A long-term evaluation of periodontal phenotypes before and after the periodontal-orthodontic-orthognathic combined treatment of lower anterior teeth in patients with skeletal Angle class Ⅲ malocclusion / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the changes of periodontal phenotype (width of keratinized gingiva, thickness and height of alveolar bone) of lower anterior teeth in patients with skeletal class Ⅲ malocclusion before and after the periodontal-orthodontic-orthognathic combined treatment.@*METHODS@#In the study, 20 patients with skeletal class Ⅲ malocclusion (6 males and 14 females) completed the periodontal-orthodontic-orthognathic combined treatment were included from March 2017 to June 2022, with 39 central incisors, 40 lateral incisors and 40 canines. The mean age was (25.40±4.27) years (20-34 years). The mean follow-up time was (3.70±1.05) years from the beginning of periodontal corticotomy regenerative surgery (PCRS) to the end of the combined treatment. Cone-beam computed tomography (CBCT) was used to measure the thickness, area and height of alveolar bone by the same researcher, taken before the PCRS (T0), 6 months after the PCRS (T1), 12 months after the PCRS (T2), before the orthognathic surgery (T3), and after the periodontal-orthodontic-orthognathic combined treatment (T4). The periodontal clinical parameters were used to evaluate changes in the soft tissue by another researcher, measured before the PCRS (T0) and after the combined treatment (T4). Changes of soft and hard tissue were evaluated by the periodontal phenotype.@*RESULTS@#The width of keratinized gingiva increased significantly (all P < 0.001) in lower anterior teeth, the central incisors, lateral incisors and canines increased by (1.82±1.57) mm, (2.03±1.48) mm and (2.05±1.27) mm, respectively. The proportion of thick periodontal biotype in the central and lateral incisors increased significantly (all P < 0.001), while the changes of periodontal biotypes in the lower canines were not obvious. The thickness of labial alveolar bone of lower anterior teeth all increased significantly after periodontal corticotomy regenerative surgery and the combined treatment (all P < 0.001). The area of labial alveolar bone of lower anterior teeth also increased significantly after the combined treatment (all P < 0.001). The whole area of labial and lingual alveolar bone of central and lateral incisors increased (P < 0.001), while the whole area of canines remained the same. All The height of the alveolar bone increased (all P < 0.001) on the labial side after the treatment.@*CONCLUSION@#The periodontal phenotypes of lower anterior teeth were significantly improved after the periodontal-orthodontic-orthognathic combined treatment in patients with skeletal Angle class Ⅲ malocclusion. The improvement was long-termly stable, and the periodontal risk was reduced.

Effect of Micro-fertilizer Containing Rare Earth on Growth，Yield and Quality of Angelica sinensis / 中国实验方剂学杂志

ObjectiveTo investigate the effects of micro-fertilizer containing rare earth of different types and concentrations on the growth，yield and quality of Angelica sinensis. MethodOn the basis of the single-factor randomized block design， the growth and index components of Angelica sinensis were determined with rare earth-containing nitrate and chloride micro-fertilizers of different concentrations as foliar fertilizers. ResultSpraying 0.8 g·mL-1 rare earth-containing chloride micro-fertilizer could increase the economic yield of A. sinensis， with the fresh yield per mu （1 mu≈667 m2） reaching 855.4 kg and the dry yield per mu 350.7 kg，which increased by 15.16% and 28.70% respectively compared with that in the control group CK1. Spraying 1.2 g·mL-1 rare earth-containing nitrate micro-fertilizer could promote the growth and development of A. sinensis and significantly increase the content of index components， with the plant height reaching 93.05 cm，the stem diameter 15.60 mm，the root diameter 16.10 mm，the main root length 36.5 cm，and the number of leaves 11.25 pieces per plant， which increased by 32.76%，31.98%，41.98%，53.36%，and 45.16%， respectively， compared with those in the control group CK2. Besides， the content of ferulic acid，volatile oil，ligustilide， and extract was 0.96%，0.41%，0.30% and 48.76%，respectively，which increased by 12.94%，17.14%，11.11%， and 12.07%，respectively，compared with that in the control group CK2. ConclusionSpraying 0.8 g·mL-1 rare earth-containing chloride micro-fertilizer and 1.2 g·mL-1 rare earth-containing nitrate micro-fertilizer can promote the growth and development of A. sinensis，improve the medicinal properties，and increase yield and quality. Rare earth-containing micro-fertilizers can be applied in the standardization of A. sinensis cultivation， which can change the production status of A. sinensis that depends on chemical fertilizers and single fertilization， and promote the green， organic and ecological cultivation of A. sinensis.

