Microbiome of periodontitis and peri-implantitis before and after therapy: Long-read 16S rRNA gene amplicon sequencing.

AIMS: The microbial profiles of peri-implantitis and periodontitis (PT) are inconclusive. The controversies mainly arise from the differences in sampling sites, targeted gene fragment, and microbiome analysis techniques. The objective of this study was to explore the microbiomes of peri-implantitis (PI), control implants (CI), PT and control teeth (CT), and the microbial change of PI after nonsurgical treatment (PIAT). METHODS: Twenty-two patients diagnosed with both PT and peri-implantitis were recruited. Clinical periodontal parameters and radiographic bone levels were recorded. In each patient, the subgingival and submucosal plaque samples were collected from sites with PI, CI, PT, CT, and PIAT. Microbiome diversity was analyzed by high-throughput amplicon sequencing using full-length of 16S rRNA gene by next generation sequencing. RESULTS: The 16S rRNA gene sequencing analysis revealed 512 OTUs in oral microbiome and 377 OTUs reached strain levels. The PI and PT groups possessed their own unique core microbiome. Treponema denticola was predominant in PI with probing depth of 8-10 mm. Interestingly, Thermovirga lienii DSM 17291 and Dialister invisus DSM 15470 were found to associate with PI. Nonsurgical treatment for peri-implantitis did not significantly alter the microbiome, except Rothia aeria. CONCLUSION: Our study suggests Treponemas species may play a pivotal role in peri-implantitis. Nonsurgical treatment did not exert a major influence on the peri-implantitis microbiome in short-term follow-up. PT and peri-implantitis possess the unique microbiome profiles, and different therapeutic strategies may be suggested in the future.

Structure, functional and physicochemical properties of lotus seed protein under different pH environments.

BACKGROUND: The present study investigated the structure, functional and physicochemical properties of lotus seed protein (LSP) under different pH environments. The structures of LSP were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Fourier transform infrared spectroscopy (FTIR), zeta potential, particle size distributions, free sulfhydryl and rheological properties. The functional and physicochemical properties of LSP were characterized by color, foaming property, emulsification property, solubility, oil holding capacity, water holding capacity, differential scanning calorimetry analysis and surface hydrophobicity. RESULTS: LSP was mainly composed of eight subunits (18, 25, 31, 47, 51, 56, 65 and 151 kDa), in which the richest band was 25 kDa. FTIR results showed that LSP had high total contents of α-helix and ß-sheet (44.81-46.85%) in acidic environments. Meanwhile, there was more ß-structure and random structure in neutral and alkaline environments (pH 7.0 and 9.0). At pH 5.0, LSP had large particle size (1576.98 nm), high emulsion stability index (91.43 min), foaming stability (75.69%) and water holding capacity (2.21 g g-1), but low solubility (35.98%), free sulfhydryl content (1.95 µmol g-1) and surface hydrophobicity (780). DSC analysis showed the denaturation temperatures (82.23 °C) of LSP at pH 5.0 was higher than those (80.10, 80.52 and 71.82 °C) at pH 3.0, 7.0 and 9.0. The analysis of rheological properties showed that LSP gel had high stability and great strength in an alkaline environment. CONCLUSION: The findings of the present study are anticipated to serve as a valuable reference for the implementation of LSP in the food industry. © 2024 Society of Chemical Industry.

Surgical repair of external cervical resorption - Prognosis and prognostic factors.

