The 3D implants were allocated into following groups: not treated (3D-None), sandblasted with a large grit and acid-etched (3D-SLA), and target-ion-induced plasma-sputtered surface (3D-TIPS). Two holes were drilled in each tibia of rabbit, and the three groups of implants were randomly placed with a mallet. Rabbits were sacrificed at two, four, and twelve weeks after the surgery. Histologic and histomorphometric analyses were performed for the evaluation of mineralized bone-to-implant contact (mBIC), osteoid-to-implant contact (OIC), total bone-to-implant contact (tBIC), mineralized bone area fraction occupancy (mBAFO), osteoid area fraction occupancy (OAFO), and total bone area fraction occupancy (tBAFO) in the inner and outer areas of lattice structure. At two weeks, 3D-TIPS showed significantly higher inner and outer tBIC and inner tBAFO compared with other groups. At four weeks, 3D-TIPS showed significantly higher outer OIC than 3D-SLA, but there were no significant differences in other variables. At twelve weeks, there were no significant differences. The surface treatment with TIPS in 3D-printed implants could enhance the osseointegration process in the rabbit tibia model, meaning that earlier osseointegration could be achieved.

In ovariectomy-induced osteoporotic rat models, BMP-2 substantially reversed an impaired alveolar bone regeneration whereas PDGF-BB failed.

OBJECTIVES: We previously suggested an ovariectomy (OVX)-induced osteoporotic rat model showing an impaired alveolar bone defect healing. This study aimed to evaluate and compare the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) and recombinant human platelet-derived growth factor-BB (rhPDGF-BB) on alveolar bone defect healing in OVX-induced osteoporotic rats. MATERIALS AND METHODS: A total of forty-one female rats were divided into four groups: a collagen group (n=10), a PDGF-BB group (n=11), a BMP-2 group (n=10), and a control group (n=10). Four months after OVX, alveolar bone drill-hole defects were created and grafted with collagen gel, rhPDGF-BB/collagen gel, or rhBMP-2/collagen gel. The defects in the control group were not grafted with any material. Defect healing was evaluated by histological, histomorphometric, and microcomputed tomographic (micro-CT) analyses at 2 and 4 weeks. RESULTS: According to the micro-CT analysis, the BMP-2 group exhibited the greatest bone volume fraction among all groups, while the PDGF-BB group did not show significant differences compared with the collagen group. The histomorphometric analysis showed a significantly larger amount of new bone area in the BMP-2 group than in the control and collagen groups at 4 weeks; however, the PDGF-BB group did not reach significant superiority compared with the other groups. CONCLUSIONS: Alveolar bone regeneration was significantly enhanced by the local use of rhBMP-2/collagen gel compared with the use of rhPDGF-BB/collagen gel in OVX-induced osteoporotic rats. CLINICAL RELEVANCE: A treatment modality using rhBMP-2 may be a promising approach to promote alveolar bone regeneration in patients suffering from postmenopausal osteoporosis.

Comparison of 3D-Printed Dental Implants with Threaded Implants for Osseointegration: An Experimental Pilot Study.

This study aimed to compare bone healing and implant stability for three types of dental implants: a threaded implant, a three-dimensional (3D)-printed implant without spikes, and a 3D-printed implant with spikes. In four beagle dogs, left and right mandibular premolars (2nd, 3rd, and 4th) and 1st molars were removed. Twelve weeks later, three types of titanium implants (threaded implant, 3D-printed implant without spikes, and 3D-printed implant with spikes) were randomly inserted into the edentulous ridges of each dog. Implant stability measurements and radiographic recordings were taken every two weeks following implant placement. Twelve weeks after implant surgery, the dogs were sacrificed and bone-to-implant contact (BIC) and bone area fraction occupied (BAFO) were compared between groups. At implant surgery, the primary stability was lower for the 3D-printed implant with spikes (74.05 ± 5.61) than for the threaded implant (83.71 ± 2.90) (p = 0.005). Afterwards, no significant difference in implants' stability was observed between groups up to post-surgery week 12. Histomorphometrical analysis did not reveal a significant difference between the three implants for BIC (p = 0.101) or BAFO (p = 0.288). Within the limits of this study, 3D-printed implants without spikes and threaded implants showed comparable implant stability measurements, BIC, and BAFO.

Periodontal Regeneration Using Recombinant Human Bone Morphogenetic Protein-2 and a Bilayer Collagen Matrix.