Expression and distribution of calprotectin in healthy and inflamed periodontal tissues / 北京大学学报(医学版)

OBJECTIVE@#Calprotectin, the heterdimer of S100A8 and S100A9, is the major cytoplasmic protein of neutrophils, which is also expressed or induced in gingival epithelial cells, activated mononuclear macrophages and vascular endothelial cells. Calprotectin is intimately associated with the initiation and progression of periodontitis, but the in vivo expression patterns of calprotectin in healthy and inflamed periodontal tissue are not fully understood. To observe the expression, distribution and cellular localization of calprotectin in the samples of healthy periodontal tissues and experimental periodontitis tissues of Beagles and to explore their relationship with periodontal inflammation and possible effect.@*METHODS@#Experimental periodontitis model was established by ligation around the mandibular second molar of the Beagle dogs, while the contralateral teeth were healthy controls. Induction duration was 12 weeks, before the dogs were executed. Tissue specimens were demineralized and serial sections were made conventionally. The in vivo expression of calprotectin in the healthy and inflamed periodontal tissues were examined by immunohistochemistry. The in vitro expression of calprotectin in human primary gingival fibroblasts (GFs) and periodontal ligament (PDL) cells were detected by immunocytochemistry.@*RESULTS@#Immunohistochemistry analysis indicated that calprotectin was expressed in gingival epithelial cells and infiltrated neutrophils in the healthy periodontium within the gingival epithelium, S100A8/A9 was most strongly expressed in the junctional epithelium, followed by surface epithelium, and least expressed in the sulcular epithelium. The S100A8/A9 expression levels were sharply defined at the junction between the junctional epithelium and the sulcular epithelium. In periodontal inflammatory lesions, the expression level of calprotectin in sulcular epithelium and junctional epithelium was up-regulated than that in the healthy gingival epithelium. Calprotectin was inducibly expressed in fibroblast-like cells in gingival connective tissue and periodontal ligament tissue, microvascular endothelial cells (ECs) and bone marrow fibroblasts under inflammatory conditions. Additionally, the expression of calprotectin in primary human GFs and PDL cells was confirmed by immunnocytochemistry staining.@*CONCLUSION@#Constitutively expressed in neutrophils and gingival epithelial cells, and calprotectin might maintain the homeostasis and integrity of periodontium. Inflammation-induced expression of calprotectin in GFs, PDL cells, microvascular ECs and bone marrow fibroblasts might process anti-microbial function and promote leukocytes transmigration to defend the host against the microorganisms.

Comparison of residual cement between CAD/CAM customized abutments and stock abutments via digital measurement in vitro / 北京大学学报(医学版)

OBJECTIVE@#To compare the residual cement between computer aided design/computer aided manufacturing customized abutments (CCA) and stock abutments (SA), and to evaluate the feasibility of digital measurement for residual cement volume by three-dimensional scanning.@*METHODS@#Twenty master models needed in this study were all taken from one 47-year-old patient with arrested periodontitis, who had already had an implant placed at his right upper central incisor site in the Department of Periodonto-logy, Peking University School and Hospital of Stomatology. After 4 weeks of soft tissue conditioning by means of customized healing abutment, the height of peri-implant soft tissue was measured, from the implant platform to mucosal margin, as 5 mm. Using customized impression coping, the impression was taken and twenty models were fabricated and allocated to 4 groups according to the type of abutments: CCA1 (5 mm transmucosal height CCA, with margin at tissue level), CCA2 (4 mm transmucosal height CCA, with 1 mm submucosal margin), SA1 (3 mm transmucosal height SA, with 2 mm submucosal margin) and SA2 (1 mm transmucosal height SA, with 4 mm submucosal margin). Crowns were cemented to the abutments, which were seated on the working models. Excess cement was removed by a prosthodontic specialist. Thereafter, the volume of residual cement was evaluated by using three-dimensional scanning technique. The area proportion of residual cement was calculated on photographs taken by a single lens reflex camera. The weight of residual cement was weighed by an analytical balance. And the correlation of residual cement volume data with residual cement area proportion or weight of residual cement acquired by traditional methods was analyzed.@*RESULTS@#Residual cement was observed on all the experiment samples. The residual cement volume of CCA was significantly less than that of SA [(0.635 3±0.535 4) mm3 vs. (2.293 8±0.943 8) mm3, P < 0.001]. Consistently, CCA had less residual cement area proportion and weight than those of SA [area proportion: 7.57%±2.99% vs. 22.68%±10.06%, P < 0.001; weight: (0.001 5±0.001 0) g vs. (0.003 7±0.001 4) g, P < 0.001]. The residual cement volume was strongly correlated with the residual cement area proportion and residual cement weight (r>0.75, P < 0.001).@*CONCLUSION@#These in vitro results suggest that CCA minimized the residual cement more effectively than SA. The method to digitally evaluate the residual cement volume is feasible, but its validity and reliability need to be further studied.