Background/purpose: External cervical resorption (ECR) is an aggressive form of root resorption, which etiology is unclear and its prognosis remains unpredictable. The purpose of this study was to investigate the prognosis and potential prognostic factors of ECR-affected teeth after surgical intervention for external repair with/without root canal treatment. Materials and methods: Treated ECR cases from 2009 to 2019 were collected retrospectively. The survival of the teeth and the status of root resorption were assessed during the follow-up period. Potential prognostic factors were analyzed with log-rank test and Kaplan-Meier statistics. Results: A total of 42 treated ECR-affected teeth were enrolled. The two-year survival rate was 71.20% [54.16%, 93.59%]. Persistent root resorption was the main complication after treatment. Patients with multiple ECR-affected teeth had greater recurrent potential than patients with solitary ECR-affected teeth. Prolonged calcium hydroxide dressing may contribute to a more favorable clinical outcome. Gender, age, tooth position and the need for root canal treatment did not show statistically significant effect on the prognosis. Conclusion: The current surgical method was able to arrest ECR in most cases. However, the case type (the number of ECR-affected teeth per patient) could highly affect the prognosis of the teeth. Clinicians should consider long-term calcium hydroxide dressing in case of pulp involvement to achieve better results.

Structure, physicochemical properties, and biological activities of protein hydrolysates from Zanthoxylum seed.

BACKGROUND: Zanthoxylum seed, as a low-cost and easily accessible plant protein resource, has good potential in the food industry. But protein and its hydrolysates from Zanthoxylum seed are underutilized due to the dearth of studies on them. This study aimed to investigate the structure and physicochemical and biological activities of Zanthoxylum seed protein (ZSP) hydrolysates prepared using Protamex®, Alcalase®, Neutrase®, trypsin, or pepsin. RESULTS: Hydrolysis using each of the five enzymes diminished average particle size and molecular weight of ZSP but increased random coil content. ZSP hydrolysate prepared using pepsin had the highest degree of hydrolysis (24.07%) and the smallest molecular weight (<13 kDa) and average particle size (129.80 nm) with the highest solubility (98.9%). In contrast, ZSP hydrolysate prepared using Alcalase had the highest surface hydrophobicity and foaming capacity (88.89%), as well as the lowest foam stability (45.00%). Moreover, ZSP hydrolysate prepared using Alcalase exhibited the best hydroxyl-radical scavenging (half maximal inhibitory concentration (IC50 ) 1.94 mg mL-1 ) and ferrous-ion chelating (IC50 0.61 mg mL-1 ) activities. Additionally, ZSP hydrolysate prepared using pepsin displayed the highest angiotensin-converting enzyme inhibition activity (IC50 0.54 mg mL-1 ). CONCLUSION: These data showed that enzyme hydrolysis improved the physicochemical properties of ZSP, and enzymatic hydrolysates of ZSP exhibited significant biological activity. These results provided validation for application of ZSP enzymatic hydrolysates as antioxidants and antihypertensive agents in the food or medicinal industries. © 2023 Society of Chemical Industry.

The structural characteristics and physicochemical properties of mung bean protein hydrolysate of protamex induced by ultrasound.

BACKGROUND: The limited physicochemical properties (such as low foaming and emulsifying capacity) of mung bean protein hydrolysate restrict its application in the food industry. Ultrasound treatment could change the structures of protein hydrolysate to accordingly affect its physicochemical properties. The aim of this study was to investigate the effects of ultrasound treatment on the structural and physicochemical properties of mung bean protein hydrolysate of protamex (MBHP). The structural characteristics of MBHP were evaluated using tricine sodium dodecylsulfate-polyacrylamide gel electrophoresis, laser scattering, fluorescence spectrometry, etc. Solubility, fat absorption capacity and foaming, emulsifying and thermal properties were determined to characterize the physicochemical properties of MBHP. RESULTS: MBHP and ultrasonicated-MBHPs (UT-MBHPs) all contained five main bands of 25.8, 12.1, 5.6, 4.8 and 3.9 kDa, illustrating that ultrasound did not change the subunits of MBHP. Ultrasound treatment increased the contents of α-helix, ß-sheet and random coil and enhanced the intrinsic fluorescence intensity of MBHP, but decreased the content of ß-turn, which demonstrated that ultrasound modified the secondary and tertiary structures of MBHP. UT-MBHPs exhibited higher solubility, foaming capacity and emulsifying properties than MBHP, among which MBHP-330 W had the highest solubility (97.32%), foaming capacity (200%), emulsification activity index (306.96 m2 g-1 ) and emulsion stability index (94.80%) at pH 9.0. CONCLUSION: Ultrasound treatment enhanced the physicochemical properties of MBHP, which could broaden its application as a vital ingredient in the food industry. © 2023 Society of Chemical Industry.