Previous studies demonstrated that recombinant human bone morphogenetic protein-2 (rhBMP-2) delivered using a collagen sponge could be a candidate for periodontal regeneration therapy. However, there is little evidence related to rhBMP-2 delivered with a bilayer collagen matrix. The aim of this study was to investigate the proper dose of rhBMP-2 using a bilayer collagen matrix for periodontal regeneration in a 1-wall defect. The mandibular first premolars and first molars of 6 beagle dogs were extracted, and an 8-week healing period was allowed. One-wall intrabony defects (4 mm in width and 5 mm in height) were made on the mesial side of the 2nd premolar and/or the distal side of the 4th premolar bilaterally. Subsequently, a bilayer collagen matrix containing 0 µg (C), 200 µg (T1), or 500 µg (T2) of lyophilized rhBMP-2 was randomly applied to the defect area. Calcein and xylenol orange were injected at 4 and 8 weeks following the surgery, respectively, to label periodic bone formation. After a 12-week healing period, the animals were sacrificed for micro-computed tomography and histomorphometric analysis. Bone mineral density and bone volume density showed statistically significant differences between the control group and group T1, while no significant differences were observed between the control group and group T2 or between groups T1 and T2. The bone height in groups T1 and T2 was smaller than that in the control group. Low doses of rhBMP-2 delivered using a bilayer collagen matrix in 1-wall intrabony defects can promote periodontal regeneration compared to no or high doses of rhBMP-2.

Comparison of experimental peri-implantitis models after application of ex vivo BMP2 gene therapy using periodontal ligament stem cells.

Peri-implantitis is an inflammatory disease that results in bone destruction around dental implants. A preclinical study using beagle models is frequently performed prior to clinical application in dentistry. Previously, we proposed an immediate peri-implantitis experimental model with a shorter experimental duration and less expense than the conventional experimental model. However, the differences in the regenerative outcomes between the immediate and conventional models were not fully revealed. In this study, we aimed to compare the regenerative outcomes between both models when ex vivo BMP2 gene therapy using autologous periodontal ligament stem cells (B2/PDLSCs) was applied to peri-implantitis defects. The results showed that the defect depths were significantly different between both models. New bone formation occurred in both models, but there were significant differences between the models. More than 70% of the defects were filled with newly formed bone in the conventional model, whereas 30-40% of the defects were filled in the immediate model. However, after adjustment for the differences in the defect depths between the models, the statistically significant differences in the regenerative outcomes between the models were lost. In conclusion, the inferior regenerative outcome of an immediate peri-implantitis model at B2/PDLSCs transplantation resulted from the defect depths, not the model itself.

Ovariectomy and timing of impaired maxillary alveolar bone regeneration: An experimental study in rats.

BACKGROUND: The aim of this study was to seek the critical time for impairment of alveolar bone regeneration after ovariectomy (OVX) in rats. METHODS: A total of 32 female rats were used. Test group rats were divided into a 2M group (n = 8), a 3M group (n = 8) and a 4M group (n = 8) according to the duration from OVX to defect creation. Bilateral OVX was performed in all test groups, and a sham operation was performed in the control group (n = 8). Drill-hole defects (1.5 mm diameter, 2 mm length) were created on both sides of the maxilla. All rats were euthanized 2 and 4 weeks after the surgery. Microcomputed tomographic (micro-CT), histological, and histomorphometric analyses and in vitro experiments were performed. RESULTS: The 4M group showed significantly less new bone formation and a lower bone mineral density than the other groups in the micro-CT analysis. The histomorphometric analysis also revealed that the 4M group showed significantly less new bone formation than the control and 2M groups. The rats in the 4M group showed significantly higher alkaline phosphatase expression levels and a larger number of calcified nodules than rats in the other groups, whereas osteoclastic activity was significantly lower in the 4M group than in the other groups. CONCLUSIONS: The critical time for impairment of alveolar bone regeneration was 4 months after OVX in rats.

Transcriptome and methylome analysis of periodontitis and peri-implantitis with tobacco use.

Peri-implantitis is similar to periodontitis in both symptoms and treatment; however, their level of similarity remains controversial. Here, we compared multiple cases of periodontitis and peri-implantitis through transcriptome and methylome profiling, and analyzed the effects of smoking as a typical risk factor. Human gingival tissues were obtained from 20 patients with periodontitis or peri-implantitis via periodontal surgical procedures. Total RNA and genomic DNA were isolated, and transcriptome and methylome datasets were generated. Comprehensive analysis of differential gene expression, DNA methylation, and protein-protein interactions indicated that periodontitis and peri-implantitis share biological similarities; however, hierarchical clustering between the two disease groups revealed distinct molecular characteristics. These differences might be related to structural differences in natural tooth-bone and implant-bone. Additionally, smoking differentially affected periodontitis and peri-implantitis in terms of host-defense mechanism impairment. Within the limitations of this study, the results provide evidence that peri-implantitis is distinct from periodontitis and that smoking potentially affects disease progression. Our study provides a foundation for the rational design of a large-scale study in the future for a more comprehensive analysis that includes microbiome and clinical data.

Enhanced Bone Regeneration by Diabetic Cell-Based Adenoviral BMP-2 Gene Therapy in Diabetic Animals.