Effect of initial periodontal therapy on blood parameters related to erythrocyte and platelet in patients with type 2 diabetes mellitus and chronic periodontitis / 北京大学学报(医学版)

OBJECTIVE@#To compare the blood parameters related to erythrocyte and platelet between baseline and 3 months after initial periodontal therapy in patients with both type 2 diabetes mellitus and chronic periodontitis (DM-P).@*METHODS@#According to the International Symposium on Classification of Periodontal Diseases and Conditions in 1999 and the diagnostic criteria of type 2 diabetes mellitus proposed by the World Health Organization in 1999, 35 patients with DM-P were recruited. All the participants received initial periodontal therapy, including oral hygiene instruction, scaling, and root planning provided by one senior periodontist. Original diet, exercise, and medication for blood glucose control were unchanged for all the participants. At baseline and 3 months after initial periodontal therapy, the clinical periodontal parameters, including probing depth (PD), bleeding index (BI) and clinical attachment loss (CAL); erythrocyte-related indexes, including red blood cell (RBC) count, hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), RBC volume distribution width (RDW); platelet-related indexes, including platelet (PLT) count, mean platelet volume (MPV), platelet distribution width (PDW), plateletocrit (PCT) were measured and compared.@*RESULTS@#Compared with baseline, the periodontal parameters, including PD [(3.370±0.601) mm vs. (2.729±0.431) mm], BI [2.160 (1.550~3.410) vs. 1.420 (1.000~2.970)] and CAL [(3.307±1.577) mm vs. (2.990±1.587) mm], were significantly reduced (P < 0.001) three months after the initial periodontal therapy; the erythrocyte-related indexes, including RBC count [(4.727±0.392)×1012/L vs. (4.825±0.394)×1012/L, P=0.010], HGB [(145.886±11.792) g/L vs. (149.200±12.979) g/L, P=0.007] and HCT [43.40% (37.50%~48.50%) vs. 43.80% (38.50%~53.20%), P=0.003], were significantly increased three months after the initial periodontal therapy; PLT count [(216.714±61.900)×109/L vs. (205.886±62.051)×109/L, P=0.016] was significantly reduced 3 months after the initial periodontal therapy.@*CONCLUSIONS@#The initial periodontal therapy can significantly improve blood parameters related to RBC and PLT, which might decrease the risk of vascular complications in DM-P patients.

Alveolar crest and relevant analysis of labial side of anterior teeth on skeletal Angle class III patients / 北京大学学报(医学版)

OBJECTIVE@#To measure the distance from cemento-enamel junction (CEJ) to alveolar crest (AC) of labial side of anterior teeth on skeletal Angle class III patients under direct vision during periodontal bone augmentation surgery and to make relevant analysis to find the relevant factors.@*METHODS@#In the study, 46 skeletal Angle class III patients (10 males and 36 females) received periodontal bone augmentation surgery of anterior teeth were included, with 67 jaws (27 maxillae and 40 mandibles) and 400 anterior teeth (161 maxillary anterior teeth and 239 mandibular anterior teeth). The mean age was 23.65 years. Maxillary anterior teeth consisted of 54 central incisors, 53 lateral incisors and 54 canines. Mandibular anterior teeth consisted of 79 central incisors, 80 lateral incisors and 80 canines. CEJ-AC was measured in three sites (mesial sites, central sites and distal sites) by Williams periodontal probes during periodontal bone augmentation surgery under direct vision by the same researcher.@*RESULTS@#The average CEJ-AC of 400 anterior teeth was (2.21±1.48) mm. The average CEJ-AC of maxillary anterior teeth was (1.72±1.13) mm, more than (2.54±1.60) mm of mandibular anterior teeth (P<0.05). The average CEJ-AC of canines was (2.42±1.78) mm, more than (2.06±1.27) mm of central incisors or (2.16±1.32) mm of lateral incisors (P<0.05). The average CEJ-AC of central sites was (3.04±2.01) mm, more than (1.79±0.86) mm of mesial sites or (1.81±0.89) mm of distal sites (P<0.05). CEJ-AC of 233 anterior teeth was more than 2 mm, accounting for 58.25%, and 117 anterior teeth with dehiscence were found, accounting for 29.25%. Multilevel and multivariate Logistic regression showed age, jaw, tooth and site were the relevant factors to the position of alveolar crest.@*CONCLUSION@#The position of alveolar crest of skeletal Angle class III patients who received periodontal bone augmentation surgery was lower than that of the general population, causing periodontal risks during decompensation orthodontics therapy before orthognathic surgery. The position of alveolar crest was lower in older patients than in younger patients, in mandibular teeth than in maxillary teeth, in canines than in central incisors or lateral incisors, and in central sites than in mesial sites or distal sites of labial side, which showed much higher risk.