Epigallocatechin gallate-immobilized antimicrobial resin with rechargeable fluorinated synergistic composite for enhanced caries control.

OBJECTIVES: Given the global prevalence of dental caries, impacting 2.5 billion individuals, the development of sophisticated prevention filling materials is crucial. Streptococcus mutans, the principal caries-causing strain, produces acids that demineralize teeth and initiate dental caries. To address this issue, we aimed to develop a synergistic resin-based composite for enhancing caries control. METHODS: The synergistic resin composite incorporates fluorinated kaolinite and silanized Al2O3 nanoparticle fillers into an epigallocatechin gallate (EGCG) immobilized urethane-modified epoxy acrylate (U-EA) resin matrix, referred to the as-prepared resin composite. The EGCG-modified TPGDA/U-EA network was synthesized by preparing methacrylate-functionalized isocyanate (HI), reacting it with EGCG to form HI-EGCG, and then incorporating HI-EGCG into the TPGDA/U-EA matrix. The lamellar space within the kaolinite layer was expanded through the intercalation of acrylamide into kaolinite, enhancing its capability to adsorb and release fluoride ions (F-). The layered structure of acrylamide/ kaolinite in the U-EA resin composite acts as a F- reservoir. RESULTS: The physico-mechanical properties of the as-prepared resin composites are comparable to those of commercial products, exhibiting lower polymerization shrinkage, substantial F- release and recharge and favorable diametral tensile strength. The immobilized EGCG in the composite exhibits potent antimicrobial properties, effectively reducing the biofilm biomass. Furthermore, the synergistic effect of EGCG and fluorinated kaolinite efficiently counteracts acid-induced hydroxyapatite dissolution, thereby suppressing demineralization and promoting enamel remineralization. SIGNIFICANCE: Our innovative EGCG and fluoride synergistic composite provides enhanced antimicrobial properties, durable anti-demineralization, and tooth remineralization effects, positioning it as a promising solution for effective caries control and long-term dental maintenance.

[Cloning and expression analysis of U6 promoters in Panax quinquefolius].

The U6 promoter is an important element driving sgRNA transcription in the CRISPR/Cas9 system. Seven PqU6 promo-ter sequences were cloned from the gDNA of Panax quinquefolium, and the transcriptional activation ability of the seven promoters was studied. In this study, seven PqU6 promoter sequences with a length of about 1 300 bp were cloned from the adventitious roots of P. quinquefolium cultivated for 5 weeks. Bioinformatics tools were used to analyze the sequence characteristics of PqU6 promoters, and the fusion expression vectors of GUS gene driven by PqU6-P were constructed. Tobacco leaves were transformed by Agrobacterium tumefaciens-mediated method for activity detection. The seven PqU6 promoters were truncated from the 5'-end to reach 283, 287, 279, 289, 295, 289, and 283 bp, respectively. The vectors for detection of promoter activity were constructed with GUS as a reported gene and used to transform P. quinquefolium callus and tobacco leaves. The results showed that seven PqU6 promoter sequences(PqU6-1P to PqU6-7P) were cloned from the gDNA of P. quinquefolium, with the length ranged from 1 246 bp to 1 308 bp. Sequence comparison results showed that the seven PqU6 promoter sequences and the AtU6-P promoter all had USE and TATA boxes, which are essential elements affecting the transcriptional activity of the U6 promoter. The results of GUS staining and enzyme activity test showed that all the seven PqU6 promoters had transcriptional activity. The PqU6-7P with a length of 1 269 bp had the highest transcriptional activity, 1.31 times that of the positive control P-35S. When the seven PqU6 promoters were truncated from the 5'-end(PqU6-1PA to PqU6-7PA), their transcriptional activities were different in tobacco leaves and P. quinquefolium callus. The transcriptional activity of PqU6-7PA promoter(283 bp) was 1.59 times that of AtU6-P promoter(292 bp) when the recipient material was P. quinquefolium callus. The findings provide more ideal endogenous U6 promoters for CRISPR/Cas9 technology in ginseng and other medicinal plants.