The application of bone morphogenetic protein 2 (BMP-2) has been extensively investigated to improve diabetes-impaired bone healing; however, the delivery of BMP-2 by gene therapy for bone regeneration has rarely been investigated in diabetic animals. In this study, we aimed to evaluate which cells induce more new bone formation in diabetic animals when cell-based BMP2 gene therapy is applied. For this purpose, we harvested bone marrow stromal cells (BMSCs) twice in the same animal before (non-diabetic BMSCs; nBMSCs) and after diabetes induction (diabetic BMSCs; dBMSCs) using modified bone marrow ablation methods. And then, cells were transduced by adenoviral vectors carrying the BMP2 gene (AdBMP2). In in vitro, AdBMP2-transfected dBMSCs (B2/dBMSCs) produced higher BMP-2 mRNA levels over 48 h, whereas AdBMP2-transfected nBMSCs (B2/nBMSCs) exhibited a transient increase in BMP-2 mRNA followed by a decrease to the baseline level within 48 h. Both B2/dBMSCs and B2/nBMSCs induced secretion of BMP-2 for 3 weeks. However, B2/dBMSC BMP-2 secretion peaked from day 3 to 10, whereas B2/nBMSC BMP-2 secretion peaked from day 1 to 7. The analysis of osteogenic activity revealed that mineralization nodule formation and the expression levels of osteogenic genes were significantly higher in B2/dBMSCs than B2/nBMSCs and were accompanied by upregulation of canonical Wnt/ß-catenin and Smad signaling. AdBMP2-transfected autologous cells were implanted into critical-sized calvarial defects in diabetic animals and induced significantly more bone regeneration than non-AdBMP2-transfected cells. In addition, B2/dBMSCs led to significantly more new bone formation than B2/nBMSCs. Thus, BMP2 gene therapy using diabetic cells effectively supported diabetic bone healing and it was related to the enhanced responses to AdBMP2 of dBMSCs.

Transcriptomics and methylomics in chronic periodontitis with tobacco use: a pilot study.

BACKGROUND: Accumulating evidence suggests that tobacco smoking affects the susceptibility to and severity of chronic periodontitis. Epigenetics may explain the role of smoking in the development and progress of periodontal disease. In this study, we performed transcriptomic and methylomic analyses of non-periodontitis and periodontitis-affected gingival tissues according to smoking status. METHODS: Human gingival tissues were obtained from 20 patients, including non-smokers with and without periodontitis (n = 5 per group) and smokers with and without periodontitis (n = 5 per group). Total RNA and genomic DNA were isolated, and their quality was validated according to strict standards. The Illumina NextSeq500 sequencing system was used to generate transcriptome and methylome datasets. RESULTS: Comprehensive analysis, including between-group correlation, differential gene expression, DNA methylation, gene set enrichment, and protein-protein interaction, indicated that smoking may change the transcription and methylation states of extracellular matrix (ECM) organization-related genes, which exacerbated the periodontal condition. CONCLUSIONS: Our results suggest that smoking-related changes in DNA methylation patterns and subsequent alterations in the expression of genes coding for ECM components may be causally related to the increased susceptibility to periodontitis in smokers as they could influence ECM organization, which in turn may have an effect on disease characteristics.

Fibronectin-Derived Oligopeptide Stimulates Osteoblast Differentiation Through a Bone Morphogenic Protein 2-Like Signaling Pathway.

BACKGROUND: In previous studies by the authors, it was demonstrated that a fibronectin (FN)-derived oligopeptide, termed F20, stimulates osteoblast differentiation in vitro and bone formation in vivo. However, the fundamental molecular mechanism by which F20 stimulates osteogenesis remains unknown. Therefore, in this study the molecular mechanism underlying the effect of F20 in osteoblast differentiation is investigated. METHODS: The role of F20 in osteoblast differentiation was examined using mouse bone-marrow-derived ST2 cell line. The effect of Smad1/5 was determined following small interfering RNA knockdown. Runt-related transcription factor (Runx) 2, alkaline phosphatase (Alp), and osteocalcin (Oc) mRNA levels were determined by quantitative real-time polymerase chain reaction, and their transcriptional activation was assessed using luciferase reporter assays. Extracellular signal-regulated kinase (ERK) phosphorylation was visualized via immunoblotting. RESULTS: Synthetic oligopeptide F20 stimulated expression of bone marker genes Runx2, Alp, and Oc in ST2 cells via Smad and ERK or mitogen-activated protein kinase signaling pathways as did bone morphogenic protein 2 (BMP2). Furthermore, Runx2 acted as a transcription factor during F20-induced osteoblast differentiation. CONCLUSIONS: Collectively, these results indicate that F20 induces osteoblast differentiation with a pattern similar to that mediated by BMP2 signaling pathway. The authors' previous data also showed that FN-derived oligopeptide improved wound healing, and it is suggested that F20 might serve as a therapeutic biomolecule to facilitate periodontal tissue regeneration.