Clinical outcomes of ultrasonic subgingival debridement combined with manual root planing in severe periodontitis / 北京大学学报(医学版)

OBJECTIVE@#To compare the clinical effects of ultrasonic subgingival debridement and ultrasonic subgingival debridement combined with manual root planing on severe periodontitis and then to investigate the necessity and significance of manual root planing.@*METHODS@#Twenty-three patients with severe periodontitis participated in this split-mouth randomized-controlled clinical trial. Baseline examination and randomization were performed after supragingival scaling: each of the upper and lower jaws had a quadrant as the test group treated with ultrasonic subgingival debridement combined with manual root planing, whereas the other two quadrants were the control group treated with ultrasonic subgingival debridement. Treatment of each patient was at intervals of one week and completed in two visits. Clinical indicators concerning probing depth (PD), clinical attachment loss (CAL) and bleeding index (BI) were recorded at baseline and 1 month, 3 months, 6 months after treatment.@*RESULTS@#There was no significant difference of periodontal indicators between the test group and the control group at baseline. Both the test group and control group resulted in significant improvement of PD, CAL and BI. One and three months after treatment, reduction of PD in the test group was higher than that in the control group [1 month: (2.13±1.31) mm vs. (1.79±1.33) mm, P<0.01; 3 months: (2.46±1.33) mm vs. (2.17±1.38) mm, P<0.01] and reduction of CAL in the test group was higher than that in the control group [1 month: (1.89±2.03) mm vs. (1.65±1.93) mm, P<0.01; 3 months: (2.03±2.05) mm vs. (1.83±1.97) mm, P<0.05]. Six months after treatment, PD in the test group and the control group decreased by (2.52±1.40) mm and (2.35±1.37) mm respectively, and the improvement in the test group was significantly better than that in the control group (P<0.01). CAL in the test group and the control group decreased by (1.89±2.14) mm and (1.77±2.00) mm respectively, and there was no statistical difference between the groups. There was no significant difference in the changes of BI between the two groups 1, 3 and 6 months after treatment.@*CONCLUSION@#Ultrasonic subgingival debridement combined with manual root planing has more reduction in PD and CAL compared with ultrasonic subgingival debridement. Therefore, it is still necessary to use manual instruments for root planing following ultrasonic subgingival debridement.

Influence of vitamin D receptor FokI polymorphism on expression of CYP24A1 in periodontal cells / 北京大学学报(医学版)