The multi-factor analysis of computer-based test scores in objective structured clinical examination of clerkship in school of dentistry, National Taiwan University.

Background/purpose: The objective structured clinical examination (OSCE) is a test designed to test technical, clinical, and practical competency in medicine teaching. This study tried to evaluate the influencing factors of the computer-based tests (CBT) results for dental students. Materials and methods: The 32 5th-year dental students of the dental school of National Taiwan University (NTU) joined preclinical CBT. The contents of CBT included clinical knowledge and practical skills of nine dental specialties. Twenty single-choice questions were played with the PowerPoint 2016 on a desktop, and the playing time for each question was 30 s. Several factors including the gender, age, and dental student admission ways (with or without the skill tests) were evaluated. Also, the comparison between the clerk's grade and CBT scores of nine dental specialties was performed. The students also filled the feedback questionnaire about the examination questions and suggestions for the CBT. Results: There were no significant differences in the mean CBT scores between male and female dental students, between two groups of dental students with or without the skill tests in the admission process, and among three different age groups of dental students. The clerk's scores were not directly related to the CBT scores. Conclusion: The CBT has been well-intentioned in dental education since 2022 in Taiwan. The CBT results revealed that the conjunction of knowledge from the textbook and clinical observation should be enhanced. The guideline should be established to integrate CBT into preclinical education and promote the performance of clinical work of dental students.

The analysis and feedback from the performance of pre-clinical operation skills tests for clerkship in School of Dentistry, National Taiwan University in 2022.

Background/purpose: The simulated pre-clinical operation skill test (PCOST) is part of the objective structured clinical examination (OSCE) and designed for clinical practice using manikins, physical typodonts, and instruments in the laboratory. This study tried to evaluate the influencing factors of the PCOST results for dental students. Materials and methods: The 32 5th-year dental students of the dental school of National Taiwan University (NTU) joined the PCOST. The contents of PCOST included two specialties: operative dentistry (OD) and prosthodontics (PS). The examination content of OD was tooth 36 mesio-occluso-distal (MOD) cavity preparation, and that of PS was crown preparation and temporary crown fabrication of tooth 11. Several factors including the gender, age, and dental student admission ways (with or without the skill tests) were evaluated. Each scoring item was calculated. The students also filled the feedback questionnaire about the examination questions and suggestions for the PCOST. Results: For both OD and PS, there were no significant differences in the mean PCOST scores between male and female dental students, between two groups of dental students with or without the skill tests in the admission process, and among three different age groups of dental students. According to the feedback questionnaire from the participated dental students, they were satisfied with the PCOST. Conclusion: The PCOST is a good simulated examination in clinical practice. However, we still need to collect more PCOST data and feedback to revise the guideline of the PCOST, and to promote the performance of clinical works of dental students.

Chlorhexidine gel topical application ameliorates inflammatory bone loss in experimental periodontitis.

OBJECTIVES: This study aimed to evaluate the impact of chlorhexidine (CHX) gel on inflammation-induced periodontal tissue destruction, osteoclastogenesis, subgingival microbiota, and on the modulation of the RANKL/OPG as well as inflammatory mediators during bone remodeling in vivo. MATERIALS AND METHODS: Ligation- and LPS injection-induced experimental periodontitis were created to investigate the effect of topical application of CHX gel in vivo. Alveolar bone loss, osteoclast number and gingival inflammation was evaluated by micro-CT, histological, immunohistochemistry and biochemical analysis. The composition of the subgingival microbiota was characterized by 16S rRNA gene sequencing. RESULTS: Data shows significant decreases in the alveolar bone destruction in rats from ligation-plus-CHX gel group compared to ligation group. In addition, significant decreases in the number of osteoclasts on bone surface and the protein level of receptor activator of nuclear factor κB ligand (RANKL) in gingival tissue were observed in rats from ligation-plus-CHX gel group. Moreover, data shows significantly decreased inflammatory cell infiltration and decreased expression of cyclooxygenase (COX-2) and inducible NO synthase (iNOS) in gingival tissue from ligation-plus-CHX gel group versus ligation group. Assessment of the subgingival microbiota revealed changes in rats with CHX gel application treatment. CONCLUSION: HX gel presents protective effect on gingival tissue inflammation, osteoclastogenesis, RANKL/OPG expression, inflammatory mediators, and alveolar bone loss in vivo, which may have a translational impact on the adjunctive use in the management of inflammation-induced alveolar bone loss.

Stimulation phosphatidylinositol 3-kinase/protein kinase B signaling by Porphyromonas gingivalis lipopolysacch aride mediates interleukin-6 and interleukin-8 mRNA/protein expression in pulpal inflammation.

BACKGROUND/PURPOSE: The signaling mechanisms for Porphyromonas gingivalis lipopolysaccharide (PgLPS)-induced inflammation in human dental pulp cells are not fully clarified. This in vitro study aimed to evaluate the involvement of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway in PgLPS-induced pulpal inflammation. METHODS: Human dental pulp cells (HDPCs) were challenged with PgLPS with or without pretreatment and coincubation with a PI3K/Akt inhibitor (LY294002). The gene or protein levels of PI3K, Akt, interleukin (IL)-6, IL-8, alkaline phosphatase (ALP), osteocalcin and osteonectin were analyzed by reverse transcription polymerase chain reaction (PCR), real-time PCR, western blotting, and immunofluorescent staining. In addition, an enzyme-linked immunosorbent assay was used to analyze IL-6 and IL-8 levels in culture medium. RESULTS: In response to 5 µg/ml PgLPS, IL-6, IL-8, and PI3K, but not Akt mRNA expression of HDPCs, was upregulated. IL-6, IL-8, PI3K, and p-Akt protein levels were stimulated by 10-50 µg/ml of PgLPS in HDPCs. PgLPS also induced IL-6 and IL-8 secretion at concentrations higher than 5 µg/ml. Pretreatment and co-incubation by LY294002 attenuated PgLPS-induced IL-6 and IL-8 mRNA expression in HDPCs. The mRNA expression of ALP, but not osteocalcin and osteonectin, was inhibited by higher concentrations of PgLPS in HDPCs. CONCLUSION: P. gingivalis contributes to pulpal inflammation in HDPCs by dysregulating PI3K/Akt signaling pathway to stimulate IL-6 and IL-8 mRNA/protein expression and secretion. These results are useful for understanding the pulpal inflammation and possible biomarkers of inflamed pulp diagnosis and treatment.

Dental technology of Taiwan during the Japanese colonial period.

Background/purpose: During the Japanese colonial period, the Taipei Hospital had already provided complete dental services with a fixed price per treatment. This study tried to compare the differences in the prices of various dental treatments between the Japanese colonial period and today. Materials and methods: This study used the "Dental Treatment Fees in the 27th Annual Report of Taipei Hospital (Taisho 12)" as the study materials to compare the differences in the prices of various dental treatments between the Japanese colonial period and today using the monthly salary of a novice elementary school teacher as an income benchmark. Results: A hundred years ago, the Taipei Hospital had already provided the dental treatments such as scaling and endodontic, operative dentistry, prosthodontic, and orthodontic treatments. Of these treatment items, the prices for prosthodontic and orthodontic treatments were more expensive. After a century of development, the costs of scaling and operative dentistry treatments dropped, while the costs of endodontic, prosthodontic, and orthodontic treatments increased. Conclusion: During the Japanese colonial period, Taiwan's dental treatment technology had already had the forms of modern dentistry with several dental specialties. At that time, the costs of dental treatments are quite expensive. Today's dental treatment items are more detailed and diverse. The health insurance system provides Taiwanese people with convenient and cheap general dental treatments such as scaling and treatments related to operative dentistry, endodontics, periodontics, and oral surgery. However, the costs of prosthodontic and orthodontic treatments increase and have to be paid by the patients themselves.