OBJECTIVE@#There is asingle nucleotide polymorphism (SNP) in the exon 2 of the vitamin D receptor (VDR) gene that can be distinguished using the restriction endonuclease FokI, and accordingly divided into three genotypes: FF, Ff and ff. VDR-FokI polymorphism was the only known SNP that could alter the protein structure of VDR. CYP24A1 is the gene encoding vitamin D 24 hydroxylase and is a vitamin D responsive gene. The influence of rs2228570 on transcriptional activation by VDR in human gingival fibroblasts (hGF) and periodontal ligament cells (hPDLC) was investigated in this study.@*METHODS@#hGF and hPDLC of 12 donors' were primarily cultured and genomic DNA was extracted. A part of genomic DNA with the length of 267 bp was obtained using PCR, which contained the SNP. VDR-Fok I genotypes were determined according to the results of restriction fragment length polymorphism. hGF and hPDLC were stimulated with 10 nmol/L 1α,25 dihydroxy vitamin D3 (1,25OH2D3) or 1 000 nmol/L 25 hydroxy vitamin D3 (25OHD3) for 48 h before RNA was extracted. Then VDR antagonist ZK159222 was used or not used during 1,25OH2D3 or 25OHD3 stimulation with hGF and hPDLC. After 1,25OH2D3 stimulation for 48 h, the proteins in hGF and hPDLC were also collected. The protein expressions of CYP24A1 and VDR were detected using Western blot.@*RESULTS@#Among the 12 donors' cell cultures, the number of FF, ff and Ff genotypes was 4, 3 and 5, respectively.After stimulation with 1,25OH2D3 or 25OHD3 for 48 h,CYP24A1 mRNA levels in FF-hGF were significantly higher than those in other hGF genotypes(1,25OH2D3: F=31.147, P<0.01; 25OHD3: F= 32.061,P <0.01), as was in FF-hPDLC (1,25OH2D3: F=23.347, P<0.01; 25OHD3: F=32.569,P<0.01). When ZK159222 was used before 1,25OH2D3 stimulation, this statistically significant difference disappeared (hGF: F=0.246, P=0.787; hPDLC: F=0.574, P=0.583). When ZK159222 was used before 25OHD3 stimulation, the trend was similar (hGF: F=1.636, P=0.248; hPDLC: F=0.582, P=0.578).After stimulation with 1,25OH2D3 for 48 h, CYP24A1 protein levels in FF-hGF were significantly higher than those in the other hGF genotypes (F=12.368, P <0.01), as was in FF-hPDLC (F=15.749, P <0.01). In hGF and hPDLC, the mRNA or protein expression of VDR of different genotypes was not significantly different under different stimulation conditions.The paired comparison showed that there was no statistically significant difference between the expression of CYP24A1 in hGF and that in hPDLC under all the stimulation conditions, as was the expression of VDR.@*CONCLUSION@#In hGF and hPDLC, the FF-VDR genotype is associated with the more remarkable up-regulation of CYP24A1than the other genotypes, indicating that transcriptional activation of FF-VDR might be higher than those of other vitamin D receptors.

Hypoxia regulates osteogenic differentiation of human periodontal ligament stem cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of hypoxia on isolated human periodontal ligament stem cells (hPDLSC) in vitro.</p><p><b>METHODS</b>hPDLSC were exposed to normoxia (20% O(2)) and hypoxia (1.5% - 2% O(2)). Total cellular RNA and protein were collected on day 1, 2, 4 and 6 after culture. Western analysis and semi-quantitative RT-PCR were used to analyze osteogenic differentiation of hPDLSC, including alkaline phosphatase (ALP), osteocalcin (OCN), secreted protein acidic and rich in cysteine (SPARC) and bone morphogenetic protein-2 (BMP-2).</p><p><b>RESULTS</b>In the first two days hypoxia slightly increased the growth of hPDLSC [A value was 0.697(hypoxia) vs 0.617 (nomoxia)] and after the third day hypoxia dramatically decreased the growth of the hPDLSC [A value was 0.870(hypoxia) vs 1.242 (nomoxia)]. Up to 90% reduction of ALP activity was observed in hPDLSC after 5 days of hypoxia [A value was 0.004(hypoxia) vs 0.049(nomoxia)]. Hypoxia decreased SPARC expression at protein level and down-regulated ALP, OCN and BMP-2 expression at mRNA level in comparison with nomoxia.</p><p><b>CONCLUSIONS</b>Hypoxia inhibited proliferation of hPDLSC and down-regulated BMP-2 mRNA expression, then down-regulated their target genes such as ALP, OCN, and SPARC, thus inhibiting critical steps in osteogenic differentiation of hPDLSC.</p>

The in vivo formation of cementum-like tissue by bovine cementoblasts / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To test the bovine cementoblasts (CBs) cementum-forming ability in vivo.</p><p><b>METHODS</b>Root fragments of newborn bovine freshly extracted mandibular incisor were cultured routinely and 4th-5th passages of CBs were harvested. CBs were then cultured in the medium supplemented with 50 mg/L alpha-ascorbic acid and 10 mmol/l beta-glycerolphosphate to form a thick layer as tissue engineering scaffold for cementum formation. Collagen membrane was used as control scaffold. 2 x 10(6) cells were attached to the CBs-made carrier as well as collagen membrane scaffolds and transplanted subcutaneously into immunodeficient mice. Transplants were harvested at 7th week. Histological sections were stained with HE, alizarin red S and van Kossa methods as well as monoclonal Ab against bovine cementum attachment protein (CAP).</p><p><b>RESULTS</b>CBs-made scaffold supported more cementum-like tissue (CLT) formation than collagen-made scaffold. The CLT formed on CBs scaffold was partly calcified with embedded cells. Uncalcified cementoid-like material could be seen on the surface and was encircled by cubical CB-like cells. The CLT was also positive to CAP and van Kossa staining.</p><p><b>CONCLUSIONS</b>These results suggest that the bovine CBs can form cementum-like tissue. The cell-made carrier is a better scaffold than collagen membrane.</p>