Physicochemical, functional and antioxidant properties of mung bean protein enzymatic hydrolysates.

This study aimed to investigate physicochemical, functional and antioxidant properties of mung bean protein (MBP) enzymatic hydrolysates (MBPEHs) by alcalase, neutrase, protamex, flavourzyme and papain. Physicochemical properties were evaluated by SDS-PAGE, particle size distribution, FTIR, ultraviolet visible and fluorescence spectrophotometries. ABTS, hydroxyl scavenging, Fe2+ chelating activity were used to evaluate antioxidant activity. Enzymolysis with five proteases decreased average particle size, α-helix, ß-sheet, surface hydrophobicity of hydrolysates. Alcalase hydrolysate had the highest degree of hydrolysis (23.55%), absolute zeta potential (33.73 mV) and the lowest molecular weight (<10 kDa). Protamex and papain hydrolysates had higher foaming capacities, emulsification activity indexes, emulsion stability indexes (235.00%, 123.07 m2/g, 132.54 min; 200.10%, 105.39 m2/g, 190.67 min) than MBP (135.03%, 20.03 m2/g, 30.88 min). Alcalase hydrolysate demonstrated the lowest IC50 (mg/mL) in ABTS (0.12), hydroxyl (2.98), Fe2+ chelating (0.22). These results provide support for application of MBPEHs as foaming agent, emulsifier and antioxidant in food industry.

Influence of ultrasound treatment on the physicochemical and antioxidant properties of mung bean protein hydrolysate.

This study aimed to investigate influence of ultrasonic treatment on physicochemical and antioxidant properties of mung bean protein hydrolysate (MPH). Physicochemical properties of MPH were evaluated by Tricine-SDS-PAGE, particle size distribution, fourier transform infrared spectroscopy (FTIR) and fluorescence spectroscopy, among others. Radicals scavenging activities of ABTS, hydroxyl, superoxide anion, Fe2+ chelating ability and reducing power characterized antioxidant activities of MPH. MPH contained four bands of 25.6, 12.8, 10.6 and 4.9 kDa, in which 4.9 kDa was the most abundant. Ultrasonic treatment increased the contents of aromatic and hydrophobic amino acids in MPH. Ultrasonic treatment decreased the content of α-helix of MPH and increased ß-sheet and ß-turn compared to MPH. MPH-546 W (ultrasonic treatment 546 W, 20 min) had the lowest average particle size (290.13 nm), zeta potential (-36.37 mV) and surface hydrophobicity (367.95 A.U.). Antioxidant activities of ultrasonicated-MPH increased with the ultrasonic power, achieving the lowest IC50 (mg/mL) of 0.1087 (ABTS), 1.796 (hydroxyl), 1.003 (superoxide anion) and 0.185 (Fe2+ chelating ability) in 546 W power. These results indicated ultrasonic treatment would be a promising method to improve the antioxidant properties of MPH, which would broaden the application scope of MPH as bioactive components in the food industry.

Dental manpower and treated dental diseases in department of dentistry, Taipei Hospital (the predecessor of National Taiwan University Hospital) in 1923.

BACKGROUND/PURPOSE: In 1906, there was a dental treatment room established in the department of surgery of Taipei Hospital to introduce modern dental services to Taiwan. It is undoubtedly the earliest hospital dentistry in Taiwan. This study tried to assess the dental manpower and treated dental diseases in Taipei Hospital, and hence to explore the appearance of Taiwan's early hospital dentistry. MATERIALS AND METHODS: This study utilized the secondary data analysis to evaluate the dental manpower and treated dental diseases of dental outpatients in the 27th Annual Report of Taipei Hospital (Taisho 12th year or 1923). RESULTS: Our results found that there were 3 dentists and one dental technician in the dental department of Taipei Hospital in 1923. These 3 dentists totally treated 3 inpatients and 1759 outpatients with 4511 outpatient visits (1959 for male patients and 2552 for female patients) in that year. Dental caries and retained root were the two most common dental diseases in Japanese and Taiwanese dental outpatients, and followed in a descending order by missing tooth, pulp necrosis, and pulpitis for Japanese outpatients as well as pulpitis, suppurative periodontitis, and acute or chronic periodontitis for Taiwanese outpatients. CONCLUSION: We conclude that the dentistry of Taipei Hospital in 1923 does have a modern form of dentistry with 3 dentists and one dental technician. Its dental patients are mainly Japanese and minorly Taiwanese. Moreover, the treated dental diseases are mainly tooth diseases for Japanese outpatients and both tooth and periodontal diseases for Taiwanese outpatients.

Er:YAG laser irradiation enhances bacterial and lipopolysaccharide clearance and human gingival fibroblast adhesion on titanium discs.

To investigate the effect of Er:YAG laser treatment on lipopolysaccharide (LPS) clearance and fibroblast adhesion on titanium disks. Grade IV titanium discs (n = 216) were used and allocated to 6 groups. Group 1 was the negative control without Porphyromonas gingivalis inoculation. Discs in Groups 2-6 were incubated with P. gingivalis to form a biofilm. Group 3 received 0.12% chlorhexidine irrigation and Group 4 received titanium curettage to remove the biofilm. Group 5 was treated with Er:YAG laser irradiation and Group 6 was treated with titanium curettage plus Er:YAG laser irradiation. The contact angle and surface roughness were measured after the various treatments. The surface microstructure and residual bacteria were examined using scanning electron microscopy and confocal laser scanning microscopy, respectively. Residual LPS was examined using a limulus amoebocyte lysate assay and human gingival fibroblast adhesion was quantified using fluorescent microscopy. Curettage plus Er:YAG laser irradiation was the most effective method for removing bacteria and LPS. No significant difference in the amount of fibroblast adhesion was found between the control and Group 6. Combined use of Er:YAG laser irradiation and curettage optimizes LPS clearance and fibroblast adhesion on titanium discs.

Biofabrication of Gingival Fibroblast Cell-Laden Collagen/Strontium-Doped Calcium Silicate 3D-Printed Bi-Layered Scaffold for Osteoporotic Periodontal Regeneration.

Periodontal disease is a chronic disease that can lead to lose teeth and even tooth loss if left untreated. Osteoporosis and periodontal disease share similar characteristics and associated factors. Current regenerative techniques for periodontal diseases are ineffective in restoring complete function and structural integrity of periodontium due to unwanted migration of cells. In this study, we applied the concept of guided tissue regeneration (GTR) and 3D fabricated gingival fibroblast cell-laden collagen/strontium-doped calcium silicate (SrCS) bi-layer scaffold for periodontal regeneration. The results revealed that the bioactive SrCS had a hydroxyapatite formation on its surface after 14 days of immersion and that SrCS could release Sr and Si ions even after 6 months of immersion. In addition, in vitro results showed that the bi-layer scaffold enhanced secretion of FGF-2, BMP-2, and VEGF from human gingival fibroblasts and increased secretion of osteogenic-related proteins ALP, BSP, and OC from WJMSCs. In vivo studies using animal osteoporotic models showed that the 3D-printed cell-laden collagen/SrCS bi-layer scaffold was able to enhance osteoporotic bone regeneration, as seen from the increased Tb.Th and BV/TV ratio and the histological stains. In conclusion, it can be seen that the bi-layer scaffolds enhanced osteogenesis and further showed that guided periodontal regeneration could be achieved using collagen/SrCS scaffolds, thus making it a potential candidate for future clinical applications.